Family- and genus-level 16S rRNA-targeted oligonucleotide probes for ecological studies of methanotrophic bacteria.

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Gulledge, J; Ahmad, A; Steudler, P A; Pomerantz, W J; Cavanaugh, C M

2001-10-01

Methanotrophic bacteria play a major role in the global carbon cycle, degrade xenobiotic pollutants, and have the potential for a variety of biotechnological applications. To facilitate ecological studies of these important organisms, we developed a suite of oligonucleotide probes for quantitative analysis of methanotroph-specific 16S rRNA from environmental samples. Two probes target methanotrophs in the family Methylocystaceae (type II methanotrophs) as a group. No oligonucleotide signatures that distinguish between the two genera in this family, Methylocystis and Methylosinus, were identified. Two other probes target, as a single group, a majority of the known methanotrophs belonging to the family Methylococcaceae (type I/X methanotrophs). The remaining probes target members of individual genera of the Methylococcaceae, including Methylobacter, Methylomonas, Methylomicrobium, Methylococcus, and Methylocaldum. One of the family-level probes also covers all methanotrophic endosymbionts of marine mollusks for which 16S rRNA sequences have been published. The two known species of the newly described genus Methylosarcina gen. nov. are covered by a probe that otherwise targets only members of the closely related genus Methylomicrobium. None of the probes covers strains of the newly proposed genera Methylocella and "Methylothermus," which are polyphyletic with respect to the recognized methanotrophic families. Empirically determined midpoint dissociation temperatures were 49 to 57 degrees C for all probes. In dot blot screening against RNA from positive- and negative-control strains, the probes were specific to their intended targets. The broad coverage and high degree of specificity of this new suite of probes will provide more detailed, quantitative information about the community structure of methanotrophs in environmental samples than was previously available.

Bacteriohopanepolyol signatures as markers for methanotrophic bacteria in peat moss

NARCIS (Netherlands)

van Winden, J.F.; Talbot, H.M.; Kip, N.; Reichart, G.J.; Pol, A.; McNamara, N.P.; Jetten, M.S.M.; Op den Camp, H.J.M.; Sinninghe Damsté, J.S.

2012-01-01

Bacteriohopanepolyols (BHPs) are bacterial biomarkers with a likely potential to identify present and past methanotrophic communities. To unravel the methanotrophic community in peat bogs, we report the BHP signatures of type I and type II methanotrophs isolated from Sphagnum mosses and of an

Bacteriohopanepolyol signatures as markers for methanotrophic bacteria in peat moss

NARCIS (Netherlands)

Winden, J.F. van; Talbot, H.M.; Kip, N.; Reichart, G.-J.; Pol, A.; McNamara, N.P.; Jetten, M.S.M.; Camp, H.J.M. Op den; Sinninghe Damsté, J.S.

2012-01-01

Bacteriohopanepolyols (BHPs) are bacterial biomarkers with a likely potential to identify present and past methanotrophic communities. To unravel the methanotrophic community inpeat bogs, we report the BHP signatures of type I and type II methanotrophs isolated from Sphagnummosses and of an extreme

Enrichment culture and identification of endophytic methanotrophs isolated from peatland plants.

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Stępniewska, Zofia; Goraj, Weronika; Kuźniar, Agnieszka; Łopacka, Natalia; Małysza, Magdalena

2017-09-01

Aerobic methane-oxidizing bacteria (MOB) are an environmentally significant group of microorganisms due to their role in the global carbon cycle. Research conducted over the past few decades has increased the interest in discovering novel genera of methane-degrading bacteria, which efficiently utilize methane and decrease the global warming effect. Moreover, methanotrophs have more promising applications in environmental bioengineering, biotechnology, and pharmacy. The investigations were undertaken to recognize the variety of endophytic methanotrophic bacteria associated with Carex nigra, Vaccinium oxycoccus, and Eriophorum vaginatum originating from Moszne peatland (East Poland). Methanotrophic bacteria were isolated from plants by adding sterile fragments of different parts of plants (roots and stems) to agar mineral medium (nitrate mineral salts (NMS)) and incubated at different methane values (1-20% CH4). Single colonies were streaked on new NMS agar media and, after incubation, transferred to liquid NMS medium. Bacterial growth dynamics in the culture solution was studied by optical density-OD600 and methane consumption. Changes in the methane concentration during incubation were controlled by the gas chromatography technique. Characterization of methanotrophs was made by fluorescence in situ hybridization (FISH) with Mg705 and Mg84 for type I methanotrophs and Ma450 for type II methanotrophs. Identification of endophytes was performed after 16S ribosomal RNA (rRNA) and mmoX gene amplification. Our study confirmed the presence of both types of methanotrophic bacteria (types I and II) with the predominance of type I methanotrophs. Among cultivable methanotrophs, there were different strains of the genus Methylomonas and Methylosinus. Furthermore, we determined the potential of the examined bacteria for methane oxidation, which ranged from 0.463 ± 0.067 to 5.928 ± 0.169 μmol/L CH4/mL/day.

Differential detection of type II methanotrophic bacteria in acidic peatlands using newly developed 16S rRNA-targeted fluorescent oligonucleotide probes.

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Dedysh, Svetlana N; Dunfield, Peter F; Derakshani, Manigee; Stubner, Stephan; Heyer, Jürgen; Liesack, Werner

2003-04-01

Abstract Based on an extensive 16S rRNA sequence database for type II methanotrophic bacteria, a set of 16S rRNA-targeted oligonucleotide probes was developed for differential detection of specific phylogenetic groups of these bacteria by fluorescence in situ hybridisation (FISH). This set of oligonucleotides included a genus-specific probe for Methylocystis (Mcyst-1432) and three species-specific probes for Methylosinus sporium (Msins-647), Methylosinus trichosporium (Msint-1268) and the recently described acidophilic methanotroph Methylocapsa acidiphila (Mcaps-1032). These novel probes were applied to further characterise the type II methanotroph community that was detected in an acidic Sphagnum peat from West Siberia in a previous study (Dedysh et al. (2001) Appl. Environ. Microbiol. 67, 4850-4857). The largest detectable population of indigenous methanotrophs simultaneously hybridised with a group-specific probe targeting all currently known Methylosinus/Methylocystis spp. (M-450), with a genus-specific probe for Methylocystis spp. (Mcyst-1432), and with an additional probe (Mcyst-1261) that had been designed to target a defined phylogenetic subgroup of Methylocystis spp. The same subgroup of Methylocystis was also detected in acidic peat sampled from Sphagnum-dominated wetland in northern Germany. The population size of this peat-inhabiting Methylocystis subgroup was 2.0+/-0.1x10(6) cells g(-1) (wet weight) of peat from Siberia and 5.5+/-0.5x10(6) cells g(-1) of peat from northern Germany. This represented 60 and 95%, respectively, of the total number of methanotroph cells detected by FISH in these two wetland sites. Other major methanotroph populations were M. acidiphila and Methylocella palustris. Type I methanotrophs accounted for not more than 1% of total methanotroph cells. Neither M. trichosporium nor M. sporium were detected in acidic Sphagnum peat.

Characterization of methanotrophic bacteria on the basis of intact phospholipid profiles.

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Fang, J; Barcelona, M J; Semrau, J D

2000-08-01

The intact phospholipid profiles (IPPs) of seven species of methanotrophs from all three physiological groups, type I, II and X, were determined using liquid chromatography/electrospray ionization/mass spectrometry. In these methanotrophs, two major classes of phospholipids were found, phosphatidylglycerol (PG) and phosphatidylethanolamine (PE) as well as its derivatives phosphatidylmethylethanolamine (PME) and phosphatidyldimethylethanolamine (PDME). Specifically, the type I methanotrophs, Methylomonas methanica, Methylomonas rubra and Methylomicrobium album BG8 were characterized by PE and PG phospholipids with predominantly C16:1 fatty acids. The type II methanotrophs, Methylosinus trichosporium OB3b and CSC1 were characterized by phospholipids of PG, PME and PDME with predominantly C18:1 fatty acids. Methylococcus capsulatus Bath, a representative of type X methanotrophs, contained mostly PE (89% of the total phospholipids). Finally, the IPPs of a recently isolated acidophilic methanotroph, Methylocella palustris, showed it had a preponderance of PME phospholipids with 18:1 fatty acids (94% of total). Principal component analysis showed these methanotrophs could be clearly distinguished based on phospholipid profiles. Results from this study suggest that IPP can be very useful in bacterial chemotaxonomy.

[Methanotrophs of the psychrophilic microbial community of the Russian Arctic tundra].

Science.gov (United States)

Berestovskaia, Iu Iu; Vasil'eva, L V; Chestnykh, O V; Zavarzin, G A

2002-01-01

In tundra, at a low temperature, there exists a slowly developing methanotrophic community. Methane-oxidizing bacteria are associated with plants growing at high humidity, such as sedge and sphagnum; no methonotrophs were found in polytrichous and aulacomnious mosses and lichens, typical of more arid areas. The methanotrophic bacterial community inhabits definite soil horizons, from moss dust to peat formed from it. Potential ability of the methanotrophic community to oxidize methane at 5 degrees C enhances with the depth of the soil profile in spite of the decreasing soil temperature. The methanotrophic community was found to gradually adapt to various temperatures due to the presence of different methane-oxidizing bacteria in its composition. Depending on the temperature and pH, different methanotrophs occupy different econiches. Within a temperature range from 5 to 15 degrees C, three morphologically distinct groups of methanotrophs could be distinguished. At pH 5-7 and 5-15 degrees C, forms morphologically similar to Methylobacter psychrophilus predominated, whereas at the acidic pH 4-6 and 10-15 degrees C, bipolar cells typical of Methylocella palustris were mostly found. The third group of methanotrophic bacteria growing at pH 5-7 and 5-10 degrees C was represented by a novel methanotroph whole large coccoid cells had a thick mucous capsule.

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

Digital Repository Service at National Institute of Oceanography (India)

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

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

Intercellular wiring enables electron transfer between methanotrophic archaea and bacteria.

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Wegener, Gunter; Krukenberg, Viola; Riedel, Dietmar; Tegetmeyer, Halina E; Boetius, Antje

2015-10-22

The anaerobic oxidation of methane (AOM) with sulfate controls the emission of the greenhouse gas methane from the ocean floor. In marine sediments, AOM is performed by dual-species consortia of anaerobic methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB) inhabiting the methane-sulfate transition zone. The biochemical pathways and biological adaptations enabling this globally relevant process are not fully understood. Here we study the syntrophic interaction in thermophilic AOM (TAOM) between ANME-1 archaea and their consortium partner SRB HotSeep-1 (ref. 6) at 60 °C to test the hypothesis of a direct interspecies exchange of electrons. The activity of TAOM consortia was compared to the first ANME-free culture of an AOM partner bacterium that grows using hydrogen as the sole electron donor. The thermophilic ANME-1 do not produce sufficient hydrogen to sustain the observed growth of the HotSeep-1 partner. Enhancing the growth of the HotSeep-1 partner by hydrogen addition represses methane oxidation and the metabolic activity of ANME-1. Further supporting the hypothesis of direct electron transfer between the partners, we observe that under TAOM conditions, both ANME and the HotSeep-1 bacteria overexpress genes for extracellular cytochrome production and form cell-to-cell connections that resemble the nanowire structures responsible for interspecies electron transfer between syntrophic consortia of Geobacter. HotSeep-1 highly expresses genes for pili production only during consortial growth using methane, and the nanowire-like structures are absent in HotSeep-1 cells isolated with hydrogen. These observations suggest that direct electron transfer is a principal mechanism in TAOM, which may also explain the enigmatic functioning and specificity of other methanotrophic ANME-SRB consortia.

Secretion of Flavins by Three Species of Methanotrophic Bacteria▿ †

OpenAIRE

Balasubramanian, Ramakrishnan; Levinson, Benjamin T.; Rosenzweig, Amy C.

2010-01-01

We detected flavins in the growth medium of the methanotrophic bacterium Methylocystis species strain M. Flavin secretion correlates with growth stage and increases under iron starvation conditions. Two other methanotrophs, Methylosinus trichosporium OB3b and Methylococcus capsulatus (Bath), secrete flavins, suggesting that flavin secretion may be common to many methanotrophic bacteria.

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

Directory of Open Access Journals (Sweden)

Bayu Agung Prahardika

2013-04-01

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

Facultative and obligate methanotrophs how to identify and differentiate them.

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Dedysh, Svetlana N; Dunfield, Peter F

2011-01-01

Aerobic methanotrophs are metabolically unique bacteria that are able to utilize methane and some other C1-compounds as sole sources of carbon and energy. A defining characteristic of these organisms is the use of methane monooxygenase (MMO) enzymes to catalyze the oxidation of methane to methanol. For a long time, all methanotrophs were considered to be obligately methylotrophic, that is, unable to grow on compounds containing C-C bonds. This notion has recently been revised. Some members of the genera Methylocella, Methylocystis, and Methylocapsa are now known to be facultative methanotrophs, that is, capable of growing on methane as well as on some multicarbon substrates. The diagnosis of facultative methanotrophy in new isolates requires a great degree of caution since methanotrophic cultures are frequently contaminated by heterotrophic bacteria that survive on metabolic by-products of methanotrophs. The presence of only a few satellite cells in a culture may lead to false conclusions regarding substrate utilization, and several early reports of facultative methanotrophy are likely attributable to impure cultures. Another recurring mistake is the misidentification of nonmethanotrophic facultative methylotrophs as facultative methanotrophs. This chapter was prepared as an aid to avoid both kinds of confusion when examining methanotrophic isolates. Copyright © 2011 Elsevier Inc. All rights reserved.

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

NARCIS (Netherlands)

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

2002-01-01

The toxic effects caused by the chemicals contained in wastewaters generated by laboratories involved in raw milk analyses were assessed using batch assays. These assays were carried out separately with methanogenic, ammonium-oxidizing, nitrite-oxidizing and denitrifying bacteria. Since sodium azide

Isolation and Physiological Characterization of Psychrophilic Denitrifying Bacteria from Permanently Cold Arctic Fjord Sediments (Svalbard, Norway)

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Canion, Andy; Prakash, Om; Green, Stefan J.; Jahnke, Linda; Kuypers, Marcel M. M.; Kostka, Joel E.

2013-01-01

A large proportion of reactive nitrogen loss from polar sediments is mediated by denitrification, but microorganisms mediating denitrification in polar environments remain poorly characterized. A combined approach of most-probable-number (MPN) enumeration, cultivation and physiological characterization was used to describe psychrophilic denitrifying bacterial communities in sediments of three Arctic fjords in Svalbard (Norway). A MPN assay showed the presence of 10(sup 3)-10(sup 6) cells of psychrophilic nitrate-respiring bacteria g(sup -1) of sediment. Fifteen strains within the Proteobacteria were isolated using a systematic enrichment approach with organic acids as electron donors and nitrate as an electron acceptor. Isolates belonged to five genera, including Shewanella, Pseudomonas, Psychromonas (Gammaproteobacteria), Arcobacter (Epsilonproteobacteria) and Herminiimonas (Betaproteobacteria). All isolates were denitrifiers, except Shewanella, which exhibited the capacity for dissimilatory nitrate reduction to ammonium (DNRA). Growth from 0 to 40 degC demonstrated that all genera except Shewanella were psychrophiles with optimal growth below 15 degC, and adaptation to low temperature was demonstrated as a shift from primarily C16:0 saturated fatty acids to C16:1 monounsaturated fatty acids at lower temperatures. This study provides the first targeted enrichment and characterization of psychrophilic denitrifying bacteria from polar sediments, and two genera, Arcobacter and Herminiimonas, are isolated for the first time from permanently cold marine sediments.

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

Digital Repository Service at National Institute of Oceanography (India)

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

> cfu/ml on minimal media containing hexadecane as the sole carbon source. Highest bacterial counts were obtained during the monsoons. 22% of bacteria capable of hexadecane utilization were nitrate reducing and 12% were denitrifying. 29...

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

DEFF Research Database (Denmark)

Krieger, Bärbel; Schwermer, Carsten U.; Rezakhani, Nastaran

2006-01-01

with Alphaproteobacteria but also including Beta- and Gammaproteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria. The diversity of the isolates was compared to the cultivation-independent diversity of nitrate-reducing and denitrifying bacteria based on narG and nosZ as functional marker genes. Growth experiments...

Substrate-dependent denitrification of abundant probe-defined denitrifying bacteria in activated sludge.

Science.gov (United States)

Morgan-Sagastume, Fernando; Nielsen, Jeppe Lund; Nielsen, Per Halkjaer

2008-11-01

The denitrification capacity of different phylogenetic bacterial groups was investigated on addition of different substrates in activated sludge from two nutrient-removal plants. Nitrate/nitrite consumption rates (CRs) were calculated from nitrate and nitrite biosensor, in situ measurements. The nitrate/nitrite CRs depended on the substrate added, and acetate alone or combined with other substrates yielded the highest rates (3-6 mg N gVSS(-1) h(-1)). The nitrate CRs were similar to the nitrite CRs for most substrates tested. The structure of the active denitrifying population was investigated using heterotrophic CO2 microautoradiography (HetCO2-MAR) and FISH. Probe-defined denitrifiers appeared as specialized substrate utilizers despite acetate being preferentially used by most of them. Azoarcus and Accumulibacter abundance in the two different sludges was related to differences in their substrate-specific nitrate/nitrite CRs. Aquaspirillum-related bacteria were the most abundant potential denitrifiers (c. 20% of biovolume); however, Accumulibacter (3-7%) and Azoarcus (2-13%) may have primarily driven denitrification by utilizing pyruvate, ethanol, and acetate. Activated sludge denitrification was potentially conducted by a diverse, versatile population including not only Betaproteobacteria (Aquaspirillum, Thauera, Accumulibacter, and Azoarcus) but also some Alphaproteobacteria and Gammaproteobacteria, as indicated by the assimilation of 14CO2 by these probe-defined groups with a complex substrate mixture as an electron donor and nitrite as an electron acceptor in HetCO2-MAR-FISH tests.

Effect of nutrient and selective inhibitor amendments on methane oxidation, nitrous oxide production, and key gene presence and expression in landfill cover soils: characterization of the role of methanotrophs, nitrifiers, and denitrifiers.

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Lee, Sung-Woo; Im, Jeongdae; Dispirito, Alan A; Bodrossy, Levente; Barcelona, Michael J; Semrau, Jeremy D

2009-11-01

Methane and nitrous oxide are both potent greenhouse gasses, with global warming potentials approximately 25 and 298 times that of carbon dioxide. A matrix of soil microcosms was constructed with landfill cover soils collected from the King Highway Landfill in Kalamazoo, Michigan and exposed to geochemical parameters known to affect methane consumption by methanotrophs while also examining their impact on biogenic nitrous oxide production. It was found that relatively dry soils (5% moisture content) along with 15 mg NH (4) (+) (kg soil)(-1) and 0.1 mg phenylacetylene(kg soil)(-1) provided the greatest stimulation of methane oxidation while minimizing nitrous oxide production. Microarray analyses of pmoA showed that the methanotrophic community structure was dominated by Type II organisms, but Type I genera were more evident with the addition of ammonia. When phenylacetylene was added in conjunction with ammonia, the methanotrophic community structure was more similar to that observed in the presence of no amendments. PCR analyses showed the presence of amoA from both ammonia-oxidizing bacteria and archaea, and that the presence of key genes associated with these cells was reduced with the addition of phenylacetylene. Messenger RNA analyses found transcripts of pmoA, but not of mmoX, nirK, norB, or amoA from either ammonia-oxidizing bacteria or archaea. Pure culture analyses showed that methanotrophs could produce significant amounts of nitrous oxide, particularly when expressing the particulate methane monooxygenase (pMMO). Collectively, these data suggest that methanotrophs expressing pMMO played a role in nitrous oxide production in these microcosms.

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.

New molecular method to detect denitrifying anaerobic methane oxidation bacteria from different environmental niches.

Science.gov (United States)

Xu, Sai; Lu, Wenjing; Muhammad, Farooq Mustafa; Liu, Yanting; Guo, Hanwen; Meng, Ruihong; Wang, Hongtao

2018-03-01

The denitrifying anaerobic methane oxidation is an ecologically important process for reducing the potential methane emission into the atmosphere. The responsible bacterium for this process was Candidatus Methylomirabilis oxyfera belonging to the bacterial phylum of NC10. In this study, a new pair of primers targeting all the five groups of NC10 bacteria was designed to amplify NC10 bacteria from different environmental niches. The results showed that the group A was the dominant NC10 phylum bacteria from the sludges and food waste digestate while in paddy soil samples, group A and group B had nearly the same proportion. Our results also indicated that NC10 bacteria could exist in a high pH environment (pH9.24) from the food waste treatment facility. The Pearson relationship analysis showed that the pH had a significant positive relationship with the NC10 bacterial diversity (pbacteria. Copyright © 2017. Published by Elsevier B.V.

Test Plan for Methanotrophic Bioreactor at Savannah River Site-TNX

International Nuclear Information System (INIS)

Berry, C.J.

1994-01-01

The primary purpose of this project is to demonstrate the feasibility and practicality of operating a methanotrophic mobile trickle filter bioreactor (MMB) unit to effectively reduce or eliminate trichloroethylene (TCE) and associated hydrocarbons from contaminated groundwater. The two-column trickle filter system can process 1.67 gallons per minute (gpm) of contaminated groundwater. During this project, the pilot system will evaluate, optimize, and demonstrate methanotrophic treatment technology (MTT). The mobile system will receive a 1--4% methane to air mixture for stimulating the methanotrophic TCE degrading bacteria, thereby increasing the rates of degradation of these contaminants. This project will also evaluate the efficacy of different bacteria for degrading TCE for use in the system at the laboratory-scale sample groundwater monitoring wells at TNX and set up the system for continued operation. The trickle filter system may be used to inexpensively treat other small-scale organic waste streams at SRS after the initial start-up. The MTT was demonstrated as an effective and efficient method of degrading TCE in the laboratory and during a field-scale in situ demonstration for degrading TCE in a groundwater plume at SRS. The methanotrophic bacteria increase significantly in population numbers and in the production of methane monooxygenase (MMO), an extremely powerful oxidizer. MMO was demonstrated as effective in oxidizing TCE and other recalcitrant compounds in laboratory studies. In the presence of MMO, TCE is oxidized to TCE-epoxide, which breaks down spontaneously into simple, easily degraded, daughter compounds. The system will receive a 1--4% methane to air mixture, which will effectively grow and maintain the methanotrophic bacteria that will degrade TCE. This demonstration will have broad applications to bioremediating contaminated groundwater systems where in situ bioremediation is not practical

Methylocella Species Are Facultatively Methanotrophic

OpenAIRE

Dedysh, Svetlana N.; Knief, Claudia; Dunfield, Peter F.

2005-01-01

All aerobic methanotrophic bacteria described to date are unable to grow on substrates containing carbon-carbon bonds. Here we demonstrate that members of the recently discovered genus Methylocella are an exception to this. These bacteria are able to use as their sole energy source the one-carbon compounds methane and methanol, as well as the multicarbon compounds acetate, pyruvate, succinate, malate, and ethanol. To conclusively verify facultative growth, acetate and methane were used as mod...

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

Directory of Open Access Journals (Sweden)

Michela eLangone

2014-02-01

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

Bacteria that degrade hazardous waste: The isolation of trichloroethylene-degrading methanotrophic bacteria and development of monoclonal antibodies specific to them

International Nuclear Information System (INIS)

Little, C.D.

1988-01-01

Trichloroethylene (TCE), a suspected carcinogen, is one of the most frequently reported groundwater contaminants at hazardous waste sites in the US. An aerobic, methane-oxidizing bacterium was isolated that degrades TCE in pure culture at concentrations commonly observed in contaminated groundwater. Strain 46-1, a Type I methanotrophic bacterium, degraded TCE when growing on methane or methanol, producing CO 2 and water-soluble products. Gas chromatography and 14 C radiotracer techniques were used to determine the rate, methane dependence, and mechanism of TCE biodegradation. TCE biodegradation by strain 46-1 appears to be a co-metabolic process that occurs when the organism is actively metabolizing a suitable growth substrate such as methane or methanol. Five mouse monoclonal antibodies (MABS) that specifically bind strain 46-1 were prepared by conventional hybridoma technology. These MABS are apparently biochemically distinct and were used to develop enzyme-linked and fluorescent immunoassays to detect strain 46-1 cells in environmental samples. A fluorescent immunoassay utilizing four of these MABS easily distinguished laboratory-grown 46-1 cells from other methanotrophic and heterotrophic bacteria, but failed to detect 46-1 cells in groundwater samples and cultures

Potential roles of anaerobic ammonium and methane oxidation in the nitrogen cycle of wetland ecosystems.

Science.gov (United States)

Zhu, Guibing; Jetten, Mike S M; Kuschk, Peter; Ettwig, Katharina F; Yin, Chengqing

2010-04-01

Anaerobic ammonium oxidation (anammox) and anaerobic methane oxidation (ANME coupled to denitrification) with nitrite as electron acceptor are two of the most recent discoveries in the microbial nitrogen cycle. Currently the anammox process has been relatively well investigated in a number of natural and man-made ecosystems, while ANME coupled to denitrification has only been observed in a limited number of freshwater ecosystems. The ubiquitous presence of anammox bacteria in marine ecosystems has changed our knowledge of the global nitrogen cycle. Up to 50% of N(2) production in marine sediments and oxygen-depleted zones may be attributed to anammox bacteria. However, there are only few indications of anammox in natural and constructed freshwater wetlands. In this paper, the potential role of anammox and denitrifying methanotrophic bacteria in natural and artificial wetlands is discussed in relation to global warming. The focus of the review is to explore and analyze if suitable environmental conditions exist for anammox and denitrifying methanotrophic bacteria in nitrogen-rich freshwater wetlands.

Evaluation of in-situ methanotrophic bioremediation for contaminated groundwater, St. Joseph, Michigan. Final report, January 1989-December 1989

International Nuclear Information System (INIS)

McCarty, P.L.; Semprini, L.; Dolan, M.E.; Harmon, T.C.; Just, S.

1990-09-01

A feasibility study of utilizing indigenous bacteria that use methane as a source of cell carbon and energy (methanotrophs) for in-situ bioremediation of groundwater contaminants at the St. Joseph site is summarized. The contaminants, compounds, can be biotransformed by methanotrophic bacteria, which are found in some locations of the site in adequate populations. The process involves stimulating the growth of native populations of methanotrophs by injecting water containing dissolved methane and oxygen into the aquifer. The stimulated population of methanotrophs in turn has the capability to degrade trichloroethylene, 1,2-cis-dichloroethylene, 1,2-trans-dichloroethylene, and vinyl chloride

Illumina sequencing-based analysis of a microbial community enriched under anaerobic methane oxidation condition coupled to denitrification revealed coexistence of aerobic and anaerobic methanotrophs.

Science.gov (United States)

Siniscalchi, Luciene Alves Batista; Leite, Laura Rabelo; Oliveira, Guilherme; Chernicharo, Carlos Augusto Lemos; de Araújo, Juliana Calabria

2017-07-01

Methane is produced in anaerobic environments, such as reactors used to treat wastewaters, and can be consumed by methanotrophs. The composition and structure of a microbial community enriched from anaerobic sewage sludge under methane-oxidation condition coupled to denitrification were investigated. Denaturing gradient gel electrophoresis (DGGE) analysis retrieved sequences of Methylocaldum and Chloroflexi. Deep sequencing analysis revealed a complex community that changed over time and was affected by methane concentration. Methylocaldum (8.2%), Methylosinus (2.3%), Methylomonas (0.02%), Methylacidiphilales (0.45%), Nitrospirales (0.18%), and Methanosarcinales (0.3%) were detected. Despite denitrifying conditions provided, Nitrospirales and Methanosarcinales, known to perform anaerobic methane oxidation coupled to denitrification (DAMO) process, were in very low abundance. Results demonstrated that aerobic and anaerobic methanotrophs coexisted in the reactor together with heterotrophic microorganisms, suggesting that a diverse microbial community was important to sustain methanotrophic activity. The methanogenic sludge was a good inoculum to enrich methanotrophs, and cultivation conditions play a selective role in determining community composition.

Diversity and activity of methanotrophs in alkaline soil from a Chinese coal mine.

Science.gov (United States)

Han, Bing; Chen, Yin; Abell, Guy; Jiang, Hao; Bodrossy, Levente; Zhao, Jiangang; Murrell, J Colin; Xing, Xin-Hui

2009-11-01

Culture-independent molecular biological techniques, including 16S rRNA gene and functional gene clone libraries and microarray analyses using pmoA (encoding a key subunit of particulate methane monooxygenase), were applied to investigate the methanotroph community structure in alkaline soil from a Chinese coal mine. This environment contained a high diversity of methanotrophs, including the type II methanotrophs Methylosinus/Methylocystis, type I methanotrophs related to Methylobacter/Methylosoma and Methylococcus, and a number of as yet uncultivated methanotrophs. In order to identify the metabolically active methane-oxidizing bacteria from this alkaline environment, DNA stable isotope probing (DNA-SIP) experiments using (13)CH(4) were carried out. This showed that both type I and type II methanotrophs were active, together with methanotrophs related to Methylocella, which had previously been found only in acidic environments. Methylotrophs, including Methylopila and Hyphomicrobium, were also detected in soil DNA and after DNA-SIP experiments. DNA sequence information on the most abundant, active methanotrophs in this alkaline soil will facilitate the design of oligonucleotide probes to monitor enrichment cultures when isolating key alkaliphilic methanotrophs from such environments.

Diversity and activity of methanotrophs in alkaline soil from a Chinese coal mine

Energy Technology Data Exchange (ETDEWEB)

Han, Bing; Chen, Yin; Abell, Guy; Jiang, Hao; Bodrossy, Levente; Zhao, Jiangang; Murrell, J. Colin; Xing, Xin-Hui [Tsinghua University, Beijing (China). Dept. of Chemical Engineering

2009-11-15

Culture-independent molecular biological techniques, including 16S rRNA gene and functional gene clone libraries and microarray analyses using pmoA (encoding a key subunit of particulate methane monooxygenase), were applied to investigate the methanotroph community structure in alkaline soil from a Chinese coal mine. This environment contained a high diversity of methanotrophs, including the type II methanotrophs Methylosinus/Methylocystis, type I methanotrophs related to Methylobacter/Methylosoma and Methylococcus, and a number of as yet uncultivated methanotrophs. In order to identify the metabolically active methane-oxidizing bacteria from this alkaline environment, DNA stable isotope probing (DNA-SIP) experiments using {sup 13}CH{sub 4} were carried out. This showed that both type I and type II methanotrophs were active, together with methanotrophs related to Methylocella, which had previously been found only in acidic environments. Methylotrophs, including Methylopila and Hyphomicrobium, were also detected in soil DNA and after DNA-SIP experiments. DNA sequence information on the most abundant, active methanotrophs in this alkaline soil will facilitate the design of oligonucleotide probes to monitor enrichment cultures when isolating key alkaliphilic methanotrophs from such environments.

Production of NO and N(inf2)O by Pure Cultures of Nitrifying and Denitrifying Bacteria during Changes in Aeration

NARCIS (Netherlands)

Kester, R.A.; De Boer, W.; Laanbroek, H.J.

1997-01-01

Peak emissions of NO and N2O are often observed after wetting of soil, The reactions to sudden changes in the aeration of cultures of nitrifying and denitrifying bacteria with respect to NO and N2O emissions were compared to obtain more information about the microbiological aspects of peak

Co-existence of Anaerobic Ammonium Oxidation Bacteria and Denitrifying Anaerobic Methane Oxidation Bacteria in Sewage Sludge: Community Diversity and Seasonal Dynamics

DEFF Research Database (Denmark)

Xu, Sai; Lu, Wenjing; Mustafa, Muhammad Farooq

2017-01-01

Anaerobic ammonium oxidation (ANAMMOX) and denitrifying anaerobic methane oxidation (DAMO) have been recently discovered as relevant processes in the carbon and nitrogen cycles of wastewater treatment plants. In this study, the seasonal dynamics of ANAMMOX and DAMO bacterial community structures......, and an unknown cluster was primarily detected in autumn and winter. Similar patterns of seasonal variation in the community structure of DAMO bacteria were also observed. Group B was the dominant in spring and summer, whereas in autumn and winter, group A and group B presented almost the same proportion...

Effects of Selected Root Exudate Components on Nitrogen Removal and Development of Denitrifying Bacteria in Constructed Wetlands

Directory of Open Access Journals (Sweden)

Hailu Wu

2017-06-01

Full Text Available Root exudates, particularly low molecular weight carbon (LMWC substrates, are major drivers of bacterial diversity and activity in the rhizosphere environment. However, it is not well understood how specific LMWC compounds—such as organic acids, soluble sugars, and amino acids—influence the community structures of denitrifying bacteria or if there are specific functions of LMWC substrates that preferentially respond to nitrogen (N removal in constructed wetlands (CWs. To address these knowledge gaps, we added mixtures of artificial exudates to CW microcosms containing N pollutant. N removal efficiency was observed over a 48-h experimental period, and at the end of the experiment, DNA was extracted from microbial samples for assessment of the bacterial community. The removal efficiencies of TN for the exudates treatments were higher than for control groups by 47.1–58.67%. Organic acid and soluble sugar treatments increased N removal, while amino acids were negative to N removal. The microbial community was changed when artificial exudates were added, but there were no significant relationships between LMWC compounds and bacterial community composition. These results indicate that although the responses of community structures of denitrifying bacteria to LMWC additions are still uncertain, there is evidence for N removal in response to exudate additions across LMWC types.

Detection, isolation, and characterization of acidophilic methanotrophs from Sphagnum mosses.

Science.gov (United States)

Kip, Nardy; Ouyang, Wenjing; van Winden, Julia; Raghoebarsing, Ashna; van Niftrik, Laura; Pol, Arjan; Pan, Yao; Bodrossy, Levente; van Donselaar, Elly G; Reichart, Gert-Jan; Jetten, Mike S M; Damsté, Jaap S Sinninghe; Op den Camp, Huub J M

2011-08-15

Sphagnum peatlands are important ecosystems in the methane cycle. Methane-oxidizing bacteria in these ecosystems serve as a methane filter and limit methane emissions. Yet little is known about the diversity and identity of the methanotrophs present in and on Sphagnum mosses of peatlands, and only a few isolates are known. The methanotrophic community in Sphagnum mosses, originating from a Dutch peat bog, was investigated using a pmoA microarray. A high biodiversity of both gamma- and alphaproteobacterial methanotrophs was found. With Sphagnum mosses as the inoculum, alpha- and gammaproteobacterial acidophilic methanotrophs were isolated using established and newly designed media. The 16S rRNA, pmoA, pxmA, and mmoX gene sequences showed that the alphaproteobacterial isolates belonged to the Methylocystis and Methylosinus genera. The Methylosinus species isolated are the first acid-tolerant members of this genus. Of the acidophilic gammaproteobacterial strains isolated, strain M5 was affiliated with the Methylomonas genus, and the other strain, M200, may represent a novel genus, most closely related to the genera Methylosoma and Methylovulum. So far, no acidophilic or acid-tolerant methanotrophs in the Gammaproteobacteria class are known. All strains showed the typical features of either type I or II methanotrophs and are, to the best of our knowledge, the first isolated (acidophilic or acid-tolerant) methanotrophs from Sphagnum mosses.

Facultative methanotrophs are abundant at terrestrial natural gas seeps.

Science.gov (United States)

Farhan Ul Haque, Muhammad; Crombie, Andrew T; Ensminger, Scott A; Baciu, Calin; Murrell, J Colin

2018-06-28

Natural gas contains methane and the gaseous alkanes ethane, propane and butane, which collectively influence atmospheric chemistry and cause global warming. Methane-oxidising bacteria, methanotrophs, are crucial in mitigating emissions of methane as they oxidise most of the methane produced in soils and the subsurface before it reaches the atmosphere. Methanotrophs are usually obligate, i.e. grow only on methane and not on longer chain alkanes. Bacteria that grow on the other gaseous alkanes in natural gas such as propane have also been characterised, but they do not grow on methane. Recently, it was shown that the facultative methanotroph Methylocella silvestris grew on ethane and propane, other components of natural gas, in addition to methane. Therefore, we hypothesised that Methylocella may be prevalent at natural gas seeps and might play a major role in consuming all components of this potent greenhouse gas mixture before it is released to the atmosphere. Environments known to be exposed to biogenic methane emissions or thermogenic natural gas seeps were surveyed for methanotrophs. 16S rRNA gene amplicon sequencing revealed that Methylocella were the most abundant methanotrophs in natural gas seep environments. New Methylocella-specific molecular tools targeting mmoX (encoding the soluble methane monooxygenase) by PCR and Illumina amplicon sequencing were designed and used to investigate various sites. Functional gene-based assays confirmed that Methylocella were present in all of the natural gas seep sites tested here. This might be due to its ability to use methane and other short chain alkane components of natural gas. We also observed the abundance of Methylocella in other environments exposed to biogenic methane, suggesting that Methylocella has been overlooked in the past as previous ecological studies of methanotrophs often used pmoA (encoding the alpha subunit of particulate methane monooxygenase) as a marker gene. New biomolecular tools designed in

Identity of active methanotrophs in landfill cover soil as revealed by DNA-stable isotope probing.

Science.gov (United States)

Cébron, Aurélie; Bodrossy, Levente; Chen, Yin; Singer, Andrew C; Thompson, Ian P; Prosser, James I; Murrell, J Colin

2007-10-01

A considerable amount of methane produced during decomposition of landfill waste can be oxidized in landfill cover soil by methane-oxidizing bacteria (methanotrophs) thus reducing greenhouse gas emissions to the atmosphere. The identity of active methanotrophs in Roscommon landfill cover soil, a slightly acidic peat soil, was assessed by DNA-stable isotope probing (SIP). Landfill cover soil slurries were incubated with (13)C-labelled methane and under either nutrient-rich nitrate mineral salt medium or water. The identity of active methanotrophs was revealed by analysis of (13)C-labelled DNA fractions. The diversity of functional genes (pmoA and mmoX) and 16S rRNA genes was analyzed using clone libraries, microarrays and denaturing gradient gel electrophoresis. 16S rRNA gene analysis revealed that the cover soil was mainly dominated by Type II methanotrophs closely related to the genera Methylocella and Methylocapsa and to Methylocystis species. These results were supported by analysis of mmoX genes in (13)C-DNA. Analysis of pmoA gene diversity indicated that a significant proportion of active bacteria were also closely related to the Type I methanotrophs, Methylobacter and Methylomonas species. Environmental conditions in the slightly acidic peat soil from Roscommon landfill cover allow establishment of both Type I and Type II methanotrophs.

Relative Contribution of nirK- and nirS- Bacterial Denitrifiers as Well as Fungal Denitrifiers to Nitrous Oxide Production from Dairy Manure Compost.

Science.gov (United States)

Maeda, Koki; Toyoda, Sakae; Philippot, Laurent; Hattori, Shohei; Nakajima, Keiichi; Ito, Yumi; Yoshida, Naohiro

2017-12-19

The relative contribution of fungi, bacteria, and nirS and nirK denirifiers to nitrous oxide (N 2 O) emission with unknown isotopic signature from dairy manure compost was examined by selective inhibition techniques. Chloramphenicol (CHP), cycloheximide (CYH), and diethyl dithiocarbamate (DDTC) were used to suppress the activity of bacteria, fungi, and nirK-possessing denitrifiers, respectively. Produced N 2 O were surveyed to isotopocule analysis, and its 15 N site preference (SP) and δ 18 O values were compared. Bacteria, fungi, nirS, and nirK gene abundances were compared by qPCR. The results showed that N 2 O production was strongly inhibited by CHP addition in surface pile samples (82.2%) as well as in nitrite-amended core samples (98.4%), while CYH addition did not inhibit the N 2 O production. N 2 O with unknown isotopic signature (SP = 15.3-16.2‰), accompanied by δ 18 O (19.0-26.8‰) values which were close to bacterial denitrification, was also suppressed by CHP and DDTC addition (95.3%) indicating that nirK denitrifiers were responsible for this N 2 O production despite being less abundant than nirS denitrifiers. Altogether, our results suggest that bacteria are important for N 2 O production with different SP values both from compost surface and pile core. However, further work is required to decipher whether N 2 O with unknown isotopic signature is mostly due to nirK denitrifiers that are taxonomically different from the SP-characterized strains and therefore have different SP values rather than also being interwoven with the contribution of the NO-detoxifying pathway and/or of co-denitrification.

Active methanotrophs in two contrasting North American peatland ecosystems revealed using DNA-SIP.

Science.gov (United States)

Gupta, Varun; Smemo, Kurt A; Yavitt, Joseph B; Basiliko, Nathan

2012-02-01

The active methanotroph community was investigated in two contrasting North American peatlands, a nutrient-rich sedge fen and nutrient-poor Sphagnum bog using in vitro incubations and (13)C-DNA stable-isotope probing (SIP) to measure methane (CH(4)) oxidation rates and label active microbes followed by fingerprinting and sequencing of bacterial and archaeal 16S rDNA and methane monooxygenase (pmoA and mmoX) genes. Rates of CH(4) oxidation were slightly, but significantly, faster in the bog and methanotrophs belonged to the class Alphaproteobacteria and were similar to other methanotrophs of the genera Methylocystis, Methylosinus, and Methylocapsa or Methylocella detected in, or isolated from, European bogs. The fen had a greater phylogenetic diversity of organisms that had assimilated (13)C, including methanotrophs from both the Alpha- and Gammaproteobacteria classes and other potentially non-methanotrophic organisms that were similar to bacteria detected in a UK and Finnish fen. Based on similarities between bacteria in our sites and those in Europe, including Russia, we conclude that site physicochemical characteristics rather than biogeography controlled the phylogenetic diversity of active methanotrophs and that differences in phylogenetic diversity between the bog and fen did not relate to measured CH(4) oxidation rates. A single crenarchaeon in the bog site appeared to be assimilating (13)C in 16S rDNA; however, its phylogenetic similarity to other CO(2)-utilizing archaea probably indicates that this organism is not directly involved in CH(4) oxidation in peat.

Monitoring Methanotrophic Bacteria in Hybrid Anaerobic-Aerobic Reactors with PCR and a Catabolic Gene Probe

Science.gov (United States)

Miguez, Carlos B.; Shen, Chun F.; Bourque, Denis; Guiot, Serge R.; Groleau, Denis

1999-01-01

We attempted to mimic in small upflow anaerobic sludge bed (UASB) bioreactors the metabolic association found in nature between methanogens and methanotrophs. UASB bioreactors were inoculated with pure cultures of methanotrophs, and the bioreactors were operated by using continuous low-level oxygenation in order to favor growth and/or survival of methanotrophs. Unlike the reactors in other similar studies, the hybrid anaerobic-aerobic bioreactors which we used were operated synchronously, not sequentially. Here, emphasis was placed on monitoring various methanotrophic populations by using classical methods and also a PCR amplification assay based on the mmoX gene fragment of the soluble methane monooxygenase (sMMO). The following results were obtained: (i) under the conditions used, Methylosinus sporium appeared to survive better than Methylosinus trichosporium; (ii) the PCR method which we used could detect as few as about 2,000 sMMO gene-containing methanotrophs per g (wet weight) of granular sludge; (iii) inoculation of the bioreactors with pure cultures of methanotrophs contributed greatly to increases in the sMMO-containing population (although the sMMO-containing population decreased gradually with time, at the end of an experiment it was always at least 2 logs larger than the initial population before inoculation); (iv) in general, there was a good correlation between populations with the sMMO gene and populations that exhibited sMMO activity; and (v) inoculation with sMMO-positive cultures helped increase significantly the proportion of sMMO-positive methanotrophs in reactors, even after several weeks of operation under various regimes. At some point, anaerobic-aerobic bioreactors like those described here might be used for biodegradation of various chlorinated pollutants. PMID:9925557

Effect of carbon dioxide and bicarbonate as inorganic carbon sources on growth and adaptation of autohydrogenotrophic denitrifying bacteria

International Nuclear Information System (INIS)

Ghafari, Shahin; Hasan, Masitah; Aroua, Mohamed Kheireddine

2009-01-01

Acclimation of autohydrogenotrophic denitrifying bacteria using inorganic carbon source (CO 2 and bicarbonate) and hydrogen gas as electron donor was performed in this study. In this regard, activated sludge was used as the seed source and sequencing batch reactor (SBR) technique was applied for accomplishing the acclimatization. Three distinct strategies in feeding of carbon sources were applied: (I) continuous sparging of CO 2 , (II) bicarbonate plus continuous sparging of CO 2 , and (III) only bicarbonate. The pH-reducing nature of CO 2 showed an unfavorable impact on denitrification rate; however bicarbonate resulted in a buffered environment in the mixed liquor and provided a suitable mean to maintain the pH in the desirable range of 7-8.2. As a result, bicarbonate as the only carbon source showed a faster adaptation, while carbon dioxide as the only carbon source as well as a complementary carbon source added to bicarbonate resulted in longer acclimation period. Adapted hydrogenotrophic denitrifying bacteria, using bicarbonate and hydrogen gas in the aforementioned pH range, caused denitrification at a rate of 13.33 mg NO 3 - -N/g MLVSS/h for degrading 20 and 30 mg NO 3 - -N/L and 9.09 mg NO 3 - -N/g MLVSS/h for degrading 50 mg NO 3 - -N/L

Methylocella species are facultatively methanotrophic.

Science.gov (United States)

Dedysh, Svetlana N; Knief, Claudia; Dunfield, Peter F

2005-07-01

All aerobic methanotrophic bacteria described to date are unable to grow on substrates containing carbon-carbon bonds. Here we demonstrate that members of the recently discovered genus Methylocella are an exception to this. These bacteria are able to use as their sole energy source the one-carbon compounds methane and methanol, as well as the multicarbon compounds acetate, pyruvate, succinate, malate, and ethanol. To conclusively verify facultative growth, acetate and methane were used as model substrates in growth experiments with the type strain Methylocella silvestris BL2. Quantitative real-time PCR targeting the mmoX gene, which encodes a subunit of soluble methane monooxygenase, showed that copies of this gene increased in parallel with cell counts during growth on either acetate or methane as the sole substrate. This verified that cells possessing the genetic basis of methane oxidation grew on acetate as well as methane. Cloning of 16S rRNA genes and fluorescence in situ hybridization with strain-specific and genus-specific oligonucleotide probes detected no contaminants in cultures. The growth rate and carbon conversion efficiency were higher on acetate than on methane, and when both substrates were provided in excess, acetate was preferably used and methane oxidation was shut down. Our data demonstrate that not all methanotrophic bacteria are limited to growing on one-carbon compounds. This could have major implications for understanding the factors controlling methane fluxes in the environment.

Co-existence of Anaerobic Ammonium Oxidation Bacteria and Denitrifying Anaerobic Methane Oxidation Bacteria in Sewage Sludge: Community Diversity and Seasonal Dynamics.

Science.gov (United States)

Xu, Sai; Lu, Wenjing; Mustafa, Muhammad Farooq; Caicedo, Luis Miguel; Guo, Hanwen; Fu, Xindi; Wang, Hongtao

2017-11-01

Anaerobic ammonium oxidation (ANAMMOX) and denitrifying anaerobic methane oxidation (DAMO) have been recently discovered as relevant processes in the carbon and nitrogen cycles of wastewater treatment plants. In this study, the seasonal dynamics of ANAMMOX and DAMO bacterial community structures and their abundance in sewage sludge collected from wastewater treatment plants were analysed. Results indicated that ANAMMOX and DAMO bacteria co-existed in sewage sludge in different seasons and their abundance was positively correlated (P bacteria in autumn and winter indicated that these seasons were the preferred time to favour the growth of ANAMMOX and DAMO bacteria. The community structure of ANNAMOX and DAMO bacteria could also shift with seasonal changes. The "Candidatus Brocadia" genus of ANAMMOX bacteria was mainly recovered in spring and summer, and an unknown cluster was primarily detected in autumn and winter. Similar patterns of seasonal variation in the community structure of DAMO bacteria were also observed. Group B was the dominant in spring and summer, whereas in autumn and winter, group A and group B presented almost the same proportion. The redundancy analysis revealed that pH and nitrate were the most significant factors affecting community structures of these two groups (P < 0.01). This study reported the diversity of ANAMMOX and DAMO in wastewater treatment plants that may be the basis for new nitrogen removal technologies.

Detection, Isolation, and Characterization of Acidophilic Methanotrophs from

NARCIS (Netherlands)

Kip, N.; Ouyang, W.J.; van Winden, J.; Raghoebarsing, A.; van Niftrik, L.; Pol, A.; Pan, Y.; Bodrossy, L.; van Donselaar, E.G.; Reichart, G.J.; Jetten, M.S.M.; Sinninghe Damsté, J.S.; Op den Camp, H.J.M.

2011-01-01

Sphagnum peatlands are important ecosystems in the methane cycle. Methane-oxidizing bacteria in these ecosystems serve as a methane filter and limit methane emissions. Yet little is known about the diversity and identity of the methanotrophs present in and on Sphagnum mosses of peatlands, and only a

Denitrifying bacteria from the genus Rhodanobacter dominate bacterial communities in the highly contaminated subsurface of a nuclear legacy waste site.

Science.gov (United States)

Green, Stefan J; Prakash, Om; Jasrotia, Puja; Overholt, Will A; Cardenas, Erick; Hubbard, Daniela; Tiedje, James M; Watson, David B; Schadt, Christopher W; Brooks, Scott C; Kostka, Joel E

2012-02-01

The effect of long-term mixed-waste contamination, particularly uranium and nitrate, on the microbial community in the terrestrial subsurface was investigated at the field scale at the Oak Ridge Integrated Field Research Challenge (ORIFRC) site in Oak Ridge, TN. The abundance, community composition, and distribution of groundwater microorganisms were examined across the site during two seasonal sampling events. At representative locations, subsurface sediment was also examined from two boreholes, one sampled from the most heavily contaminated area of the site and another from an area with low contamination. A suite of DNA- and RNA-based molecular tools were employed for community characterization, including quantitative PCR of rRNA and nitrite reductase genes, community composition fingerprinting analysis, and high-throughput pyrotag sequencing of rRNA genes. The results demonstrate that pH is a major driver of the subsurface microbial community structure and that denitrifying bacteria from the genus Rhodanobacter (class Gammaproteobacteria) dominate at low pH. The relative abundance of bacteria from this genus was positively correlated with lower-pH conditions, and these bacteria were abundant and active in the most highly contaminated areas. Other factors, such as the concentration of nitrogen species, oxygen level, and sampling season, did not appear to strongly influence the distribution of Rhodanobacter bacteria. The results indicate that these organisms are acid-tolerant denitrifiers, well suited to the acidic, nitrate-rich subsurface conditions, and pH is confirmed as a dominant driver of bacterial community structure in this contaminated subsurface environment.

Differential Transcriptional Activation of Genes Encoding Soluble Methane Monooxygenase in a Facultative Versus an Obligate Methanotroph

OpenAIRE

Angela V. Smirnova; Peter F. Dunfield

2018-01-01

Methanotrophs are a specialized group of bacteria that can utilize methane (CH4) as a sole energy source. A key enzyme responsible for methane oxidation is methane monooxygenase (MMO), of either a soluble, cytoplasmic type (sMMO), or a particulate, membrane-bound type (pMMO). Methylocella silvestris BL2 and Methyloferula stellata AR4 are closely related methanotroph species that oxidize methane via sMMO only. However, Methyloferula stellata is an obligate methanotroph, while Methylocella silv...

Effect of carbon dioxide and bicarbonate as inorganic carbon sources on growth and adaptation of autohydrogenotrophic denitrifying bacteria

Energy Technology Data Exchange (ETDEWEB)

Ghafari, Shahin; Hasan, Masitah [Department of Chemical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur (Malaysia); Aroua, Mohamed Kheireddine [Department of Chemical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur (Malaysia)], E-mail: mk_aroua@um.edu.my

2009-03-15

Acclimation of autohydrogenotrophic denitrifying bacteria using inorganic carbon source (CO{sub 2} and bicarbonate) and hydrogen gas as electron donor was performed in this study. In this regard, activated sludge was used as the seed source and sequencing batch reactor (SBR) technique was applied for accomplishing the acclimatization. Three distinct strategies in feeding of carbon sources were applied: (I) continuous sparging of CO{sub 2}, (II) bicarbonate plus continuous sparging of CO{sub 2}, and (III) only bicarbonate. The pH-reducing nature of CO{sub 2} showed an unfavorable impact on denitrification rate; however bicarbonate resulted in a buffered environment in the mixed liquor and provided a suitable mean to maintain the pH in the desirable range of 7-8.2. As a result, bicarbonate as the only carbon source showed a faster adaptation, while carbon dioxide as the only carbon source as well as a complementary carbon source added to bicarbonate resulted in longer acclimation period. Adapted hydrogenotrophic denitrifying bacteria, using bicarbonate and hydrogen gas in the aforementioned pH range, caused denitrification at a rate of 13.33 mg NO{sub 3}{sup -}-N/g MLVSS/h for degrading 20 and 30 mg NO{sub 3}{sup -}-N/L and 9.09 mg NO{sub 3}{sup -}-N/g MLVSS/h for degrading 50 mg NO{sub 3}{sup -}-N/L.

The oxidative TCA cycle operates during methanotrophic growth of the Type I methanotroph Methylomicrobium buryatense 5GB1.

Science.gov (United States)

Fu, Yanfen; Li, Yi; Lidstrom, Mary

2017-07-01

Methanotrophs are a group of bacteria that use methane as sole carbon and energy source. Type I methanotrophs are gamma-proteobacterial methanotrophs using the ribulose monophosphate cycle (RuMP) cycle for methane assimilation. In order to facilitate metabolic engineering in the industrially promising Type I methanotroph Methylomicrobium buryatense 5GB1, flux analysis of cellular metabolism is needed and 13 C tracer analysis is a foundational tool for such work. This biological system has a single-carbon input and a special network topology that together pose challenges to the current well-established methodology for 13 C tracer analysis using a multi-carbon input such as glucose, and to date, no 13 C tracer analysis of flux in a Type I methanotroph has been reported. In this study, we showed that by monitoring labeling patterns of several key intermediate metabolites in core metabolism, it is possible to quantitate the relative flux ratios for important branch points, such as the malate node. In addition, it is possible to assess the operation of the TCA cycle, which has been thought to be incomplete in Type I methanotrophs. Surprisingly, our analysis provides direct evidence of a complete, oxidative TCA cycle operating in M. buryatense 5GB1 using methane as sole carbon and energy substrate, contributing about 45% of the total flux for de novo malate production. Combined with mutant analysis, this method was able to identify fumA (METBUDRAFT_1453/MBURv2__60244) as the primary fumarase involved in the oxidative TCA cycle, among 2 predicted fumarases, supported by 13 C tracer analysis on both fumA and fumC single knockouts. Interrupting the oxidative TCA cycle leads to a severe growth defect, suggesting that the oxidative TCA cycle functions to not only provide precursors for de novo biomass synthesis, but also to provide reducing power to the system. This information provides new opportunities for metabolic engineering of M. buryatense for the production of

Detection, Isolation, and Characterization of Acidophilic Methanotrophs from Sphagnum Mosses ▿ †

Science.gov (United States)

Kip, Nardy; Ouyang, Wenjing; van Winden, Julia; Raghoebarsing, Ashna; van Niftrik, Laura; Pol, Arjan; Pan, Yao; Bodrossy, Levente; van Donselaar, Elly G.; Reichart, Gert-Jan; Jetten, Mike S. M.; Sinninghe Damsté, Jaap S.; Op den Camp, Huub J. M.

2011-01-01

Sphagnum peatlands are important ecosystems in the methane cycle. Methane-oxidizing bacteria in these ecosystems serve as a methane filter and limit methane emissions. Yet little is known about the diversity and identity of the methanotrophs present in and on Sphagnum mosses of peatlands, and only a few isolates are known. The methanotrophic community in Sphagnum mosses, originating from a Dutch peat bog, was investigated using a pmoA microarray. A high biodiversity of both gamma- and alphaproteobacterial methanotrophs was found. With Sphagnum mosses as the inoculum, alpha- and gammaproteobacterial acidophilic methanotrophs were isolated using established and newly designed media. The 16S rRNA, pmoA, pxmA, and mmoX gene sequences showed that the alphaproteobacterial isolates belonged to the Methylocystis and Methylosinus genera. The Methylosinus species isolated are the first acid-tolerant members of this genus. Of the acidophilic gammaproteobacterial strains isolated, strain M5 was affiliated with the Methylomonas genus, and the other strain, M200, may represent a novel genus, most closely related to the genera Methylosoma and Methylovulum. So far, no acidophilic or acid-tolerant methanotrophs in the Gammaproteobacteria class are known. All strains showed the typical features of either type I or II methanotrophs and are, to the best of our knowledge, the first isolated (acidophilic or acid-tolerant) methanotrophs from Sphagnum mosses. PMID:21724892

Denitrifying bacteria from the genus Rhodanobacter dominate bacterial communities in the highly contaminated subsurface of a nuclear legacy waste site

Energy Technology Data Exchange (ETDEWEB)

Green, Stefan [Florida State University; Prakash, Om [Florida State University; Jasrotia, Puja [Florida State University; Overholt, Will [Florida State University; Cardenas, Erick [Michigan State University, East Lansing; Hubbard, Daniela [Florida State University; Tiedje, James M. [Michigan State University, East Lansing; Watson, David B [ORNL; Schadt, Christopher Warren [ORNL; Brooks, Scott C [ORNL; Kostka, Joel [Florida State University

2011-01-01

The effect of long-term mixed-waste contamination, particularly uranium and nitrate, on the microbial community in the terrestrial subsurface was investigated at the field scale at the Oak Ridge Integrated Field Research Challenge (ORIFRC) site in Oak Ridge, TN. The abundance, community composition, and distribution of groundwater microorganisms were examined across the site during two seasonal sampling events. At representative locations, subsurface sediment was also examined from two boreholes, one sampled from the most heavily contaminated area of the site and another from an area with low contamination. A suite of DNA- and RNA-based molecular tools were employed for community characterization, including quantitative PCR of ribosomal RNA and nitrite reductase genes, community composition fingerprinting analysis, and high-throughput pyrotag sequencing of rRNA genes. The results demonstrate that pH is a major driver of the subsurface microbial community structure, and denitrifying bacteria from the genus Rhodanobacter (class Gammaproteobacteria) dominate at low pH. The relative abundance of bacteria from this genus was positively correlated with lower pH conditions, and these bacteria were abundant and active in the most highly contaminated areas. Other factors, such as concentration of nitrogen species, oxygen and sampling season did not appear to strongly influence the distribution of Rhodanobacter. Results indicate that these organisms are acid-tolerant denitrifiers, well suited to the acidic, nitrate-rich subsurface conditions, and pH is confirmed as a dominant driver of bacterial community structure in this contaminated subsurface environment.

Production and consumption of nitric oxide by three methanotrophic bacteria.

Science.gov (United States)

Ren, T; Roy, R; Knowles, R

2000-09-01

We studied nitrogen oxide production and consumption by methanotrophs Methylobacter luteus (group I), Methylosinus trichosporium OB3b (group II), and an isolate from a hardwood swamp soil, here identified by 16S ribosomal DNA sequencing as Methylobacter sp. strain T20 (group I). All could consume nitric oxide (nitrogen monoxide, NO), and produce small amounts of nitrous oxide (N(2)O). Only Methylobacter strain T20 produced large amounts of NO (>250 parts per million by volume [ppmv] in the headspace) at specific activities of up to 2.0 x 10(-17) mol of NO cell(-1) day(-1), mostly after a culture became O(2) limited. Production of NO by strain T20 occurred mostly in nitrate-containing medium under anaerobic or nearly anaerobic conditions, was inhibited by chlorate, tungstate, and O(2), and required CH(4). Denitrification (methanol-supported N(2)O production from nitrate in the presence of acetylene) could not be detected and thus did not appear to be involved in the production of NO. Furthermore, cd(1) and Cu nitrite reductases, NO reductase, and N(2)O reductase could not be detected by PCR amplification of the nirS, nirK, norB, and nosZ genes, respectively. M. luteus and M. trichosporium produced some NO in ammonium-containing medium under aerobic conditions, likely as a result of methanotrophic nitrification and chemical decomposition of nitrite. For Methylobacter strain T20, arginine did not stimulate NO production under aerobiosis, suggesting that NO synthase was not involved. We conclude that strain T20 causes assimilatory reduction of nitrate to nitrite, which then decomposes chemically to NO. The production of NO by methanotrophs such as Methylobacter strain T20 could be of ecological significance in habitats near aerobic-anaerobic interfaces where fluctuating O(2) and nitrate availability occur.

Observation of high seasonal variation in community structure of denitrifying bacteria in arable soil receiving artificial fertilizer and cattle manure by determining T-RFLP of nir gene fragments

DEFF Research Database (Denmark)

Priemé, Anders; Wolsing, Martin

2004-01-01

Temporal and spatial variation of communities of soil denitrifying bacteria at sites receiving mineral fertilizer (60 and 120 kg N ha-1 year-1) and cattle manure (75 and 150 kg N ha-1 year-1) were explored using terminal restriction fragment length polymorphism (T-RFLP) analyses of PCR amplified...... nitrite reductase (nirK and nirS) gene fragments. The analyses were done three times during the year: in March, July and October. nirK gene fragments could be amplified in all three months, whereas nirS gene fragments could be amplified only in March. Analysis of similarities in T-RFLP patterns revealed...... a significant seasonal shift in the community structure of nirK-containing bacteria. Also, sites treated with mineral fertilizer or cattle manure showed different communities of nirK-containing denitrifying bacteria, since the T-RFLP patterns of soils treated with these fertilizers were significantly different...

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.

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

Science.gov (United States)

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.

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

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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.

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

Science.gov (United States)

Sharp, Christine E; Stott, Matthew B; Dunfield, Peter F

2012-01-01

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 (13)CH(4)-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 labeling with (13)CO(2) and (13)CH(4), individually and in combination. Testing the protocol in "M. infernorum" strain V4 resulted in assimilation of (13)CO(2) but not (13)CH(4), verifying its autotrophic lifestyle. To specifically detect methanotrophs (as opposed to other autotrophs) via (13)CO(2)-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 (13)CH(4) + (12)CO(2) caused little shift in the density distribution of verrucomicrobial-pmoA genes relative to controls. However, labeling with (13)CO(2) in combination with (12)CH(4) or (13)CH(4) 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

Nitrogen source effects on the denitrifying anaerobic methane oxidation culture and anaerobic ammonium oxidation bacteria enrichment process.

Science.gov (United States)

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

2017-05-01

The co-culture system of denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (Anammox) has a potential application in wastewater treatment plant. This study explored the effects of permutation and combination of nitrate, nitrite, and ammonium on the culture enrichment from freshwater sediments. The co-existence of NO 3 - , NO 2 - , and NH 4 + shortened the enrichment time from 75 to 30 days and achieved a total nitrogen removal rate of 106.5 mg/L/day on day 132. Even though ammonium addition led to Anammox bacteria increase and a higher nitrogen removal rate, DAMO bacteria still dominated in different reactors with the highest proportion of 64.7% and the maximum abundance was 3.07 ± 0.25 × 10 8 copies/L (increased by five orders of magnitude) in the nitrite reactor. DAMO bacteria showed greater diversity in the nitrate reactor, and one was similar to M. oxyfera; DAMO bacteria in the nitrite reactor were relatively unified and similar to M. sinica. Interestingly, no DAMO archaea were found in the nitrate reactor. This study will improve the understanding of the impact of nitrogen source on DAMO and Anammox co-culture enrichment.

Niche differentiation in nitrogen metabolism among methanotrophs within an operational taxonomic unit.

Science.gov (United States)

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

2014-04-04

The currently accepted thesis on nitrogenous fertilizer additions on methane oxidation activity assumes niche partitioning among methanotrophic species, with activity responses to changes in nitrogen content being dependent on the in situ methanotrophic community structure Unfortunately, widely applied tools for microbial community assessment only have a limited phylogenetic resolution mostly restricted to genus level diversity, and not to species level as often mistakenly assumed. As a consequence, intragenus or intraspecies metabolic versatility in nitrogen metabolism was never evaluated nor considered among methanotrophic bacteria as a source of differential responses of methane oxidation to nitrogen amendments. We demonstrated that fourteen genotypically different Methylomonas strains, thus distinct below the level at which most techniques assign operational taxonomic units (OTU), show a versatile physiology in their nitrogen metabolism. Differential responses, even among strains with identical 16S rRNA or pmoA gene sequences, were observed for production of nitrite and nitrous oxide from nitrate or ammonium, nitrogen fixation and tolerance to high levels of ammonium, nitrate, and hydroxylamine. Overall, reduction of nitrate to nitrite, nitrogen fixation, higher tolerance to ammonium than nitrate and tolerance and assimilation of nitrite were general features. Differential responses among closely related methanotrophic strains to overcome inhibition and toxicity from high nitrogen loads and assimilation of various nitrogen sources yield competitive fitness advantages to individual methane-oxidizing bacteria. Our observations proved that community structure at the deepest phylogenetic resolution potentially influences in situ functioning.

Community Structure of Active Aerobic Methanotrophs in Red Mangrove (Kandelia obovata) Soils Under Different Frequency of Tides.

Science.gov (United States)

Shiau, Yo-Jin; Cai, Yuanfeng; Lin, Yu-Te; Jia, Zhongjun; Chiu, Chih-Yu

2018-04-01

Methanotrophs are important microbial communities in coastal ecosystems. They reduce CH 4 emission in situ, which is influenced by soil conditions. This study aimed to understand the differences in active aerobic methanotrophic communities in mangrove forest soils experiencing different inundation frequency, i.e., in soils from tidal mangroves, distributed at lower elevations, and from dwarf mangroves, distributed at higher elevations. Labeling of pmoA gene of active methanotrophs using DNA-based stable isotope probing (DNA-SIP) revealed that methanotrophic activity was higher in the dwarf mangrove soils than in the tidal mangrove soils, possibly because of the more aerobic soil conditions. Methanotrophs affiliated with the cluster deep-sea-5 belonging to type Ib methanotrophs were the most dominant methanotrophs in the fresh mangrove soils, whereas type II methanotrophs also appeared in the fresh dwarf mangrove soils. Furthermore, Methylobacter and Methylosarcina were the most important active methanotrophs in the dwarf mangrove soils, whereas Methylomonas and Methylosarcina were more active in the tidal mangrove soils. High-throughput sequencing of the 16S ribosomal RNA (rRNA) gene also confirmed similar differences in methanotrophic communities at the different locations. However, several unclassified methanotrophic bacteria were found by 16S rRNA MiSeq sequencing in both fresh and incubated mangrove soils, implying that methanotrophic communities in mangrove forests may significantly differ from the methanotrophic communities documented in previous studies. Overall, this study showed the feasibility of 13 CH 4 DNA-SIP to study the active methanotrophic communities in mangrove forest soils and revealed differences in the methanotrophic community structure between coastal mangrove forests experiencing different tide frequencies.

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

Science.gov (United States)

Lau, Evan; Nolan, Edward J.; Dillard, Zachary W.; Dague, Ryan D.; Semple, Amanda L.; Wentzell, Wendi L.

2015-01-01

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 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. PMID:27682082

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

Science.gov (United States)

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. Copyright © 2015 Elsevier Ltd. All rights reserved.

Nitrous oxide emission and denitrifier communities in drip-irrigated calcareous soil as affected by chemical and organic fertilizers.

Science.gov (United States)

Tao, Rui; Wakelin, Steven A; Liang, Yongchao; Hu, Baowei; Chu, Guixin

2018-01-15

The effects of consecutive application of chemical fertilizer with or without organic fertilizer on soil N 2 O emissions and denitrifying community structure in a drip-irrigated field were determined. The four fertilizer treatments were (i) unfertilized, (ii) chemical fertilizer, (iii) 60% chemical fertilizer plus cattle manure, and (iv) 60% chemical fertilizer plus biofertilizer. The treatments with organic amendments (i.e. cattle manure and biofertilizer) reduced cumulative N 2 O emissions by 4.9-9.9%, reduced the N 2 O emission factor by 1.3-42%, and increased denitrifying enzyme activities by 14.3-56.2%. The nirK gene copy numbers were greatest in soil which received only chemical fertilizer. In contrast, nirS- and nosZ-copy numbers were greatest in soil amended with chemical fertilizer plus biofertilizer. Chemical fertilizer application with or without organic fertilizer significantly changed the community structure of nirK-type denitrifiers relative to the unfertilized soil. In comparison, the nirS- and nosZ-type denitrifier genotypes varied in treatments receiving organic fertilizer but not chemical fertilizer alone. The changes in the denitrifier communities were closely associated with soil organic carbon (SOC), NO 3 - , NH 4 + , water holding capacity, and soil pH. Modeling indicated that N 2 O emissions in this soil were primarily associated with the abundance of nirS type denitrifying bacteria, SOC, and NO 3 - . Overall, our findings indicate that (i) the organic fertilizers increased denitrifying enzyme activity, increased denitrifying-bacteria gene copy numbers, but reduced N 2 O emissions, and (ii) nirS- and nosZ-type denitrifiers were more sensitive than nirK-type denitrifiers to the organic fertilizers. Copyright © 2017. Published by Elsevier B.V.

Amplification of marine methanotrophic enrichment DNA with 16S rDNA PCR primers for type II alpha proteobacteria methanotrophs.

Science.gov (United States)

Rockne, Karl J; Strand, Stuart E

2003-09-01

Type II alpha proteobacteria methanotrophs are capable of a wide range of cometabolic transformations of chlorinated solvents and polycyclic aromatic hydrocarbons (PAHs), and this activity has been exploited in many terrestrial bioremediation systems. However, at present, all known obligately marine methanotrophic isolates are Type I gamma proteobacteria which do not have this activity to the extent of Type II methanotrophs. In previous work in our laboratory, determining the presence of Type II alpha proteobacteria methanotrophs in marine enrichment cultures that co-metabolized PAHs required a more sensitive assay. 16S rDNA PCR primers were designed based on oligonucleotide probes for serine pathway methanotrophs and serine pathway methylotrophs with an approximate amplification fragment size of 870 base pairs. Comparison of the primers using double primer BLAST searches in established nucleotide databases showed potential amplification with all Methylocystis and Methylosinus spp., as well as potential amplification with Methylocella palustrus. DNA from Methylosinus trichosporium OB3b, a Type II methanotroph, amplified with the primers with a fragment size of approximately 850 base pairs, whereas DNA extracted from Methylomonas methanica, a Type I methanotroph, did not. The primers were used to amplify DNA extracted from two marine methanotrophic enrichment cultures: a low nitrogen/low copper enrichment to select for Type II methanotrophs and a high nitrogen/high copper enrichment to select for Type I methanotrophs. Although DNA from both cultures amplified with the PCR primers, amplification was stronger in cultures that were specifically enriched for Type II methanotrophs, suggesting the presence of higher numbers of Type II methanotrophs. These results provide further evidence for the existence of Type II marine methanotrophs, suggesting the possibility of exploiting cometabolic activity in marine systems.

Identification of the autotrophic denitrifying community in nitrate removal reactors by DNA-stable isotope probing.

Science.gov (United States)

Xing, Wei; Li, Jinlong; Cong, Yuan; Gao, Wei; Jia, Zhongjun; Li, Desheng

2017-04-01

Autotrophic denitrification has attracted increasing attention for wastewater with insufficient organic carbon sources. Nevertheless, in situ identification of autotrophic denitrifying communities in reactors remains challenging. Here, a process combining micro-electrolysis and autotrophic denitrification with high nitrate removal efficiency was presented. Two batch reactors were fed organic-free nitrate influent, with H 13 CO 3 - and H 12 CO 3 - as inorganic carbon sources. DNA-based stable-isotope probing (DNA-SIP) was used to obtain molecular evidence for autotrophic denitrifying communities. The results showed that the nirS gene was strongly labeled by H 13 CO 3 - , demonstrating that the inorganic carbon source was assimilated by autotrophic denitrifiers. High-throughput sequencing and clone library analysis identified Thiobacillus-like bacteria as the most dominant autotrophic denitrifiers. However, 88% of nirS genes cloned from the 13 C-labeled "heavy" DNA fraction showed low similarity with all culturable denitrifiers. These findings provided functional and taxonomical identification of autotrophic denitrifying communities, facilitating application of autotrophic denitrification process for wastewater treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

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

DEFF Research Database (Denmark)

Mcllroy, Simon; Starnawska, Anna; Starnawski, Piotr

2015-01-01

Denitrification is essential to the removal of nitrogen from wastewater during treatment, yet an understanding of the diversity of the active denitrifying bacteria responsible in full-scale wastewater treatment plants (WWTPs) is lacking. In this study, stable-isotope probing (SIP) was applied......-labelled complex substrate was used for SIP incubations, under nitrite-reducing conditions, in order to maximize the capture of the potentially metabolically diverse denitrifiers likely present. Members of the Rhodoferax, Dechloromonas, Sulfuritalea, Haliangium and Thermomonas were represented in the 16S rRNA gene...

Differential Transcriptional Activation of Genes Encoding Soluble Methane Monooxygenase in a Facultative Versus an Obligate Methanotroph.

Science.gov (United States)

Smirnova, Angela V; Dunfield, Peter F

2018-03-06

Methanotrophs are a specialized group of bacteria that can utilize methane (CH₄) as a sole energy source. A key enzyme responsible for methane oxidation is methane monooxygenase (MMO), of either a soluble, cytoplasmic type (sMMO), or a particulate, membrane-bound type (pMMO). Methylocella silvestris BL2 and Methyloferula stellata AR4 are closely related methanotroph species that oxidize methane via sMMO only. However, Methyloferula stellata is an obligate methanotroph, while Methylocella silvestris is a facultative methanotroph able to grow on several multicarbon substrates in addition to methane. We constructed transcriptional fusions of the mmo promoters of Methyloferula stellata and Methylocella silvestris to a promoterless gfp in order to compare their transcriptional regulation in response to different growth substrates, in the genetic background of both organisms. The following patterns were observed: (1) The mmo promoter of the facultative methanotroph Methylocella silvestris was either transcriptionally downregulated or repressed by any growth substrate other than methane in the genetic background of Methylocella silvetris ; (2) Growth on methane alone upregulated the mmo promoter of Methylocella silvetris in its native background but not in the obligate methanotroph Methyloferula stellata ; (3) The mmo promoter of Methyloferula stellata was constitutive in both organisms regardless of the growth substrate, but with much lower promoter activity than the mmo promoter of Methylocella silvetris . These results support a conclusion that a different mode of transcriptional regulation of sMMO contributes to the facultative lifestyle of Methylocella silvetris compared to the obligate methanotroph Methyloferula stellata .

Resilience of (seed bank) aerobic methanotrophs and methanotrophic activity to desiccation and heat stress.

NARCIS (Netherlands)

Ho, A.; Lueke, C.; Reim, A.; Frenzel, P.

2016-01-01

In seasonally changing environments, aerobic methanotrophs are exposed to elevated temperatures and drought. Prior exposure to adverse conditions (site history) may leave an imprint on the methanotrophic community composition in the form of a seed bank. While the significance of a microbial seed

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

Differential Transcriptional Activation of Genes Encoding Soluble Methane Monooxygenase in a Facultative Versus an Obligate Methanotroph

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Angela V. Smirnova

2018-03-01

Full Text Available Methanotrophs are a specialized group of bacteria that can utilize methane (CH4 as a sole energy source. A key enzyme responsible for methane oxidation is methane monooxygenase (MMO, of either a soluble, cytoplasmic type (sMMO, or a particulate, membrane-bound type (pMMO. Methylocella silvestris BL2 and Methyloferula stellata AR4 are closely related methanotroph species that oxidize methane via sMMO only. However, Methyloferula stellata is an obligate methanotroph, while Methylocella silvestris is a facultative methanotroph able to grow on several multicarbon substrates in addition to methane. We constructed transcriptional fusions of the mmo promoters of Methyloferula stellata and Methylocella silvestris to a promoterless gfp in order to compare their transcriptional regulation in response to different growth substrates, in the genetic background of both organisms. The following patterns were observed: (1 The mmo promoter of the facultative methanotroph Methylocella silvestris was either transcriptionally downregulated or repressed by any growth substrate other than methane in the genetic background of Methylocella silvetris; (2 Growth on methane alone upregulated the mmo promoter of Methylocella silvetris in its native background but not in the obligate methanotroph Methyloferula stellata; (3 The mmo promoter of Methyloferula stellata was constitutive in both organisms regardless of the growth substrate, but with much lower promoter activity than the mmo promoter of Methylocella silvetris. These results support a conclusion that a different mode of transcriptional regulation of sMMO contributes to the facultative lifestyle of Methylocella silvetris compared to the obligate methanotroph Methyloferula stellata.

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

Science.gov (United States)

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

2017-01-01

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

Microbial characterization of toluene-degrading denitrifying consortia obtained from terrestrial and marine ecosystems.

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An, Y-J; Joo, Y-H; Hong, I-Y; Ryu, H-W; Cho, K-S

2004-10-01

The degradation characteristics of toluene coupled to nitrate reduction were investigated in enrichment culture and the microbial communities of toluene-degrading denitrifying consortia were characterized by denaturing gradient gel electrophoresis (DGGE) technique. Anaerobic nitrate-reducing bacteria were enriched from oil-contaminated soil samples collected from terrestrial (rice field) and marine (tidal flat) ecosystems. Enriched consortia degraded toluene in the presence of nitrate as a terminal electron acceptor. The degradation rate of toluene was affected by the initial substrate concentration and co-existence of other hydrocarbons. The types of toluene-degrading denitrifying consortia depended on the type of ecosystem. The clone RS-7 obtained from the enriched consortium of the rice field was most closely related to a toluene-degrading and denitrifying bacterium, Azoarcus denitrificians (A. tolulyticus sp. nov.). The clone TS-11 detected in the tidal flat enriched consortium was affiliated to Thauera sp. strain S2 (T. aminoaromatica sp. nov.) that was able to degrade toluene under denitrifying conditions. This indicates that environmental factors greatly influence microbial communities obtained from terrestrial (rice field) and marine (tidal flat) ecosystems.

Feasibility of atmospheric methane removal using methanotrophic biotrickling filters.

Science.gov (United States)

Yoon, Sukhwan; Carey, Jeffrey N; Semrau, Jeremy D

2009-07-01

Methane is a potent greenhouse gas with a global warming potential ~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(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(2) credits is increased, can also be economically attractive.

Oxidation of methane in biotrickling filters inoculated with methanotrophic bacteria.

Science.gov (United States)

Cáceres, Manuel; Dorado, Antonio D; Gentina, Juan C; Aroca, Germán

2017-11-01

The oxidation of methane (CH 4 ) using biofilters has been proposed as an alternative to mitigate anthropogenic greenhouse gas emissions with a low concentration of CH 4 that cannot be used as a source of energy. However, conventional biofilters utilize organic packing materials that have a short lifespan, clogging problems, and are commonly inoculated with non-specific microorganisms leading to unpredictable CH 4 elimination capacities (EC) and removal efficiencies (RE). The main objective of this work was to characterize the oxidation of CH 4 in two biotrickling filters (BTFs) packed with polyethylene rings and inoculated with two methanotrophic bacteria, Methylomicrobium album and Methylocystis sp., in order to determine EC and CO 2 production (pCO 2 ) when using a specific inoculum. The repeatability of the results in both BTFs was determined when they operated at the same inlet load of CH 4 . A dynamic mathematical model that describes the CH 4 abatement in the BTFs was developed and validated using mass transfer and kinetic parameters estimated independently. The results showed that EC and pCO 2 of the BTFs are not identical but very similar for all the conditions tested. The use of specific inoculum has shown a faster startup and higher EC per unit area (0.019 gCH 4  m -2  h -1 ) in comparison to most of the previous studies at the same CH 4 load rate (23.2 gCH 4  m -3  h -1 ). Global mass balance showed that the maximum reduction of CO 2 equivalents was 98.5 gCO 2eq  m -3  h -1 . The developed model satisfactorily described CH 4 abatement in BTFs for a wide range of conditions.

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 13CH4 and incubated under ambient and elevated ammonium concentrations. After 7 days, methanotroph activity was assessed by

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

International Nuclear Information System (INIS)

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

2016-01-01

Highlights: • It is the first time to evaluate the effect of Cu"2"+ on denitrifying biogranules. • A high level of Cu(II) was investigated during batch assays and continuous tests. • Mechanisms of the effects of Cu"2"+ on denitrifying biogranules were discussed. • Effects of pre-exposure to Cu"2"+ and starvation treatments were investigated. - Abstract: This study aimed to investigate the short- and long-term effects of Cu"2"+ on the activity and performance of denitrifying bacteria. The short-term effects of various concentrations of Cu"2"+ on the denitrifying bacteria were evaluated using batch assays. The specific denitrifying activity (SDA) decreased from 14.3 ± 2.2 (without Cu"2"+) to 6.1 ± 0.1 mg N h"−"1 g"−"1 VSS (100 mgCu"2"+ L"−"1) when Cu"2"+ increased from 0 to 100 mg L"−"1 with an increment of 10 mgCu"2"+ L"−"1. A non-competitive inhibition model was used to calculate the 50% inhibition concentration (IC_5_0) of Cu"2"+ on denitrifying sludge (30.6 ± 2.5 mg L"−"1). Monod and Luong models were applied to investigate the influence of the initial substrate concentration, and the results suggested that the maximum substrate removal rate would be reduced with Cu"2"+ supplementation. Pre-exposure to Cu"2"+ could lead to an 18.2–46.2% decrease in the SDA and decreasing percentage of the SDA increased with both exposure time and concentration. In the continuous-flow test, Cu"2"+ concentration varied from 1 to 75 mg L"−"1; however, no clear deterioration was observed in the reactor, and the reactor was kept stable, with the total nitrogen removal efficiency and total organic carbon efficiency greater than 89.0 and 85.0%, respectively. The results demonstrated the short-term inhibition of Cu"2"+ upon denitrification, and no notable adversity was observed during the continuous-flow test after long-term acclimation.

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

KAUST Repository

Meulepas, Roel J W; Jagersma, Christian G; Khadem, Ahmad F; Stams, Alfons J M; Lens, Piet N L

2010-01-01

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

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

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

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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.

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.

Oxygen-limited metabolism in the methanotroph Methylomicrobium buryatense 5GB1C

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Alexey Gilman

2017-10-01

Full Text Available The bacteria that grow on methane aerobically (methanotrophs support populations of non-methanotrophs in the natural environment by excreting methane-derived carbon. One group of excreted compounds are short-chain organic acids, generated in highest abundance when cultures are grown under O2-starvation. We examined this O2-starvation condition in the methanotroph Methylomicrobium buryatense 5GB1. The M. buryatense 5GB1 genome contains homologs for all enzymes necessary for a fermentative metabolism, and we hypothesize that a metabolic switch to fermentation can be induced by low-O2 conditions. Under prolonged O2-starvation in a closed vial, this methanotroph increases the amount of acetate excreted about 10-fold, but the formate, lactate, and succinate excreted do not respond to this culture condition. In bioreactor cultures, the amount of each excreted product is similar across a range of growth rates and limiting substrates, including O2-limitation. A set of mutants were generated in genes predicted to be involved in generating or regulating excretion of these compounds and tested for growth defects, and changes in excretion products. The phenotypes and associated metabolic flux modeling suggested that in M. buryatense 5GB1, formate and acetate are excreted in response to redox imbalance. Our results indicate that even under O2-starvation conditions, M. buryatense 5GB1 maintains a metabolic state representing a combination of fermentation and respiration metabolism.

Metagenomic identification of active methanogens and methanotrophs in serpentinite springs of the Voltri Massif, Italy

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William J. Brazelton

2017-01-01

Full Text Available The production of hydrogen and methane by geochemical reactions associated with the serpentinization of ultramafic rocks can potentially support subsurface microbial ecosystems independent of the photosynthetic biosphere. Methanogenic and methanotrophic microorganisms are abundant in marine hydrothermal systems heavily influenced by serpentinization, but evidence for methane-cycling archaea and bacteria in continental serpentinite springs has been limited. This report provides metagenomic and experimental evidence for active methanogenesis and methanotrophy by microbial communities in serpentinite springs of the Voltri Massif, Italy. Methanogens belonging to family Methanobacteriaceae and methanotrophic bacteria belonging to family Methylococcaceae were heavily enriched in three ultrabasic springs (pH 12. Metagenomic data also suggest the potential for hydrogen oxidation, hydrogen production, carbon fixation, fermentation, and organic acid metabolism in the ultrabasic springs. The predicted metabolic capabilities are consistent with an active subsurface ecosystem supported by energy and carbon liberated by geochemical reactions within the serpentinite rocks of the Voltri Massif.

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

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Javier Vilcáez

2009-01-01

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

Termites facilitate methane oxidation and shape the methanotrophic community.

Science.gov (United States)

Ho, Adrian; Erens, Hans; Mujinya, Basile Bazirake; Boeckx, Pascal; Baert, Geert; Schneider, Bellinda; Frenzel, Peter; Boon, Nico; Van Ranst, Eric

2013-12-01

Termite-derived methane contributes 3 to 4% to the total methane budget globally. Termites are not known to harbor methane-oxidizing microorganisms (methanotrophs). However, a considerable fraction of the methane produced can be consumed by methanotrophs that inhabit the mound material, yet the methanotroph ecology in these environments is virtually unknown. The potential for methane oxidation was determined using slurry incubations under conditions with high (12%) and in situ (∼0.004%) methane concentrations through a vertical profile of a termite (Macrotermes falciger) mound and a reference soil. Interestingly, the mound material showed higher methanotrophic activity. The methanotroph community structure was determined by means of a pmoA-based diagnostic microarray. Although the methanotrophs in the mound were derived from populations in the reference soil, it appears that termite activity selected for a distinct community. Applying an indicator species analysis revealed that putative atmospheric methane oxidizers (high-indicator-value probes specific for the JR3 cluster) were indicative of the active nest area, whereas methanotrophs belonging to both type I and type II were indicative of the reference soil. We conclude that termites modify their environment, resulting in higher methane oxidation and selecting and/or enriching for a distinct methanotroph population.

Termites facilitate methane oxidation and shape the methanotrophic community

NARCIS (Netherlands)

Ho, A.; Erens, H.; Mujinya, B.B.; Boeckx, P.; Baert, G.; Schneider, B.; Frenzel, P.; Boon, N.; Van Ranst, E.

2013-01-01

Termite-derived methane contributes 3-4% to the total methane budget globally. Termites are not known to harbor methane-oxidizing microorganisms (methanotrophs). However, a considerable fraction of methane produced can be consumed by methanotrophs that inhabit the mound material. Yet, methanotroph

Solubility of methane in water and in a medium for the cultivation of methanotrophs bacteria

International Nuclear Information System (INIS)

Serra, Maria Celeste C.; Pessoa, F.L.P.; Palavra, A.M.F.

2006-01-01

Solubility of methane in water and in an aqueous growth medium for the cultivation of methanotrophs bacteria was determined over the temperature range 293.15 to 323.15 K and at atmospheric pressure. The measurements were carried out in a Ben-Naim/Baer type apparatus with a precision of about ±0.3%. The experimental results were determined using accurate thermodynamic relations. The mole fractions of the dissolved gas at the gas partial pressure of 101.325 kPa, the Henry coefficients at the water vapour pressure and the Ostwald coefficients at infinite dilution were obtained. A comparison between the solubility of methane in water and those observed in fermentation medium over the temperature range of 298.15 to 308.15 K has shown that this gas is about ±2.3% more soluble in water. The temperature dependence of the mole fractions of methane was also determined using the Clarke-Glew-Weiss equation and the thermodynamic quantities, Gibbs energy, enthalpy and entropy changes, associated with the dissolution process were calculated. Furthermore, the experimental Henry coefficients for methane in water are compared with those calculated by the scaled particle theory. The estimated Henry coefficients are about ±4% lower than the experimental ones

Direct immunofluorescence and enzyme-linked immunosorbent assays for evaluating chlorinated hydrocarbon degrading bacteria

Energy Technology Data Exchange (ETDEWEB)

Brigmon, R.L.; Franck, M.M.; Brey, J.; Fliermans, C.B. [Westinghouse Savannah River, Aiken, SC (United States). Environmental Biotechnology Section; Scott, D.; Lanclos, K. [Medical Coll. of Georgia, Augusta, GA (United States)

1997-06-01

Immunological procedures were developed to enumerate chlorinated hydrocarbon degrading bacteria. Polyclonal antibodies (Pabs) were produced by immunizing New Zealand white rabbits against 18 contaminant-degrading bacteria. These included methanotrophic and chlorobenzene (CB) degrading species. An enzyme-linked immunosorbent assay (ELISA) was used to test for specificity and sensitivity of the Pabs. Direct fluorescent antibodies (DFAs) were developed with these Pabs against select methanotrophic bacteria isolated from a trichloroethylene (TCE) contaminated landfill at the Savannah River Site (SRS) and cultures from the American Type Culture Collection (ATCC). Analysis of cross reactivity testing data showed some of the Pabs to be group specific while others were species specific. The threshold of sensitivity for the ELISA is 105 bacteria cells/ml. The DFA can detect as few as one bacterium per ml after concentration. Results from the DFA and ELISA techniques for enumeration of methanotrophic bacteria in groundwater were higher but not significantly different (P < 0.05) compared to indirect microbiological techniques such as MPN. These methods provide useful information on in situ community structure and function for bioremediation applications within 1--4 hours of sampling.

Microbial Community Composition and Ultrastructure of Granules from a Full-Scale Anammox Reactor

KAUST Repository

Gonzalez-Gil, Graciela

2014-12-11

Granules in anammox reactors contain besides anammox bacteria other microbial communities whose identity and relationship with the anammox bacteria are not well understood. High calcium concentrations are often supplied to anammox reactors to obtain sufficient bacterial aggregation and biomass retention. The aim of this study was to provide the first characterization of bacterial and archaeal communities in anammox granules from a full-scale anammox reactor and to explore on the possible role of calcium in such aggregates. High magnification imaging using backscattered electrons revealed that anammox bacteria may be embedded in calcium phosphate precipitates. Pyrosequencing of 16S rRNA gene fragments showed, besides anammox bacteria (Brocadiacea, 32 %), substantial numbers of heterotrophic bacteria Ignavibacteriacea (18 %) and Anaerolinea (7 %) along with heterotrophic denitrifiers Rhodocyclacea (9 %), Comamonadacea (3 %), and Shewanellacea (3 %) in the granules. It is hypothesized that these bacteria may form a network in which heterotrophic denitrifiers cooperate to achieve a well-functioning denitrification system as they can utilize the nitrate intrinsically produced by the anammox reaction. This network may provide a niche for the proliferation of archaea. Hydrogenotrophic methananogens, which scavenge the key fermentation product H2, were the most abundant archaea detected. Cells resembling the polygon-shaped denitrifying methanotroph Candidatus Methylomirabilis oxyfera were observed by electron microscopy. It is hypothesized that the anammox process in a full-scale reactor triggers various reactions overall leading to efficient denitrification and a sink of carbon as biomass in anammox granules.

Microbial community composition and ultrastructure of granules from a full-scale anammox reactor.

Science.gov (United States)

Gonzalez-Gil, Graciela; Sougrat, Rachid; Behzad, Ali R; Lens, Piet N L; Saikaly, Pascal E

2015-07-01

Granules in anammox reactors contain besides anammox bacteria other microbial communities whose identity and relationship with the anammox bacteria are not well understood. High calcium concentrations are often supplied to anammox reactors to obtain sufficient bacterial aggregation and biomass retention. The aim of this study was to provide the first characterization of bacterial and archaeal communities in anammox granules from a full-scale anammox reactor and to explore on the possible role of calcium in such aggregates. High magnification imaging using backscattered electrons revealed that anammox bacteria may be embedded in calcium phosphate precipitates. Pyrosequencing of 16S rRNA gene fragments showed, besides anammox bacteria (Brocadiacea, 32%), substantial numbers of heterotrophic bacteria Ignavibacteriacea (18%) and Anaerolinea (7%) along with heterotrophic denitrifiers Rhodocyclacea (9%), Comamonadacea (3%), and Shewanellacea (3%) in the granules. It is hypothesized that these bacteria may form a network in which heterotrophic denitrifiers cooperate to achieve a well-functioning denitrification system as they can utilize the nitrate intrinsically produced by the anammox reaction. This network may provide a niche for the proliferation of archaea. Hydrogenotrophic methananogens, which scavenge the key fermentation product H2, were the most abundant archaea detected. Cells resembling the polygon-shaped denitrifying methanotroph Candidatus Methylomirabilis oxyfera were observed by electron microscopy. It is hypothesized that the anammox process in a full-scale reactor triggers various reactions overall leading to efficient denitrification and a sink of carbon as biomass in anammox granules.

Submerged macrophytes shape the abundance and diversity of bacterial denitrifiers in bacterioplankton and epiphyton in the Shallow Fresh Lake Taihu, China.

Science.gov (United States)

Fan, Zhou; Han, Rui-Ming; Ma, Jie; Wang, Guo-Xiang

2016-07-01

nirK and nirS genes are important functional genes involved in the denitrification pathway. Recent studies about these two denitrifying genes are focusing on sediment and wastewater microbe. In this study, we conducted a comparative analysis of the abundance and diversity of denitrifiers in the epiphyton of submerged macrophytes Potamogeton malaianus and Ceratophyllum demersum as well as in bacterioplankton in the shallow fresh lake Taihu, China. Results showed that nirK and nirS genes had significant different niches in epiphyton and bacterioplankton. Bacterioplankton showed greater abundance of nirK gene in terms of copy numbers and lower abundance of nirS gene. Significant difference in the abundance of nirK and nirS genes also existed between the epiphyton from different submerged macrophytes. Similar community diversity yet different community abundance was observed between epiphytic bacteria and bacterioplankton. No apparent seasonal variation was found either in epiphytic bacteria or bacterioplankton; however, environmental parameters seemed to have direct relevancy with nirK and nirS genes. Our study suggested that submerged macrophytes have greater influence than seasonal parameters in shaping the presence and abundance of bacterial denitrifiers. Further investigation needs to focus on the potential contact and relative contribution between denitrifiers and environmental factors.

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.

Metaproteomic identification of diazotrophic methanotrophs and their localization in root tissues of field-grown rice plants.

Science.gov (United States)

Bao, Zhihua; Okubo, Takashi; Kubota, Kengo; Kasahara, Yasuhiro; Tsurumaru, Hirohito; Anda, Mizue; Ikeda, Seishi; Minamisawa, Kiwamu

2014-08-01

In a previous study by our group, CH4 oxidation and N2 fixation were simultaneously activated in the roots of wild-type rice plants in a paddy field with no N input; both processes are likely controlled by a rice gene for microbial symbiosis. The present study examined which microorganisms in rice roots were responsible for CH4 oxidation and N2 fixation under the field conditions. Metaproteomic analysis of root-associated bacteria from field-grown rice (Oryza sativa Nipponbare) revealed that nitrogenase complex-containing nitrogenase reductase (NifH) and the alpha subunit (NifD) and beta subunit (NifK) of dinitrogenase were mainly derived from type II methanotrophic bacteria of the family Methylocystaceae, including Methylosinus spp. Minor nitrogenase proteins such as Methylocella, Bradyrhizobium, Rhodopseudomonas, and Anaeromyxobacter were also detected. Methane monooxygenase proteins (PmoCBA and MmoXYZCBG) were detected in the same bacterial group of the Methylocystaceae. Because these results indicated that Methylocystaceae members mediate both CH4 oxidation and N2 fixation, we examined their localization in rice tissues by using catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). The methanotrophs were localized around the epidermal cells and vascular cylinder in the root tissues of the field-grown rice plants. Our metaproteomics and CARD-FISH results suggest that CH4 oxidation and N2 fixation are performed mainly by type II methanotrophs of the Methylocystaceae, including Methylosinus spp., inhabiting the vascular bundles and epidermal cells of rice roots. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Methanotrophic abundance and community fingerprint in pine and ...

African Journals Online (AJOL)

methanotrophs) is important to assess the microbial oxidation of the greenhouse gas methane (CH4) in soil under different land uses. Soil samples were collected from two plantation plots of pine and tea in southern China. Methanotrophic abundance ...

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

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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.

NifH and NifD phylogenies: an evolutionary basis for understanding nitrogen fixation capabilities of methanotrophic bacteria.

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Dedysh, Svetlana N; Ricke, Peter; Liesack, Werner

2004-05-01

The ability to utilize dinitrogen as a nitrogen source is an important phenotypic trait in most currently known methanotrophic bacteria (MB). This trait is especially important for acidophilic MB, which inhabit acidic oligotrophic environments, highly depleted in available nitrogen compounds. Phylogenetically, acidophilic MB are most closely related to heterotrophic dinitrogen-fixing bacteria of the genus BEIJERINCKIA: To further explore the phylogenetic linkage between these metabolically different organisms, the sequences of nifH and nifD gene fragments from acidophilic MB of the genera Methylocella and Methylocapsa, and from representatives of Beijerinckia, were determined. For reference, nifH and nifD sequences were also obtained from some type II MB of the alphaproteobacterial Methylosinus/Methylocystis group and from gammaproteobacterial type I MB. The trees constructed for the inferred amino acid sequences of nifH and nifD were highly congruent. The phylogenetic relationships among MB in the NifH and NifD trees also agreed well with the corresponding 16S rRNA-based phylogeny, except for two distinctive features. First, different methods used for phylogenetic analysis grouped the NifH and NifD sequences of strains of the gammaproteobacterial MB Methylococcus capsulatus within a clade mainly characterized by Alphaproteobacteria, including acidophilic MB and type II MB of the Methylosinus/Methylocystis group. From this and other genomic data from Methylococcus capsulatus Bath, it is proposed that an ancient event of lateral gene transfer was responsible for this aberrant branching. Second, the identity values of NifH and NifD sequences between Methylocapsa acidiphila B2 and representatives of Beijerinckia were clearly higher (98.5 and 96.6 %, respectively) than would be expected from their 16S rRNA-based relationships. Possibly, these two bacteria originated from a common acidophilic dinitrogen-fixing ancestor, and were subject to similar evolutionary pressure

Relationships Between Denitrifier Abundance, Denitrifier Diversity and Denitrification in Gulf of Mexico Hypoxic Zone Sediments.

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Proctor, L. M.; Childs, C.; MacAuley, S.

2002-12-01

The largest zone of anthropogenic bottom water hypoxia in the Western Hemisphere occurs seasonally in the northern Gulf of Mexico. This hypoxic zone reaches its greatest extent in the summer months and is a consequence of seasonal stratification of the water column combined with the decomposition of organic matter derived from accelerated rates of primary production. The enhanced primary production is driven by inorganic nitrogen input from the Mississippi River and these conditions would seem ideal for supporting high levels of denitrification. Yet sediment denitrification exhibited a wide range, even at the height of the seasonal hypoxia. Therefore, we compared benthic denitrifier abundances and denitrifier diversity at several stations over two seasons exhibiting extremes in denitrification to evaluate the relationship between abundances, diversity and denitrification levels. Sediment denitrification ranged from 20 to 100 umol m-2 h-1, with rates in July, 2000 approximately half that observed in July, 2001. The highest rates were generally observed at stations with bottom water DO concentrations between 1 and 3 mg l-1. Relative denitrifier abundances, using nirS and nirK as proxies for denitrifiers, suggested a direct relationship between abundances and denitrification while denitrifier diversity, measured by T-RFLPs of nirS and nirK, suggested an inverse relationship between diversity and denitrification. These results suggest that several factors are important in understanding what controls denitrification in Gulf of Mexico hypoxic zone sediments.

Continuously Monocropped Jerusalem Artichoke Changed Soil Bacterial Community Composition and Ammonia-Oxidizing and Denitrifying Bacteria Abundances.

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Zhou, Xingang; Wang, Zhilin; Jia, Huiting; Li, Li; Wu, Fengzhi

2018-01-01

Soil microbial communities have profound effects on the growth, nutrition and health of plants in agroecosystems. Understanding soil microbial dynamics in cropping systems can assist in determining how agricultural practices influence soil processes mediated by microorganisms. In this study, soil bacterial communities were monitored in a continuously monocropped Jerusalem artichoke (JA) system, in which JA was successively monocropped for 3 years in a wheat field. Soil bacterial community compositions were estimated by amplicon sequencing of the 16S rRNA gene. Abundances of ammonia-oxidizing and denitrifying bacteria were estimated by quantitative PCR analysis of the amoA , nirS , and nirK genes. Results showed that 1-2 years of monocropping of JA did not significantly impact the microbial alpha diversity, and the third cropping of JA decreased the microbial alpha diversity ( P < 0.05). Principal coordinates analysis and permutational multivariate analysis of variance analyses revealed that continuous monocropping of JA changed soil bacterial community structure and function profile ( P < 0.001). At the phylum level, the wheat field was characterized with higher relative abundances of Latescibacteria , Planctomycetes , and Cyanobacteria , the first cropping of JA with Actinobacteria , the second cropping of JA with Acidobacteria , Armatimonadetes , Gemmatimonadetes , and Proteobacteria . At the genus level, the first cropping of JA was enriched with bacterial species with pathogen-antagonistic and/or plant growth promoting potentials, while members of genera that included potential denitrifiers increased in the second and third cropping of JA. The first cropping of JA had higher relative abundances of KO terms related to lignocellulose degradation and phosphorus cycling, the second cropping of JA had higher relative abundances of KO terms nitrous-oxide reductase and nitric-oxide reductase, and the third cropping of JA had higher relative abundances of KO terms

Continuously Monocropped Jerusalem Artichoke Changed Soil Bacterial Community Composition and Ammonia-Oxidizing and Denitrifying Bacteria Abundances

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Xingang Zhou

2018-04-01

Full Text Available Soil microbial communities have profound effects on the growth, nutrition and health of plants in agroecosystems. Understanding soil microbial dynamics in cropping systems can assist in determining how agricultural practices influence soil processes mediated by microorganisms. In this study, soil bacterial communities were monitored in a continuously monocropped Jerusalem artichoke (JA system, in which JA was successively monocropped for 3 years in a wheat field. Soil bacterial community compositions were estimated by amplicon sequencing of the 16S rRNA gene. Abundances of ammonia-oxidizing and denitrifying bacteria were estimated by quantitative PCR analysis of the amoA, nirS, and nirK genes. Results showed that 1–2 years of monocropping of JA did not significantly impact the microbial alpha diversity, and the third cropping of JA decreased the microbial alpha diversity (P < 0.05. Principal coordinates analysis and permutational multivariate analysis of variance analyses revealed that continuous monocropping of JA changed soil bacterial community structure and function profile (P < 0.001. At the phylum level, the wheat field was characterized with higher relative abundances of Latescibacteria, Planctomycetes, and Cyanobacteria, the first cropping of JA with Actinobacteria, the second cropping of JA with Acidobacteria, Armatimonadetes, Gemmatimonadetes, and Proteobacteria. At the genus level, the first cropping of JA was enriched with bacterial species with pathogen-antagonistic and/or plant growth promoting potentials, while members of genera that included potential denitrifiers increased in the second and third cropping of JA. The first cropping of JA had higher relative abundances of KO terms related to lignocellulose degradation and phosphorus cycling, the second cropping of JA had higher relative abundances of KO terms nitrous-oxide reductase and nitric-oxide reductase, and the third cropping of JA had higher relative abundances of KO

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

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

2015-08-01

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

Effects of mechanical disintegration of activated sludge on the activity of nitrifying and denitrifying bacteria and phosphorus accumulating organisms.

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Zubrowska-Sudol, Monika; Walczak, Justyna

2014-09-15

The purpose of the study was to analyse the impact of hydrodynamic disintegration of thickened excess activated sludge, performed at different levels of energy density (70, 140 and 210 kJ/L), on the activity of microorganisms involved in nutrient removal from wastewater, i.e. nitrifiers, denitrifiers and phosphorus accumulating organisms (PAOs). Ammonium and nitrogen utilisation rates and phosphorus release rates for raw and disintegrated sludge were determined using batch tests. The experiment also included: 1) analysis of organic and nutrient compound release from activated sludge flocs, 2) determination of the sludge disintegration degree (DD), and 3) evaluation of respiratory activity of the biomass by using the oxygen uptake rate (OUR) batch test. It was shown that the activity degree of the examined groups of microorganisms depended on energy density and related sludge disintegration degree, and that inactivation of individual groups of microorganisms occurred at different values of DD. Least resistant to the destruction of activated sludge flocs turned out to be phosphorus accumulating organisms, while the most resistant were denitrifiers. A decrease of 20-40% in PAO activity was noted already at DD equal to 3-5%. The threshold values of DD, after crossing which the inactivation of nitrifiers and denitrifiers occurred, were equal to 8% and 10%, respectively. At lesser DD values an increase in the activity of these groups of microorganisms was observed, averaging 20.2-41.7% for nitrifiers and 9.98-36.3% for denitrifiers. Copyright © 2014 Elsevier Ltd. All rights reserved.

The genes and enzymes of sucrose metabolism in moderately thermophilic methanotroph Methylocaldum szegediense O12.

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But, Sergey Y; Solntseva, Natalia P; Egorova, Svetlana V; Mustakhimov, Ildar I; Khmelenina, Valentina N; Reshetnikov, Alexander; Trotsenko, Yuri A

2018-05-01

Four enzymes involved in sucrose metabolism: sucrose phosphate synthase (Sps), sucrose phosphate phosphatase (Spp), sucrose synthase (Sus) and fructokinase (FruK), were obtained as his-tagged proteins from the moderately thermophilic methanotroph Methylocaldum szegediense O12. Sps, Spp, FruK and Sus demonstrated biochemical properties similar to those of other bacterial counterparts, but the translated amino acid sequences of Sps and Spp displayed high divergence from the respective microbial enzymes. The Sus of M. szegediense O12 catalyzed the reversible reaction of sucrose cleavage in the presence of ADP or UDP and preferred ADP as a substrate, thus implying a connection between sucrose and glycogen metabolism. Sus-like genes were found only in a few methanotrophs, whereas amylosucrase was generally used in sucrose cleavage in this group of bacteria. Like other microbial fructokinases, FruK of M. szegediense O12 showed a high specificity to fructose.

[Research progress and discovery process of facultative methanotrophs--a review].

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Zhao, Tiantao; Xing, Zhilin; Zhang, Lijie

2013-08-04

Facultative methanotrophs are a group of phylogenetically diverse microorganisms characterized by their ability to use methane and some other compounds containing C-C bond as their sole source of carbon and energy. Recently, which belong to the facultative methanotrophs in the genera Methylocella, Methylocapsa and Methylocystis, which belong to the Alphaproteobacteria, have been reported that can grow on larger organic acids or ethanol for some species, as well as methane. In this paper, the research history of facultative methanotrophs was summarized systematically, some other facultative methane-oxidizing microorganisms were introduced, the metabolic mechanisms of utilizing multi-carbon compounds by facultative methanotrophs were analyzed, and the current problems and the future engineering applications were discussed.

Anaerobic bacteria that dechlorinate perchloroethene.

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Fathepure, B Z; Nengu, J P; Boyd, S A

1987-01-01

In this study, we identified specific cultures of anaerobic bacteria that dechlorinate perchlorethene (PCE). The bacteria that significantly dechlorinated PCE were strain DCB-1, an obligate anaerobe previously shown to dechlorinate chlorobenzoate, and two strains of Methanosarcina. The rate of PCE dechlorination by DCB-1 compared favorably with reported rates of trichloroethene bio-oxidation by methanotrophs. Even higher PCE dechlorination rates were achieved when DCB-1 was grown in a methanogenic consortium. PMID:3426224

Site-specific variability in BTEX biodegradation under denitrifying conditions

International Nuclear Information System (INIS)

Kao, C.M.; Borden, R.C.

1997-01-01

Laboratory microcosm experiments were conducted to evaluate the feasibility of benzene, toluene, ethylbenzene, m-xylene, and o-xylene (BTEX) biodegradation under denitrifying conditions. Nine different sources of inocula, including contaminated and uncontaminated soil cores from four different sites and activated sludge, were used to establish microcosms. BTEX was not degraded under denitrifying conditions in microcosms inoculated with aquifer material from Rocky Point and Traverse City. However, rapid depletion of glucose under denitrifying conditions was observed in microcosms containing Rocky Point aquifer material. TEX degradation was observed in microcosms containing Rocky Point aquifer material. TEX degradation was observed in microcosms containing aquifer material from Fort Bragg and Sleeping Bear Dunes and sewage sludge. Benzene was recalcitrant in all microcosms tested. The degradation of o-xylene ceased after toluene, ethylbenzene, and m-xylene were depleted in the Fort Bragg and sludge microcosms, but o-xylene continued to degrade in microcosms with contaminated Sleeping Bear Dunes soil. The most probable number (MPN) of denitrifiers in these nine different inocula were measured using a microtiter technique. There was no correlation between the MPN of denitrifiers and the TEX degradation rate under denitrifying conditions. Experimental results indicate that the degradation sequence and TEX degradation rate under denitrifying conditions may differ among sites. Results also indicate that denitrification alone may not be a suitable bioremediation technology for gasoline-contaminated aquifers because of the inability of denitrifiers to degrade benzene

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

Science.gov (United States)

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

1986-01-01

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

Effect of land use on the density of nitrifying and denitrifying bacteria in the Colombian Coffee Region

Directory of Open Access Journals (Sweden)

Vallejo Quintero Victoria Eugenia

2011-12-01

Full Text Available

Soil microbial communities involved in the cycling of nitrogen (N are essential to maintaining and improving soil fertility, productivity and functionality of natural and agricultural ecosystems. However, some compounds generated during the metabolic processes performed by nitrifying (NB and denitrifying (DB bacteria are associated with the production of greenhouse gases, groundwater pollution and acidification. Therefore, the study of these bacteria is essential for economic and environmental sustainability. This study evaluated the effect of different land uses in two river basins (La Vieja and Otun on NB and DB densities. Two sampling events (SE were conducted by selecting the most representative land uses. Physicochemical (T °, pH, moisture and nitrate and microbiological properties (NB and DB densities were evaluated. In both SEs, significantly higher densities of NB and DB were observed in the land uses: pasture, guadua (DB only and unshaded coffee (La Vieja and onion (Otun. These land uses, excluding guadua, are dependent on nitrogen fertilizers, which together with the activities of grazing livestock on pastures may lead to greater availability of substrates for the NB. The use of agricultural machinery and overgrazing in pasture and onion uses generate compacted soil and other physical disturbances, encouraging the growth of DB. Forests had the lowest densities of NB and DB possibly due to a reduced availability of N and the releasing of allelopathic compounds from certain plants. Finally, the densities of ammonium-oxidizing bacteria had the greatest differences between the land uses evaluated, demonstrating its high sensitivity to agricultural management practices and livestock. We suggest that changes in the abundance of this community could

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

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

2014-10-01

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

Detection and enumeration of methanotrophs in acidic Sphagnum peat by 16S rRNA fluorescence in situ hybridization, including the use of newly developed oligonucleotide probes for Methylocella palustris.

Science.gov (United States)

Dedysh, S N; Derakshani, M; Liesack, W

2001-10-01

Two 16S rRNA-targeted oligonucleotide probes, Mcell-1026 and Mcell-181, were developed for specific detection of the acidophilic methanotroph Methylocella palustris using fluorescence in situ hybridization (FISH). The fluorescence signal of probe Mcell-181 was enhanced by its combined application with the oligonucleotide helper probe H158. Mcell-1026 and Mcell-181, as well as 16S rRNA oligonucleotide probes with reported group specificity for either type I methanotrophs (probes M-84 and M-705) or the Methylosinus/Methylocystis group of type II methanotrophs (probes MA-221 and M-450), were used in FISH to determine the abundance of distinct methanotroph groups in a Sphagnum peat sample of pH 4.2. M. palustris was enumerated at greater than 10(6) cells per g of peat (wet weight), while the detectable population size of type I methanotrophs was three orders of magnitude below the population level of M. palustris. The cell counts with probe MA-221 suggested that only 10(4) type II methanotrophs per g of peat (wet weight) were present, while the use of probe M-450 revealed more than 10(6) type II methanotroph cells per g of the same samples. This discrepancy was due to the fact that probe M-450 targets almost all currently known strains of Methylosinus and Methylocystis, whereas probe MA-221, originally described as group specific, does not detect a large proportion of Methylocystis strains. The total number of methanotrophic bacteria detected by FISH was 3.0 (+/-0.2) x 10(6) cells per g (wet weight) of peat. This was about 0.8% of the total bacterial cell number. Thus, our study clearly suggests that M. palustris and a defined population of Methylocystis spp. were the predominant methanotrophs detectable by FISH in an acidic Sphagnum peat bog.

[Identification and function test of an alkali-tolerant denitrifying bacterium].

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Wang, Ru; Zheng, Ping; Li, Wei; Chen, Hui; Chen, Tingting; Ghulam, Abbas

2013-04-04

We obtained an alkali-tolerant denitrifying bacterium, and determined its denitrifying activity and alkali-tolerance. An alkali-tolerant denitrifying bacterial strain was obtained by isolation and purification. We identified the bacterial strain by morphological observation, physiological test and 16S rRNA analysis. We determined the denitrifying activity and alkali-tolerance by effects of initial nitrate concentration and initial pH on denitrification. An alkali-tolerant denitrifier strain R9 was isolated from the lab-scale high-rate denitrifying reactor, and it was identified as Diaphorobater nitroreducens. The strain R9 grew heterotrophically with methanol as the electron donor and nitrate as the electron acceptor. The nitrate conversion was 93.25% when strain R9 was cultivated for 288 h with initial nitrate concentration 50 mg/L and initial pH 9.0. The denitrification activity could be inhibited at high nitrate concentration with a half inhibition constant of 202.73 mg N/L. Strain R9 showed a good alkali tolerance with the nitrate removal rate at pH 11.0 remained 86% of that at pH 9.0. Strain R9 was identified as Diaphorobater nitroreducens, and it was an alkali-tolerant denitrifying bacterium with optimum pH value of 9.0.

Aquatic plant surface as a niche for methanotrophs

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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.

Mapping spatial patterns of denitrifiers at large scales (Invited)

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Philippot, L.; Ramette, A.; Saby, N.; Bru, D.; Dequiedt, S.; Ranjard, L.; Jolivet, C.; Arrouays, D.

2010-12-01

Little information is available regarding the landscape-scale distribution of microbial communities and its environmental determinants. Here we combined molecular approaches and geostatistical modeling to explore spatial patterns of the denitrifying community at large scales. The distribution of denitrifrying community was investigated over 107 sites in Burgundy, a 31 500 km2 region of France, using a 16 X 16 km sampling grid. At each sampling site, the abundances of denitrifiers and 42 soil physico-chemical properties were measured. The relative contributions of land use, spatial distance, climatic conditions, time and soil physico-chemical properties to the denitrifier spatial distribution were analyzed by canonical variation partitioning. Our results indicate that 43% to 85% of the spatial variation in community abundances could be explained by the measured environmental parameters, with soil chemical properties (mostly pH) being the main driver. We found spatial autocorrelation up to 739 km and used geostatistical modelling to generate predictive maps of the distribution of denitrifiers at the landscape scale. Studying the distribution of the denitrifiers at large scale can help closing the artificial gap between the investigation of microbial processes and microbial community ecology, therefore facilitating our understanding of the relationships between the ecology of denitrifiers and N-fluxes by denitrification.

Denitrification as an adaptive trait in soil and groundwater bacteria

Energy Technology Data Exchange (ETDEWEB)

Bergwall, C

1997-09-01

The focus of this thesis is on selection and adaptation processes in bacteria with emphasis on denitrifying bacteria in groundwater. Other nitrogen transformation processes such as dissimilatory nitrate reduction to ammonium (nitrate ammonification) and nitrification of forest soil bacteria are briefly discussed. Microcosms with sterile sediment and groundwater were inoculated with single denitrifying strains isolated from three groundwater aquifers, two of which are agricultural aquifers (in situ NO{sub 3}{sup -}-N was 24.1 and 35.2 mg1{sup -1}) and the third which is a pristine lake water infiltration aquifer (in situ NO{sub 3}{sup -}-N was 6.3 mg1{sup -1}). The average denitrification activity for strains from the nitrate contaminated sites were twice as high as the activity of the strains from the pristine site. Denitrification were carbon limited and glucose amendment increased the denitrification activity about a 2-fold for all strains. The strain specific differences in denitrification rates increased to a 2.5-fold after carbon addition indicating that the differences in reduction rates cannot be explained by different carbon utilisation rates but rather reflect innate differences in the reductases of the strains. A preliminary identification of the molecular target for adaptation was performed with artificial electron donors and electron acceptors for all enzymatic steps in the denitrification pathway. Nitrous oxide reductase activity was significantly higher in denitrifiers from the nitrate contaminated sites. This suggests that nos genes may be the molecular target, possibly by mutation or gene duplication for adaptation to high nitrate concentrations. Two anaerobic denitrifiers from each of the contaminated sites were capable of aerobic denitrification indicating that high nitrate concentrations may select for strains that denitrifies in the presence of both oxygen and nitrate. Microcosm experiments with fertilized coniferous forest soil showed that the

Denitrifying sulfide removal process on high-salinity wastewaters.

Science.gov (United States)

Liu, Chunshuang; Zhao, Chaocheng; Wang, Aijie; Guo, Yadong; Lee, Duu-Jong

2015-08-01

Denitrifying sulfide removal (DSR) process comprising both heterotrophic and autotrophic denitrifiers can simultaneously convert nitrate, sulfide, and acetate into nitrogen gas, elemental sulfur (S(0)), and carbon dioxide, respectively. Sulfide- and nitrate-laden wastewaters at 2-35 g/L NaCl were treated by DSR process. A C/N ratio of 3:1 was proposed to maintain high S(0) conversion rate. The granular sludge with a compact structure and smooth outer surface was formed. The microbial communities of DSR consortium via high-throughput sequencing method suggested that salinity shifts the predominating heterotrophic denitrifiers at 10 g/L NaCl.

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...... was the only nitrogen source. The observed Monod constant for methane utilization increased with increasing concentration of ammonia. This shows that ammonia is a weak competitive inhibitor as observed by other researchers. Relatively high levels of both ammonia (70 mg/l) and copper (300 mu-g/l) inhibited...... the methane degradation, probably due to the toxic effect of copper-amine complexes....

Responses of bacterial community structure and denitrifying bacteria in biofilm to submerged macrophytes and nitrate

Science.gov (United States)

Zhang, Songhe; Pang, Si; Wang, Peifang; Wang, Chao; Guo, Chuan; Addo, Felix Gyawu; Li, Yi

2016-10-01

Submerged macrophytes play important roles in constructed wetlands and natural water bodies, as these organisms remove nutrients and provide large surfaces for biofilms, which are beneficial for nitrogen removal, particularly from submerged macrophyte-dominated water columns. However, information on the responses of biofilms to submerged macrophytes and nitrogen molecules is limited. In the present study, bacterial community structure and denitrifiers were investigated in biofilms on the leaves of four submerged macrophytes and artificial plants exposed to two nitrate concentrations. The biofilm cells were evenly distributed on artificial plants but appeared in microcolonies on the surfaces of submerged macrophytes. Proteobacteria was the most abundant phylum in all samples, accounting for 27.3-64.8% of the high-quality bacterial reads, followed by Chloroflexi (3.7-25.4%), Firmicutes (3.0-20.1%), Acidobacteria (2.7-15.7%), Actinobacteria (2.2-8.7%), Bacteroidetes (0.5-9.7%), and Verrucomicrobia (2.4-5.2%). Cluster analysis showed that bacterial community structure can be significantly different on macrophytes versus from those on artificial plants. Redundancy analysis showed that electrical conductivity and nitrate concentration were positively correlated with Shannon index and operational taxonomic unit (OTU) richness (log10 transformed) but somewhat negatively correlated with microbial density. The relative abundances of five denitrifying genes were positively correlated with nitrate concentration and electrical conductivity but negatively correlated with dissolved oxygen.

IDENTIFICATION AND ECOPHYSIOLOGY OF ACTIVE DENITRIFIERS IN ACTIVATED SLUDGE

DEFF Research Database (Denmark)

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

reactor studies. To obtain better identification of active denitrifying communities in full-scale wastewater treatment plants (WWTPs) we applied DNA-SIP with 13C-labelled substrates, and RT-PCR of expressed denitrification genes (nirS, nirK and nosZ) upon various substrate-inductions. To come around...... were determined with quantitative FISH, while their active metabolic pathways were investigated directly in activated sludge with a tag-based metatranscriptomic approach under acetate-utilizing and denitrifying conditions. The different methods revealed a majority of denitrifiers in all WWTPs belonging...

Enrichment of marine anammox bacteria from seawater-related samples and bacterial community study.

Science.gov (United States)

Kawagoshi, Y; Nakamura, Y; Kawashima, H; Fujisaki, K; Furukawa, K; Fujimoto, A

2010-01-01

Anaerobic ammonium oxidation (anammox) is a novel nitrogen pathway catalyzed by anammox bacteria which are obligate anaerobic chemoautotrophs. In this study, enrichment culture of marine anammox bacteria (MAAOB) from the samples related to seawater was conducted. Simultaneous removal of ammonium and nitrite was confirmed in continuous culture inoculated with sediment of a sea-based waste disposal site within 50 days. However, no simultaneous nitrogen removal was observed in cultures inoculated with seawater-acclimated denitrifying sludge or with muddy sediment of tideland even during 200 days. Nitrogen removal rate of 0.13 kg/m(3)/day was achieved at nitrogen loading rate of 0.16 kg/m(3)/day after 320th days in the culture inoculated with the sediment of waste disposal site. The nitrogen removal ratio between ammonium nitrogen and nitrite nitrogen was 1:1.07. Denaturing gradient gel electrophoresis (DGGE) analysis indicated that an abundance of the bacteria close to MAAOB and coexistence of ammonium oxidizing bacteria and denitrifying bacteria in the culture.

Characterization of bacterial consortia capable of degrading 4-chlorobenzoate and 4-bromobenzoate under denitrifying conditions.

Science.gov (United States)

Song, Bongkeun; Kerkhof, Lee J; Häggblom, Max M

2002-08-06

4-Chlorobenzoate and 4-bromobenzoate were readily degraded in denitrifying enrichment cultures established with river sediment, estuarine sediment or agricultural soil as inoculum. Stable denitrifying consortia were obtained and maintained by serial dilution and repeated feeding of substrates. Microbial community analyses were performed to characterize the 4-chlorobenzoate and 4-bromobenzoate degrading consortia with terminal restriction fragment length polymorphism (T-RFLP) and cloning of 16S rRNA genes from the cultures. Interestingly, two major terminal restriction fragments (T-RFs) in the 4-chlorobenzoate degrading consortia and one T-RF in the 4-bromobenzoate utilizing consortium were observed from T-RFLP analysis regardless of their geographical and ecological origins. The two T-RFs (clones 4CB1 and 4CB2) in 4-chlorobenzoate degrading consortia were identified as members of the beta-subunit of the Proteobacteria on the basis of 16S rRNA sequencing analysis. Phylogenetic analysis of 16S rRNA genes showed that clone 4CB1 was closely related to Thauera aromatica while clone 4CB2 was distantly related to the genera Limnobacter and Ralstonia. The 4-bromobenzoate utilizing consortium mainly consisted of one T-RF, which was identical to clone 4CB2 in spite of different enrichment substrate. This suggests that degradation of 4-chlorobenzoate and 4-bromobenzoate under denitrifying conditions was mediated by bacteria belonging to the beta-subunit of the Proteobacteria.

Mathematical Modeling of Nitrous Oxide Production during Denitrifying Phosphorus Removal Process.

Science.gov (United States)

Liu, Yiwen; Peng, Lai; Chen, Xueming; Ni, Bing-Jie

2015-07-21

A denitrifying phosphorus removal process undergoes frequent alternating anaerobic/anoxic conditions to achieve phosphate release and uptake, during which microbial internal storage polymers (e.g., Polyhydroxyalkanoate (PHA)) could be produced and consumed dynamically. The PHA turnovers play important roles in nitrous oxide (N2O) accumulation during the denitrifying phosphorus removal process. In this work, a mathematical model is developed to describe N2O dynamics and the key role of PHA consumption on N2O accumulation during the denitrifying phosphorus removal process for the first time. In this model, the four-step anoxic storage of polyphosphate and four-step anoxic growth on PHA using nitrate, nitrite, nitric oxide (NO), and N2O consecutively by denitrifying polyphosphate accumulating organisms (DPAOs) are taken into account for describing all potential N2O accumulation steps in the denitrifying phosphorus removal process. The developed model is successfully applied to reproduce experimental data on N2O production obtained from four independent denitrifying phosphorus removal study reports with different experimental conditions. The model satisfactorily describes the N2O accumulation, nitrogen reduction, phosphate release and uptake, and PHA dynamics for all systems, suggesting the validity and applicability of the model. The results indicated a substantial role of PHA consumption in N2O accumulation due to the relatively low N2O reduction rate by using PHA during denitrifying phosphorus removal.

Nitrous oxide emission by the non-denitrifying, nitrate ammonifier Bacillus licheniformis.

Science.gov (United States)

Sun, Yihua; De Vos, Paul; Heylen, Kim

2016-01-19

Firmicutes have the capacity to remove excess nitrate from the environment via either denitrification, dissimilatory nitrate reduction to ammonium or both. The recent renewed interest in their nitrogen metabolism has revealed many interesting features, the most striking being their wide variety of dissimilatory nitrate reduction pathways. In the present study, nitrous oxide production from Bacillus licheniformis, a ubiquitous Gram-positive, spore-forming species with many industrial applications, is investigated. B. licheniformis has long been considered a denitrifier but physiological experiments on three different strains demonstrated that nitrous oxide is not produced from nitrate in stoichiometric amounts, rather ammonium is the most important end-product, produced during fermentation. Significant strain dependency in end-product ratios, attributed to nitrite and ammonium, and medium dependency in nitrous oxide production were also observed. Genome analyses confirmed the lack of a nitrite reductase to nitric oxide, the key enzyme of denitrification. Based on the gene inventory and building on knowledge from other non-denitrifying nitrous oxide emitters, hypothetical pathways for nitrous oxide production, involving NarG, NirB, qNor and Hmp, are proposed. In addition, all publically available genomes of B. licheniformis demonstrated similar gene inventories, with specific duplications of the nar operon, narK and hmp genes as well as NarG phylogeny supporting the evolutionary separation of previously described distinct BALI1 and BALI2 lineages. Using physiological and genomic data we have demonstrated that the common soil bacterium B. licheniformis does not denitrify but is capable of fermentative dissimilatory nitrate/nitrite reduction to ammonium (DNRA) with concomitant production of N2O. Considering its ubiquitous nature and non-fastidious growth in the lab, B. licheniformis is a suitable candidate for further exploration of the actual mechanism of N2O

Microbial Community Composition and Ultrastructure of Granules from a Full-Scale Anammox Reactor

KAUST Repository

Gonzalez-Gil, Graciela; Sougrat, Rachid; Behzad, Ali Reza; Lens, Piet Nl L; Saikaly, Pascal

2014-01-01

. Hydrogenotrophic methananogens, which scavenge the key fermentation product H2, were the most abundant archaea detected. Cells resembling the polygon-shaped denitrifying methanotroph Candidatus Methylomirabilis oxyfera were observed by electron microscopy

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.

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

Science.gov (United States)

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

2010-12-01

Nitrous oxide (N2O) fluxes from soils result from its production by nitrification and denitrification and reduction during denitrification. The structure of the denitrifying microbial community contributes to the control of net N2O fluxes. Although molecular techniques are promising for identifying the active community of N2O producers, there are few data until now because methods to explore gene expression of N2O production are laborious and disregard regulation of activity at the enzyme level. The isotopologue signatures of N2O including δ18O, average δ15N (δ15Nbulk) and 15N site preference (SP = difference in δ15N between the central and peripheral N positions of the asymmetric N2O molecule) have been used to estimate the contribution of partial processes to net N2O fluxes to the atmosphere. However, the use of this approach to study N2O dynamics in soils requires knowledge of isotopic signatures of N2O precursors and isotopologue fractionation factors (ɛ) of all processes of N2O production and consumption. In contrast to δ18O and δ15Nbulk, SP is independent of precursor signatures and hence is a promising parameter here. It is assumed that SP of produced N2O is almost exclusively controlled by the enzymatic isotope effects of NO reductases (NOR). These enzymes are known to be structurally different between certain classes of N2O producers with each class causing different isotope effects (Schmidt et al., 2004). The NH2OH-to-N2O step of nitrifiers and the NO3-to-N2O step of fungal denitrifiers are associated with large site-specific 15N effects with SP of 33 to 37 ‰ (Sutka et al., 2006, 2008) while the few tested species of gram-negative bacterial denitrifiers (cNOR group) exhibited low SP of -5 to 0‰ (Sutka et al., 2006; Toyoda et al., 2005). The aim of our study was to determine site-specific fractionation factors of the NO3-to-N2O step (ɛSP) for several species of denitrifiers representing each of the known NOR-types of bacteria, i.e. cNOR, q

The lipid response of aerobic marine methanotroph communities under changing environmental conditions.

Science.gov (United States)

Rush, D.; Villanueva, L.; van der Meer, M.; S Sinninghe Damsté, J.

2017-12-01

Methane (CH4) originating from marine environments accounts for a significant amount of atmospheric greenhouse gas. Aerobic methanotrophs, which convert CH4 to CO­2, are responsible for quenching a part of this methane before its release. Modern-day climate projections show a rapid shift towards a warmer, more acidic ocean. How do these important methanotrophic communities respond to such changes to their environment? Here, we present the results of microcosm experiments from three marine regions influenced by CH4. Particulate organic matter and sediment were collected from the Black Sea, the Baltic Sea, and the North Sea, at depths ideal for aerobic methanotroph communities at the time of sampling (e.g. oxic, in area of active CH4 release). These were incubated under different temperatures, pHs, and labelled 13CH4 concentrations. We monitored methane concentration in these microcosms as an indication of 13CH4 consumption by methanotrophs. Once the methane concentration was lipids of the organisms oxidising methane in order to elucidate which organisms are performing methane oxidation and whether they synthesize specific biomarker lipids. Particular attention will be paid to the abundances and diversity of bacteriohopanepolyol lipids, known methanotroph biomarkers. The ultimate goal of our investigation is to determine the effect changes in these environmental parameters have on aerobic methanotroph community structures and their lipid fingerprints. By establishing reliable biomarker lipids for aerobic methanotrophy at certain conditions, we will then be able to investigate the contribution of aerobic methanotrophy throughout Earth's history, especially at times when CH4 concentrations were higher than they are at present.

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.

Starvation and recovery in the deep-sea methanotroph Methyloprofundus sedimenti

OpenAIRE

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...

Mapping the distribution of the denitrifier community at large scales (Invited)

Science.gov (United States)

Philippot, L.; Bru, D.; Ramette, A.; Dequiedt, S.; Ranjard, L.; Jolivet, C.; Arrouays, D.

2010-12-01

Little information is available regarding the landscape-scale distribution of microbial communities and its environmental determinants. Here we combined molecular approaches and geostatistical modeling to explore spatial patterns of the denitrifying community at large scales. The distribution of denitrifrying community was investigated over 107 sites in Burgundy, a 31 500 km2 region of France, using a 16 X 16 km sampling grid. At each sampling site, the abundances of denitrifiers and 42 soil physico-chemical properties were measured. The relative contributions of land use, spatial distance, climatic conditions, time and soil physico-chemical properties to the denitrifier spatial distribution were analyzed by canonical variation partitioning. Our results indicate that 43% to 85% of the spatial variation in community abundances could be explained by the measured environmental parameters, with soil chemical properties (mostly pH) being the main driver. We found spatial autocorrelation up to 740 km and used geostatistical modelling to generate predictive maps of the distribution of denitrifiers at the landscape scale. Studying the distribution of the denitrifiers at large scale can help closing the artificial gap between the investigation of microbial processes and microbial community ecology, therefore facilitating our understanding of the relationships between the ecology of denitrifiers and N-fluxes by denitrification.

Spatial Variability of Cyanobacteria and Heterotrophic Bacteria in Lake Taihu (China).

Science.gov (United States)

Qian, Haifeng; Lu, Tao; Song, Hao; Lavoie, Michel; Xu, Jiahui; Fan, Xiaoji; Pan, Xiangliang

2017-09-01

Cyanobacterial blooms frequently occur in Lake Taihu (China), but the intertwined relationships between biotic and abiotic factors modulating the frequency and duration of the blooms remain enigmatic. To better understand the relationships between the key abiotic and biotic factors and cyanobacterial blooms, we measured the abundance and diversity of prokaryotic organisms by high-throughput sequencing, the abundance of key genes involved in microcystin production and nitrogen fixation or loss as well as several physicochemical parameters at several stations in Lake Taihu during a cyanobacterial bloom of Microcystis sp.. Measurements of the copy number of denitrification-related genes and 16S rRNA analyses show that denitrification potential and denitrifying bacteria abundance increased in concert with non-diazotrophic cyanobacteria (Microcystis sp.), suggesting limited competition between cyanobacteria and heterotrophic denitrifiers for nutrients, although potential bacteria-mediated N loss may hamper Microcystis growth. The present study provides insight into the importance of different abiotic and biotic factors in controlling cyanobacteria and heterotrophic bacteria spatial variability in Lake Taihu.

Responses of mixed methanotrophic consortia to variable Cu2+/Fe2+ ratios.

Science.gov (United States)

Chidambarampadmavathy, Karthigeyan; Karthikeyan, Obulisamy Parthiba; Huerlimann, Roger; Maes, Gregory E; Heimann, Kirsten

2017-07-15

Methane mitigation in landfill top cover soils is mediated by methanotrophs whose optimal methane (CH 4 ) oxidation capacity is governed by environmental and complex microbial community interactions. Optimization of CH 4 remediating bio-filters need to take microbial responses into account. Divalent copper (Cu 2+ ) and iron (Fe 2+ ) are present in landfills at variable ratios and play a vital role in methane oxidation capacity and growth of methanotrophs. This study, as a first of its kind, therefore quantified effects of variable Cu 2+ and Fe 2+ (5:5, 5:25 and 5:50 μM) ratios on mixed methanotrophic communities enriched from landfill top cover (LB) and compost soils (CB). CH 4 oxidation capacity, CH 4 removal efficiencies, fatty acids content/profiles and polyhydroxybutyrate (PHB; a biopolymer) contents were also analysed to quantify performance and potential co-product development. Mixed methanotroph cultures were raised in 10 L continuous stirred tank reactors (CSTRs, Bioflo ® & Celligen ® 310 Fermentor/Bioreactor; John Morris Scientific, Chatswood, NSW, Australia). Community structure was determined by amplifying the V3-V4 region of 16s rRNA gene. Community structure and, consequently, fatty acid-profiles changed significantly with increasing Cu 2+ /Fe 2+ ratios, and responses were different for LB and CB. Effects on methane oxidation capacities and PHB content were similar in the LB- and CB-CSTR, decreasing with increasing Cu 2+ /Fe 2+ ratios, while biomass growth was unaffected. In general, high Fe 2+ concentration favored growth of the type -II methanotroph Methylosinus in the CB-CSTR, but methanotroph abundances decreased in the LB-CSTR. Increase in Cu 2+ /Fe 2+ ratio increased the growth of Sphingopyxis in both systems, while Azospirllum was co-dominant in the LB- but absent in the CB-CSTR. After 13 days, methane oxidation capacities and PHB content decreased by ∼50% and more in response to increasing Fe 2+ concentrations. Although methanotroph

Modelling the growth of a methanotrophic biofilm

DEFF Research Database (Denmark)

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

1999-01-01

This article discusses the growth of methanotrophic biofilms. Several independent biofilm growths scenarios involving different inocula were examined. Biofilm growth, substrate removal and product formation were monitored throughout the experiments. Based on the oxygen consumption it was concluded...... 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...... was performed on this model. It indicated that the most influential parameters were those related to the biofilm (i.e. density; solid-volume fraction; thickness). This suggests that in order to improve the model, further research regarding the biofilm structure and composition is needed....

[Methanotrophs and methylobacteria are found in woody plant tissues within a winter period].

Science.gov (United States)

Doronina, N V; Ivanova, E G; Suzina, N F; Trotsenko, Iu A

2004-01-01

Samples of tree seeds, buds, and needles collected within a winter period at ambient temperatures from -11 to -17 degrees C were analyzed for the presence of methylotrophic microflora. Thin sections of blue spruce needles were found to contain bacteria morphologically close to pink-pigmented methylobacteria. The methylobacteria that were isolated in pure cultures from samples of linden seeds and buds, pine and blue spruce needles, as well as of lilac, maple, and apple buds, were classified into the genera Methylobacterium and Paracoccus based on the data of morphological studies, enzyme assay, and DNA-DNA hybridization analysis. The methanotrophs that were isolated in pure cultures from samples of linden buds and blue spruce needles were identified into the genus Methylocystis based on the data of morphological studies, enzyme assay, DNA-DNA hybridization, and the phylogenetic analysis of the particulate methane monooxygenase gene pmoA sequences. The inference is made that aerobic methylotrophic bacteria are permanently associated with plants. At the beginning of the vegetative period in spring, the phyllosphere of coniferous and deciduous trees is colonized by the methylotrophic bacteria that have wintered inside plant tissues.

Diversity of active aerobic methanotrophs along depth profiles of arctic and subarctic lake water column and sediments

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) emitted from high-latitude lakes accounts for 2–6% of the global atmospheric CH4 budget. Methanotrophs in lake sediments and water columns mitigate the amount of CH4 that enters the atmosphere, yet their identity and activity in arctic and subarctic lakes are poorly understood. We used stable isotope probing (SIP), quantitative PCR (Q-PCR), pyrosequencing and enrichment cultures to determine the identity and diversity of active aerobic methanotrophs in the water columns and sediments (0–25 cm) from an arctic tundra lake (Lake Qalluuraq) on the north slope of Alaska and a subarctic taiga lake (Lake Killarney) in Alaska's interior. The water column CH4 oxidation potential for these shallow (~2m deep) lakes was greatest in hypoxic bottom water from the subarctic lake. The type II methanotroph, Methylocystis, was prevalent in enrichment cultures of planktonic methanotrophs from the water columns. In the sediments, type I methanotrophs (Methylobacter, Methylosoma and Methylomonas) at the sediment-water interface (0–1 cm) were most active in assimilating CH4, whereas the type I methanotroph Methylobacter and/or type II methanotroph Methylocystis contributed substantially to carbon acquisition in the deeper (15–20 cm) sediments. In addition to methanotrophs, an unexpectedly high abundance of methylotrophs also actively utilized CH4-derived carbon. This study provides new insight into the identity and activity of methanotrophs in the sediments and water from high-latitude lakes.

Denitrifying Bioreactors Resist Disturbance from Fluctuating Water Levels

Directory of Open Access Journals (Sweden)

Sarah K. Hathaway

2017-06-01

Full Text Available Nitrate can be removed from wastewater streams, including subsurface agricultural drainage systems, using woodchip bioreactors to promote microbial denitrification. However, the variations in water flow in these systems could make reliable performance from this microbially-mediated process a challenge. In the current work, the effects of fluctuating water levels on nitrate removal, denitrifying activity, and microbial community composition in laboratory-scale bioreactors were investigated. The performance was sensitive to changing water level. An average of 31% nitrate was removed at high water level and 59% at low water level, despite flow adjustments to maintain a constant theoretical hydraulic retention time. The potential activity, as assessed through denitrifying enzyme assays, averaged 0.0008 mg N2O-N/h/dry g woodchip and did not show statistically significant differences between reactors, sampling depths, or operational conditions. In the denitrifying enzyme assays, nitrate removal consistently exceeded nitrous oxide production. The denitrifying bacterial communities were not significantly different from each other, regardless of water level, meaning that the denitrifying bacterial community did not change in response to disturbance. The overall bacterial communities, however, became more distinct between the two reactors when one reactor was operated with periodic disturbances of changing water height, and showed a stronger effect at the most severely disturbed location. The communities were not distinguishable, though, when comparing the same location under high and low water levels, indicating that the communities in the disturbed reactor were adapted to fluctuating conditions rather than to high or low water level. Overall, these results describe a biological treatment process and microbial community that is resistant to disturbance via water level fluctuations.

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

International Nuclear Information System (INIS)

Coyne, P.; Smith, G.

1995-01-01

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

Transformation of nitrogen and distribution of nitrogen-related bacteria in a polluted urban stream.

Science.gov (United States)

Jiao, Y; Jin, W B; Zhao, Q L; Zhang, G D; Yan, Y; Wan, J

2009-01-01

Most researchers focused on either nitrogen species or microbial community for polluted urban stream while ignoring the interaction between them and its effect on nitrogen transformation, which restricted the rational selection of an effective and feasible remediation technology. Taking Buji stream in Shenzhen (China) as target stream, the distribution of nitrogen-related bacteria was investigated by most probable number (MPN) besides analysis of nitrogen species etc. The nitrogen-related bacteria in sediment were 10(2) times richer than those in water. Owing to their faster growth, the MPN of ammonifying bacteria and denitrifying bacteria were 10(5) and 10(2) times higher than those of nitrifying bacteria, respectively. The ammonifying bacteria numbers were significantly related to BOD5 in water, while nitrifying bacteria in sediment correlated well with nitrate in water. Thus, nitrification occurred mainly in sediment surface and was limited by low proportion of nitrifying bacteria. The denitrifying bacteria in sediment had good relationship with BOD5 and nitrite and nitrate in water. Low DO and rich organic compounds were beneficial to denitrification but unfavourable to nitrification. Denitrification was restricted by low nitrite and nitrate concentration. These results could be served as a reference for implementing the remediation scheme of nitrogen polluted urban stream.

Community size and composition of ammonia oxidizers and denitrifiers in an alluvial intertidal wetland ecosystem

Directory of Open Access Journals (Sweden)

Ziye eHu

2014-07-01

Full Text Available Global nitrogen cycling is mainly mediated by the activity of microorganisms. Nitrogen cycle processes are mediated by functional groups of microorganisms that are affected by constantly changing environmental conditions and substrate availability. In this study, we investigated the temporal and spatial patterns of nitrifier and denitrifier communities in an intertidal wetland. Soil samples were collected over four distinct seasons from three locations with different vegetative cover. Multiple environmental factors and process rates were measured and analyzed together with the community size and composition profiles. We observed that the community size and composition of the nitrifiers and denitrifiers are affected significantly by seasonal factors, while vegetative cover affected the community composition. The seasonal impacts on the community size of ammonia oxidizing archaea (AOA are much higher than that of ammonia oxidizing bacteria (AOB. The seasonal change was a more important indicator for AOA community composition patterns, while vegetation was more important for the AOB community patterns. The microbial process rates were correlated with both the community size and composition.

Application of fluorescent antibody and enzyme-linked immunosorbent assays for TCE and PAH degrading bacteria

Energy Technology Data Exchange (ETDEWEB)

Brigmon, R.L.; Franck, M.; Brey, J.; Scott, D.; Lanclos, K.; Fliermans, C.

1996-07-01

Historically, methods used to identify methanotrophic and polyaromatic hydrocarbon-degrading (PAH) bacteria in environmental samples have been inadequate because isolation and identification procedures are time-consuming and often fail to separate specific bacteria from other environmental microorganisms. Methanotrophic bacteria have been isolated and characterized from TCE-contaminated soils (Bowman et al. 1993; Fliermans et al., 1988). Fliermans et al., (1988) and others demonstrated that cultures enriched with methane and propane could cometabolically degrade a wide variety of chlorinated aliphatic hydrocarbons including ethylene; 1,2-cisdichloroethylene (c-DCE); 1,2-trans-dichloroethylene (t-DCE); vinyl chloride (VC); toluene; phenol and cresol. Characterization of select microorganisms in the natural setting is important for the evaluation of bioremediation potential and its effectiveness. This realization has necessitated techniques that are selective, sensitive and easily applicable to soils, sediments, and groundwater (Fliermans, et al., 1994). Additionally these techniques can identify and quantify microbial types in situ in real time

Insight into the effects of biochar on manure composting: evidence supporting the relationship between N2O emission and denitrifying community.

Science.gov (United States)

Wang, Cheng; Lu, Haohao; Dong, Da; Deng, Hui; Strong, P J; Wang, Hailong; Wu, Weixiang

2013-07-02

Although nitrous oxide (N2O) emissions from composting contribute to the accelerated greenhouse effect, it is difficult to implement practical methods to mitigate these emissions. In this study, the effects of biochar amendment during pig manure composting were investigated to evaluate the inter-relationships between N2O emission and the abundance of denitrifying bacteria. Analytical results from two pilot composting treatments with (PWSB, pig manure + wood chips + sawdust + biochar) or without (PWS, pig manure + wood chips + sawdust) biochar (3% w/w) demonstrated that biochar amendment not only lowered NO2(-)-N concentrations but also lowered the total N2O emissions from pig manure composting, especially during the later stages. Quantification of functional genes involved in denitrification and Spearman rank correlations matrix revealed that the N2O emission rates correlated with the abundance of nosZ, nirK, and nirS genes. Biochar-amended pig manure had a higher pH and a lower moisture content. Biochar amendment altered the abundance of denitrifying bacteria significantly; less N2O-producing and more N2O-consuming bacteria were present in the PWSB, and this significantly lowered N2O emissions in the maturation phase. Together, the results demonstrate that biochar amendment could be a novel greenhouse gas mitigation strategy during pig manure composting.

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.

Effects of nitrogen load on the function and diversity of methanotrophs in the littoral wetland of a boreal lake

Directory of Open Access Journals (Sweden)

Henri MP Siljanen

2012-02-01

Full Text Available Methane is the second most abundant greenhouse gas in the atmosphere. A major part of the total methane emissions from lake ecosystems are emitted from littoral wetlands. Methane emissions are significantly reduced by methanotrophs as they use methane as the sole energy and carbon source. Function of methanotrophs can be either activated or suppressed by nitrogen. However, the effects of nitrogen on methanotrophs in littoral wetlands are unknown. Here we report how nitrogen loading in situ affected the function and diversity of methanotrophs in a littoral wetland. Methanotrophic community composition and functional diversity were analyzed with particulate methane monooxygenase (pmoA gene targeted microarray. Nitrogen load had no effects on methane oxidation potential and methane fluxes. Nitrogen load activated pmoA gene transcription of type I (Methylobacter, Methylomonas and LW21-freshwater phylotypes methanotrophs, but decreased the relative abundance of type II (Methylocystis, Methylosinus trichosporium and Methylosinus phylotypes methanotrophs. Hence, the overall activity of a methanotroph community in littoral wetlands is unlikely to be affected by nitrogen leached from the catchment area.

Influencing factors analysis of anammox bacteria cultured by mixing denitrifying-anammox

OpenAIRE

Sihui WANG; Yuanyuan SONG; Yunman LIU; Yankai GUO; Jing LIAN; Jianbo GUO

2017-01-01

In order to get the optimal growth conditions of anammox bacteria, the mature-cultured anammox granule sludge is used to investigate the influencing factors. The effects of temperature, pH value, COD and influent substrate (NO-2-N and NH+4-N) on anammox bacteria activity are investigated. The results demonstrate that the optimal temperature is 40 ℃ and the optimal pH value is between 7.0~8.0 for anammox bacteria. The anammox bacteria activity is not inhibited severely when COD concentration i...

Nutrient recovery from industrial wastewater as single cell protein by a co-culture of green microalgae and methanotrophs

DEFF Research Database (Denmark)

Rasouli, Zahra; Valverde Pérez, Borja; D'Este, Martina

2018-01-01

wastewater and upcycle them to feed grade single cell protein. Results demonstrated that both algae and bacteria could remove or assimilate most of the organic carbon present in the wastewater (~95% removal for monocultures and 91% for the algal-bacterial consortium). However, their growth stopped before......, for all cultures the protein content (45% of dry weight, DW, for methanotrophs; 52.5% of DW for algae; and 27.6% of DW for consortium) and amino acid profile was suitable for substitution of conventional protein sources. Further research should focus on increasing productivity of biomass grown...

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

Energy Technology Data Exchange (ETDEWEB)

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

2014-01-01

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

Physical disturbance to ecological niches created by soil structure alters community composition of methanotrophs.

Science.gov (United States)

Kumaresan, Deepak; Stralis-Pavese, Nancy; Abell, Guy C J; Bodrossy, Levente; Murrell, J Colin

2011-10-01

Aggregates of different sizes and stability in soil create a composite of ecological niches differing in terms of physico-chemical and structural characteristics. The aim of this study was to identify, using DNA-SIP and mRNA-based microarray analysis, whether shifts in activity and community composition of methanotrophs occur when ecological niches created by soil structure are physically perturbed. Landfill cover soil was subject to three treatments termed: 'control' (minimal structural disruption), 'sieved' (sieved soil using 2 mm mesh) and 'ground' (grinding using mortar and pestle). 'Sieved' and 'ground' soil treatments exhibited higher methane oxidation potentials compared with the 'control' soil treatment. Analysis of the active community composition revealed an effect of physical disruption on active methanotrophs. Type I methanotrophs were the most active methanotrophs in 'sieved' and 'ground' soil treatments, whereas both Type I and Type II methanotrophs were active in the 'control' soil treatment. The result emphasize that changes to a particular ecological niche may not result in an immediate change to the active bacterial composition and change in composition will depend on the ability of the bacterial communities to respond to the perturbation. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

Propionate oxidation by and methanol inhibition of anaerobic ammonium-oxidizing bacteria.

Science.gov (United States)

Güven, Didem; Dapena, Ana; Kartal, Boran; Schmid, Markus C; Maas, Bart; van de Pas-Schoonen, Katinka; Sozen, Seval; Mendez, Ramon; Op den Camp, Huub J M; Jetten, Mike S M; Strous, Marc; Schmidt, Ingo

2005-02-01

Anaerobic ammonium oxidation (anammox) is a recently discovered microbial pathway and a cost-effective way to remove ammonium from wastewater. Anammox bacteria have been described as obligate chemolithoautotrophs. However, many chemolithoautotrophs (i.e., nitrifiers) can use organic compounds as a supplementary carbon source. In this study, the effect of organic compounds on anammox bacteria was investigated. It was shown that alcohols inhibited anammox bacteria, while organic acids were converted by them. Methanol was the most potent inhibitor, leading to complete and irreversible loss of activity at concentrations as low as 0.5 mM. Of the organic acids acetate and propionate, propionate was consumed at a higher rate (0.8 nmol min(-1) mg of protein(-1)) by Percoll-purified anammox cells. Glucose, formate, and alanine had no effect on the anammox process. It was shown that propionate was oxidized mainly to CO(2), with nitrate and/or nitrite as the electron acceptor. The anammox bacteria carried out propionate oxidation simultaneously with anaerobic ammonium oxidation. In an anammox enrichment culture fed with propionate for 150 days, the relative amounts of anammox cells and denitrifiers did not change significantly over time, indicating that anammox bacteria could compete successfully with heterotrophic denitrifiers for propionate. In conclusion, this study shows that anammox bacteria have a more versatile metabolism than previously assumed.

Diversity and activity of denitrifiers of Chilean arid soil ecosystems

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Julieta eOrlando

2012-04-01

Full Text Available The Chilean sclerophyllous matorral is a Mediterranean semiarid ecosystem affected by erosion, with low soil fertility and limited by nitrogen. However, limitation of resources is even more severe for desert soils such as from the Atacama Desert, one of the most extreme arid deserts on Earth. Topsoil organic matter, nitrogen and moisture content were significantly higher in the semiarid soil compared to the desert soil. Although the most significant loss of biologically preferred nitrogen from terrestrial ecosystems occurs via denitrification, virtually nothing is known on the activity and composition of denitrifier communities thriving in arid soils. In this study, we explored denitrifier communities from two soils with profoundly distinct edaphic factors. While denitrification activity in the desert soil was below detection limit, the semiarid soil sustained denitrification activity. To elucidate the genetic potential of the soils to sustain denitrification processes we performed community analysis of denitrifiers based on nitrite reductase (nirK and nirS genes as functional marker genes for this physiological group. Presence of nirK-type denitrifiers in both soils was demonstrated but failure to amplify nirS from the desert soil suggests very low abundance of nirS-type denitrifiers shedding light on the lack of denitrification activity. Phylogenetic analysis showed a very low diversity of nirK with only three distinct genotypes in the desert soil which conditions presumably exert a high selection pressure. While nirK diversity was also limited to only few, albeit distinct genotypes, the semiarid matorral soil showed a surprisingly broad genetic variability of the nirS gene. The Chilean matorral is a shrub land plant community which form vegetational patches stabilizing the soil and increasing its nitrogen and carbon content. These islands of fertility may sustain the development and activity of the overall microbial community and of

Diversity and activity of denitrifiers of chilean arid soil ecosystems.

Science.gov (United States)

Orlando, Julieta; Carú, Margarita; Pommerenke, Bianca; Braker, Gesche

2012-01-01

The Chilean sclerophyllous matorral is a Mediterranean semiarid ecosystem affected by erosion, with low soil fertility, and limited by nitrogen. However, limitation of resources is even more severe for desert soils such as from the Atacama Desert, one of the most extreme arid deserts on Earth. Topsoil organic matter, nitrogen and moisture content were significantly higher in the semiarid soil compared to the desert soil. Although the most significant loss of biologically preferred nitrogen from terrestrial ecosystems occurs via denitrification, virtually nothing is known on the activity and composition of denitrifier communities thriving in arid soils. In this study we explored denitrifier communities from two soils with profoundly distinct edaphic factors. While denitrification activity in the desert soil was below detection limit, the semiarid soil sustained denitrification activity. To elucidate the genetic potential of the soils to sustain denitrification processes we performed community analysis of denitrifiers based on nitrite reductase (nirK and nirS) genes as functional marker genes for this physiological group. Presence of nirK-type denitrifiers in both soils was demonstrated but failure to amplify nirS from the desert soil suggests very low abundance of nirS-type denitrifiers shedding light on the lack of denitrification activity. Phylogenetic analysis showed a very low diversity of nirK with only three distinct genotypes in the desert soil which conditions presumably exert a high selection pressure. While nirK diversity was also limited to only few, albeit distinct genotypes, the semiarid matorral soil showed a surprisingly broad genetic variability of the nirS gene. The Chilean matorral is a shrub land plant community which form vegetational patches stabilizing the soil and increasing its nitrogen and carbon content. These islands of fertility may sustain the development and activity of the overall microbial community and of denitrifiers in particular.

Denitrification and Biodiversity of Denitrifiers in a High-Mountain Mediterranean Lake

Directory of Open Access Journals (Sweden)

Antonio Castellano-Hinojosa

2017-10-01

Full Text Available Wet deposition of reactive nitrogen (Nr species is considered a main factor contributing to N inputs, of which nitrate (NO3− is usually the major component in high-mountain lakes. The microbial group of denitrifiers are largely responsible for reduction of nitrate to molecular dinitrogen (N2 in terrestrial and aquatic ecosystems, but the role of denitrification in removal of contaminant nitrates in high-mountain lakes is not well understood. We have used the oligotrophic, high-altitude La Caldera lake in the Sierra Nevada range (Spain as a model to study the role of denitrification in nitrate removal. Dissolved inorganic Nr concentration in the water column of la Caldera, mainly nitrate, decreased over the ice-free season which was not associated with growth of microbial plankton or variations in the ultraviolet radiation. Denitrification activity, estimated as nitrous oxide (N2O production, was measured in the water column and in sediments of the lake, and had maximal values in the month of August. Relative abundance of denitrifying bacteria in sediments was studied by quantitative polymerase chain reaction of the 16S rRNA and the two phylogenetically distinct clades nosZI and nosZII genes encoding nitrous oxide reductases. Diversity of denitrifiers in sediments was assessed using a culture-dependent approach and after the construction of clone libraries employing the nosZI gene as a molecular marker. In addition to genera Polymorphum, Paracoccus, Azospirillum, Pseudomonas, Hyphomicrobium, Thauera, and Methylophaga, which were present in the clone libraries, Arthrobacter, Burkholderia, and Rhizobium were also detected in culture media that were not found in the clone libraries. Analysis of biological activities involved in the C, N, P, and S cycles from sediments revealed that nitrate was not a limiting nutrient in the lake, allowed N2O production and determined denitrifiers’ community structure. All these results indicate that

Denitrification and Biodiversity of Denitrifiers in a High-Mountain Mediterranean Lake

Science.gov (United States)

Castellano-Hinojosa, Antonio; Correa-Galeote, David; Carrillo, Presentación; Bedmar, Eulogio J.; Medina-Sánchez, Juan M.

2017-01-01

Wet deposition of reactive nitrogen (Nr) species is considered a main factor contributing to N inputs, of which nitrate (NO3−) is usually the major component in high-mountain lakes. The microbial group of denitrifiers are largely responsible for reduction of nitrate to molecular dinitrogen (N2) in terrestrial and aquatic ecosystems, but the role of denitrification in removal of contaminant nitrates in high-mountain lakes is not well understood. We have used the oligotrophic, high-altitude La Caldera lake in the Sierra Nevada range (Spain) as a model to study the role of denitrification in nitrate removal. Dissolved inorganic Nr concentration in the water column of la Caldera, mainly nitrate, decreased over the ice-free season which was not associated with growth of microbial plankton or variations in the ultraviolet radiation. Denitrification activity, estimated as nitrous oxide (N2O) production, was measured in the water column and in sediments of the lake, and had maximal values in the month of August. Relative abundance of denitrifying bacteria in sediments was studied by quantitative polymerase chain reaction of the 16S rRNA and the two phylogenetically distinct clades nosZI and nosZII genes encoding nitrous oxide reductases. Diversity of denitrifiers in sediments was assessed using a culture-dependent approach and after the construction of clone libraries employing the nosZI gene as a molecular marker. In addition to genera Polymorphum, Paracoccus, Azospirillum, Pseudomonas, Hyphomicrobium, Thauera, and Methylophaga, which were present in the clone libraries, Arthrobacter, Burkholderia, and Rhizobium were also detected in culture media that were not found in the clone libraries. Analysis of biological activities involved in the C, N, P, and S cycles from sediments revealed that nitrate was not a limiting nutrient in the lake, allowed N2O production and determined denitrifiers’ community structure. All these results indicate that denitrification could be a

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

International Nuclear Information System (INIS)

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 ∼ 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

Methylovulum psychrotolerans sp. nov., a cold-adapted methanotroph from low-temperature terrestrial environments, and emended description of the genus Methylovulum.

Science.gov (United States)

Oshkin, Igor Y; Belova, Svetlana E; Danilova, Olga V; Miroshnikov, Kirill K; Rijpstra, W Irene C; Sinninghe Damsté, Jaap S; Liesack, Werner; Dedysh, Svetlana N

2016-06-01

Two isolates of aerobic methanotrophic bacteria, strains Sph1T and Sph2, were obtained from cold methane seeps in a floodplain of the river Mukhrinskaya, Irtysh basin, West Siberia. Another morphologically and phenotypically similar methanotroph, strain OZ2, was isolated from a sediment of a subarctic freshwater lake, Archangelsk region, northern Russia. Cells of these three strains were Gram-stain-negative, light-pink-pigmented, non-motile, encapsulated, large cocci that contained an intracytoplasmic membrane system typical of type I methanotrophs. They possessed a particulate methane monooxygenase enzyme and utilized only methane and methanol. Strains Sph1T, Sph2 and OZ2 were able to grow at a pH range of 4.0-8.9 (optimum at pH 6.0-7.0) and at temperatures between 2 and 36 °C. Although their temperature optimum was at 20-25 °C, these methanotrophs grew well at lower temperatures, down to 4 °C. The major cellular fatty acids were C16 : 1ω5c, C16 : 1ω6c, C16 : 1ω7c, C16 : 1ω8c, C16 : 0 and C14 : 0; the DNA G+C content was 51.4-51.9 mol%. Strains Sph1T, Sph2 and OZ2 displayed nearly identical (99.1-99.7 % similarity) 16S rRNA gene sequences and belonged to the family Methylococcaceae of the class Gammaproteobacteria. The most closely related organism was Methylovulum miyakonense HT12T (96.0-96.5 % 16S rRNA gene sequence similarity and 90 % pmoA sequence similarity). The novel isolates, however, differed from Methylovulum miyakonense HT12T by cell morphology, pigmentation, absence of soluble methane monooxygenase, more active growth at low temperatures, growth over a broader pH range and higher DNA G+C content. On the basis of these differences, we propose a novel species, Methylovulum psychrotolerans sp. nov., to accommodate these methanotrophs. Strain Sph1T (=LMG 29227T=VKM B-3018T) is the type strain.

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

International Nuclear Information System (INIS)

Chen Chuan; Ren Nanqi; Wang Aijie; Liu Lihong; Lee, Duu-Jong

2010-01-01

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

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.

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

Energy Technology Data Exchange (ETDEWEB)

Maeda, Koki, E-mail: k_maeda@affrc.go.jp [Dairy Research Division, National Agricultural Research Center for Hokkaido Region, National Agricultural and Food Research Organization, 1 Hitsujigaoka, Sapporo 062-8555 (Japan); Department of Environmental Chemistry and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502 (Japan); Toyoda, Sakae [Department of Environmental Chemistry and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502 (Japan); Hanajima, Dai [Dairy Research Division, National Agricultural Research Center for Hokkaido Region, National Agricultural and Food Research Organization, 1 Hitsujigaoka, Sapporo 062-8555 (Japan); Yoshida, Naohiro [Department of Environmental Chemistry and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502 (Japan)

2013-03-15

Highlights: ► Nitrous oxide (N{sub 2}O) productions of each compost zones were compared. ► The pile surface emitted significant fluxes of N{sub 2}O. ► The isotopic signature of N{sub 2}O from surface and NO{sub 2}{sup −} amended core were different. ► The denitrifying gene abundance was significantly higher in pile surface than the pile core. -- Abstract: During the dairy manure composting process, significant nitrous oxide (N{sub 2}O) emissions occur just after the pile turnings. To understand the characteristics of this N{sub 2}O emission, samples were taken from the compost surface and core independently, and the N{sub 2}O production was monitored in laboratory incubation experiments. Equal amounts of surface and core samples were mixed to simulate the turning, and the {sup 15}N isotope ratios within the molecules of produced N{sub 2}O were analyzed by isotopomer analysis. The results showed that the surface samples emitted significant levels of N{sub 2}O, and these emissions were correlated with NO{sub x}{sup −}-N accumulation. Moreover, the surface samples and surface-core mixed samples incubated at 30 °C produced N{sub 2}O with a low site preference (SP) value (−0.9 to 7.0‰) that was close to bacteria denitrification (0‰), indicating that denitrifiers in the surface samples are responsible for this N{sub 2}O production. On the other hand, N{sub 2}O produced by NO{sub 2}{sup −}-amended core samples and surface samples incubated at 60 °C showed unrecognized isotopic signatures (SP = 11.4–20.3‰). From these results, it was revealed that the N{sub 2}O production occurring just after the turnings was mainly derived from bacterial denitrification (including nitrifier denitrification) of NO{sub x}{sup −}-N under mesophilic conditions, and surface denitrifying bacteria appeared to be the main contributor to this process.

Complete genome sequence of the aerobic facultative methanotroph Methylocella silvestris BL2.

Science.gov (United States)

Chen, Yin; Crombie, Andrew; Rahman, M Tanvir; Dedysh, Svetlana N; Liesack, Werner; Stott, Matthew B; Alam, Maqsudul; Theisen, Andreas R; Murrell, J Colin; Dunfield, Peter F

2010-07-01

Methylocella silvestris BL2 is an aerobic methanotroph originally isolated from an acidic forest soil in Germany. It is the first fully authenticated facultative methanotroph. It grows not only on methane and other one-carbon (C(1)) substrates, but also on some compounds containing carbon-carbon bonds, such as acetate, pyruvate, propane, and succinate. Here we report the full genome sequence of this bacterium.

Effects of Long-Term CO2 Enrichment on Soil-Atmosphere CH4 Fluxes and the Spatial Micro-Distribution of Methanotrophic Bacteria.

Science.gov (United States)

Karbin, Saeed; Guillet, Cécile; Kammann, Claudia I; Niklaus, Pascal A

2015-01-01

Effects of elevated atmospheric CO2 concentrations on plant growth and associated C cycling have intensively been studied, but less is known about effects on the fluxes of radiatively active trace gases other than CO2. Net soil-atmosphere CH4 fluxes are determined by the balance of soil microbially-driven methane (CH4) oxidation and methanogenesis, and both might change under elevated CO2. Here, we studied CH4 dynamics in a permanent grassland exposed to elevated CO2 for 14 years. Soil-atmosphere fluxes of CH4 were measured using large static chambers, over a period of four years. The ecosystem was a net sink for atmospheric CH4 for most of the time except summer to fall when net CH4 emissions occurred. We did not detect any elevated CO2 effects on CH4 fluxes, but emissions were difficult to quantify due to their discontinuous nature, most likely because of ebullition from the saturated zone. Potential methanotrophic activity, determined by incubation of fresh sieved soil under standardized conditions, also did not reveal any effect of the CO2 treatment. Finally, we determined the spatial micro-distribution of methanotrophic activity at less than 5× atmospheric (10 ppm) and elevated (10000 ppm) CH4 concentrations, using a novel auto-radiographic technique. These analyses indicated that domains of net CH4 assimilation were distributed throughout the analyzed top 15 cm of soils, with no dependence on CH4 concentration or CO2 treatment. Our investigations suggest that elevated CO2 exerts no or only minor effects on CH4 fluxes in the type of ecosystem we studied, at least as long as soil moisture differences are small or absent as was the case here. The autoradiographic analyses further indicate that the spatial niche of CH4 oxidation does not shift in response to CO2 enrichment or CH4 concentration, and that the same type of methanotrophs may oxidize CH4 from atmospheric and soil-internal sources.

Biological Methanol Production by a Type II Methanotroph Methylocystis bryophila.

Science.gov (United States)

Patel, Sanjay K S; Mardina, Primata; Kim, Sang-Yong; Lee, Jung-Kul; Kim, In-Won

2016-04-28

Methane (CH₄) is the most abundant component in natural gas. To reduce its harmful environmental effect as a greenhouse gas, CH₄ can be utilized as a low-cost feed for the synthesis of methanol by methanotrophs. In this study, several methanotrophs were examined for their ability to produce methanol from CH₄; including Methylocella silvestris, Methylocystis bryophila, Methyloferula stellata, and Methylomonas methanica. Among these methanotrophs, M. bryophila exhibited the highest methanol production. The optimum process parameters aided in significant enhancement of methanol production up to 4.63 mM. Maximum methanol production was observed at pH 6.8, 30°C, 175 rpm, 100 mM phosphate buffer, 50 mM MgCl₂ as a methanol dehydrogenase inhibitor, 50% CH₄ concentration, 24 h of incubation, and 9 mg of dry cell mass ml(-1) inoculum load, respectively. Optimization of the process parameters, screening of methanol dehydrogenase inhibitors, and supplementation with formate resulted in significant improvements in methanol production using M. bryophila. This report suggests, for the first time, the potential of using M. bryophila for industrial methanol production from CH₄.

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

Science.gov (United States)

Chen, Chuan; Ren, Nanqi; Wang, Aijie; Liu, Lihong; Lee, Duu-Jong

2010-07-15

The denitrifying sulfide removal (DSR) process with bio-granules comprising both heterotrophic and autotrophic denitrifiers can simultaneously convert nitrate, sulfide and acetate into di-nitrogen gas, elementary sulfur and carbon dioxide, respectively, at high loading rates. This study determines the reaction rate of sulfide oxidized into sulfur, as well as the reduction of nitrate to nitrite, would be enhanced under a micro-aerobic condition. The presence of limited oxygen mitigated the inhibition effects of sulfide on denitrifier activities, and enhanced the performance of DSR granules. The advantages and disadvantages of applying the micro-aerobic condition to the DSR process are discussed. 2010 Elsevier B.V. All rights reserved.

ESTANDARIZACIÓN DE CONDICIONES PARA LA PRUEBA CUANTITATIVA DEL NMP CON BACTERIAS NITRIFICANTES Y DENITRIFICANTES USANDO COMO MATRIZ COMPOST

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N. Rodríguez-Moreno

2007-12-01

Full Text Available In order to standardize methodologies that allow to determine the concentration of nitrifying and denitrifyingbacteria in compost, bacteria growth was evaluated at two different concentrations of nitrogen substrate(NH 42 SO4 and KNO3, with and without calcium carbonate. Technique used was the most probablenumber (MPN, revealing the tubes by means Griess reagent (nitrite, Nessler reagent (ammonium, andthe zinc powder (nitrate. Four treatments for nitrifying and denitrifying bacteria were applied respectively,with concentrations of 0,5 or 0,33 g of (NH4 2SO4 and 0,5 or 2g of KNO3. Biomass production wasevaluated. No significant differences were evidenced in biomass of days14 th,15th and 16th for nitrifyingbacteria; however, 15 th day 115.88 MPN/g of compost was obtained for nitrifying bacteria, and 31.42MPN/g for denitrifying bacteria. Treatment 1 (0.5g of (NH42SO4 with 1g of CaCO3 in nitrifying and 0.5gof KNO3 with 5g of CaCO3 in denitrifying group represent the best biomass condition: 112.67 MPN/g and27.53 MPN/g of compost. 15 days were necessary for incubation because it is the time needed to activateenzymes regulating pH with addition of the CaCO3.

Enrichments of methanotrophic-heterotrophic cultures with high poly-β-hydroxybutyrate (PHB) accumulation capacities.

Science.gov (United States)

Zhang, Tingting; Wang, Xiaowei; Zhou, Jiti; Zhang, Yu

2018-03-01

Methanotrophic-heterotrophic communities were selectively enriched from sewage sludge to obtain a mixed culture with high levels of poly-β-hydroxybutyrate (PHB) accumulation capacity from methane. Methane was used as the carbon source, N 2 as sole nitrogen source, and oxygen and Cu content were varied. Copper proved essential for PHB synthesis. All cultures enriched with Cu could accumulate high content of PHB (43.2%-45.9%), while only small amounts of PHB were accumulated by cultures enriched without Cu (11.9%-17.5%). Batch assays revealed that communities grown with Cu and a higher O 2 content synthesized more PHB, which had a wider optimal CH 4 :O 2 range and produced a high PHB content (48.7%) even though in the presence of N 2 . In all methanotrophic-heterotrophic communities, both methanotrophic and heterotrophic populations showed the ability to accumulate PHB. Although methane was added as the sole carbon source, heterotrophs dominated with abundances between 77.2% and 85.6%. All methanotrophs detected belonged to type II genera, which formed stable communities with heterotrophs of different PHB production capacities. Copyright © 2017. Published by Elsevier B.V.

Identification of the dominant sulfate-reducing bacterial partner of anaerobic methanotrophs of the ANME-2 clade.

Science.gov (United States)

Schreiber, Lars; Holler, Thomas; Knittel, Katrin; Meyerdierks, Anke; Amann, Rudolf

2010-08-01

The anaerobic oxidation of methane (AOM) with sulfate as terminal electron acceptor is mediated by consortia of methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB). Whereas three clades of ANME have been repeatedly studied with respect to phylogeny, key genes and genomic capabilities, little is known about their sulfate-reducing partner. In order to identify the partner of anaerobic methanotrophs of the ANME-2 clade, bacterial 16S rRNA gene libraries were constructed from cultures highly enriched for ANME-2a and ANME-2c in consortia with Deltaproteobacteria of the Desulfosarcina/Desulfococcus group (DSS). Phylogenetic analysis of those and publicly available sequences from AOM sites supported the hypothesis by Knittel and colleagues that the DSS partner belongs to the diverse SEEP-SRB1 cluster. Six subclusters of SEEP-SRB1, SEEP-SRB1a to SEEP-SRB1f, were proposed and specific oligonucleotide probes were designed. Using fluorescence in situ hybridization on samples from six different AOM sites, SEEP-SRB1a was identified as sulfate-reducing partner in up to 95% of total ANME-2 consortia. SEEP-SRB1a cells exhibited a rod-shaped, vibrioid, or coccoid morphology and were found to be associated with subgroups ANME-2a and ANME-2c. Moreover, SEEP-SRB1a was also detected in 8% to 23% of ANME-3 consortia in Haakon Mosby Mud Volcano sediments, previously described to be predominantly associated with SRB of the Desulfobulbus group. SEEP-SRB1a contributed to only 0.3% to 0.7% of all single cells in almost all samples indicating that these bacteria are highly adapted to a symbiotic relationship with ANME-2. © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.

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.

Complete Genome Sequence of the Aerobic Facultative Methanotroph Methylocella silvestris BL2▿

Science.gov (United States)

Chen, Yin; Crombie, Andrew; Rahman, M. Tanvir; Dedysh, Svetlana N.; Liesack, Werner; Stott, Matthew B.; Alam, Maqsudul; Theisen, Andreas R.; Murrell, J. Colin; Dunfield, Peter F.

2010-01-01

Methylocella silvestris BL2 is an aerobic methanotroph originally isolated from an acidic forest soil in Germany. It is the first fully authenticated facultative methanotroph. It grows not only on methane and other one-carbon (C1) substrates, but also on some compounds containing carbon-carbon bonds, such as acetate, pyruvate, propane, and succinate. Here we report the full genome sequence of this bacterium. PMID:20472789

Complete Genome Sequence of the Aerobic Facultative Methanotroph Methylocella silvestris BL2▿

OpenAIRE

Chen, Yin; Crombie, Andrew; Rahman, M. Tanvir; Dedysh, Svetlana N.; Liesack, Werner; Stott, Matthew B.; Alam, Maqsudul; Theisen, Andreas R.; Murrell, J. Colin; Dunfield, Peter F.

2010-01-01

Methylocella silvestris BL2 is an aerobic methanotroph originally isolated from an acidic forest soil in Germany. It is the first fully authenticated facultative methanotroph. It grows not only on methane and other one-carbon (C1) substrates, but also on some compounds containing carbon-carbon bonds, such as acetate, pyruvate, propane, and succinate. Here we report the full genome sequence of this bacterium.

Sulphur bacteria mediated formation of Palaeoproterozoic phosphorites

Science.gov (United States)

Joosu, Lauri; Lepland, Aivo; Kirsimäe, Kalle

2014-05-01

Modern phosphorite formation is typically associated with high productivity in upwelling areas where apatite (Ca-phosphate) precipitation is mediated by sulphur oxidising bacteria [1]. They inhabit the oxic/anoxic interface within the upper few centimetres of sediment column, accumulating phosphate in their cells under oxic conditions and releasing it rapidly when conditions become anoxic. Sulphur bacteria are known to live in close association with a consortium of anaerobic methane oxidising archaea and syntrophic sulphate-reducing bacteria. Paleoproterozoic, c. 2.0 Ga Zaonega Formation in Karelia, Russia contains several P-rich intervals in the upper part of 1500 m thick succession of organic-rich sedimentary rocks interlayered with mafic tuffs and lavas. Apatite in these P-rich intervals forms impure laminae, lenses and round-oval nodules which diameters typically range from 300 to 1000 μm. Individual apatite particles in P-rich laminae and nodules commonly occur as cylinders that are 1-8 μm long and have diameters of 0.5-4 μm. Cross-sections of best preserved cylindrical apatite particles reveal a thin outer rim whereas the internal parts consist of small anhedral elongated crystallites, intergrown with carbonaceous material. During recrystallization the outer rim thickens towards interior and cylinders may attain hexagonal crystal habit, but their size and shape remains largely unchanged [2]. The sizes of Zaonega nodules are similar to giant sulphide-oxidising bacteria known from modern and ancient settings [3, 4]. Individual apatite cylinders and aggregates have shapes and sizes similar to the methanotrophic archaea that inhabit microbial mats in modern seep/vent areas where they operate in close associations with sulphur-oxidising microbial communities [5]. Seep/vent influence during the Zaonega phosphogenesis is indicated by variable, though positive Eu anomaly, expected in magmatically active sedimentary environment experiencing several lava flows

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.

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

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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.

Peatland succession induces a shift in the community composition of Sphagnum-associated active methanotrophs.

Science.gov (United States)

Putkinen, Anuliina; Larmola, Tuula; Tuomivirta, Tero; Siljanen, Henri M P; Bodrossy, Levente; Tuittila, Eeva-Stiina; Fritze, Hannu

2014-06-01

Sphagnum-associated methanotrophs (SAM) are an important sink for the methane (CH4) formed in boreal peatlands. We aimed to reveal how peatland succession, which entails a directional change in several environmental variables, affects SAM and their activity. Based on the pmoA microarray results, SAM community structure changes when a peatland develops from a minerotrophic fen to an ombrotrophic bog. Methanotroph subtypes Ia, Ib, and II showed slightly contrasting patterns during succession, suggesting differences in their ecological niche adaptation. Although the direct DNA-based analysis revealed a high diversity of type Ib and II methanotrophs throughout the studied peatland chronosequence, stable isotope probing (SIP) of the pmoA gene indicated they were active mainly during the later stages of succession. In contrast, type Ia methanotrophs showed active CH4 consumption in all analyzed samples. SIP-derived (13)C-labeled 16S rRNA gene clone libraries revealed a high diversity of SAM in every succession stage including some putative Methylocella/Methyloferula methanotrophs that are not detectable with the pmoA-based approach. In addition, a high diversity of 16S rRNA gene sequences likely representing cross-labeled nonmethanotrophs was discovered, including a significant proportion of Verrucomicrobia-related sequences. These results help to predict the effects of changing environmental conditions on SAM communities and activity. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Co-composting of municipal solid waste mixed with matured sewage sludge: The relationship between N2O emissions and denitrifying gene abundance.

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Bian, Rongxing; Sun, Yingjie; Li, Weihua; Ma, Qiang; Chai, Xiaoli

2017-12-01

Aerobic composting is an alternative measure to the disposal of municipal solid waste (MSW). However, it produces nitrous oxide (N 2 O), a highly potent greenhouse via microbial nitrification and denitrification. In this study, the effects of matured sewage sludge (MSS) amendment on N 2 O emissions and the inter-relationships between N 2 O emissions and the abundance of denitrifying bacteria were investigated during aerobic composting of MSW. The results demonstrated that MSW composting with MSS amendments (C1, and C2, with a MSW to MSS ratio of 2:1 and 4:1, (v/v), respectively) significantly increased N 2 O emissions during the initial stage, yet contributed to the mitigation of N 2 O emissions during the cooling and maturation stage. MSS amended composting emitted a total of 18.4%-25.7% less N 2 O than the control treatment without MSS amendment (CK). Matured sewage sludge amendment also significantly altered the abundance of denitrifying bacteria. The quantification of denitrifying functional genes revealed that the N 2 O emission rate had a significant positive correlation with the abundance of the nirS, nirK genes in both treatments with MSS amendment. The nosZ/(nirS + nirK) ratio could be a good indicator for predicting N 2 O emissions. The higher N 2 O emission rate during the initial stage of composting mixed with MSS was characterized by lower nosZ/(nirS + nirK) ratios, compared to CK treatment. Higher ratios of nosZ/(nirS + nirK) were measured during the cooling and maturation stage in treatments with MSS which resulted in a reduction of the N 2 O emissions. These results demonstrated that MSS amendment could be a valid strategy for mitigating N 2 O emissions during MSW composting. Copyright © 2017 Elsevier Ltd. All rights reserved.

The potential of methane-oxidizing bacteria for applications in environmental biotechnology

Energy Technology Data Exchange (ETDEWEB)

Wendlandt, Karin-Dagmar; Stottmeister, Ulrich; Jechorek, Mirko [Helmholtz-Center for Environmental Research UFZ, Leipzig (Germany); Helm, Jana [School of Physics and Astronomy, The University of Edinburgh, Edinburgh (United Kingdom); Soltmann, Bettina [Institute for Materials Science, Dresden University of Technology, Dresden (Germany); Beck, Matthias [Oncotec, Pharma Production GmbH, Dessau-Rosslau (Germany)

2010-04-15

Methanotrophic bacteria possess a unique set of enzymes enabling them to oxidize, degrade and transform organic molecules and synthesize new compounds. Therefore, they have great potential in environmental biotechnology. The application of these unique properties was demonstrated in three case studies: (i) Methane escaping from leaky gas pipes may lead to massive mortality of trees in urban areas. Lack of oxygen within the soil surrounding tree roots caused by methanotrophic activity was identified as one of the reasons for this phenomenon. The similarity between metabolic reactions performed by the key enzymes of methanotrophs (methane monooxygenase) and ammonium oxidizers (ammonium monooxygenase) might offer a solution to this problem by applying commercially available nitrification and urease inhibitors. (ii) Methanotrophs are able to co-metabolically degrade contaminants such as low-molecular-weight-chlorinated hydrocarbons in soil and water in the presence of methane. Batch and continuous trichloroethylene degradation experiments in laboratory-scale reactors using Methylocystis sp. GB 14 were performed, partly with cells entrapped in a polymer matrix. (iii) Using a short, two-stage pilot-scale process, the intracellular polymer accumulation of poly-{beta}-hydroxybutyrate (PHB) in methanotrophs reached a maximum of 52%. Interestingly, an ultra-high-molecular-weight PHB of 3.1 MDa was accumulated under potassium deficiency. Under strictly controlled conditions (temperature, pH and methane supply) this process can be nonsterile because of the establishment of a stable microbial community (dominant species Methylocystis sp. GB 25 {>=}86% by biomass). The possibility to substitute methane with biogas from renewable sources facilitates the development of a methane-based PHB production process that yields a high-quality biopolymer at competitive costs. (Abstract Copyright [2010], Wiley Periodicals, Inc.)

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.

Anaerobic methane oxidation coupled to denitrification is the dominant methane sink in a deep lake

DEFF Research Database (Denmark)

Deutzmann, Joerg S.; Stief, Peter; Brandes, Josephin

2014-01-01

Anaerobic methane oxidation coupled to denitrification, also known as “nitrate/nitrite-dependent anaerobic methane oxidation” (n-damo), was discovered in 2006. Since then, only a few studies have identified this process and the associated microorganisms in natural environments. In aquatic sediments......, the close proximity of oxygen- and nitrate-consumption zones can mask n-damo as aerobic methane oxidation. We therefore investigated the vertical distribution and the abundance of denitrifying methanotrophs related to Candidatus Methylomirabilis oxyfera with cultivation-independent molecular techniques...... in the sediments of Lake Constance. Additionally, the vertical distribution of methane oxidation and nitrate consumption zones was inferred from high-resolution microsensor profiles in undisturbed sediment cores. M. oxyfera-like bacteria were virtually absent at shallow-water sites (littoral sediment) and were...

Windrow composting mitigated CH4 emissions: characterization of methanogenic and methanotrophic communities in manure management.

Science.gov (United States)

Chen, Ruirui; Wang, Yiming; Wei, Shiping; Wang, Wei; Lin, Xiangui

2014-12-01

With increasing livestock breeding, methane (CH4 ) emissions from manure management will increasingly contribute more to atmospheric CH4 concentration. The dynamics of methanogens and methanotrophs have not yet been studied in the manure environment. The current study combines surface CH4 emissions with methanogenic and methanotrophic community analyses from two management practices, windrow composting (WCOM) and solid storage (SSTO). Our results showed that there was an c. 50% reduction of CH4 emissions with WCOM compared with SSTO over a 50-day period. A sharp decrease in the quantities of both methanogens and methanotrophs in WCOM suggested that CH4 mitigation was mainly due to decreased CH4 production rather than increased CH4 oxidation. Pyrosequencing analysis demonstrated that aeration caused a clear shift of dominant methanogens in the manure, with specifically a significant decrease in Methanosarcina and increase in Methanobrevibacter. The composition of methanogenic community was influenced by manure management and regulated CH4 production. A sharp increase in the quantity of methanotrophs in SSTO suggested that microbial CH4 oxidation is an important sink for the CH4 produced. The increased abundance of Methylococcaceae in SSTO suggested that Type I methanotrophs have an advantage in CH4 oxidation in occupying niches under low CH4 and high O2 conditions. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Molecular characterization of diazotrophic and denitrifying bacteria associated with mangrove roots.

Science.gov (United States)

Flores-Mireles, Ana L; Winans, Stephen C; Holguin, Gina

2007-11-01

An analysis of the molecular diversity of N(2) fixers and denitrifiers associated with mangrove roots was performed using terminal restriction length polymorphism (T-RFLP) of nifH (N(2) fixation) and nirS and nirK (denitrification), and the compositions and structures of these communities among three sites were compared. The number of operational taxonomic units (OTU) for nifH was higher than that for nirK or nirS at all three sites. Site 3, which had the highest organic matter and sand content in the rhizosphere sediment, as well as the lowest pore water oxygen concentration, had the highest nifH diversity. Principal component analysis of biogeochemical parameters identified soil texture, organic matter content, pore water oxygen concentration, and salinity as the main variables that differentiated the sites. Nonmetric multidimensional scaling (MDS) analyses of the T-RFLP data using the Bray-Curtis coefficient, group analyses, and pairwise comparisons between the sites clearly separated the OTU of site 3 from those of sites 1 and 2. For nirS, there were statistically significant differences in the composition of OTU among the sites, but the variability was less than for nifH. OTU defined on the basis of nirK were highly similar, and the three sites were not clearly separated on the basis of these sequences. The phylogenetic trees of nifH, nirK, and nirS showed that most of the cloned sequences were more similar to sequences from the rhizosphere isolates than to those from known strains or from other environments.

Molecular Characterization of Diazotrophic and Denitrifying Bacteria Associated with Mangrove Roots▿

Science.gov (United States)

Flores-Mireles, Ana L.; Winans, Stephen C.; Holguin, Gina

2007-01-01

An analysis of the molecular diversity of N2 fixers and denitrifiers associated with mangrove roots was performed using terminal restriction length polymorphism (T-RFLP) of nifH (N2 fixation) and nirS and nirK (denitrification), and the compositions and structures of these communities among three sites were compared. The number of operational taxonomic units (OTU) for nifH was higher than that for nirK or nirS at all three sites. Site 3, which had the highest organic matter and sand content in the rhizosphere sediment, as well as the lowest pore water oxygen concentration, had the highest nifH diversity. Principal component analysis of biogeochemical parameters identified soil texture, organic matter content, pore water oxygen concentration, and salinity as the main variables that differentiated the sites. Nonmetric multidimensional scaling (MDS) analyses of the T-RFLP data using the Bray-Curtis coefficient, group analyses, and pairwise comparisons between the sites clearly separated the OTU of site 3 from those of sites 1 and 2. For nirS, there were statistically significant differences in the composition of OTU among the sites, but the variability was less than for nifH. OTU defined on the basis of nirK were highly similar, and the three sites were not clearly separated on the basis of these sequences. The phylogenetic trees of nifH, nirK, and nirS showed that most of the cloned sequences were more similar to sequences from the rhizosphere isolates than to those from known strains or from other environments. PMID:17827324

The activity and community structure of total bacteria and denitrifying bacteria across soil depths and biological gradients in estuary ecosystem.

Science.gov (United States)

Lee, Seung-Hoon; Kang, Hojeong

2016-02-01

The distribution of soil microorganisms often shows variations along soil depth, and even in the same soil layer, each microbial group has a specific niche. In particular, the estuary soil is intermittently flooded, and the characteristics of the surface soil layer are different from those of other terrestrial soils. We investigated the microbial community structure and activity across soil depths and biological gradients composed of invasive and native plants in the shallow surface layer of an estuary ecosystem by using molecular approaches. Our results showed that the total and denitrifying bacterial community structures of the estuarine wetland soil differed according to the short depth gradient. In growing season, gene copy number of 16S rRNA were 1.52(±0.23) × 10(11), 1.10(±0.06) × 10(11), and 4.33(±0.16) × 10(10) g(-1) soil; nirS were 5.41(±1.25) × 10(8), 4.93(±0.94) × 10(8), and 2.61(±0.28) × 10(8) g(-1) soil; and nirK were 9.67(±2.37) × 10(6), 3.42(±0.55) × 10(6), and 2.12(±0.19) × 10(6) g(-1) soil in 0 cm, 5 cm, and 10 cm depth layer, respectively. The depth-based difference was distinct in the vegetated sample and in the growing season, evidencing the important role of plants in structuring the microbial community. In comparison with other studies, we observed differences in the microbial community and functions even across very short depth gradients. In conclusion, our results suggested that (i) in the estuary ecosystem, the denitrifying bacterial community could maintain its abundance and function within shallow surface soil layers through facultative anaerobiosis, while the total bacterial community would be both quantitatively and qualitatively affected by the soil depth, (ii) the nirS gene community, rather than the nirK one, should be the first candidate used as an indicator of the microbial denitrification process in the estuary system, and (iii) as the microbial community is distributed and plays a certain niche role according to

Abundance and diversity of bacterial nitrifiers and denitrifiers and their functional genes in tannery wastewater treatment plants revealed by high-throughput sequencing.

Directory of Open Access Journals (Sweden)

Zhu Wang

Full Text Available Biological nitrification/denitrification is frequently used to remove nitrogen from tannery wastewater containing high concentrations of ammonia. However, information is limited about the bacterial nitrifiers and denitrifiers and their functional genes in tannery wastewater treatment plants (WWTPs due to the low-throughput of the previously used methods. In this study, 454 pyrosequencing and Illumina high-throughput sequencing, combined with molecular methods, were used to comprehensively characterize structures and functions of nitrification and denitrification bacterial communities in aerobic and anaerobic sludge of two full-scale tannery WWTPs. Pyrosequencing of 16S rRNA genes showed that Proteobacteria and Synergistetes dominated in the aerobic and anaerobic sludge, respectively. Ammonia-oxidizing bacteria (AOB amoA gene cloning revealed that Nitrosomonas europaea dominated the ammonia-oxidizing community in the WWTPs. Metagenomic analysis showed that the denitrifiers mainly included the genera of Thauera, Paracoccus, Hyphomicrobium, Comamonas and Azoarcus, which may greatly contribute to the nitrogen removal in the two WWTPs. It is interesting that AOB and ammonia-oxidizing archaea had low abundance although both WWTPs demonstrated high ammonium removal efficiency. Good correlation between the qPCR and metagenomic analysis is observed for the quantification of functional genes amoA, nirK, nirS and nosZ, indicating that the metagenomic approach may be a promising method used to comprehensively investigate the abundance of functional genes of nitrifiers and denitrifiers in the environment.

Diversity of the active methanotrophic community in acidic peatlands as assessed by mRNA and SIP-PLFA analyses.

Science.gov (United States)

Chen, Yin; Dumont, Marc G; McNamara, Niall P; Chamberlain, Paul M; Bodrossy, Levente; Stralis-Pavese, Nancy; Murrell, J Colin

2008-02-01

The active methanotroph community was investigated for the first time in heather (Calluna)-covered moorlands and Sphagnum/Eriophorum-covered UK peatlands. Direct extraction of mRNA from these soils facilitated detection of expression of methane monooxygenase genes, which revealed that particulate methane monooxygenase and not soluble methane monooxygenase was probably responsible for CH(4) oxidation in situ, because only pmoA transcripts (encoding a subunit of particulate methane monooxygenase) were readily detectable. Differences in methanotroph community structures were observed between the Calluna-covered moorland and Sphagnum/Eriophorum-covered gully habitats. As with many other Sphagnum-covered peatlands, the Sphagnum/Eriophorum-covered gullies were dominated by Methylocystis. Methylocella and Methylocapsa-related species were also present. Methylobacter-related species were found as demonstrated by the use of a pmoA-based diagnostic microarray. In Calluna-covered moorlands, in addition to Methylocella and Methylocystis, a unique group of peat-associated type I methanotrophs (Gammaproteobacteria) and a group of uncultivated type II methanotrophs (Alphaproteobacteria) were also found. The pmoA sequences of the latter were only distantly related to Methylocapsa and also to the RA-14 group of methanotrophs, which are believed to be involved in oxidation of atmospheric concentrations of CH(4). Soil samples were also labelled with (13)CH(4), and subsequent analysis of the (13)C-labelled phospholipid fatty acids (PLFAs) showed that 16:1 omega 7, 18:1 omega 7 and 18:1 omega 9 were the major labelled PLFAs. The presence of (13)C-labelled 18:1 omega 9, which was not a major PLFA of any extant methanotrophs, indicated the presence of novel methanotrophs in this peatland.

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

Science.gov (United States)

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

2011-12-01

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

Functional consortium for denitrifying sulfide removal process.

Science.gov (United States)

Chen, Chuan; Ren, Nanqi; Wang, Aijie; Liu, Lihong; Lee, Duu-Jong

2010-03-01

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

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.

Co-effects of pyrene and nitrate on the activity and abundance of soil denitrifiers under anaerobic condition.

Science.gov (United States)

Zhou, Zhi-Feng; Yao, Yan-Hong; Wang, Ming-Xia; Zuo, Xiao-Hu

2017-10-01

It has previously been confirmed that polycyclic aromatic hydrocarbons (PAHs) could be degraded by soil microbes coupling with denitrification, but the relationships among soil denitrifiers, PAHs, and nitrate under obligate anaerobic condition are still unclear. Here, co-effects of pyrene and nitrate on the activity and abundance of soil denitrifiers were investigated through a 45-day incubation experiment. Two groups of soil treatments with (N 30 ) and without (N 0 ) nitrate (30 mg kg -1 dry soil) amendment were conducted, and each group contained three treatments with different pyrene concentrations (0, 30, and 60 mg kg -1 dry soil denoted as P 0 , P 30 , and P 60 , respectively). The pyrene content, abundances of denitrification concerning genes (narG, periplasmic nitrate reductase gene; nirS, cd 1 -nitrite reductase gene; nirK, copper-containing nitrite reductase gene), and productions of N 2 O and CO 2 were measured at day 3, 14, 28, and 45, and the bacterial community structures in four represented treatments (N 0 P 0 , N 0 P 60 , N 30 P 0 , and N 30 P 60 ) were analyzed at day 45. The results indicated that the treatments with higher pyrene concentration had higher final pyrene removal rates than the treatments with lower pyrene concentration. Additionally, intensive emission of N 2 O was detected in all treatments only at day 3, but a continuous production of CO 2 was measured in each treatment during the incubation. Nitrate amendment could enhance the activity of soil denitrifiers, and be helpful for soil microbes to sustain their activity. While pyrene seemed had no influence on the productions of N 2 O and CO 2 , and amendment with pyrene or nitrate both had no obvious effect on abundances of denitrification concerning genes. Furthermore, it was nitrate but not pyrene had an obvious influence on the community structure of soil bacteria. These results revealed that, under anaerobic condition, the activity and abundance of soil denitrifiers both were

Methyloferula stellata gen. nov., sp. nov., an acidophilic, obligately methanotrophic bacterium that possesses only a soluble methane monooxygenase.

Science.gov (United States)

Vorobev, Alexey V; Baani, Mohamed; Doronina, Nina V; Brady, Allyson L; Liesack, Werner; Dunfield, Peter F; Dedysh, Svetlana N

2011-10-01

Two strains of aerobic methanotrophic bacteria, AR4(T) and SOP9, were isolated from acidic (pH 3.8-4.0) Sphagnum peat bogs in Russia. Another phenotypically similar isolate, strain LAY, was obtained from an acidic (pH 4.0) forest soil in Germany. Cells of these strains were Gram-negative, non-pigmented, non-motile, thin rods that multiplied by irregular cell division and formed rosettes or amorphous cell conglomerates. Similar to Methylocella species, strains AR4(T), SOP9 and LAY possessed only a soluble form of methane monooxygenase (sMMO) and lacked intracytoplasmic membranes. Growth occurred only on methane and methanol; the latter was the preferred growth substrate. mRNA transcripts of sMMO were detectable in cells when either methane or both methane and methanol were available. Carbon was assimilated via the serine and ribulose-bisphosphate (RuBP) pathways; nitrogen was fixed via an oxygen-sensitive nitrogenase. Strains AR4(T), SOP9 and LAY were moderately acidophilic, mesophilic organisms capable of growth between pH 3.5 and 7.2 (optimum pH 4.8-5.2) and at 4-33 °C (optimum 20-23 °C). The major cellular fatty acid was 18 : 1ω7c and the quinone was Q-10. The DNA G+C content was 55.6-57.5 mol%. The isolates belonged to the family Beijerinckiaceae of the class Alphaproteobacteria and were most closely related to the sMMO-possessing methanotrophs of the genus Methylocella (96.4-97.0 % 16S rRNA gene sequence similarity), particulate MMO (pMMO)-possessing methanotrophs of the genus Methylocapsa (96.1-97.0 %), facultative methylotrophs of the genus Methylovirgula (96.1-96.3 %) and non-methanotrophic organotrophs of the genus Beijerinckia (96.5-97.0 %). Phenotypically, strains AR4(T), SOP9 and LAY were most similar to Methylocella species, but differed from members of this genus by cell morphology, greater tolerance of low pH, detectable activities of RuBP pathway enzymes and inability to grow on multicarbon compounds. Therefore, we propose a novel

The use of {sup 13}C labelling of bacterial lipids in the characterisation of ambient methane-oxidising bacteria in soils

Energy Technology Data Exchange (ETDEWEB)

Crossman, Z.M.; Evershed, R.P. [Bristol Univ., Organic Geochemistry Unit, Biogeochemistry Research Centre, Bristol (United Kingdom); Ineson, P. [York Univ., Dept. of Biology, York (United Kingdom)

2005-05-15

The occurrence of methane-oxidising bacteria in soils has received increasing attention because of their role as a sink for atmospheric methane. However, such bacteria are not amenable to modern culturing techniques and hence the widespread interest in the development of methods of cultivation-independent analysis. In the following investigation, a combination of stable isotope labelling with phospholipid fatty acid (PLFA) and bacteriohopanoid analysis was employed in an effort to characterise this functional group of bacteria. Results suggest a novel population of methane-oxidising bacteria related to type II culturable methanotrophs, in particular, the Methylocapsa and Methylocella genera of bacteria. (Author)

Functional-Structural Analysis of Nitrogen-Cycle Bacteria in a Hypersaline Mat from the Omani Desert

DEFF Research Database (Denmark)

Abed, Raeid M M; de Beer, Dirk; Stief, Peter

2015-01-01

to sequences from the Rhizobiales group. Sequences of the nosZ gene were the most diverse and clustered with sequences from various genera. Our results demonstrate that the hypersaline mat from Oman harbors nitrifying and denitrifying bacteria with the potential to perform respective processes at detectable...

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

Energy Technology Data Exchange (ETDEWEB)

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

2011-05-15

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

Draft Genome Sequence of Methyloferula stellata AR4, an Obligate Methanotroph Possessing Only a Soluble Methane Monooxygenase

OpenAIRE

Dedysh, Svetlana N.; Naumoff, Daniil G.; Vorobev, Alexey V.; Kyrpides, Nikos; Woyke, Tanja; Shapiro, Nicole; Crombie, Andrew T.; Murrell, J. Colin; Kalyuzhnaya, Marina G.; Smirnova, Angela V.; Dunfield, Peter F.

2015-01-01

Methyloferula stellata AR4 is an aerobic acidophilic methanotroph, which, in contrast to most known methanotrophs but similar to Methylocella spp., possesses only a soluble methane monooxygenase. However, it differs from Methylocella spp. by its inability to grow on multicarbon substrates. Here, we report the draft genome sequence of this bacterium.

Soil environmental conditions and microbial build-up mediate the effect of plant diversity on soil nitrifying and denitrifying enzyme activities in temperate grasslands.

Directory of Open Access Journals (Sweden)

Xavier Le Roux

Full Text Available Random reductions in plant diversity can affect ecosystem functioning, but it is still unclear which components of plant diversity (species number - namely richness, presence of particular plant functional groups, or particular combinations of these and associated biotic and abiotic drivers explain the observed relationships, particularly for soil processes. We assembled grassland communities including 1 to 16 plant species with a factorial separation of the effects of richness and functional group composition to analyze how plant diversity components influence soil nitrifying and denitrifying enzyme activities (NEA and DEA, respectively, the abundance of nitrifiers (bacterial and archaeal amoA gene number and denitrifiers (nirK, nirS and nosZ gene number, and key soil environmental conditions. Plant diversity effects were largely due to differences in functional group composition between communities of identical richness (number of sown species, though richness also had an effect per se. NEA was positively related to the percentage of legumes in terms of sown species number, the additional effect of richness at any given legume percentage being negative. DEA was higher in plots with legumes, decreased with increasing percentage of grasses, and increased with richness. No correlation was observed between DEA and denitrifier abundance. NEA increased with the abundance of ammonia oxidizing bacteria. The effect of richness on NEA was entirely due to the build-up of nitrifying organisms, while legume effect was partly linked to modified ammonium availability and nitrifier abundance. Richness effect on DEA was entirely due to changes in soil moisture, while the effects of legumes and grasses were partly due to modified nitrate availability, which influenced the specific activity of denitrifiers. These results suggest that plant diversity-induced changes in microbial specific activity are important for facultative activities such as denitrification

Mechanisms of nitrous oxide (N2 O) formation and reduction in denitrifying biofilms.

Science.gov (United States)

Sabba, Fabrizio; Picioreanu, Cristian; Nerenberg, Robert

2017-12-01

Nitrous oxide (N 2 O) is a potent greenhouse gas that can be formed in wastewater treatment processes by ammonium oxidizing and denitrifying microorganisms. While N 2 O emissions from suspended growth systems have been extensively studied, and some recent studies have addressed emissions from nitrifying biofilms, much less is known about N 2 O emissions from denitrifying biofilm processes. This research used modeling to evaluate the mechanisms of N 2 O formation and reduction in denitrifying biofilms. The kinetic model included formation and consumption of key denitrification species, including nitrate (NO3-), nitrite (NO2-), nitric oxide (NO), and N 2 O. The model showed that, in presence of excess of electron donor, denitrifying biofilms have two distinct layers of activity: an outer layer where there is net production of N 2 O and an inner layer where there is net consumption. The presence of oxygen (O 2 ) had an important effect on N 2 O emission from suspended growth systems, but a smaller effect on biofilm systems. The effects of NO3- and O 2 differed significantly based on the biofilm thickness. Overall, the effects of biofilm thickness and bulk substrate concentrations on N 2 O emissions are complex and not always intuitive. A key mechanism for denitrifying biofilms is the diffusion of N 2 O and other intermediates from one zone of the biofilm to another. This leads to zones of N 2 O formation or consumption transformations that would not exist in suspended growth systems. © 2017 Wiley Periodicals, Inc.

Soil C and N statuses determine the effect of maize inoculation by plant growth-promoting rhizobacteria on nitrifying and denitrifying communities.

Science.gov (United States)

Florio, Alessandro; Pommier, Thomas; Gervaix, Jonathan; Bérard, Annette; Le Roux, Xavier

2017-08-21

Maize inoculation by Azospirillum stimulates root growth, along with soil nitrogen (N) uptake and root carbon (C) exudation, thus increasing N use efficiency. However, inoculation effects on soil N-cycling microbial communities have been overlooked. We hypothesized that inoculation would (i) increase roots-nitrifiers competition for ammonium, and thus decrease nitrifier abundance; and (ii) increase roots-denitrifiers competition for nitrate and C supply to denitrifiers by root exudation, and thus limit or benefit denitrifiers depending on the resource (N or C) mostly limiting these microorganisms. We quantified (de)nitrifiers abundance and activity in the rhizosphere of inoculated and non-inoculated maize on 4 sites over 2 years, and ancillary soil variables. Inoculation effects on nitrification and nitrifiers (AOA, AOB) were not consistent between the three sampling dates. Inoculation influenced denitrifiers abundance (nirK, nirS) differently among sites. In sites with high C limitation for denitrifiers (i.e. limitation of denitrification by C > 66%), inoculation increased nirS-denitrifier abundance (up to 56%) and gross N 2 O production (up to 84%), likely due to increased root C exudation. Conversely, in sites with low C limitation (<47%), inoculation decreased nirS-denitrifier abundance (down to -23%) and gross N 2 O production (down to -18%) likely due to an increased roots-denitrifiers competition for nitrate.

Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments

Directory of Open Access Journals (Sweden)

Elizabeth Trembath-Reichert

2016-04-01

Full Text Available Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM. This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME and sulfate-reducing bacteria (SRB. These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range of Deltaproteobacteria diversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seep sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag. Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. Many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to

Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments.

Science.gov (United States)

Trembath-Reichert, Elizabeth; Case, David H; Orphan, Victoria J

2016-01-01

Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM). This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range of Deltaproteobacteria diversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seep sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag). Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP) iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases) and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. Many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to many co

Draft Genome Sequence of Methyloferula stellata AR4, an Obligate Methanotroph Possessing Only a Soluble Methane Monooxygenase.

Science.gov (United States)

Dedysh, Svetlana N; Naumoff, Daniil G; Vorobev, Alexey V; Kyrpides, Nikos; Woyke, Tanja; Shapiro, Nicole; Crombie, Andrew T; Murrell, J Colin; Kalyuzhnaya, Marina G; Smirnova, Angela V; Dunfield, Peter F

2015-03-05

Methyloferula stellata AR4 is an aerobic acidophilic methanotroph, which, in contrast to most known methanotrophs but similar to Methylocella spp., possesses only a soluble methane monooxygenase. However, it differs from Methylocella spp. by its inability to grow on multicarbon substrates. Here, we report the draft genome sequence of this bacterium. Copyright © 2015 Dedysh et al.

[Analysis of phylogenetic criteria for estimation of the rank of taxa in methane-oxidizing bacteria].

Science.gov (United States)

Romanovskaia, V A; Rokitko, P V

2011-01-01

To determine a possibility of application of phylogenetic criteria for estimating the taxa rank, the intra- and interspecies, as well as intergeneric relatedness of methanotrophs on the basis of 16S rRNA gene sequences was estimated. We used sequences of 16S rRNA genes of the studied isolates of obligate methanotrophs which have been deposited in UCM (Ukrainian Collection of Microorganisms), and of type strains of other obligate methanotrophs species (from GenBank database). It is shown, that the levels of interspecies and intergeneric relatedness in different families of methanotrophs are not identical, and therefore they can be used for differentiation of taxa only within one family. The carried out analysis has shown, that it is necessary to reconsider taxonomic position: (1) of two phenotypically similar species of Methylomonas (M. aurantiaca and M. fodinarum), similarity of 16S rRNA genes which is 99.4%, similarity of their total DNA--up to 80% that rather testifies to strain differences, than to species differences; (2) of species Methylomicrobium agile and M album which are phylogenetically more related to genus Methylobacter (97% of affinity), than Methylomicrobium (94% of affinity); (3) of genera of the family Beijerinckiaceae (Methylocella and Methylocapsa), and also genera of the family Methylocystaceae (Methylosinus and Methylocystis), whereas high level of relatedness (97% and more) of these bacteria with other methanotrophic genera (within one family) practically corresponds to a range of relatedness of species (within some genera) in the family Methylococcaceae. When determining phylogenetic criteria which can characterize the ranks of taxa, it was revealed, that the levels of interspecies relatedness of methanotrophic genera of the families Methylocystaceae and Beijerinckiaceae (97.8-99.1% and 97.8%, accordingly) considerably exceed the level of genera formation in the family Methylococcaceae (94.0-98.2%) and, moreover, approach the value of

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

Science.gov (United States)

Frostegard, A.; Bakken, L. R.

2010-12-01

Emissions of N2O from agricultural soils are largely caused by denitrifying bacteria. Field measurements of N2O fluxes show large variations and depend on several environmental factors, and possibly also on the composition of the denitrifying microbial community. The temporal and spatial variation of fluxes are not adequately captured by biogeochemical models, and few options for mitigations have been invented, which underscores the need to understand the mechanisms underlying the emissions of N2O. Analyses of denitrification genes and transcripts extracted from soils are important for describing the system, but may have limited value for prediction of N2O emissions. In contrast, phenotypic analyses are direct measures of the organisms’ responses to changing environmental conditions. Our approach is to combine phenotypic characterizations using high-resolution gas kinetics, with gene transcription analyses to study denitrification regulatory phenotypes (DRP) of bacterial strains or complex microbial communities. The rich data sets obtained provide a basis for refinement of biochemical and physiological research on this key process in the nitrogen cycle. The strength of this combined approach is illustrated by a series of experiments investigating effects of soil pH on denitrification. Soil pH emerges as a master variable determining the microbial community composition as well as its denitrification product ratio (N2O/N2), with higher ratio in acid than in alkaline soil. It is therefore likely that emissions of N2O from agro-ecosystems will increase in large parts of the world where soil pH is decreasing due to intensified management and increased use of chemical fertilizers. Considering its immense implications, surprisingly few attempts have been made to unravel the mechanisms involved in the pH-control of the product stoichiometry of denitrification. We investigated the kinetics of gas transformations (O2, NO, N2O and N2) and transcription of functional genes

Trace-gas metabolic versatility of the facultative methanotroph Methylocella silvestris

Science.gov (United States)

Crombie, Andrew T.; Murrell, J. Colin

2014-06-01

The climate-active gas methane is generated both by biological processes and by thermogenic decomposition of fossil organic material, which forms methane and short-chain alkanes, principally ethane, propane and butane. In addition to natural sources, environments are exposed to anthropogenic inputs of all these gases from oil and gas extraction and distribution. The gases provide carbon and/or energy for a diverse range of microorganisms that can metabolize them in both anoxic and oxic zones. Aerobic methanotrophs, which can assimilate methane, have been considered to be entirely distinct from utilizers of short-chain alkanes, and studies of environments exposed to mixtures of methane and multi-carbon alkanes have assumed that disparate groups of microorganisms are responsible for the metabolism of these gases. Here we describe the mechanism by which a single bacterial strain, Methylocella silvestris, can use methane or propane as a carbon and energy source, documenting a methanotroph that can utilize a short-chain alkane as an alternative to methane. Furthermore, during growth on a mixture of these gases, efficient consumption of both gases occurred at the same time. Two soluble di-iron centre monooxygenase (SDIMO) gene clusters were identified and were found to be differentially expressed during bacterial growth on these gases, although both were required for efficient propane utilization. This report of a methanotroph expressing an additional SDIMO that seems to be uniquely involved in short-chain alkane metabolism suggests that such metabolic flexibility may be important in many environments where methane and short-chain alkanes co-occur.

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

The Effect of Biofertilizer on The Diversity of N2O Reducing Bacteria in Paddy Fields of Sukabumi, Indonesia

Directory of Open Access Journals (Sweden)

Alfan Cahyadi

2017-12-01

Full Text Available Some of the methanotrophic bacteria and N2O reducing bacteria have been proven to be able to support the plant growth and increase the productivity of paddy. However effect of the methanotrophic and N2O reducing bacteria application as a biofertilizer to indigenous N2O reducing bacteria is still not well known yet. The aim of this study was to analyze the diversity of N2O reducing bacteria in lowland paddy soil based on a nosZ gene. Soil samples were taken from lowland paddy soils in Pelabuhan Ratu Sukabumi, West Java, Indonesia. There were two treatments for the paddy field soil, ie. biofertilizer-treated field 20% fertilizer (50 kg/ha with the addition of biofertilizer and 100% fertilizer. PCR amplification of nosZ gene was successfully conducted using nosZF and nosZR primer pair. Denaturing Gradient Gel Electrophoresis (DGGE process was conducted at 150 V for 5.5h. There were three differences nosZ bands were sequenced. The phylogenetic analysis showed that they were close to uncultured bacteria. Microbial diversity in the biofertilizer-treated field was higher than that of in the 100% fertilizer-treated field. The biofertilizer treatment has higher in microbial diversity than that of applied non-biofertilizer paddy fields. This research might have impact in the application of biofertilizers due to the emission of N2O as a green house gas from paddy fields farming activity. The biofertilizer has great potential application in sustainable environmental friendly agriculture systems.

Methylocella palustris gen. nov., sp. nov., a new methane-oxidizing acidophilic bacterium from peat bogs, representing a novel subtype of serine-pathway methanotrophs.

Science.gov (United States)

Dedysh, S N; Liesack, W; Khmelenina, V N; Suzina, N E; Trotsenko, Y A; Semrau, J D; Bares, A M; Panikov, N S; Tiedje, J M

2000-05-01

A new genus, Methylocella, and a new species, Methylocella palustris, are proposed for three strains of methane-oxidizing bacteria isolated from acidic Sphagnum peat bogs. These bacteria are aerobic, Gram-negative, colourless, non-motile, straight and curved rods that utilize the serine pathway for carbon assimilation, multiply by normal cell division and contain intracellular poly-beta-hydroxybutyrate granules (one at each pole). These strains use methane and methanol as sole sources of carbon and energy and are moderately acidophilic organisms with growth between pH 4.5 and pH 7.0, the optimum being at pH 5.0-5.5. The temperature range for growth is 10-28 degrees C with the optimum at 15-20 degrees C. The intracytoplasmic membrane system is different from those of type I and II methanotrophs. Cells contain an extensive periplasmic space and a vesicular membrane system connected to the cytoplasmic membrane. The strains grew only on media with a low salt content (0.2-0.5 g l(-1)). All three strains were found to possess soluble methane monooxygenase and are able to fix atmospheric nitrogen via an oxygen-sensitive nitrogenase. No products were observed in a PCR with particulate methane monooxygenase-targeted primers; hybridization with a pmoA probe was also negative. The major phospholipid fatty acids are 18:1 acids. The G+C content of the DNA is 61.2 mol%. The three strains share identical 16S rRNA gene sequences and represent a novel lineage of methane-oxidizing bacteria within the alpha-subclass of the class Proteobacteria and are only moderately related to type II methanotrophs of the Methylocystis-Methylosinus group. The three strains are most closely related to the acidophilic heterotrophic bacterium Beijerinckia indica subsp. indica (96.5% 16S rDNA sequence similarity). Collectively, these strains comprise a new species and genus Methylocella palustris gen. nov., sp. nov.; strain KT (= ATCC 700799T) is the type strain.

Reduction of produced elementary sulfur in denitrifying sulfide removal process.

Science.gov (United States)

Zhou, Xu; Liu, Lihong; Chen, Chuan; Ren, Nanqi; Wang, Aijie; Lee, Duu-Jong

2011-05-01

Denitrifying sulfide removal (DSR) processes simultaneously convert sulfide, nitrate, and chemical oxygen demand from industrial wastewater into elemental sulfur, dinitrogen gas, and carbon dioxide, respectively. The failure of a DSR process is signaled by high concentrations of sulfide in reactor effluent. Conventionally, DSR reactor failure is blamed for overcompetition for heterotroph to autotroph communities. This study indicates that the elementary sulfur produced by oxidizing sulfide that is a recoverable resource from sulfide-laden wastewaters can be reduced back to sulfide by sulfur-reducing Methanobacterium sp. The Methanobacterium sp. was stimulated with excess organic carbon (acetate) when nitrite was completely consumed by heterotrophic denitrifiers. Adjusting hydraulic retention time of a DSR reactor when nitrite is completely consumed provides an additional control variable for maximizing DSR performance.

Denitrifying woodchip bioreactor and phosphorus filter pairing to minimize pollution swapping.

Science.gov (United States)

Christianson, Laura E; Lepine, Christine; Sibrell, Philip L; Penn, Chad; Summerfelt, Steven T

2017-09-15

Pairing denitrifying woodchip bioreactors and phosphorus-sorbing filters provides a unique, engineered approach for dual nutrient removal from waters impaired with both nitrogen (N) and phosphorus (P). This column study aimed to test placement of two P-filter media (acid mine drainage treatment residuals and steel slag) relative to a denitrifying system to maximize N and P removal and minimize pollution swapping under varying flow conditions (i.e., woodchip column hydraulic retention times (HRTs) of 7.2, 18, and 51 h; P-filter HRTs of 7.6-59 min). Woodchip denitrification columns were placed either upstream or downstream of P-filters filled with either medium. The configuration with woodchip denitrifying systems placed upstream of the P-filters generally provided optimized dissolved P removal efficiencies and removal rates. The P-filters placed upstream of the woodchip columns exhibited better P removal than downstream-placed P-filters only under overly long (i.e., N-limited) retention times when highly reduced effluent exited the woodchip bioreactors. The paired configurations using mine drainage residuals provided significantly greater P removal than the steel slag P-filters (e.g., 25-133 versus 8.8-48 g P removed m -3 filter media d -1 , respectively), but there were no significant differences in N removal between treatments (removal rates: 8.0-18 g N removed m -3 woodchips d -1 ; N removal efficiencies: 18-95% across all HRTs). The range of HRTs tested here resulted in various undesirable pollution swapping by-products from the denitrifying bioreactors: nitrite production when nitrate removal was not complete and sulfate reduction, chemical oxygen demand production and decreased pH during overly long retention times. The downstream P-filter placement provided a polishing step for removal of chemical oxygen demand and nitrite. Copyright © 2017 The Conservation Fund. Published by Elsevier Ltd.. All rights reserved.

Denitrifying woodchip bioreactor and phosphorus filter pairing to minimize pollution swapping

Science.gov (United States)

Christianson, Laura E.; Lepine, Christine; Sibrell, Philip; Penn, Chad J.; Summerfelt, Steven T.

2017-01-01

Pairing denitrifying woodchip bioreactors and phosphorus-sorbing filters provides a unique, engineered approach for dual nutrient removal from waters impaired with both nitrogen (N) and phosphorus (P). This column study aimed to test placement of two P-filter media (acid mine drainage treatment residuals and steel slag) relative to a denitrifying system to maximize N and P removal and minimize pollution swapping under varying flow conditions (i.e., woodchip column hydraulic retention times (HRTs) of 7.2, 18, and 51 h; P-filter HRTs of 7.6–59 min). Woodchip denitrification columns were placed either upstream or downstream of P-filters filled with either medium. The configuration with woodchip denitrifying systems placed upstream of the P-filters generally provided optimized dissolved P removal efficiencies and removal rates. The P-filters placed upstream of the woodchip columns exhibited better P removal than downstream-placed P-filters only under overly long (i.e., N-limited) retention times when highly reduced effluent exited the woodchip bioreactors. The paired configurations using mine drainage residuals provided significantly greater P removal than the steel slag P-filters (e.g., 25–133 versus 8.8–48 g P removed m−3 filter media d−1, respectively), but there were no significant differences in N removal between treatments (removal rates: 8.0–18 g N removed m−3 woodchips d−1; N removal efficiencies: 18–95% across all HRTs). The range of HRTs tested here resulted in various undesirable pollution swapping by-products from the denitrifying bioreactors: nitrite production when nitrate removal was not complete and sulfate reduction, chemical oxygen demand production and decreased pH during overly long retention times. The downstream P-filter placement provided a polishing step for removal of chemical oxygen demand and nitrite.

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

Czech Academy of Sciences Publication Activity Database

Čuhel, Jiří; Šimek, Miloslav

2011-01-01

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

Soil properties impacting denitrifier community size, structure, and activity in New Zealand dairy-grazed pasture

Science.gov (United States)

Jha, Neha; Saggar, Surinder; Giltrap, Donna; Tillman, Russ; Deslippe, Julie

2017-09-01

Denitrification is an anaerobic respiration process that is the primary contributor of the nitrous oxide (N2O) produced from grassland soils. Our objective was to gain insight into the relationships between denitrifier community size, structure, and activity for a range of pasture soils. We collected 10 dairy pasture soils with contrasting soil textures, drainage classes, management strategies (effluent irrigation or non-irrigation), and geographic locations in New Zealand, and measured their physicochemical characteristics. We measured denitrifier abundance by quantitative polymerase chain reaction (qPCR) and assessed denitrifier diversity and community structure by terminal restriction fragment length polymorphism (T-RFLP) of the nitrite reductase (nirS, nirK) and N2O reductase (nosZ) genes. We quantified denitrifier enzyme activity (DEA) using an acetylene inhibition technique. We investigated whether varied soil conditions lead to different denitrifier communities in soils, and if so, whether they are associated with different denitrification activities and are likely to generate different N2O emissions. Differences in the physicochemical characteristics of the soils were driven mainly by soil mineralogy and the management practices of the farms. We found that nirS and nirK communities were strongly structured along gradients of soil water and phosphorus (P) contents. By contrast, the size and structure of the nosZ community was unrelated to any of the measured soil characteristics. In soils with high water content, the richnesses and abundances of nirS, nirK, and nosZ genes were all significantly positively correlated with DEA. Our data suggest that management strategies to limit N2O emissions through denitrification are likely to be most important for dairy farms on fertile or allophanic soils during wetter periods. Finally, our data suggest that new techniques that would selectively target nirS denitrifiers may be the most effective for limiting N2O

In Situ Bioremediation of 1,4-Dioxane by Methane Oxidizing Bacteria in Coupled Anaerobic-Aerobic Zones

Science.gov (United States)

2016-02-11

FINAL REPORT In Situ Bioremediation of 1,4-Dioxane by Methane Oxidizing Bacteria in Coupled Anaerobic-Aerobic Zones SERDP Project ER-2306...volatile organic compound (CVOCs), ethene and ethane in groundwater at Raritan Arsenal Area 18C after in situ bioremediation . 4 List of...aquifers, the bioremediation approach most commonly used for chlorinated solvents. The ability of methanotrophs to biodegrade 1,4-dioxane was

Identification of active methanotrophs in a landfill cover soil through detection of expression of 16S rRNA and functional genes.

Science.gov (United States)

Chen, Yin; Dumont, Marc G; Cébron, Aurélie; Murrell, J Colin

2007-11-01

Active methanotrophs in a landfill soil were revealed by detecting the 16S rRNA of methanotrophs and the mRNA transcripts of key genes involved in methane oxidation. New 16S rRNA primers targeting type I and type II methanotrophs were designed and optimized for analysis by denaturing gradient gel electrophoresis. Direct extraction of RNA from soil enabled the analysis of the expression of the functional genes: mmoX, pmoA and mxaF, which encode subunits of soluble methane monooxygenase, particulate methane monooxygenase and methanol dehydrogenase respectively. The 16S rRNA polymerase chain reaction (PCR) primers for type I methanotrophs detected Methylomonas, Methylosarcina and Methylobacter sequences from both soil DNA and cDNA which was generated from RNA extracted directly from the landfill cover soil. The 16S rRNA primers for type II methanotrophs detected primarily Methylocella and some Methylocystis 16S rRNA genes. Phylogenetic analysis of mRNA recovered from the soil indicated that Methylobacter, Methylosarcina, Methylomonas, Methylocystis and Methylocella were actively expressing genes involved in methane and methanol oxidation. Transcripts of pmoA but not mmoX were readily detected by reverse transcription polymerase chain reaction (RT-PCR), indicating that particulate methane monooxygenase may be largely responsible for methane oxidation in situ.

Community Structure of Denitrifiers, Bacteria, and Archaea along Redox Gradients in Pacific Northwest Marine Sediments by Terminal Restriction Fragment Length Polymorphism Analysis of Amplified Nitrite Reductase (nirS) and 16S rRNA Genes

Science.gov (United States)

Braker, Gesche; Ayala-del-Río, Héctor L.; Devol, Allan H.; Fesefeldt, Andreas; Tiedje, James M.

2001-01-01

Steep vertical gradients of oxidants (O2 and NO3−) in Puget Sound and Washington continental margin sediments indicate that aerobic respiration and denitrification occur within the top few millimeters to centimeters. To systematically explore the underlying communities of denitrifiers, Bacteria, and Archaea along redox gradients at distant geographic locations, nitrite reductase (nirS) genes and bacterial and archaeal 16S rRNA genes (rDNAs) were PCR amplified and analyzed by terminal restriction fragment length polymorphism (T-RFLP) analysis. The suitablility of T-RFLP analysis for investigating communities of nirS-containing denitrifiers was established by the correspondence of dominant terminal restriction fragments (T-RFs) of nirS to computer-simulated T-RFs of nirS clones. These clones belonged to clusters II, III, and IV from the same cores and were analyzed in a previous study (G. Braker, J. Zhou, L. Wu, A. H. Devol, and J. M. Tiedje, Appl. Environ. Microbiol. 66:2096–2104, 2000). T-RFLP analysis of nirS and bacterial rDNA revealed a high level of functional and phylogenetic diversity, whereas the level of diversity of Archaea was lower. A comparison of T-RFLPs based on the presence or absence of T-RFs and correspondence analysis based on the frequencies and heights of T-RFs allowed us to group sediment samples according to the sampling location and thus clearly distinguish Puget Sound and the Washington margin populations. However, changes in community structure within sediment core sections during the transition from aerobic to anaerobic conditions were minor. Thus, within the top layers of marine sediments, redox gradients seem to result from the differential metabolic activities of populations of similar communities, probably through mixing by marine invertebrates rather than from the development of distinct communities. PMID:11282647

Soil denitrifier community size changes with land use change to perennial bioenergy cropping systems

Science.gov (United States)

Thompson, Karen A.; Deen, Bill; Dunfield, Kari E.

2016-10-01

Dedicated biomass crops are required for future bioenergy production. However, the effects of large-scale land use change (LUC) from traditional annual crops, such as corn-soybean rotations to the perennial grasses (PGs) switchgrass and miscanthus, on soil microbial community functioning is largely unknown. Specifically, ecologically significant denitrifying communities, which regulate N2O production and consumption in soils, may respond differently to LUC due to differences in carbon (C) and nitrogen (N) inputs between crop types and management systems. Our objective was to quantify bacterial denitrifying gene abundances as influenced by corn-soybean crop production compared to PG biomass production. A field trial was established in 2008 at the Elora Research Station in Ontario, Canada (n  =  30), with miscanthus and switchgrass grown alongside corn-soybean rotations at different N rates (0 and 160 kg N ha-1) and biomass harvest dates within PG plots. Soil was collected on four dates from 2011 to 2012 and quantitative PCR was used to enumerate the total bacterial community (16S rRNA) and communities of bacterial denitrifiers by targeting nitrite reductase (nirS) and N2O reductase (nosZ) genes. Miscanthus produced significantly larger yields and supported larger nosZ denitrifying communities than corn-soybean rotations regardless of management, indicating large-scale LUC from corn-soybean to miscanthus may be suitable in variable Ontario climatic conditions and under varied management, while potentially mitigating soil N2O emissions. Harvesting switchgrass in the spring decreased yields in N-fertilized plots, but did not affect gene abundances. Standing miscanthus overwinter resulted in higher 16S rRNA and nirS gene copies than in fall-harvested crops. However, the size of the total (16S rRNA) and denitrifying bacterial communities changed differently over time and in response to LUC, indicating varying controls on these communities.

nirS-type denitrifying bacterial assemblages respond to environmental conditions of a shallow estuary.

Science.gov (United States)

Lisa, Jessica A; Jayakumar, Amal; Ward, Bess B; Song, Bongkeun

2017-12-01

Molecular analysis of dissimilatory nitrite reductase genes (nirS) was conducted using a customized microarray containing 165 nirS probes (archetypes) to identify members of sedimentary denitrifying communities. The goal of this study was to examine denitrifying community responses to changing environmental variables over spatial and temporal scales in the New River Estuary (NRE), NC, USA. Multivariate statistical analyses revealed three denitrifier assemblages and uncovered 'generalist' and 'specialist' archetypes based on the distribution of archetypes within these assemblages. Generalists, archetypes detected in all samples during at least one season, were commonly world-wide found in estuarine and marine ecosystems, comprised 8%-29% of the abundant NRE archetypes. Archetypes found in a particular site, 'specialists', were found to co-vary based on site specific conditions. Archetypes specific to the lower estuary in winter were designated Cluster I and significantly correlated by sediment Chl a and porewater Fe 2+ . A combination of specialist and more widely distributed archetypes formed Clusters II and III, which separated based on salinity and porewater H 2 S respectively. The co-occurrence of archetypes correlated with different environmental conditions highlights the importance of habitat type and niche differentiation among nirS-type denitrifying communities and supports the essential role of individual community members in overall ecosystem function. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Respiratory transformation of nitrous oxide (N2O) to dinitrogen by Bacteria and Archaea.

Science.gov (United States)

Zumft, Walter G; Kroneck, Peter M H

2007-01-01

N2O is a potent greenhouse gas and stratospheric reactant that has been steadily on the rise since the beginning of industrialization. It is an obligatory inorganic metabolite of denitrifying bacteria, and some production of N2O is also found in nitrifying and methanotrophic bacteria. We focus this review on the respiratory aspect of N2O transformation catalysed by the multicopper enzyme nitrous oxide reductase (N2OR) that provides the bacterial cell with an electron sink for anaerobic growth. Two types of Cu centres discovered in N2OR were both novel structures among the Cu proteins: the mixed-valent dinuclear Cu(A) species at the electron entry site of the enzyme, and the tetranuclear Cu(Z) centre as the first catalytically active Cu-sulfur complex known. Several accessory proteins function as Cu chaperone and ABC transporter systems for the biogenesis of the catalytic centre. We describe here the paradigm of Z-type N2OR, whose characteristics have been studied in most detail in the genera Pseudomonas and Paracoccus. Sequenced bacterial genomes now provide an invaluable additional source of information. New strains harbouring nos genes and capability of N2O utilization are being uncovered. This reveals previously unknown relationships and allows pattern recognition and predictions. The core nos genes, nosZDFYL, share a common phylogeny. Most principal taxonomic lineages follow the same biochemical and genetic pattern and share the Z-type enzyme. A modified N2OR is found in Wolinella succinogenes, and circumstantial evidence also indicates for certain Archaea another type of N2OR. The current picture supports the view of evolution of N2O respiration prior to the separation of the domains Bacteria and Archaea. Lateral nos gene transfer from an epsilon-proteobacterium as donor is suggested for Magnetospirillum magnetotacticum and Dechloromonas aromatica. In a few cases, nos gene clusters are plasmid borne. Inorganic N2O metabolism is associated with a diversity of

Deep nirS amplicon sequencing of San Francisco Bay sediments enables prediction of geography and environmental conditions from denitrifying community composition.

Science.gov (United States)

Lee, Jessica A; Francis, Christopher A

2017-12-01

Denitrification is a dominant nitrogen loss process in the sediments of San Francisco Bay. In this study, we sought to understand the ecology of denitrifying bacteria by using next-generation sequencing (NGS) to survey the diversity of a denitrification functional gene, nirS (encoding cytchrome-cd 1 nitrite reductase), along the salinity gradient of San Francisco Bay over the course of a year. We compared our dataset to a library of nirS sequences obtained previously from the same samples by standard PCR cloning and Sanger sequencing, and showed that both methods similarly demonstrated geography, salinity and, to a lesser extent, nitrogen, to be strong determinants of community composition. Furthermore, the depth afforded by NGS enabled novel techniques for measuring the association between environment and community composition. We used Random Forests modelling to demonstrate that the site and salinity of a sample could be predicted from its nirS sequences, and to identify indicator taxa associated with those environmental characteristics. This work contributes significantly to our understanding of the distribution and dynamics of denitrifying communities in San Francisco Bay, and provides valuable tools for the further study of this key N-cycling guild in all estuarine systems. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Salinity shifts in marine sediment: Importance of number of fluctuation rather than their intensities on bacterial denitrifying community.

Science.gov (United States)

Zaghmouri, Imen; Michotey, Valerie D; Armougom, Fabrice; Guasco, Sophie; Bonin, Patricia C

2018-05-01

The sensitivity of denitrifying community to salinity fluctuations was studied in microcosms filled with marine coastal sediments subjected to different salinity disturbances over time (sediment under frequent salinity changes vs sediment with "stable" salinity pattern). Upon short-term salinity shift, denitrification rate and denitrifiers abundance showed high resistance whatever the sediment origin is. Denitrifying community adapted to frequent salinity changes showed high resistance when salinity increases, with a dynamic nosZ relative expression level. Marine sediment denitrifying community, characterized by more stable pattern, was less resistant when salinity decreases. However, after two successive variations of salinity, it shifted toward the characteristic community of fluctuating conditions, with larger proportion of Pseudomonas-nosZ, exhibiting an increase of nosZ relative expression level. The impact of long-term salinity variation upon bacterial community was confirmed at ribosomal level with a higher percentage of Pseudomonas and lower proportion of nosZII clade genera. Copyright © 2018 Elsevier Ltd. All rights reserved.

Fermentative Bacteria Influence the Competition between Denitrifiers and DNRA Bacteria

Directory of Open Access Journals (Sweden)

Eveline M. van den Berg

2017-09-01

results of this study clearly show that not only the ratio of available substrates, but also the nature of the electron donor influences the outcome of competition between DNRA and denitrification. Apparently, fermentative bacteria are competitive for the electron donor and thereby alter the ratio of available substrates for nitrate reduction.

Denitrifying bacterial communities affect current production and nitrous oxide accumulation in a microbial fuel cell.

Science.gov (United States)

Vilar-Sanz, Ariadna; Puig, Sebastià; García-Lledó, Arantzazu; Trias, Rosalia; Balaguer, M Dolors; Colprim, Jesús; Bañeras, Lluís

2013-01-01

The biocathodic reduction of nitrate in Microbial Fuel Cells (MFCs) is an alternative to remove nitrogen in low carbon to nitrogen wastewater and relies entirely on microbial activity. In this paper the community composition of denitrifiers in the cathode of a MFC is analysed in relation to added electron acceptors (nitrate and nitrite) and organic matter in the cathode. Nitrate reducers and nitrite reducers were highly affected by the operational conditions and displayed high diversity. The number of retrieved species-level Operational Taxonomic Units (OTUs) for narG, napA, nirS and nirK genes was 11, 10, 31 and 22, respectively. In contrast, nitrous oxide reducers remained virtually unchanged at all conditions. About 90% of the retrieved nosZ sequences grouped in a single OTU with a high similarity with Oligotropha carboxidovorans nosZ gene. nirS-containing denitrifiers were dominant at all conditions and accounted for a significant amount of the total bacterial density. Current production decreased from 15.0 A · m(-3) NCC (Net Cathodic Compartment), when nitrate was used as an electron acceptor, to 14.1 A · m(-3) NCC in the case of nitrite. Contrarily, nitrous oxide (N2O) accumulation in the MFC was higher when nitrite was used as the main electron acceptor and accounted for 70% of gaseous nitrogen. Relative abundance of nitrite to nitrous oxide reducers, calculated as (qnirS+qnirK)/qnosZ, correlated positively with N2O emissions. Collectively, data indicate that bacteria catalysing the initial denitrification steps in a MFC are highly influenced by main electron acceptors and have a major influence on current production and N2O accumulation.

Acquisition of epibiotic bacteria along the life cycle of the hydrothermal shrimp Rimicaris exoculata.

Science.gov (United States)

Guri, Mathieu; Durand, Lucile; Cueff-Gauchard, Valérie; Zbinden, Magali; Crassous, Philippe; Shillito, Bruce; Cambon-Bonavita, Marie-Anne

2012-03-01

The caridean shrimp Rimicaris exoculata dominates the fauna at several Mid-Atlantic Ridge hydrothermal vent sites. This shrimp has an enlarged gill chamber, harboring a dense ectosymbiotic community of chemoautotrophic bacteria associated with mineral oxide deposits. Until now, their acquisition is not fully understood. At three hydrothermal vent sites, we analyzed the epibionts diversity at different moult stages and also in the first stages of the shrimp life (eggs, hatched eggs (with larvae) and juveniles). Hatched eggs associated with young larvae were collected for the first time directly from gravid females at the Logachev vent site during the Serpentine cruise. An approach using 16S rRNA clone libraries, scanning and transmission electron microscopy, and fluorescent in situ hybridization was used. Molecular results and microscope observations indicated a switch in the composition of the bacterial community between early R. exoculata life cycle stage (egg libraries dominated by the Gammaproteobacteria) and later stages (juvenile/adult libraries dominated by the Epsilonproteobacteria). We hypothesized that the epibiotic phylotype composition could vary according to the life stage of the shrimp. Our results confirmed the occurrence of a symbiosis with Gammaproteobacteria and Epsilonproteobacteria, but more complex than previously assumed. We revealed the presence of active type-I methanotrophic bacteria colonizing the cephalothorax of shrimps from the Rainbow site. They were also present on the eggs from the Logachev site. This could be the first 'epibiotic' association between methanotrophic bacteria and hydrothermal vent crustacean. We discuss possible transmission pathways for epibionts linked to the shrimp life cycle.

Functional bacteria and process metabolism of the Denitrifying Sulfur conversion-associated Enhanced Biological Phosphorus Removal (DS-EBPR) system: An investigation by operating the system from deterioration to restoration.

Science.gov (United States)

Guo, Gang; Wu, Di; Hao, Tianwei; Mackey, Hamish Robert; Wei, Li; Wang, Haiguang; Chen, Guanghao

2016-05-15

A sulfur conversion-associated Enhanced Biological Phosphorus (P) Removal (EBPR) system is being developed to cater for the increasing needs to treat saline/brackish wastewater resulting from seawater intrusion into groundwater and sewers and frequent use of sulfate coagulants during drinking water treatment, as well as to meet the demand for eutrophication control in warm climate regions. However, the major functional bacteria and metabolism in this emerging biological nutrient removal system are still poorly understood. This study was thus designed to explore the functional microbes and metabolism in this new EBPR system by manipulating the deterioration, failure and restoration of a lab-scale system. This was achieved by changing the mixed liquor suspended solids (MLSS) concentration to monitor and evaluate the relationships among sulfur conversion (including sulfate reduction and sulfate production), P removal, variation in microbial community structures, and stoichiometric parameters. The results show that the stable Denitrifying Sulfur conversion-associated EBPR (DS-EBPR) system was enriched by sulfate-reducing bacteria (SRB) and sulfide-oxidizing bacteria (SOB). These bacteria synergistically participated in this new EBPR process, thereby inducing an appropriate level of sulfur conversion crucial for achieving a stable DS-EBPR performance, i.e. maintaining sulfur conversion intensity at 15-40 mg S/L, corresponding to an optimal sludge concentration of 6.5 g/L. This range of sulfur conversion favors microbial community competition and various energy flows from internal polymers (i.e. polysulfide or elemental sulfur (poly-S(2-)/S(0)) and poly-β-hydroxyalkanoates (PHA)) for P removal. If this range was exceeded, the system might deteriorate or even fail due to enrichment of glycogen-accumulating organisms (GAOs). Four methods of restoring the failed system were investigated: increasing the sludge concentration, lowering the salinity or doubling the COD

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

DEFF Research Database (Denmark)

Chirinda, Ngonidzashe; Olesen, Jørgen Eivind; Porter, John Roy

2011-01-01

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

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

Science.gov (United States)

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

2016-01-01

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

Environmental distribution and abundance of the facultative methanotroph Methylocella

OpenAIRE

Rahman, Md Tanvir; Crombie, Andrew; Chen, Yin; Stralis-Pavese, Nancy; Bodrossy, Levente; Meir, Patrick; McNamara, Niall P; Murrell, J Colin

2010-01-01

Methylocella spp. are facultative methanotrophs, which are able to grow not only on methane but also on multicarbon substrates such as acetate, pyruvate or malate. Methylocella spp. were previously thought to be restricted to acidic soils such as peatlands, in which they may have a key role in methane oxidation. There is little information on the abundance and distribution of Methylocella spp. in the environment. New primers were designed, and a real-time quantitative PCR method was developed...

Nitrogen fixation in denitrified marine waters.

Directory of Open Access Journals (Sweden)

Camila Fernandez

Full Text Available Nitrogen fixation is an essential process that biologically transforms atmospheric dinitrogen gas to ammonia, therefore compensating for nitrogen losses occurring via denitrification and anammox. Currently, inputs and losses of nitrogen to the ocean resulting from these processes are thought to be spatially separated: nitrogen fixation takes place primarily in open ocean environments (mainly through diazotrophic cyanobacteria, whereas nitrogen losses occur in oxygen-depleted intermediate waters and sediments (mostly via denitrifying and anammox bacteria. Here we report on rates of nitrogen fixation obtained during two oceanographic cruises in 2005 and 2007 in the eastern tropical South Pacific (ETSP, a region characterized by the presence of coastal upwelling and a major permanent oxygen minimum zone (OMZ. Our results show significant rates of nitrogen fixation in the water column; however, integrated rates from the surface down to 120 m varied by ∼30 fold between cruises (7.5±4.6 versus 190±82.3 µmol m(-2 d(-1. Moreover, rates were measured down to 400 m depth in 2007, indicating that the contribution to the integrated rates of the subsurface oxygen-deficient layer was ∼5 times higher (574±294 µmol m(-2 d(-1 than the oxic euphotic layer (48±68 µmol m(-2 d(-1. Concurrent molecular measurements detected the dinitrogenase reductase gene nifH in surface and subsurface waters. Phylogenetic analysis of the nifH sequences showed the presence of a diverse diazotrophic community at the time of the highest measured nitrogen fixation rates. Our results thus demonstrate the occurrence of nitrogen fixation in nutrient-rich coastal upwelling systems and, importantly, within the underlying OMZ. They also suggest that nitrogen fixation is a widespread process that can sporadically provide a supplementary source of fixed nitrogen in these regions.

Nitrogen Fixation in Denitrified Marine Waters

Science.gov (United States)

Fernandez, Camila; Farías, Laura; Ulloa, Osvaldo

2011-01-01

Nitrogen fixation is an essential process that biologically transforms atmospheric dinitrogen gas to ammonia, therefore compensating for nitrogen losses occurring via denitrification and anammox. Currently, inputs and losses of nitrogen to the ocean resulting from these processes are thought to be spatially separated: nitrogen fixation takes place primarily in open ocean environments (mainly through diazotrophic cyanobacteria), whereas nitrogen losses occur in oxygen-depleted intermediate waters and sediments (mostly via denitrifying and anammox bacteria). Here we report on rates of nitrogen fixation obtained during two oceanographic cruises in 2005 and 2007 in the eastern tropical South Pacific (ETSP), a region characterized by the presence of coastal upwelling and a major permanent oxygen minimum zone (OMZ). Our results show significant rates of nitrogen fixation in the water column; however, integrated rates from the surface down to 120 m varied by ∼30 fold between cruises (7.5±4.6 versus 190±82.3 µmol m−2 d−1). Moreover, rates were measured down to 400 m depth in 2007, indicating that the contribution to the integrated rates of the subsurface oxygen-deficient layer was ∼5 times higher (574±294 µmol m−2 d−1) than the oxic euphotic layer (48±68 µmol m−2 d−1). Concurrent molecular measurements detected the dinitrogenase reductase gene nifH in surface and subsurface waters. Phylogenetic analysis of the nifH sequences showed the presence of a diverse diazotrophic community at the time of the highest measured nitrogen fixation rates. Our results thus demonstrate the occurrence of nitrogen fixation in nutrient-rich coastal upwelling systems and, importantly, within the underlying OMZ. They also suggest that nitrogen fixation is a widespread process that can sporadically provide a supplementary source of fixed nitrogen in these regions. PMID:21687726

Ammonia-oxidizing Bacteria and Archaea in the Rhizosphere of Freshwater Macrophytes

DEFF Research Database (Denmark)

Herrmann, Martina; Schramm, Andreas

2007-01-01

AMMONIA-OXIDIZING ARCHAEA AND BACTERIA IN THE RHIZOSPHERE OF FRESHWATER MACROPHYTES Martina Herrmann and Andreas Schramm Department of Biological Sciences, Microbiology, University of Aarhus, Denmark Aquatic macrophytes such as Littorella uniflora and Lobelia dortmanna release oxygen from...... their roots and thereby stimulate nitrification and coupled nitrification-denitrification in their rhizosphere. However, oxygen release and inorganic nitrogen concentrations differ markedly between macrophyte species. We therefore propose (i) that the rhizosphere of freshwater macrophytes harbours a species......-specific microbial community distinct from that of unvegetated sediment and (ii) that aquatic macrophytes have an impact on abundance and activity of nitrifying and denitrifying bacteria in freshwater sediment. The goal of this study was to test these hypotheses for the key functional group for coupled nitrification...

Plastic carrier polishing chamber reduces pollution swapping from denitrifying woodchip bioreactors

Science.gov (United States)

Denitrifying bioreactors with solid organic carbon sources (i.e., “woodchip bioreactors”) have proven to be relatively simple and cost effective treatment systems for nitrate-laden agricultural and aquacultural waters and wastewaters. However, because this technology is still relatively new, design ...

Draft Genome Sequence of Methylocella silvestris TVC, a Facultative Methanotroph Isolated from Permafrost

OpenAIRE

Wang, Jing; Geng, Kan; Farhan Ul Haque, Muhammad; Crombie, Andrew; Street, Lorna E.; Wookey, Philip A.; Ma, Ke; Murrell, J. Colin; Pratscher, Jennifer

2018-01-01

Permafrost environments play a crucial role in global carbon and methane cycling. We report here the draft genome sequence of Methylocella silvestris TVC, a new facultative methanotroph strain, isolated from the Siksik Creek catchment in the continuous permafrost zone of Inuvik (Northwest Territories, Canada).

Metabolism of Hydrocarbons in n-Alkane-Utilizing Anaerobic Bacteria.

Science.gov (United States)

Wilkes, Heinz; Buckel, Wolfgang; Golding, Bernard T; Rabus, Ralf

2016-01-01

The glycyl radical enzyme-catalyzed addition of n-alkanes to fumarate creates a C-C-bond between two concomitantly formed stereogenic carbon centers. The configurations of the two diastereoisomers of the product resulting from n-hexane activation by the n-alkane-utilizing denitrifying bacterium strain HxN1, i.e. (1-methylpentyl)succinate, were assigned as (2S,1'R) and (2R,1'R). Experiments with stereospecifically deuterated n-(2,5-2H2)hexanes revealed that exclusively the pro-S hydrogen atom is abstracted from C2 of the n-alkane by the enzyme and later transferred back to C3 of the alkylsuccinate formed. These results indicate that the alkylsuccinate-forming reaction proceeds with an inversion of configuration at the carbon atom (C2) of the n-alkane forming the new C-C-bond, and thus stereochemically resembles a SN2-type reaction. Therefore, the reaction may occur in a concerted manner, which may avoid the highly energetic hex-2-yl radical as an intermediate. The reaction is associated with a significant primary kinetic isotope effect (kH/kD ≥3) for hydrogen, indicating that the homolytic C-H-bond cleavage is involved in the first irreversible step of the reaction mechanism. The (1-methylalkyl)succinate synthases of n-alkane-utilizing anaerobic bacteria apparently have very broad substrate ranges enabling them to activate not only aliphatic but also alkyl-aromatic hydrocarbons. Thus, two denitrifiers and one sulfate reducer were shown to convert the nongrowth substrate toluene to benzylsuccinate and further to the dead-end product benzoyl-CoA. For this purpose, however, the modified β-oxidation pathway known from alkylbenzene-utilizing bacteria was not employed, but rather the pathway used for n-alkane degradation involving CoA ligation, carbon skeleton rearrangement and decarboxylation. Furthermore, various n-alkane- and alkylbenzene-utilizing denitrifiers and sulfate reducers were found to be capable of forming benzyl alcohols from diverse alkylbenzenes

Cytochrome cd1-containing nitrite reductase encoding gene nirS as a new functional biomarker for detection of anaerobic ammonium oxidizing (Anammox) bacteria.

Science.gov (United States)

Li, Meng; Ford, Tim; Li, Xiaoyan; Gu, Ji-Dong

2011-04-15

A newly designed primer set (AnnirS), together with a previously published primer set (ScnirS), was used to detect anammox bacterial nirS genes from sediments collected from three marine environments. Phylogenetic analysis demonstrated that all retrieved sequences were clearly different from typical denitrifiers' nirS, but do group together with the known anammox bacterial nirS. Sequences targeted by ScnirS are closely related to Scalindua nirS genes recovered from the Peruvian oxygen minimum zone (OMZ), whereas sequences targeted by AnnirS are more closely affiliated with the nirS of Candidatus 'Kuenenia stuttgartiensis' and even form a new phylogenetic nirS clade, which might be related to other genera of the anammox bacteria. Analysis demonstrated that retrieved sequences had higher sequence identities (>60%) with known anammox bacterial nirS genes than with denitrifiers' nirS, on both nucleotide and amino acid levels. Compared to the 16S rRNA and hydrazine oxidoreductase (hzo) genes, the anammox bacterial nirS not only showed consistent phylogenetic relationships but also demonstrated more reliable quantification of anammox bacteria because of the single copy of the nirS gene in the anammox bacterial genome and the specificity of PCR primers for different genera of anammox bacteria, thus providing a suitable functional biomarker for investigation of anammox bacteria.

Cytochromes c': Structure, Reactivity and Relevance to Haem-Based Gas Sensing.

Science.gov (United States)

Hough, Michael A; Andrew, Colin R

2015-01-01

Cytochromes c' are a group of class IIa cytochromes with pentacoordinate haem centres and are found in photosynthetic, denitrifying and methanotrophic bacteria. Their function remains unclear, although roles in nitric oxide (NO) trafficking during denitrification or in cellular defence against nitrosoative stress have been proposed. Cytochromes c' are typically dimeric with each c-type haem-containing monomer folding as a four-α-helix bundle. Their hydrophobic and crowded distal sites impose severe restrictions on the binding of distal ligands, including diatomic gases. By contrast, NO binds to the proximal haem face in a similar manner to that of the eukaryotic NO sensor, soluble guanylate cyclase and bacterial analogues. In this review, we focus on how structural features of cytochromes c' influence haem spectroscopy and reactivity with NO, CO and O2. We also discuss the relevance of cytochrome c' to understanding the mechanisms of gas binding to haem-based sensor proteins. © 2015 Elsevier Ltd. All rights reserved.

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2014-01-15

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

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

NARCIS (Netherlands)

Nguyen, P.D.; Ginkel, van C.G.; Plugge, C.M.

2008-01-01

The anaerobic degradation of tetradecylamine and other long-chain alkylamines by a newly isolated denitrifying bacterium was studied. Strain ZN6 was isolated from a mixture of soil and active sludge and was identified as representing Pseudomonas stutzeri, based on partial 16S rRNA gene sequence

Draft Genome Sequence of Methylocella silvestris TVC, a Facultative Methanotroph Isolated from Permafrost.

Science.gov (United States)

Wang, Jing; Geng, Kan; Farhan Ul Haque, Muhammad; Crombie, Andrew; Street, Lorna E; Wookey, Philip A; Ma, Ke; Murrell, J Colin; Pratscher, Jennifer

2018-02-22

Permafrost environments play a crucial role in global carbon and methane cycling. We report here the draft genome sequence of Methylocella silvestris TVC, a new facultative methanotroph strain, isolated from the Siksik Creek catchment in the continuous permafrost zone of Inuvik (Northwest Territories, Canada). Copyright © 2018 Wang et al.

Fractionation of Nitrogen and Oxygen Isotopes and Roles of Bacteria during Denitrification

Science.gov (United States)

Kang, J.; Buyanjargal, A.; Jeen, S. W.

2017-12-01

Nitrate in groundwater can cause health and environmental problems when not properly treated. The purpose of this study was to develop a treatment method for nitrate in groundwater using organic carbon-based reactive mixtures (i.e., wood chips and gravel) through column experiments and to evaluate reaction mechanisms responsible for the treatment. The column experiments were operated for a total of 19 months. The results from the geochemical analyses for the experiments suggest that cultures of denitrifying bacteria used organic carbon while utilizing nitrate as their electron acceptor via denitrification process. Proteobacteria was the most abundant phylum in all samples, accounting for 45.7% of the bacterial reads, followed by Firmicutes (22.6%) and Chlorobi (10.6%). Bacilli, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Actinobacteria_c consisted of 32, 30, 23, 11, and 2% of denitrifying bacteria class. The denitrification process caused fractionation of nitrogen and oxygen isotopes of nitrate while nitrate concentration decreased. When fitted to the Rayleigh's fractionation model, enrichment factors (ɛ) were 11.5‰ and 5.6‰ for 15N and 18O isotopes, respectively. Previous studies suggested that nitrogen isotope enrichment factors of denitrification are within the range of 4.7 to 40‰ and oxygen isotopic enrichment factors are between 8 and 18.3‰. This study shows that nitrate in groundwater can be effectively treated using passive treatment systems, such as permeable reactive barriers (PRBs), and denitrificaton is the dominant process reponsible for the removal of nitrate.

Bioavailability and biodegradation of weathered diesel fuel in aquifer material under denitrifying conditions

International Nuclear Information System (INIS)

Bregnard, T.P.A.; Hoehener, P.; Zeyer, J.

1998-01-01

During the in situ bioremediation of a diesel fuel-contaminated aquifer in Menziken, Switzerland, aquifer material containing weathered diesel fuel (WDF) and indigenous microorganisms was excavated. This material was used to identify factors limiting WDF biodegradation under denitrifying conditions. Incubations of this material for 360 to 390 d under denitrifying conditions resulted in degradation of 23% of the WDF with concomitant consumption of NO 3 - and production of inorganic carbon. The biodegradation of WDF and the rate of NO 3 - consumption was stimulated by agitation of the microcosms. Biodegradation was not stimulated by the addition of a biosurfactant (rhamnolipids) or a synthetic surfactant (Triton X-100) at concentrations above their critical micelle concentrations. The rhamnolipids were biodegraded preferentially to WDF, whereas Triton X-100 was not degraded. Both surfactants reduced the surface tension of the growth medium from 72 to <35 dynes/cm and enhanced the apparent aqueous solubility of the model hydrocarbon n-hexadecane by four orders of magnitude. Solvent-extracted WDF, added at a concentration equal to that already present in the aquifer material, was also biodegraded by the microcosms, but not at a higher rate than the WDF already present in the material. The results show that the denitrifying biodegradation of WDF is not necessarily limited by bioavailability but rather by the inherent recalcitrance of WDF

Edaphic Conditions Regulate Denitrification Directly and Indirectly by Altering Denitrifier Abundance in Wetlands along the Han River, China.

Science.gov (United States)

Xiong, Ziqian; Guo, Laodong; Zhang, Quanfa; Liu, Guihua; Liu, Wenzhi

2017-05-16

Riparian wetlands play a critical role in retaining nitrogen (N) from upland runoff and improving river water quality, mainly through biological processes such as soil denitrification. However, the relative contribution of abiotic and biotic factors to riparian denitrification capacity remains elusive. Here we report the spatiotemporal dynamics of potential and unamended soil denitrification rates in 20 wetlands along the Han River, an important water source in central China. We also quantified the abundance of soil denitrifying microorganisms using nirK and nirS genes. Results showed that soil denitrification rates were significantly different between riparian and reservoir shoreline wetlands, but not between mountain and lowland wetlands. In addition, soil denitrification rates showed strong seasonality, with higher values in August (summer) and April (spring) but lower values in January (winter). The potential and unamended denitrification rates were positively correlated with edaphic conditions (moisture and carbon concentration), denitrifier abundance, and plant species richness. Path analysis further revealed that edaphic conditions could regulate denitrification rates both directly and indirectly through their effects on denitrifier abundance. Our findings highlight that not only environmental factors, but also biotic factors including denitrifying microorganisms and standing vegetation, play an important role in regulating denitrification rate and N removal capacity in riparian wetlands.

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

. Anaerobic methane oxidation regulates methane emissions in marine sediments and appears to occur through a reversal of a methane-producing metabolism. We tested the assumption that ANME are obligate methanotrophs by detecting and quantifying gene transcription of ANME-1 across zones of methane oxidation...

Simultaneous pollutant removal and electricity generation in denitrifying microbial fuel cell with boric acid-borate buffer solution.

Science.gov (United States)

Chen, Gang; Zhang, Shaohui; Li, Meng; Wei, Yan

2015-01-01

A double-chamber denitrifying microbial fuel cell (MFC), using boric acid-borate buffer solution as an alternative to phosphate buffer solution, was set up to investigate the influence of buffer solution concentration, temperature and external resistance on electricity generation and pollutant removal efficiency. The result revealed that the denitrifying MFC with boric acid-borate buffer solution was successfully started up in 51 days, with a stable cell voltage of 205.1 ± 1.96 mV at an external resistance of 50 Ω. Higher concentration of buffer solution favored nitrogen removal and electricity generation. The maximum power density of 8.27 W/m(3) net cathodic chamber was obtained at a buffer solution concentration of 100 mmol/L. An increase in temperature benefitted electricity generation and nitrogen removal. A suitable temperature for this denitrifying MFC was suggested to be 25 °C. Decreasing the external resistance favored nitrogen removal and organic matter consumption by exoelectrogens.

[Phylogenetic analysis and nitrogen removal characteristics of a heterotrophic nitrifying-aerobic denitrifying bacteria strain from marine environment].

Science.gov (United States)

Sun, Xuemei; Li, Qiufen; Zhang, Yan; Liu, Huaide; Zhao, Jun; Qu, Keming

2012-06-04

We determined the phylogenetic position of a heterotrophic nitrifying-aerobic denitrifying bacterium X3, and detected its nitrogen removal characteristics for providing evidence to explain the principle of heterotrophic nitrification-aerobic denitrification and to improve the process in purification of marine-culture wastewater. The evolutionary position of the strain was determined based on its morphological, physiological, biochemical characteristics and 16SrRNA gene sequence. The nitrification-denitrification ability of this strain was detected by detecting its nitrogen removal efficiency and growth on different inorganic nitrogen source. Strain X3 was identified as Halomonas sp. It grew optimally at salinity 3%, pH 8.5, C:N 10:1 at 28 degrees C, and it could still survive at 15% salinity. The removal of NH4+ -N, NO2(-) -N and NO3(-) -N was 98.29%, 99.07%, 96.48% respectively within 24 h. When three inorganic nitrogen existed simultaneously, it always utilized ammonia firstly, and the total inorganic nitrogen removal was higher than with only one nitrogen, suggesting that strain X3 has the ability of simultaneous nitrification and denitrification and completing the whole nitrogen removing process. Strain X3 belonged to the genus of Halomonas. It had strong simultaneous nitrification and denitrification capability and could live in high-salinity environment.

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.

Denitrification of groundwater using a sulfur-oxidizing autotrophic denitrifying anaerobic fluidized-bed MBR: performance and bacterial community structure.

Science.gov (United States)

Zhang, Lili; Zhang, Chao; Hu, Chengzhi; Liu, Huijuan; Qu, Jiuhui

2015-03-01

This paper investigates a novel sulfur-oxidizing autotrophic denitrifying anaerobic fluidized bed membrane bioreactor (AnFB-MBR) that has the potential to overcome the limitations of conventional sulfur-oxidizing autotrophic denitrification systems. The AnFB-MBR produced consistent high-quality product water when fed by a synthetic groundwater with NO3 (-)-N ranging 25-80 mg/L and operated at hydraulic retention times of 0.5-5.0 h. A nitrate removal rate of up to 4.0 g NO3 (-)-N/Lreactord was attained by the bioreactor, which exceeded any reported removal capacity. The flux of AnFB-MBR was maintained in the range of 1.5-15 L m(-2) h(-1). Successful membrane cleaning was practiced with cleaning cycles of 35-81 days, which had no obvious effect on the AnFB-MBR performance. The (15) N-tracer analyses elucidated that nitrogen was converted into (15) N2-N and (15) N-biomass accounting for 88.1-93.1 % and 6.4-11.6 % of the total nitrogen produced, respectively. Only 0.3-0.5 % of removed nitrogen was in form of (15)N2O-N in sulfur-oxidizing autotrophic denitrification process, reducing potential risks of a significant amount of N2O emissions. The sulfur-oxidizing autotrophic denitrifying bacterial consortium was composed mainly of bacteria from Proteobacteria, Chlorobi, and Chloroflexi phyla, with genera Thiobacillus, Sulfurimonas, and Ignavibacteriales dominating the consortium. The pyrosequencing assays also suggested that the stable microbial communities corresponded to the elevated performance of the AnFB-MBR. Overall, this research described relatively high nitrate removal, acceptable flux, indicating future potential for the technology in practice.

Oxygen regulation of nitrate uptake in denitrifying Pseudomonas aeruginosa.

OpenAIRE

Hernandez, D; Rowe, J J

1987-01-01

Oxygen had an immediate and reversible inhibitory effect on nitrate respiration by denitrifying cultures of Pseudomonas aeruginosa. Inhibition of nitrate utilization by oxygen appeared to be at the level of nitrate uptake, since nitrate reduction to nitrite in cell extracts was not affected by oxygen. The degree of oxygen inhibition was dependent on the concentration of oxygen, and increasing nitrate concentrations could not overcome the inhibition. The inhibitory effect of oxygen was maximal...

Activation of Acetone and Other Simple Ketones in Anaerobic Bacteria.

Science.gov (United States)

Heider, Johann; Schühle, Karola; Frey, Jasmin; Schink, Bernhard

2016-01-01

Acetone and other ketones are activated for subsequent degradation through carboxylation by many nitrate-reducing, phototrophic, and obligately aerobic bacteria. Acetone carboxylation leads to acetoacetate, which is subsequently activated to a thioester and degraded via thiolysis. Two different types of acetone carboxylases have been described, which require either 2 or 4 ATP equivalents as an energy supply for the carboxylation reaction. Both enzymes appear to combine acetone enolphosphate with carbonic phosphate to form acetoacetate. A similar but more complex enzyme is known to carboxylate the aromatic ketone acetophenone, a metabolic intermediate in anaerobic ethylbenzene metabolism in denitrifying bacteria, with simultaneous hydrolysis of 2 ATP to 2 ADP. Obligately anaerobic sulfate-reducing bacteria activate acetone to a four-carbon compound as well, but via a different process than bicarbonate- or CO2-dependent carboxylation. The present evidence indicates that either carbon monoxide or a formyl residue is used as a cosubstrate, and that the overall ATP expenditure of this pathway is substantially lower than in the known acetone carboxylase reactions. © 2016 S. Karger AG, Basel.

Anaerobic oxidation of 2-chloroethanol under denitrifying conditions by Pseudomonas stutzeri strain JJ.

Science.gov (United States)

Dijk, J A; Stams, A J M; Schraa, G; Ballerstedt, H; de Bont, J A M; Gerritse, J

2003-11-01

A bacterium that uses 2-chloroethanol as sole energy and carbon source coupled to denitrification was isolated from 1,2-dichloroethane-contaminated soil. Its 16 S rDNA sequence showed 98% similarity with the type strain of Pseudomonas stutzeri (DSM 5190) and the isolate was tentatively identified as Pseudomonas stutzeri strain JJ. Strain JJ oxidized 2-chloroethanol completely to CO(2) with NO(3)(- )or O(2) as electron acceptor, with a preference for O(2) if supplied in combination. Optimum growth on 2-chloroethanol with nitrate occurred at 30 degrees C with a mu(max) of 0.14 h(-1) and a yield of 4.4 g protein per mol 2-chloroethanol metabolized. Under aerobic conditions, the mu(max) was 0.31 h(-1). NO(2)(-) also served as electron acceptor, but reduction of Fe(OH)(3), MnO(2), SO(4)(2-), fumarate or ClO(3)(-) was not observed. Another chlorinated compound used as sole energy and carbon source under aerobic and denitrifying conditions was chloroacetate. Various different bacterial strains, including some closely related Pseudomonas stutzeri strains, were tested for their ability to grow on 2-chloroethanol as sole energy and carbon source under aerobic and denitrifying conditions, respectively. Only three strains, Pseudomonas stutzeri strain LMD 76.42, Pseudomonas putida US2 and Xanthobacter autotrophicus GJ10, grew aerobically on 2-chloroethanol. This is the first report of oxidation of 2-chloroethanol under denitrifying conditions by a pure bacterial culture.

Denitrifying capabilities of Tetrasphaera and their contribution towards nitrous oxide production in enhanced biological phosphorus removal processes.

Science.gov (United States)

Marques, Ricardo; Ribera-Guardia, Anna; Santos, Jorge; Carvalho, Gilda; Reis, Maria A M; Pijuan, Maite; Oehmen, Adrian

2018-06-15

Denitrifying enhanced biological phosphorus removal (EBPR) systems can be an efficient means of removing phosphate (P) and nitrate (NO 3 - ) with low carbon source and oxygen requirements. Tetrasphaera is one of the most abundant polyphosphate accumulating organisms present in EBPR systems, but their capacity to achieve denitrifying EBPR has not previously been determined. An enriched Tetrasphaera culture, comprising over 80% of the bacterial biovolume was obtained in this work. Despite the denitrification capacity of Tetrasphaera, this culture achieved only low levels of anoxic P-uptake. Batch tests with different combinations of NO 3 - , nitrite (NO 2 - ) and nitrous oxide (N 2 O) revealed lower N 2 O accumulation by Tetrasphaera as compared to Accumulibacter and Competibacter when multiple electron acceptors were added. Electron competition was observed during the addition of multiple nitrogen electron acceptors species, where P uptake appeared to be slightly favoured over glycogen production in these situations. This study increases our understanding of the role of Tetrasphaera-related organisms in denitrifying EBPR systems. Copyright © 2018 Elsevier Ltd. All rights reserved.

Evidence of co-metabolic bentazone transformation by methanotrophic enrichment from a groundwater-fed rapid sand filter

DEFF Research Database (Denmark)

Hedegaard, Mathilde Jørgensen; Deliniere, Hélène; Prasse, Carsten

2018-01-01

from 58 to 158, well within the range for methanotrophic co-metabolic degradation of trace contaminants calculated from the literature, with normalized substrate preferences varying from 3 to 400. High-resolution mass spectrometry revealed formation of the transformation products (TPs) 6-OH, 8-OH......The herbicide bentazone is recalcitrant in aquifers and is therefore frequently detected in wells used for drinking water production. However, bentazone degradation has been observed in filter sand from a rapid sand filter at a waterworks with methane-rich groundwater. Here, the association between...... and bentazone at concentrations below 2 mg/L showed methanotrophic co-metabolic bentazone transformation: The culture removed 53% of the bentazone in 21 days in presence of 5 mg/L of methane, while only 31% was removed in absence of methane. Addition of acetylene inhibited methane oxidation and stopped...

Denitrifying metabolism of the methylotrophic marine bacterium Methylophaga nitratireducenticrescens strain JAM1.

Science.gov (United States)

Mauffrey, Florian; Cucaita, Alexandra; Constant, Philippe; Villemur, Richard

2017-01-01

R during the growth and the N 2 O accumulation phases suggests NO production in strain JAM1 cultures. By showing that all the three denitrification reductases are active, this demonstrates that M. nitratireducenticrescens JAM1 is one of many bacteria species that maintain genes associated primarily with denitrification, but not necessarily related to the maintenance of the entire pathway. The reason to maintain such an incomplete pathway could be related to the specific role of strain JAM1 in the denitrifying biofilm of the denitrification reactor from which it originates. The production of N 2 O in strain JAM1 did not involve Nar, contrary to what was demonstrated in Escherichia coli . M. nitratireducenticrescens JAM1 is the only reported Methylophaga species that has the capacity to grow under anoxic conditions by using [Formula: see text] and N 2 O as sole electron acceptors for its growth. It is also one of a few marine methylotrophs that is studied at the physiological and genetic levels in relation to its capacity to perform denitrifying activities.

Plastic biofilm carrier after corn cobs reduces nitrate loading in laboratory denitrifying bioreactors

Science.gov (United States)

Nitrate-nitrogen removal rates can be increased substantially in denitrifying bioreactors with a corn cob bed medium compared to woodchips; however, additional organic carbon (C) is released into the effluent. This laboratory column experiment was conducted to test the performance of a post-bed cha...

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

Science.gov (United States)

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

2017-02-01

Cultivation of microbial representatives of specific functional guilds from environmental samples depends largely on the suitability of the applied growth conditions. Especially the cultivation of marine methanotrophs has received little attention, resulting in only a limited number of ex situ cultures available. In this study we investigated the effect of adhesion material and headspace composition on the methane oxidation activity in methanotrophic enrichments obtained from marine sediment. Addition of sterilized natural sediment or alternatively the addition of acid-washed silicon dioxide significantly increased methane oxidation. This positive effect was attributed to bacterial adhesion on the particles via extracellular compounds, with a minimum amount of particles required for effect. As a result, the particles were immobilized, thus creating a stratified environment in which a limited diffusive gas gradients could build up and various microniches were formed. Such diffusive gas gradient might necessitate high headspace concentrations of CH 4 and CO 2 for sufficient concentrations to reach the methane-oxidizing bacteria in the enrichment culture technique. Therefore, high concentrations of methane and carbon dioxide, in addition to the addition of adhesion material, were tested and indeed further stimulated methane oxidation. Use of adhesion material in combination with high concentrations of methane and carbon dioxide might thus facilitate the cultivation and subsequent enrichment of environmentally important members of this functional guild. The exact mechanism of the observed positive effects on methane oxidation and the differential effect on methanotrophic diversity still needs to be explored.

The role of Sphagnum mosses in the methane cycling of a boreal mire.

Science.gov (United States)

Larmola, Tuula; Tuittila, Eeva-Stiina; Tiirola, Marja; Nykänen, Hannu; Martikainen, Pertti J; Yrjälä, Kim; Tuomivirta, Tero; Fritze, Hannu

2010-08-01

Peatlands are a major natural source of atmospheric methane (CH4). Emissions from Sphagnum-dominated mires are lower than those measured from other mire types. This observation may partly be due to methanotrophic (i.e., methane-consuming) bacteria associated with Sphagnum. Twenty-three of the 41 Sphagnum species in Finland can be found in the peatland at Lakkasuo. To better understand the Sphagnum-methanotroph system, we tested the following hypotheses: (1) all these Sphagnum species support methanotrophic bacteria; (2) water level is the key environmental determinant for differences in methanotrophy across habitats; (3) under dry conditions, Sphagnum species will not host methanotrophic bacteria; and (4) methanotrophs can move from one Sphagnum shoot to another in an aquatic environment. To address hypotheses 1 and 2, we measured the water table and CH4 oxidation for all Sphagnum species at Lakkasuo in 1-5 replicates for each species. Using this systematic approach, we included Sphagnum spp. with narrow and broad ecological tolerances. To estimate the potential contribution of CH4 to moss carbon, we measured the uptake of delta13C supplied as CH4 or as carbon dioxide dissolved in water. To test hypotheses 2-4, we transplanted inactive moss patches to active sites and measured their methanotroph communities before and after transplantation. All 23 Sphagnum species showed methanotrophic activity, confirming hypothesis 1. We found that water level was the key environmental factor regulating methanotrophy in Sphagnum (hypothesis 2). Mosses that previously exhibited no CH4 oxidation became active when transplanted to an environment in which the microbes in the control mosses were actively oxidizing CH4 (hypothesis 4). Newly active transplants possessed a Methylocystis signature also found in the control Sphagnum spp. Inactive transplants also supported a Methylocystis signature in common with active transplants and control mosses, which rejects hypothesis 3. Our

Coupled effects of methane monooxygenase and nitrogen source on growth and poly-β-hydroxybutyrate (PHB) production of Methylosinus trichosporium OB3b.

Science.gov (United States)

Zhang, Tingting; Zhou, Jiti; Wang, Xiaowei; Zhang, Yu

2017-02-01

The coupled effects of nitrogen source and methane monooxygenase (MMO) on the growth and poly-β-hydroxybutyrate (PHB) accumulation capacity of methanotrophs were explored. The ammonia-supplied methanotrophs expressing soluble MMO (sMMO) grew at the highest rate, while N 2 -fixing bacteria expressing particulate MMO (pMMO) grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing pMMO, which might cause their slightly lower growth rate. The highest PHB content (51.0%) was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing pMMO. Ammonia-supplied bacteria also accumulated a higher content of PHB (45.2%) with the expression of pMMO, while N 2 -fixing bacteria containing pMMO only showed low PHB production capacity (32.1%). The maximal PHB contents of bacteria expressing sMMO were low, with no significant change under different nitrogen source conditions. The low MMO activity, low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N 2 with the expression of pMMO were greatly improved in the cyclic NO 3 - N 2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing pMMO. Copyright © 2016. Published by Elsevier B.V.

The optimal ecological factors and the denitrification populationof a denitrifying process for sulfate reducing bacteriainhibition

Science.gov (United States)

Li, Chunying

2018-02-01

SRB have great negative impacts on the oil production in Daqing Oil field. A continuous-flow anaerobic baffled reactors (ABR) are applied to investigate the feasibility and optimal ecological factors for the inhibition of SRB by denitrifying bacteria (DNB). The results showed that the SO42- to NO3- concentration ratio (SO42-/NO3-) are the most important ecological factor. The input of NO3- and lower COD can enhance the inhibition of S2-production effectively. The effective time of sulfate reduction is 6 h. Complete inhibition of SRB is obtained when the influent COD concentration is 600 mg/L, the SO42-/NO3- is 1/1 (600 mg/L for each), N is added simultaneously in the 2# and the 5# ABR chambers. By extracting the total DNA of wastewater from the effective chamber, 16SrDNA clones of a bacterium had been constructed. It is showed that the Proteobacteria accounted for eighty- four percent of the total clones. The dominant species was the Neisseria. Sixteen percent of the total clones were the Bacilli of Frimicutes. It indicated that DNB was effective and feasible for SRB inhibition.

Community structures and activity of denitrifying microbes in a forested catchment in central Japan: survey using nitrite reductase genes

Science.gov (United States)

Ohte, N.; Aoki, M.; Katsuyama, C.; Suwa, Y.; Tange, T.

2012-12-01

To elucidate the mechanisms of denitrification processes in the forested catchment, microbial ecological approaches have been applied in an experimental watershed that has previously investigated its hydrological processes. The study catchment is located in the Chiba prefecture in central Japan under the temperate Asian monsoon climate. Potential activities of denitrification of soil samples were measured by incubation experiments under anoxic condition associated with Na15NO3 addition. Existence and variety of microbes having nitrite reductase genes were investigated by PCR amplification, cloning and sequencings of nirK and nirS fragments after DNA extraction. Contrary to our early expectation that the potential denitrification activity was higher at deeper soil horizon with consistent groundwater residence than that in the surface soil, denitrification potential was higher in shallower soil horizons than deeper soils. This suggested that the deficiency of NO3- as a respiratory substrate for denitrifier occurred in deeper soils especially in the summer. However, high denitrification activity and presence of microbes having nirK and nirS in surface soils usually under aerobic condition was explainable by the fact that the majority of denitrifying bacteria have been recognized as a facultative anaerobic bacterium. This also suggests the possibility of that denitrification occurs even in the surface soils if the wet condition is provided by rainwater during and after a storm event. Community structures of microbes having nirK were different between near surface and deeper soil horizons, and ones having nirS was different between saturated zone (under groundwater table) and unsaturated soil horizons. These imply that microbial communities with nisK are sensitive to the concentration of soil organic matters and ones with nirS is sensitive to soil moisture contents.

Microbial CH4 and N2O consumption in acidic wetlands

Directory of Open Access Journals (Sweden)

Steffen eKolb

2012-03-01

Full Text Available Acidic wetlands are global sources of the atmospheric greenhouse gases methane (CH4, and nitrous oxide (N2O. Consumption of both atmospheric gases has been observed in various acidic wetlands, but information on the microbial mechanisms underlying these phenomena is scarce. A substantial amount of CH4 is consumed in sub soil by aerobic methanotrophs at anoxic–oxic interfaces (e.g., tissues of Sphagnum mosses, rhizosphere of vascular plant roots. Methylocystis-related species are likely candidates that are involved in the consumption of atmospheric CH4 in acidic wetlands. Oxygen availability regulates the activity of methanotrophs of acidic wetlands. Other parameters impacting on the methanotroph-mediated CH4 consumption have not been systematically evaluated. N2O is produced and consumed by microbial denitrification, thus rendering acidic wetlands as temporary sources or sinks for N2O. Denitrifier communities in such ecosystems are diverse, and largely uncultured and/or new, and environmental factors that control their consumption activity are unresolved. Analyses of the composition of N2O reductase genes in acidic wetlands suggest that acid-tolerant Proteobacteria have the potential to mediate N2O consumption in such soils. Thus, the fragmented current state of knowledge raises open questions concerning methanotrophs and dentrifiers that consume atmospheric CH4 and N2O in acidic wetlands.

Denitrifying metabolism of the methylotrophic marine bacterium Methylophaga nitratireducenticrescens strain JAM1

Directory of Open Access Journals (Sweden)

Florian Mauffrey

2017-11-01

-input. ${\\mathrm{NO}}_{2}^{-}$ NO 2 − was shown to be the key intermediate of this production process. Upregulation in the expression of cnorB1, cnorB2, nnrS and norR during the growth and the N2O accumulation phases suggests NO production in strain JAM1 cultures. Discussion By showing that all the three denitrification reductases are active, this demonstrates that M. nitratireducenticrescens JAM1 is one of many bacteria species that maintain genes associated primarily with denitrification, but not necessarily related to the maintenance of the entire pathway. The reason to maintain such an incomplete pathway could be related to the specific role of strain JAM1 in the denitrifying biofilm of the denitrification reactor from which it originates. The production of N2O in strain JAM1 did not involve Nar, contrary to what was demonstrated in Escherichia coli. M. nitratireducenticrescens JAM1 is the only reported Methylophaga species that has the capacity to grow under anoxic conditions by using ${\\mathrm{NO}}_{3}^{-}$ NO 3 − and N2O as sole electron acceptors for its growth. It is also one of a few marine methylotrophs that is studied at the physiological and genetic levels in relation to its capacity to perform denitrifying activities.

Regulation of methane oxidation in the facultative methanotroph Methylocella silvestris BL2.

Science.gov (United States)

Theisen, Andreas R; Ali, M Hanif; Radajewski, Stefan; Dumont, Marc G; Dunfield, Peter F; McDonald, Ian R; Dedysh, Svetlana N; Miguez, Carlos B; Murrell, J Colin

2005-11-01

The molecular regulation of methane oxidation in the first fully authenticated facultative methanotroph Methylocella silvestris BL2 was assessed during growth on methane and acetate. Problems of poor growth of Methylocella spp. in small-scale batch culture were overcome by growth in fermentor culture. The genes encoding soluble methane monooxygenase were cloned and sequenced, which revealed that the structural genes for soluble methane monooxygenase, mmoXYBZDC, were adjacent to two genes, mmoR and mmoG, encoding a sigma54 transcriptional activator and a putative GroEL-like chaperone, located downstream (3') of mmoC. Transcriptional analysis revealed that the genes were all cotranscribed from a sigma54-dependent promoter located upstream (5') of mmo X. The transcriptional start site was mapped. Transcriptional analysis of soluble methane monooxygenase genes and expression studies on fermentor grown cultures showed that acetate repressed transcription of sMMO in M. silvestris BL2. The possibility of the presence of a particulate, membrane-bound methane monooxygenase enzyme in M. silvestris BL2 and the copper-mediated regulation of soluble methane monooxygenase was investigated. Both were shown to be absent. A promoter probe vector was constructed and used to assay transcription of the promoter of the soluble methane monoxygenase genes of M. silvestris BL2 grown under various conditions and with different substrates. These data represent the first insights into the molecular physiology of a facultative methanotroph.

Draft Genome Sequence of an Active Heterotrophic Nitrifier-Denitrifier, Cupriavidus pauculus UM1

OpenAIRE

Putonti, Catherine; Polley, Nathaniel; Castignetti, Domenic

2018-01-01

ABSTRACT Here, we present the draft genome sequence of Cupriavidus pauculus UM1, a metal-resistant heterotrophic nitrifier-denitrifier capable of synthesizing nitrite from pyruvic oxime. The size of the genome is 7,402,815 bp with a GC content of 64.8%. This draft assembly consists of 38 scaffolds.

Directory of Open Access Journals (Sweden)

Mohammed A. Almalki

2017-11-01

Full Text Available Methanotrophic bacteria have a unique ability to utilize methane as their carbon and energy sources. Therefore, methanotrophs play a key role in suppressing methane emissions from different ecosystems and hence in alleviating the global climate change. Despite methanotrophs having many ecological, economical and biotechnological applications, little is known about this group of bacteria in Al-Ahsa. Therefore, the main objective of the current work was to expand our understanding of methane oxidizing bacteria in Al-Ahsa region. The specific aim was to describe a methanotrophic strain isolated from Al-Bohyriya well, Al-Ahsa using phenotypic, genotypic (such as 16S rRNA and pmoA gene sequencing and phylogenetic characterization. The results indicated that the strain belongs to the genus Methylomonas that belongs to Gammaproteobacteria as revealed by the comparative sequence analysis of the 16S rRNA and pmoA genes. There is a general agreement in the profile of the phylogenetic trees based on the sequences of 16srRNA and pmoA genes of the strain BOH1 indicating that both genes are efficient taxonomic marker in methanotrophic phylogeny. The strain possesses the particulate but not the soluble methane monooxygenase as a key enzyme for methane metabolism. Further investigation such as DNA:DNA hybridization is needed to assign the strain as a novel species of the genus Methyomonas and this will open the door to explore the talents of the strain for its potential role in alleviating global warming and biotechnological applications in Saudi Arabia such as bioremediation of toxic by-products released in oil industry. In addition, the strain enhances our knowledge of methanotrophic bacteria and their adaptation to desert ecosystems.

Fluorescent antibody application in bioremediation procedures at the Savannah River Site

International Nuclear Information System (INIS)

Brigmon, R.L.; Richardson, B.S.; Franck, M.M.

1997-01-01

Direct Fluorescent Antibodies (DFA) and Most Probable Number (MPN) techniques are currently being employed at the Savannah River Site to monitor methanotrophic bacteria for the bioremediation of trichloroethylene (TCE) in field studies. Direct Fluorescent Antibodies were developed against various methanotrophic bacteria isolated from SRS as well as methanotrophic bacteria acquired from the American Type Culture Collection (ATCC). DFA's are anticipated to be more efficient for monitoring methanotroph activity than MPN's because of shorter processing time, lower cost, and the direct nature of the assay. The DFA method is a direct technique, in that samples are processed immediately and can be enumerated within an hour. The MPN method is indirect, since samples must be cultured for 6-8 weeks before measuring methane consumption and carbon dioxide production. Indirect methods are not highly selective and have limited application. The greatest advantage of a faster assay, is that bioremediation procedures utilizing methanotrophic bacteria could be amended. These amendments would be based on environmental monitoring with results in real time (1 hour). The elimination of the MPN technique and the use of DFA's will save significantly on both materials and labor. The data obtained from the DFA's and MPN's were statistically compared to each other and to total bacterial counts (AODC). The statistical analysis used was Analysis of Variants (ANOVA). Using this analysis, groundwater samples were found to be not significantly different; whereas soil were significantly different. These methods were employed on soil samples from the Southern Sector and ground water samples from the TCE-contaminated Sanitary Landfill at SRS. Acridine Orange Direct Counts were compared to show relative differences between total bacterial and methanotroph population

Co-cultivation of Green Microalgae and Methanotrophic Bacteria for Single Cell Protein Production from Wastewater

DEFF Research Database (Denmark)

Rasouli, Zahra; Valverde Pérez, Borja; D'Este, Martina

2017-01-01

microalgae – as a means to recover nutrients from industrial wastewater and upcycle them to feed grade single cell protein. Results demonstrated that both algae and bacteria could remove or assimilate most of the organic carbon present in the wastewater. However, their growth stopped before nutrients...

Methane and Trichloroethylene Oxidation by an Estuarine Methanotroph, Methylobacter sp. Strain BB5.1

OpenAIRE

Smith, Kelly S.; Costello, Andria M.; Lidstrom, Mary E.

1998-01-01

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.

Methanotrophic activity and diversity in different Sphagnum magellanicum dominated habitats in the southernmost peat bogs of Patagonia

NARCIS (Netherlands)

Kip, N.; Fritz, C.; Langelaan, E. S.; Pan, Y.; Bodrossy, L.; Pancotto, V.; Jetten, M. S. M.; Smolders, A. J. P.; den Camp, H. J. M. Op

2012-01-01

Sphagnum peatlands are important ecosystems in the methane cycle. Methanotrophs living inside the dead hyaline cells or on the Sphagnum mosses are able to act as a methane filter and thereby reduce methane emissions. We investigated in situ methane concentrations and the corresponding activity and

Seasonality distribution of the abundance and activity of nitrification and denitrification microorganisms in sediments of surface flow constructed wetlands planted with Myriophyllum elatinoides during swine wastewater treatment.

Science.gov (United States)

Li, Xi; Zhang, Miaomiao; Liu, Feng; Chen, Liang; Li, Yuyuan; Li, Yong; Xiao, Rulin; Wu, Jinshui

2018-01-01

Surface flow constructed wetlands (SFCWs) planted with Myriophyllum elatinoides for treatment of swine wastewater were examined to evaluate the effect of season, segment (site S1, S2, and S3), and treatment (100mgL -1 TN, T1; 300mgL -1 TN, T2; 500mgL -1 TN, T3) on the activity, and abundances of nitrifying and, denitrifying microorganisms, and on the abundance of sediment bacteria. The activity and abundances of nitrifiers, denitrifiers, and the abundance of bacteria were the highest in T3 samples, especially in S1 (Pswine wastewater treatment stimulate the growth of nitrifiers, denitrifiers and bacteria in sediments. Copyright © 2017 Elsevier Ltd. All rights reserved.

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.

Detection of putatively thermophilic anaerobic methanotrophs in diffuse hydrothermal vent fluids.

Science.gov (United States)

Merkel, Alexander Y; Huber, Julie A; Chernyh, Nikolay A; Bonch-Osmolovskaya, Elizaveta A; Lebedinsky, Alexander V

2013-02-01

The anaerobic oxidation of methane (AOM) is carried out by a globally distributed group of uncultivated Euryarchaeota, the anaerobic methanotrophic arachaea (ANME). In this work, we used G+C analysis of 16S rRNA genes to identify a putatively thermophilic ANME group and applied newly designed primers to study its distribution in low-temperature diffuse vent fluids from deep-sea hydrothermal vents. We found that the G+C content of the 16S rRNA genes (P(GC)) is significantly higher in the ANME-1GBa group than in other ANME groups. Based on the positive correlation between the P(GC) and optimal growth temperatures (T(opt)) of archaea, we hypothesize that the ANME-1GBa group is adapted to thrive at high temperatures. We designed specific 16S rRNA gene-targeted primers for the ANME-1 cluster to detect all phylogenetic groups within this cluster, including the deeply branching ANME-1GBa group. The primers were successfully tested both in silico and in experiments with sediment samples where ANME-1 phylotypes had previously been detected. The primers were further used to screen for the ANME-1 microorganisms in diffuse vent fluid samples from deep-sea hydrothermal vents in the Pacific Ocean, and sequences belonging to the ANME-1 cluster were detected in four individual vents. Phylotypes belonging to the ANME-1GBa group dominated in clone libraries from three of these vents. Our findings provide evidence of existence of a putatively extremely thermophilic group of methanotrophic archaea that occur in geographically and geologically distinct marine hydrothermal habitats.

Isolation of amoebic-bacterial consortia capable of degrading trichloroethylene

International Nuclear Information System (INIS)

Tyndall, R.L.; Ironside, K.; Little, C.D.; Katz, S.; Kennedy, J.

1990-01-01

Groundwater from a waste disposal site contaminated with chlorinated alkenes was examined for the presence of amoebic-bacterial consortia capable of degrading the suspected carcinogen, trichloroethylene (TCE). Consortia were readily isolated from all of four test wells. They contained free-living amoebae, and heterotrophic and methylotrophic bacteria. Electron microscopic examination showed bacteria localized throughout the amoebic cytoplasm and an abundance of hyphomicrobium, but not Type I methanotrophs. The presence of Type II methanotrophs was indirectly indicated by lipid analysis of one consortium. The consortia have been passaged for over two years on mineral salts media in a methane atmosphere, which would not be expected to maintain the heterotrophs or amoebae separately. The methanotrophic bacteria apparently provided a stable nutrient source, allowing the persistence of the various genera. By use of 14 C-radiotracer techniques, the degradation of TCE by the consortia was observed with 14 C eventuating predominantly in CO 2 and water-soluble products. In a more detailed examination of one consortia, the amoebae and heterotrohic components did not degrade TCE, while a mixed culture of heterotrophs and methanotrophs did degrade TCE, suggesting the latter component was the primary cause for the consortium's ability to degrade TCE. Amoebic-bacterial consortia may play a role in stabilizing and preserving methylotrophic bacteria in hostile environments

Draft Genome Sequence of an Active Heterotrophic Nitrifier-Denitrifier, Cupriavidus pauculus UM1.

Science.gov (United States)

Putonti, Catherine; Polley, Nathaniel; Castignetti, Domenic

2018-02-08

Here, we present the draft genome sequence of Cupriavidus pauculus UM1, a metal-resistant heterotrophic nitrifier-denitrifier capable of synthesizing nitrite from pyruvic oxime. The size of the genome is 7,402,815 bp with a GC content of 64.8%. This draft assembly consists of 38 scaffolds. Copyright © 2018 Putonti et al.

The transcriptome of Bathymodiolus azoricus gill reveals expression of genes from endosymbionts and free-living deep-sea bacteria.

Science.gov (United States)

Egas, Conceição; Pinheiro, Miguel; Gomes, Paula; Barroso, Cristina; Bettencourt, Raul

2012-08-01

Deep-sea environments are largely unexplored habitats where a surprising number of species may be found in large communities, thriving regardless of the darkness, extreme cold, and high pressure. Their unique geochemical features result in reducing environments rich in methane and sulfides, sustaining complex chemosynthetic ecosystems that represent one of the most surprising findings in oceans in the last 40 years. The deep-sea Lucky Strike hydrothermal vent field, located in the Mid Atlantic Ridge, is home to large vent mussel communities where Bathymodiolus azoricus represents the dominant faunal biomass, owing its survival to symbiotic associations with methylotrophic or methanotrophic and thiotrophic bacteria. The recent transcriptome sequencing and analysis of gill tissues from B. azoricus revealed a number of genes of bacterial origin, hereby analyzed to provide a functional insight into the gill microbial community. The transcripts supported a metabolically active microbiome and a variety of mechanisms and pathways, evidencing also the sulfur and methane metabolisms. Taxonomic affiliation of transcripts and 16S rRNA community profiling revealed a microbial community dominated by thiotrophic and methanotrophic endosymbionts of B. azoricus and the presence of a Sulfurovum-like epsilonbacterium.

Microbial characteristics of purple paddy soil in response to Pb pollution.

Science.gov (United States)

Jiang, Qiu-Ju; Zhang, Yue-Qiang; Zhang, La-Mei; Zhou, Xin-Bin; Shi, Xiao-Jun

2014-05-01

The study focused on the change of microbial characteristics affected by Plumbum pollution with purple paddy soil in an incubation experiment. The results showed that low concentration of Plumbum had little effect on most of microbial amounts, biological activity and enzymatic activity. However, denitrifying activity was inhibited severely, and inhibition rate was up to 98%. Medium and high concentration of Plumbum significantly reduced the amounts and activity of all microorganisms and enzymatic activity, which increased with incubation time. Negative correlations were found between Plumbum concentrations and microbial amounts, biological activity and enzymatic activities except fungi and actinomyces. Thus they can be used to indicate the Plumbum pollution levels to some extent. LD(50) of denitrifying bacteria (DB) and ED50 of denitrifying activity were 852mg/kg and 33.5mg/kg. Across all test soil microbes, denitrifying bacteria was most sensitive to Plumbum pollution in purple paddy soil. Value of early warning showed that anaerobic cellulose-decomposing bacteria (ACDB) and actinomyces were also sensitive to Plumbum pollution. We concluded that denitrifying activity, actinomyces, ACDB or DB can be chosen as predictor of Plumbum contamination in purple paddy soil.

Methane distribution and oxidation around the Lena Delta in summer 2013

Science.gov (United States)

Bussmann, Ingeborg; Hackbusch, Steffen; Schaal, Patrick; Wichels, Antje

2017-11-01

The Lena River is one of the largest Russian rivers draining into the Laptev Sea. The predicted increases in global temperatures are expected to cause the permafrost areas surrounding the Lena Delta to melt at increasing rates. This melting will result in high amounts of methane reaching the waters of the Lena and the adjacent Laptev Sea. The only biological sink that can lower methane concentrations within this system is methane oxidation by methanotrophic bacteria. However, the polar estuary of the Lena River, due to its strong fluctuations in salinity and temperature, is a challenging environment for bacteria. We determined the activity and abundance of aerobic methanotrophic bacteria by a tracer method and by the quantitative polymerase chain reaction. We described the methanotrophic population with a molecular fingerprinting method (monooxygenase intergenic spacer analysis), as well as the methane distribution (via a headspace method) and other abiotic parameters, in the Lena Delta in September 2013. The median methane concentrations were 22 nmol L-1 for riverine water (salinity (S) 20). The Lena River was not the source of methane in surface water, and the methane concentrations of the bottom water were mainly influenced by the methane concentration in surface sediments. However, the bacterial populations of the riverine and polar waters showed similar methane oxidation rates (0.419 and 0.400 nmol L-1 d-1), despite a higher relative abundance of methanotrophs and a higher estimated diversity in the riverine water than in the polar water. The methane turnover times ranged from 167 days in mixed water and 91 days in riverine water to only 36 days in polar water. The environmental parameters influencing the methane oxidation rate and the methanotrophic population also differed between the water masses. We postulate the presence of a riverine methanotrophic population that is limited by sub-optimal temperatures and substrate concentrations and a polar

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

Science.gov (United States)

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

2008-11-01

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

[Influences of long-term application of organic and inorganic fertilizers on the composition and abundance of nirS-type denitrifiers in black soil].

Science.gov (United States)

Yin, Chang; Fan, Fen-Liang; Li, Zhao-Jun; Song, A-Lin; Zhu, Ping; Peng, Chang; Liang, Yong-Chao

2012-11-01

The objectives of this study were to explore the effects of long-term organic and inorganic fertilizations on the composition and abundance of nirS-type denitrifiers in black soil. Soil samples were collected from 4 treatments (i. e. no fertilizer treatment, CK; organic manure treatment, OM; chemical fertilizer treatment (NPK) and combination of organic and chemical fertilizers treatment (MNPK)) in Gongzhuling Long-term Fertilization Experiment Station. Composition and abundance of nirS-type denitrifiers were analyzed with terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR (Q-PCR), respectively. Denitrification enzyme activity (DEA) and soil properties were also measured. Application of organic fertilizers (OM and MNPK) significantly increased the DEAs of black soil, with the DEAs in OM and MNPK being 5.92 and 6.03 times higher than that in CK treatment, respectively, whereas there was no significant difference between NPK and CK. OM and MNPK treatments increased the abundances of nirS-type denitrifiers by 2.73 and 3.83 times relative to that of CK treatment, respectively. The abundance of nirS-type denitrifiers in NPK treatment was not significantly different from that of CK. The T-RFLP analysis of nirS genes showed significant differences in community composition between organic and inorganic treatments, with the emergence of a 79 bp T-RF, a significant decrease in relative abundance of the 84 bp T-RF and a loss of the 99 bp T-RF in all organic treatments. Phylogenetic analysis indicated that the airS-type denitrifiers in the black soil were mainly composed of alpha, beta and gamma-Proteobacteria. The 79 bp-type denitrifiers inhabiting exclusively in organic treatments (OM and MNPK) were affiliated to Pseudomonadaceae in gamma-Proteobacteria and Burkholderiales in beta-Proteobacteria. The 84 bp-types were related to Burkholderiales and Rhodocyclales. Correlation analysis indicated that pH, concentrations of total nitrogen

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

Science.gov (United States)

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

2015-10-01

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

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

Digital Repository Service at National Institute of Oceanography (India)

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

Denitrification often occurs in the water column, underlying zones of intense productivity and decomposition in upwelling regions. In the denitrifying zone off the southwest coast of India, high concentrations of nitrite (greater than 15 mu M...

The influence of bacteria-dominated diets on Daphnia magna somatic growth, reproduction, and lipid composition.

Science.gov (United States)

Taipale, Sami J; Brett, Michael T; Pulkkinen, Katja; Kainz, Martin J

2012-10-01

We explored how dietary bacteria affect the life history traits and biochemical composition of Daphnia magna, using three bacteria taxa with very different lipid composition. Our objectives were to (1) examine whether and how bacteria-dominated diets affect Daphnia survival, growth, and fecundity, (2) see whether bacteria-specific fatty acid (FA) biomarkers accrued in Daphnia lipids, and (3) explore the quantitative relationship between bacteria availability in Daphnia diets and the amounts of bacterial FA in their lipids. Daphnia were fed monospecific and mixed diets of heterotrophic (Micrococcus luteus) or methanotrophic bacteria (Methylomonas methanica and Methylosinus trichosporium) and two phytoplankton species (Cryptomonas ozolinii and Scenedesmus obliquus). Daphnia neonates fed pure bacteria diets died after 6-12 days and produced no viable offspring, whereas those fed pure phytoplankton diets had high survival, growth, and reproduction success. Daphnia fed a mixed diet with 80% M. luteus and 20% of either phytoplankton had high somatic growth, but low reproduction. Conversely, Daphnia fed mixed diets including 80% of either methane-oxidizing bacteria and 20% Cryptomonas had high reproduction rates, but low somatic growth. All Daphnia fed mixed bacteria and phytoplankton diets had strong evidence of both bacteria- and phytoplankton-specific FA biomarkers in their lipids. FA mixing model calculations indicated that Daphnia that received 80% of their carbon from bacteria assimilated 46 ± 25% of their FA from this source. A bacteria-phytoplankton gradient experiment showed a strong positive correlation between the proportions of the bacterial FA in the Daphnia and their diet, indicating that bacterial utilization can be traced in this keystone consumer using FA biomarkers. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

The Transcriptome of Bathymodiolus azoricus Gill Reveals Expression of Genes from Endosymbionts and Free-Living Deep-Sea Bacteria

Directory of Open Access Journals (Sweden)

Raul Bettencourt

2012-08-01

Full Text Available Deep-sea environments are largely unexplored habitats where a surprising number of species may be found in large communities, thriving regardless of the darkness, extreme cold, and high pressure. Their unique geochemical features result in reducing environments rich in methane and sulfides, sustaining complex chemosynthetic ecosystems that represent one of the most surprising findings in oceans in the last 40 years. The deep-sea Lucky Strike hydrothermal vent field, located in the Mid Atlantic Ridge, is home to large vent mussel communities where Bathymodiolus azoricus represents the dominant faunal biomass, owing its survival to symbiotic associations with methylotrophic or methanotrophic and thiotrophic bacteria. The recent transcriptome sequencing and analysis of gill tissues from B. azoricus revealed a number of genes of bacterial origin, hereby analyzed to provide a functional insight into the gill microbial community. The transcripts supported a metabolically active microbiome and a variety of mechanisms and pathways, evidencing also the sulfur and methane metabolisms. Taxonomic affiliation of transcripts and 16S rRNA community profiling revealed a microbial community dominated by thiotrophic and methanotrophic endosymbionts of B. azoricus and the presence of a Sulfurovum-like epsilonbacterium.

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

Science.gov (United States)

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

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

Water column methanotrophy controlled by a rapid oceanographic switch

NARCIS (Netherlands)

Steinle, L.; Graves, C.A.; Treude, T.; Ferré, B.; Biastoch, A.; Bussmann, I.; Berndt, C.; Krastel, S.; James, R.H.; Behrens, E.; Böning, C.W.; Greinert, J.; Sapart, C.-J.; Scheinert, M.; Sommer, S.; Lehmann, M.F.; Niemann, H.

2015-01-01

From the seabed to the water column, where it may be consumed by aerobic methanotrophic bacteria. The size and activity of methanotrophic communities, which determine the amount of methane consumed in the water column,are thought to be mainly controlled by nutrient and redoxdynamics3–7. Here, we

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

equipped with a thermal con- ductivity detector (TCD) with a CTR1 column ( Alltech Associates Inc., Deerfield, IL) with He as carrier gas. The injection...grow at this pH (Bowman, 2006). Ama- ral and Knowles (1995) reported that Type II methanotrophs had a competitive advantage when oxygen was limited and

A novel surface-enhanced Raman scattering (SERS) detection for natural gas exploration using methane-oxidizing bacteria.

Science.gov (United States)

Liang, Weiwei; Chen, Qiao; Peng, Fang; Shen, Aiguo; Hu, Jiming

2018-07-01

Methane-oxidizing bacteria (MOB), a unique group of Gram-negative bacteria utilizing methane as a sole source of carbon and energy, have been proved to be a biological indicator for gas prospecting. Field and cultivation-free detection of MOB is important but still challenging in current microbial prospecting of oil and gas (MPOG) system. Herein, SERS was used for the first time to our knowledge to investigate two species of methanotrophs and four closely relevant bacteria that universally coexisted in the upper soil of natural gas. A special but very simple approach was utilized to make silver nanoparticles (Ag NPs) sufficiently contact with every single bacterial cell, and highly strong and distinct Raman signals free from any native fluorescence have been obtained, and successfully utilized for distinguishing MOB from other species. A more convincing multi-Raman criterion based on single Raman bands, and further the entire Raman spectrum in combination with statistical analysis (e.g., principal component analysis (PCA)), which were found capable of classifying MOB related bacterial cells in soil with an accuracy of 100%. This study therefore demonstrated sensitive and rapid SERS measurement technique accompanied by complete Raman database of various gas reservoirs related bacteria could aid field exploration of natural gas reservoir. Copyright © 2018 Elsevier B.V. All rights reserved.

Changes in bacterial community structure correlate with initial operating conditions of a field-scale denitrifying fluidized bed reactor

Energy Technology Data Exchange (ETDEWEB)

Hwang, C. [Miami Univ., Oxford, OH (United States). Dept. of Microbiology; Wu, W.M. [Stanford Univ., CA (United States). Dept. of Civil and Environmental Engineering; Gentry, T.J. [Oak Ridge National Lab., TN (US). Environmental Sciences Div.] (and others)

2006-08-15

High levels of nitrate are present in groundwater migrating from the former waste disposal ponds at the Y-12 National Security Complex in Oak Ridge, TN. A field-scale denitrifying fluidized bed reactor (FBR) was designed, constructed, and operated with ethanol as an electron donor for the removal of nitrate. After inoculation, biofilms developed on the granular activated carbon particles. Changes in the bacterial community of the FBR were evaluated with clone libraries (n=500 partial sequences) of the small-subunit rRNA gene for samples taken over a 4-month start-up period. Early phases of start-up operation were characterized by a period of selection, followed by low diversity and predominance by Azoarcus-like sequences. Possible explanations were high pH and nutrient limitations. After amelioration of these conditions, diversification increased rapidly, with the appearance of Dechloromonas, Pseudomonas, and Hydrogenophaga sequences. Changes in NO{sub 3}, SO{sub 4}, and pH also likely contributed to shifts in community composition. The detection of sulfate-reducing-bacteria-like sequences closely related to Desulfovibrio and Desulfuromonas in the FBR have important implications for downstream applications at the field site. (orig.)

Stratification of Diversity and Activity of Methanogenic and Methanotrophic Microorganisms in a Nitrogen-Fertilized Italian Paddy Soil

Directory of Open Access Journals (Sweden)

Annika Vaksmaa

2017-11-01

Full Text Available Paddy fields are important ecosystems, as rice is the primary food source for about half of the world’s population. Paddy fields are impacted by nitrogen fertilization and are a major anthropogenic source of methane. Microbial diversity and methane metabolism were investigated in the upper 60 cm of a paddy soil by qPCR, 16S rRNA gene amplicon sequencing and anoxic 13C-CH4 turnover with a suite of electron acceptors. The bacterial community consisted mainly of Acidobacteria, Chloroflexi, Proteobacteria, Planctomycetes, and Actinobacteria. Among archaea, Euryarchaeota and Bathyarchaeota dominated over Thaumarchaeota in the upper 30 cm of the soil. Bathyarchaeota constituted up to 45% of the total archaeal reads in the top 5 cm. In the methanogenic community, Methanosaeta were generally more abundant than the versatile Methanosarcina. The measured maximum methane production rate was 444 nmol gdwh-1, and the maximum rates of nitrate-, nitrite-, and iron-dependent anaerobic oxidation of methane (AOM were 57 nmol, 55 nmol, and 56 nmol gdwh-1, respectively, at different depths. qPCR revealed a higher abundance of ‘Candidatus Methanoperedens nitroreducens’ than methanotrophic NC10 phylum bacteria at all depths, except at 60 cm. These results demonstrate that there is substantial potential for AOM in fertilized paddy fields, with ‘Candidatus Methanoperedens nitroreducens’ archaea as a potential important contributor.

Methylocapsa aurea sp. nov., a facultative methanotroph possessing a particulate methane monooxygenase, and emended description of the genus Methylocapsa.

Science.gov (United States)

Dunfield, Peter F; Belova, Svetlana E; Vorob'ev, Alexey V; Cornish, Sabrina L; Dedysh, Svetlana N

2010-11-01

An aerobic, methanotrophic bacterium, designated KYG(T), was isolated from a forest soil in Germany. Cells of strain KYG(T) were Gram-negative, non-motile, slightly curved rods that multiplied by binary fission and produced yellow colonies. The cells contained intracellular granules of poly-β-hydroxybutyrate at each cell pole, a particulate methane monooxygenase (pMMO) and stacks of intracytoplasmic membranes (ICMs) packed in parallel along one side of the cell envelope. Strain KYG(T) grew at pH 5.2-7.2 and 2-33 °C and could fix atmospheric nitrogen under reduced oxygen tension. The major cellular fatty acid was C(18 : 1)ω7c (81.5 %) and the DNA G+C content was 61.4 mol%. Strain KYG(T) belonged to the family Beijerinckiaceae of the class Alphaproteobacteria and was most closely related to the obligate methanotroph Methylocapsa acidiphila B2(T) (98.1 % 16S rRNA gene sequence similarity and 84.7 % pmoA sequence similarity). Unlike Methylocapsa acidiphila B2(T), which grows only on methane and methanol, strain KYG(T) was able to grow facultatively on acetate. Facultative acetate utilization is a characteristic of the methanotrophs of the genus Methylocella, but the genus Methylocella does not produce pMMO or ICMs. Strain KYG(T) differed from Methylocapsa acidiphila B2(T) on the basis of substrate utilization pattern, pigmentation, pH range, cell ultrastructure and efficiency of dinitrogen fixation. Therefore, we propose a novel species, Methylocapsa aurea sp. nov., to accommodate this bacterium. The type strain is KYG(T) (=DSM 22158(T) =VKM B-2544(T)).

Continuous measurements of nitrous oxide isotopomers during incubation experiments

DEFF Research Database (Denmark)

Winther, Malte Nordmann; Balslev-Harder, David; Christensen, Søren

2016-01-01

relevant for studies of atmospheric chemistry. We present results from continuous incubation experiments with denitrifying bacteria, Pseudomonas fluorescens (producing and reducing N2O) and P. chlororaphis (only producing N2O). The continuous position dependent measurements reveal the transient pattern....... fluorescens, the calculations results in SP values of −5.7 ‰ ± 5.6 during production of N2O and 2.3 ‰ ± 3.2 during reduction of N2O. In summary, we implemented continuous measurements of N2O isotopomers during incubation of denitrifying bacteria and believe that similar experiments will lead to a better...... understanding of denitrifying bacteria and N2O turnover in soils and sediments and ultimately hands-on knowledge on the biotic mechanisms behind greenhouse gas exchange of the Globe....

Enrichment of anaerobic nitrate-dependent methanotrophic ?Candidatus Methanoperedens nitroreducens? archaea from an Italian paddy field soil

OpenAIRE

Vaksmaa, Annika; Guerrero-Cruz, Simon; van Alen, Theo A.; Cremers, Geert; Ettwig, Katharina F.; L?ke, Claudia; Jetten, Mike S. M.

2017-01-01

Paddy fields are a significant source of methane and contribute up to 20% of total methane emissions from wetland ecosystems. These inundated, anoxic soils featuring abundant nitrogen compounds and methane are an ideal niche for nitrate-dependent anaerobic methanotrophs. After 2?years of enrichment with a continuous supply of methane and nitrate as the sole electron donor and acceptor, a stable enrichment dominated by ?Candidatus Methanoperedens nitroreducens? archaea and ?Candidatus Methylom...

Chemosynthetic bacteria found in bivalve species from mud volcanoes of the Gulf of Cadiz.

Science.gov (United States)

Rodrigues, Clara F; Webster, Gordon; Cunha, Marina R; Duperron, Sébastien; Weightman, Andrew J

2010-09-01

As in other cold seeps, the dominant bivalves in mud volcanoes (MV) from the Gulf of Cadiz are macrofauna belonging to the families Solemyidae (Acharax sp., Petrasma sp.), Lucinidae (Lucinoma sp.), Thyasiridae (Thyasira vulcolutre) and Mytilidae (Bathymodiolus mauritanicus). The delta(13)C values measured in solemyid, lucinid and thyasirid specimens support the hypothesis of thiotrophic nutrition, whereas isotopic signatures of B. mauritanicus suggest methanotrophic nutrition. The indication by stable isotope analysis that chemosynthetic bacteria make a substantial contribution to the nutrition of the bivalves led us to investigate their associated bacteria and their phylogenetic relationships based on comparative 16S rRNA gene sequence analysis. PCR-denaturing gradient gel electrophoresis analysis and cloning of bacterial 16S rRNA-encoding genes confirmed the presence of sulfide-oxidizing symbionts within gill tissues of many of the studied specimens. Phylogenetic analysis of bacterial 16S rRNA gene sequences demonstrated that most bacteria were related to known sulfide-oxidizing endosymbionts found in other deep-sea chemosynthetic environments, with the co-occurrence of methane-oxidizing symbionts in Bathymodiolus specimens. This study confirms the presence of several chemosynthetic bivalves in the Gulf of Cadiz and further highlights the importance of sulfide- and methane-oxidizing symbionts in the trophic ecology of macrobenthic communities in MV.

Highly enriched Betaproteobacteria growing anaerobically with p-xylene and nitrate

DEFF Research Database (Denmark)

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

2010-01-01

The identity of the microorganisms capable of anaerobic p-xylene degradation under denitrifying conditions is hitherto unknown. Here, we report highly enriched cultures of freshwater denitrifying bacteria that grow anaerobically with p-xylene as the sole organic carbon source and electron donor. ...

Methane Seep in Shallow-Water Permeable Sediment Harbors High Diversity of Anaerobic Methanotrophic Communities, Elba, Italy

Science.gov (United States)

Ruff, S. Emil; Kuhfuss, Hanna; Wegener, Gunter; Lott, Christian; Ramette, Alban; Wiedling, Johanna; Knittel, Katrin; Weber, Miriam

2016-01-01

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 distinct

Methane Seep in Shallow-Water Permeable Sediment Harbors High Diversity of Anaerobic Methanotrophic Communities, Elba, Italy.

Science.gov (United States)

Ruff, S Emil; Kuhfuss, Hanna; Wegener, Gunter; Lott, Christian; Ramette, Alban; Wiedling, Johanna; Knittel, Katrin; Weber, Miriam

2016-01-01

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 distinct

Expanding diversity of potential bacterial partners of the methanotrophic ANME archaea using Magneto-FISH

Science.gov (United States)

Trembath-Reichert, E.; Green-Saxena, A.; Steele, J. A.; Orphan, V. J.

2012-12-01

Sulfate-coupled anaerobic oxidation of methane (AOM) in marine sediments is the major sink for methane in the oceans. This process is believed to be catalyzed by as yet uncultured syntrophic consortia of ANME archaea (affiliated with the Methanosarcinales) and sulfate-reducing bacteria belonging to the Desulfosarcina/Desulfococcus and Desulfobulbaceae. These syntrophic consortia have been described from methane-rich habitats worldwide and appear to be most concentrated in areas of high methane flux, such as cold seeps along continental margins. The extent of the diversity and ecophysiological potential of these microbial associations is still poorly constrained. In an effort to better characterize the diversity of microorganisms forming associations with different clades of methanotrophic ANME archaea (ANME-1, ANME-2a/b/c, ANME-3) and link these organisms to potentially diagnostic metabolic genes (e.g. mcrA, dsrAB, aprA), we employed a unique culture-independent whole cell capture technique which combines Fluorescence In Situ Hybridization with immuno-magnetic cell capture (Magneto-FISH). We used Magneto-FISH for targeted enrichment of specific ANME groups and their associated bacteria directly from formalin- and ethanol-fixed methane seep sediment. The identity and metabolic gene diversity of captured microorganisms were then assessed by clone library construction and sequencing. Diversity recovered from Magneto-FISH experiments using general and clade-specific ANME targeted probes show both the expected selectivity of the FISH probes (i.e. predominately ANME-2c subclade captured with an ANME-2c probe and multiple ANME groups recovered with the general probe targeting most ANME). Follow up FISH experiments were conducted to confirm physical associations between ANME and unique bacterial members (deltaproteobacteria and other non-sulfate reducing groups) that were common to multiple Magneto-FISH capture experiments. Analyses of metabolic gene diversity for archaeal

Evidence for involvement of gut-associated denitrifying bacteria in emission of nitrous oxide (N(2)O) by earthworms obtained from garden and forest soils.

Science.gov (United States)

Matthies, C; Griesshammer, A; Schmittroth, M; Drake, H L

1999-08-01

-associated denitrifying bacteria are responsible for the in vivo emission of N(2)O by earthworms and contribute to the N(2)O that is emitted from certain terrestrial ecosystems.

Methylocapsa palsarum sp. nov., a methanotroph isolated from a subArctic discontinuous permafrost ecosystem.

Science.gov (United States)

Dedysh, Svetlana N; Didriksen, Alena; Danilova, Olga V; Belova, Svetlana E; Liebner, Susanne; Svenning, Mette M

2015-10-01

An aerobic methanotrophic bacterium was isolated from a collapsed palsa soil in northern Norway and designated strain NE2T. Cells of this strain were Gram-stain-negative, non-motile, non-pigmented, slightly curved thick rods that multiplied by normal cell division. The cells possessed a particulate methane monooxygenase enzyme (pMMO) and utilized methane and methanol. Strain NE2T grew in a wide pH range of 4.1–8.0 (optimum pH 5.2–6.5) at temperatures between 6 and 32 °C (optimum 18–25 °C), and was capable of atmospheric nitrogen fixation under reduced oxygen tension. The major cellular fatty acids were C18 : 1ω7c, C16 : 0 and C16 : 1ω7c, and the DNA G+C content was 61.7 mol%. The isolate belonged to the family Beijerinckiaceae of the class Alphaproteobacteria and was most closely related to the facultative methanotroph Methylocapsa aurea KYGT (98.3 % 16S rRNA gene sequence similarity and 84 % PmoA sequence identity). However, strain NE2T differed from Methylocapsa aurea KYGT by cell morphology, the absence of pigmentation, inability to grow on acetate, broader pH growth range, and higher tolerance to NaCl. Therefore, strain NE2T represents a novel species of the genus Methylocapsa, for which we propose the name Methylocapsa palsarum sp. nov. The type strain is NE2T ( = LMG 28715T = VKM B-2945T).

Environmental distribution and abundance of the facultative methanotroph Methylocella.

Science.gov (United States)

Rahman, Md Tanvir; Crombie, Andrew; Chen, Yin; Stralis-Pavese, Nancy; Bodrossy, Levente; Meir, Patrick; McNamara, Niall P; Murrell, J Colin

2011-06-01

Methylocella spp. are facultative methanotrophs, which are able to grow not only on methane but also on multicarbon substrates such as acetate, pyruvate or malate. Methylocella spp. were previously thought to be restricted to acidic soils such as peatlands, in which they may have a key role in methane oxidation. There is little information on the abundance and distribution of Methylocella spp. in the environment. New primers were designed, and a real-time quantitative PCR method was developed and validated targeting Methylocella mmoX (encoding the α-subunit of the soluble methane monooxygenase) that allowed the quantification of Methylocella spp. in environmental samples. We also developed and validated specific PCR assays, which target 16S rRNA genes of known Methylocella spp. These were used to investigate the distribution of Methylocella spp. in a variety of environmental samples. It was revealed that Methylocella species are widely distributed in nature and not restricted to acidic environments.

Short exposure to acetylene to distinguish between nitrifier and denitrifier nitrous oxide production in soil and sediment samples

NARCIS (Netherlands)

Kester, R.A.; Boer, W. de; Laanbroek, H.J.

1996-01-01

The contribution of nitrifiers and denitrifiers to the nitrous oxide production in slurries of calcareous silt loam and river bank sediment at different oxygen concentrations was determined using acetylene as nitrification inhibitor. The addition of 10 Pa acetylene resulted in inhibition of

Short exposure to acetylene to distinguish between nitrifier and denitrifier nitrous oxide production in soil and sediment samples

NARCIS (Netherlands)

Kester, R.A.; De Boer, W.; Laanbroek, H.J.

1996-01-01

The contribution of nitrifiers and denitrifiers to the nitrous oxide production in slurries of calcareous silt loam and river bank sediment at different oxygen concentrations was determined using acetylene as nitrification inhibitor. The addition of 10 Pa acetylene resulted in inhibition of nitrous

Diversity and activity of methanotrophic related bacteria in subsurface sediments of the Krishna-Godavari Basin, India

Digital Repository Service at National Institute of Oceanography (India)

Sujith, P.P.; Sheba, M.; Gonsalves, M.J.B.D.

system (Bio-Rad, USA). DNA sequencing and phylogenetic analysis: The ampli- fied PCR products were purified prior to sequencing according to the manufacturer’s guidelines, using the GenEluteTM PCR Cleanup Kit (Sigma, USA). The puri- fied PCR... at the interface between the reduced zones of the environment6 in the marine sediments7, where methane production and oxidation may take place. These bacteria can be obligate or facultative methano- trophs8. Based on their physiology and phylogeny...

Communities of nirS-type denitrifiers in the water column of the oxygen minimum zone in the eastern South Pacific.

Science.gov (United States)

Castro-González, Maribeb; Braker, Gesche; Farías, Laura; Ulloa, Osvaldo

2005-09-01

The major sites of water column denitrification in the ocean are oxygen minimum zones (OMZ), such as one in the eastern South Pacific (ESP). To understand the structure of denitrifying communities in the OMZ off Chile, denitrifier communities at two sites in the Chilean OMZ (Antofagasta and Iquique) and at different water depths were explored by terminal restriction fragment length polymorphism analysis and cloning of polymerase chain reaction (PCR)-amplified nirS genes. NirS is a functional marker gene for denitrification encoding cytochrome cd1-containing nitrite reductase, which catalyses the reduction of nitrite to nitric oxide, the key step in denitrification. Major differences were found between communities from the two geographic locations. Shifts in community structure occurred along a biogeochemical gradient at Antofagasta. Canonical correspondence analysis indicated that O2, NO3-, NO2- and depth were important environmental factors governing these communities along the biogeochemical gradient in the water column. Phylogenetic analysis grouped the majority of clones from the ESP in distinct clusters of genes from presumably novel and yet uncultivated denitrifers. These nirS clusters were distantly related to those found in the water column of the Arabian Sea but the phylogenetic distance was even higher compared with environmental sequences from marine sediments or any other habitat. This finding suggests similar environmental conditions trigger the development of denitrifiers with related nirS genotypes despite large geographic distances.

Methane distribution and oxidation around the Lena Delta in summer 2013

Directory of Open Access Journals (Sweden)

I. Bussmann

2017-11-01

Full Text Available The Lena River is one of the largest Russian rivers draining into the Laptev Sea. The predicted increases in global temperatures are expected to cause the permafrost areas surrounding the Lena Delta to melt at increasing rates. This melting will result in high amounts of methane reaching the waters of the Lena and the adjacent Laptev Sea. The only biological sink that can lower methane concentrations within this system is methane oxidation by methanotrophic bacteria. However, the polar estuary of the Lena River, due to its strong fluctuations in salinity and temperature, is a challenging environment for bacteria. We determined the activity and abundance of aerobic methanotrophic bacteria by a tracer method and by the quantitative polymerase chain reaction. We described the methanotrophic population with a molecular fingerprinting method (monooxygenase intergenic spacer analysis, as well as the methane distribution (via a headspace method and other abiotic parameters, in the Lena Delta in September 2013. The median methane concentrations were 22 nmol L−1 for riverine water (salinity (S  < 5, 19 nmol L−1 for mixed water (5 < S < 20 and 28 nmol L−1 for polar water (S > 20. The Lena River was not the source of methane in surface water, and the methane concentrations of the bottom water were mainly influenced by the methane concentration in surface sediments. However, the bacterial populations of the riverine and polar waters showed similar methane oxidation rates (0.419 and 0.400 nmol L−1 d−1, despite a higher relative abundance of methanotrophs and a higher estimated diversity in the riverine water than in the polar water. The methane turnover times ranged from 167 days in mixed water and 91 days in riverine water to only 36 days in polar water. The environmental parameters influencing the methane oxidation rate and the methanotrophic population also differed between the water masses. We

[Formation of microbial populations on the surface of protective coatings].

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Kopteva, Zh P; Zanina, V V; Piliashenko-Novokhatnyĭ, A I; Kopteva, A E; Kozlova, I A

2001-01-01

Formation of microbial cenosis on the surface of polyethylene-, polyurethane- and oil-bitumen-based protective coatings was studied in dynamics during 1, 3, 7, 14 and 21 days. It has been shown that the biofilm was formed on the protective materials during 14 days and consisted of ammonifying, denitrifying, hydrocarbon-oxidizing and sulphate-reducing bacteria referred to Pseudomonas, Arthrobacter, Bacillus and Kesulfovibrio genera. The bacteria which form the biofilm on coatings possess high denitrifying and sulphate-reducing activities. Corrosion inhibitors-biocydes, introduced in composition of oil-bitumen coatings suppressed growth and metabolic activity of corrosion-active bacteria.

Sources for sedimentary bacteriohopanepolyols as revealed by 16S rDNA stratigraphy.

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Coolen, Marco J L; Talbot, Helen M; Abbas, Ben A; Ward, Christopher; Schouten, Stefan; Volkman, John K; Damsté, Jaap S Sinninghe

2008-07-01

Bacteriohopanoids are widespread lipid biomarkers in the sedimentary record. Many aerobic and anaerobic bacteria are potential sources of these lipids which sometimes complicates the use of these biomarkers as proxies for ecological and environmental changes. Therefore, we applied preserved 16S ribosomal RNA genes to identify likely Holocene biological sources of bacteriohopanepolyols (BHPs) in the sulfidic sediments of the permanently stratified postglacial Ace Lake, Antarctica. A suite of intact BHPs were identified, which revealed a variety of structural forms whose composition differed through the sediment core reflecting changes in bacterial populations induced by large changes in lake salinity. Stable isotopic compositions of the hopanols formed from periodic acid-cleaved BHPs, showed that some were substantially depleted in (13)C, indicative of their methanotrophic origin. Using sensitive molecular tools, we found that Type I and II methanotrophic bacteria (respectively Methylomonas and Methylocystis) were unique to the oldest lacustrine sediments (> 9400 years BP), but quantification of fossil DNA revealed that the Type I methanotrophs, including methanotrophs related to methanotrophic gill symbionts of deep-sea cold-seep mussels, were the main precursors of the 35-amino BHPs (i.e. aminopentol, -tetrol and -triols). After isolation of the lake approximately 3000 years ago, one Type I methanotroph of the 'methanotrophic gill symbionts cluster' remained the most obvious source of aminotetrol and -triol. We, furthermore, identified a Synechococcus phylotype related to pelagic freshwater strains in the oldest lacustrine sediments as a putative source of 2-methylbacteriohopanetetrol (2-Me BHT). This combined application of advanced geochemical and paleogenomical tools further refined our knowledge about Holocene biogeochemical processes in Ace Lake.

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

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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 (CH 4 ) 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 CH 4 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% CH 4 under a very high gas flow rate (2.5 L min -1 ; 20-25% CH 4 ) in continuous-stirred tank reactors (∼10 L working volume) and the biomass contained long-chain fatty acids (i.e. C 16 and C 18 ). Cultivation of the MMC on plastic bio-balls abated ∼95-97% CH 4 in pilot-scale non-sterile outdoor-operated bio-filters (0.1 L min -1 ; 1% CH 4 ). Contamination by cyanobacteria had beneficial effects on treating low-level CH 4 , 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 CH 4 and CO 2 at industrial scale.

A doubling of microphytobenthos biomass coincides with a tenfold increase in denitrifier and total bacterial abundances in intertidal sediments of a temperate estuary.

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Helen Decleyre

Full Text Available Surface sediments are important systems for the removal of anthropogenically derived inorganic nitrogen in estuaries. They are often characterized by the presence of a microphytobenthos (MPB biofilm, which can impact bacterial communities in underlying sediments for example by secretion of extracellular polymeric substances (EPS and competition for nutrients (including nitrogen. Pyrosequencing and qPCR was performed on two intertidal surface sediments of the Westerschelde estuary characterized by a two-fold difference in MPB biomass but no difference in MPB composition. Doubling of MPB biomass was accompanied by a disproportionately (ten-fold increase in total bacterial abundances while, unexpectedly, no difference in general community structure was observed, despite significantly lower bacterial richness and distinct community membership, mostly for non-abundant taxa. Denitrifier abundances corresponded likewise while community structure, both for nirS and nirK denitrifiers, remained unchanged, suggesting that competition with diatoms for nitrate is negligible at concentrations in the investigated sediments (appr. 1 mg/l NO3-. This study indicates that MPB biomass increase has a general, significantly positive effect on total bacterial and denitrifier abundances, with stimulation or inhibition of specific bacterial groups that however do not result in a re-structured community.

Draft genome sequence of Bacillus azotoformans MEV2011, a (Co-) denitrifying strain unable to grow with oxygen.

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Nielsen, Maja; Schreiber, Lars; Finster, Kai; Schramm, Andreas

2015-01-01

Bacillus azotoformans MEV2011, isolated from soil, is a microaerotolerant obligate denitrifier, which can also produce N2 by co-denitrification. Oxygen is consumed but not growth-supportive. The draft genome has a size of 4.7 Mb and contains key genes for both denitrification and dissimilatory nitrate reduction to ammonium.

Performance of denitrifying microbial fuel cell subjected to variation in pH, COD concentration and external resistance.

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Li, Jin-Tao; Zhang, Shao-Hui; Hua, Yu-Mei

2013-01-01

The effects of pH, chemical oxygen demand (COD) concentration and external resistance on denitrifying microbial fuel cell were evaluated in terms of electricity generation characteristics and pollutant removal performance. The results showed that anodic influent with weakly alkaline or neutral pH and cathodic influent with weakly acidic pH favored pollutant removal and electricity generation. The suitable influent pH of the anode and cathode were found to be 7.5-8.0 and 6.0-6.5, respectively. In the presence of sufficient nitrate in the cathode, higher influent COD concentration led to more electricity generation and greater pollutant removal rates. With an anodic influent pH of 8.0 and a cathodic influent pH of 6.0, an influent COD concentration of 400 mg/L was deemed to be appropriate. Low external resistance favored nitrate and COD removal. The results suggest that operation of denitrifying microbial fuel cell at a lower external resistance would be desirable for pollutant removal but not electricity generation.

Degradation of organic pollutants by methane grown microbial consortia.

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Hesselsoe, Martin; Boysen, Susanne; Iversen, Niels; Jørgensen, Lars; Murrell, J Colin; McDonald, Ian; Radajewski, Stefan; Thestrup, Helle; Roslev, Peter

2005-10-01

Microbial consortia were enriched from various environmental samples with methane as the sole carbon and energy source. Selected consortia that showed a capacity for co-oxidation of naphthalene were screened for their ability to degrade methyl-tert-butyl-ether (MTBE), phthalic acid esters (PAE), benzene, xylene and toluene (BTX). MTBE was not removed within 24 h by any of the consortia examined. One consortium enriched from activated sludge ("AAE-A2"), degraded PAE, including (butyl-benzyl)phthalate (BBP), and di-(butyl)phthalate (DBP). PAE have not previously been described as substrates for methanotrophic consortia. The apparent Km and Vmax for DBP degradation by AAE-A2 at 20 degrees C was 3.1 +/- 1.2 mg l(-1) and 8.7 +/- 1.1 mg DBP (g protein x h)(-1), respectively. AAE-A2 also showed fast degradation of BTX (230 +/- 30 nmol benzene (mg protein x h)(-1) at 20 degrees C). Additionally, AAE-A2 degraded benzene continuously for 2 weeks. In contrast, a pure culture of the methanotroph Methylosinus trichosporium OB3b ceased benzene degradation after only 2 days. Experiments with methane mono-oxygenase inhibitors or competitive substrates suggested that BTX degradation was carried out by methane-oxidizing bacteria in the consortium, whereas the degradation of PAE was carried out by non-methanotrophic bacteria co-existing with methanotrophs. The composition of the consortium (AAE-A2) based on polar lipid fatty acid (PLFA) profiles showed dominance of type II methanotrophs (83-92% of biomass). Phylogeny based on a 16S-rRNA gene clone library revealed that the dominating methanotrophs belonged to Methylosinus/Methylocystis spp. and that members of at least 4 different non-methanotrophic genera were present (Pseudomonas, Flavobacterium, Janthinobacterium and Rubivivax).

Isotopologue signatures of nitrous oxide produced by nitrate-ammonifying bacteria isolated from soil

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Behrendt, Undine; Well, Reinhard; Giesemann, Anette; Ulrich, Andreas; Augustin, Jürgen

2015-04-01

Agricultural soils are the largest single source of anthropogenic N2O to the atmosphere, primarily driven by microbiological processes such as denitrification and dissimilatory nitrate reduction to ammonium (DNRA). Both processes occur under similar conditions of low oxygen concentration and therefore, source partitioning of emitted N2O is difficult. Understanding what controls the dynamics and reaction equilibrium of denitrification and DNRA is important and may allow the development of more effective mitigation strategies. 15N site preference (SP), i.e. the difference between 15N of the central and peripheral N-position of the asymmetric N2O molecule, differs depending on processes involved in N2O formation. Hence investigation of the isotopomer ratios of formed N2O potentially presents a reliable mean to identify its source. In this study, bacterial isolates obtained from organic soils were screened for their ability to reduce nitrate/nitrite to ammonium and to release N2O to the atmosphere. Taxonomic characterisation of the strains revealed that N2O formation was only detected in ammonifying strains affiliated to several genera of the family Enterobacteriaceae and strains belonging to the genus Bacillus and Paenibacillus. Sampling of N2O was conducted by incubation of strains under oxic and anoxic conditions. Investigation of the 15N site preference showed SP values in the range of 39 to 57 o . Incubation conditions had no influence on the SP. The lowest values were achieved by a strain of the species Escherichia coli which was included in this study as a DNRA reference bacterium harbouring the NrfA gene that is coding the nitrite reductase, associated with respiratory nitrite ammonification. Soil isolates showed SP-values higher than 40 o . Comparison of these results with SP-values of N2O produced by denitrifying bacteria in pure cultures (-5 to 0 o )^[1, 2]revealedsignificantdifferences.Incontrast,N_2OproducedbydenitrifyingfungidisplayedSP - valuesinarangeof

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

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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

Anaerobic Oxidization of Methane in a Minerotrophic Peatland: Enrichment of Nitrite-Dependent Methane-Oxidizing Bacteria

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Zhu, Baoli; van Dijk, Gijs; Fritz, Christian; Smolders, Alfons J. P.; Pol, Arjan; Jetten, Mike S. M.

2012-01-01

The importance of anaerobic oxidation of methane (AOM) as a methane sink in freshwater systems is largely unexplored, particularly in peat ecosystems. Nitrite-dependent anaerobic methane oxidation (n-damo) was recently discovered and reported to be catalyzed by the bacterium “Candidatus Methylomirabilis oxyfera,” which is affiliated with the NC10 phylum. So far, several “Ca. Methylomirabilis oxyfera” enrichment cultures have been obtained using a limited number of freshwater sediments or wastewater treatment sludge as the inoculum. In this study, using stable isotope measurements and porewater profiles, we investigated the potential of n-damo in a minerotrophic peatland in the south of the Netherlands that is infiltrated by nitrate-rich ground water. Methane and nitrate profiles suggested that all methane produced was oxidized before reaching the oxic layer, and NC10 bacteria could be active in the transition zone where countergradients of methane and nitrate occur. Quantitative PCR showed high NC10 bacterial cell numbers at this methane-nitrate transition zone. This soil section was used to enrich the prevalent NC10 bacteria in a continuous culture supplied with methane and nitrite at an in situ pH of 6.2. An enrichment of nitrite-reducing methanotrophic NC10 bacteria was successfully obtained. Phylogenetic analysis of retrieved 16S rRNA and pmoA genes showed that the enriched bacteria were very similar to the ones found in situ and constituted a new branch of NC10 bacteria with an identity of less than 96 and 90% to the 16S rRNA and pmoA genes of “Ca. Methylomirabilis oxyfera,” respectively. The results of this study expand our knowledge of the diversity and distribution of NC10 bacteria in the environment and highlight their potential contribution to nitrogen and methane cycles. PMID:23042166

Populasi dan aktivitas denitrifikasi serta emisi gas N2 O pada lahan pertanian organik, pertanian intensif, dan hutan

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Dwi Agustiyani

2012-10-01

Full Text Available This research investigate the population and potentials denitrification activity from three different soils, organically farmed soil, intensive farmed soil and forest soil. Our objectives were to explore spatial gradients in denitrifier populations, examine whether populations density and its potential activity was related to soil chemical properties (C and N content, and determine the potential emission of gas N2O. Results indicated biological functional differences between these three different soil ecosystems. Forest soil had the highest population density of denitrifying bacteria and also had significant potential denitrifying activities. The highest potentials denitrifying activity in the soil affected to the lowest emission of N2O gas. The lowest population and potential denitrifying activity was measured in the intensive farmed soil. Those conditions might be promoted the potentials emission of N2O.

Abundance, composition and activity of ammonia oxidizer and denitrifier communities in metal polluted rice paddies from South China.

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Yuan Liu

Full Text Available While microbial nitrogen transformations in soils had been known to be affected by heavy metal pollution, changes in abundance and community structure of the mediating microbial populations had been not yet well characterized in polluted rice soils. Here, by using the prevailing molecular fingerprinting and enzyme activity assays and comparisons to adjacent non-polluted soils, we examined changes in the abundance and activity of ammonia oxidizing and denitrifying communities of rice paddies in two sites with different metal accumulation situation under long-term pollution from metal mining and smelter activities. Potential nitrifying activity was significantly reduced in polluted paddies in both sites while potential denitrifying activity reduced only in the soils with high Cu accumulation up to 1300 mg kg-1. Copy numbers of amoA (AOA and AOB genes were lower in both polluted paddies, following the trend with the enzyme assays, whereas that of nirK was not significantly affected. Analysis of the DGGE profiles revealed a shift in the community structure of AOA, and to a lesser extent, differences in the community structure of AOB and denitrifier between soils from the two sites with different pollution intensity and metal composition. All of the retrieved AOB sequences belonged to the genus Nitrosospira, among which species Cluster 4 appeared more sensitive to metal pollution. In contrast, nirK genes were widely distributed among different bacterial genera that were represented differentially between the polluted and unpolluted paddies. This could suggest either a possible non-specific target of the primers conventionally used in soil study or complex interactions between soil properties and metal contents on the observed community and activity changes, and thus on the N transformation in the polluted rice soils.

Biotechnological conversion of methane to methanol: evaluation of progress and potential

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Charlotte E. Bjorck

2018-01-01

Full Text Available Sources of methane are numerous, and vary greatly in their use and sustainable credentials. A Jekyll and Hyde character, it is a valuable energy source present as geological deposits of natural gas, however it is also potent greenhouse gas, released during many waste management processes. Gas-to-liquid technologies are being investigated as a means to exploit and monetise non-traditional and unutilised methane sources. The product identified as having the greatest potential is methanol due to it being a robust, commercially mature conversion process from methane and its beneficial fuel characteristics. Commercial methane to methanol conversion requires high temperatures and pressures, in an energy intensive and costly process. In contrast methanotrophic bacteria perform the desired transformation under ambient conditions, using methane monooxygenase (MMO enzymes. Despite the great potential of these bacteria a number of biotechnical difficulties are hindering progress towards an industrially suitable process. We have identified five major challenges that exist as barriers to a viable conversion process that, to our knowledge, have not previously been examined as distinct process challenges. Although biotechnological applications of methanotrophic bacteria have been reviewed in part, no review has comprehensively covered progress and challenges for a methane to methanol process from an industrial perspective. All published examples to date of methanotroph catalysed conversion of methane to methanol are collated, and standardised to allow direct comparison. The focus will be on conversion of methane to methanol by whole-cell, wild type, methanotroph cultures, and the potential for their application in an industrially relevant process. A recent shift in the research community focus from a mainly biological angle to an overall engineering approach, offers potential to exploit methanotrophs in an industrially relevant biotechnological gas

Use of oligodeoxynucleotide signature probes for identification of physiological groups of methylotrophic bacteria

International Nuclear Information System (INIS)

Tsien, H.C.; Bratina, B.J.; Tsuji, K.; Hanson, R.S.

1990-01-01

Oligodeoxynucleotide sequences that uniquely complemented 16S rRNAs of each group of methylotrophs were synthesized and used as hybridization probes for the identification of methylotrophic bacteria possessing the serine and ribulose monophosphate (RuMP) pathways for formaldehyde fixation. The specificity of the probes was determined by hybridizing radiolabeled probes with slot-blotted RNAs of methylotrophs and other eubacteria followed by autoradiography. The washing temperature was determined experimentally to be 50 and 52 degrees C for 9-α (serine pathway) and 10-γ (RuMP pathway) probes, respectively. RNAs isolated from serine pathway methylotrophs bound to probe 9-α, and RNAs from RuMP pathway methylotrophs bound to probe 10-γ. Nonmethylotrophic eubacterial RNAs did not bind to either probe. The probes were also labeled with fluorescent dyes. Cells fixed to microscope slides were hybridized with these probes, washed, and examined in a fluorescence microscope equipped with appropriate filter sets. Cells of methylotrophic bacteria possessing the serine or RuMP pathway specifically bind probes designed for each group. Samples with a mixture of cells of type I and II methanotrophs were detected and differentiated with single probes or mixed probes labeled with different fluorescent dyes, which enabled the detection of both types of cells in the same microscopic field

Analyzing the Effects of Nitrogen Deficiency on the PHB Production of Methylosinus trichosporium OB3b

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Kyauk, E.

2011-12-01

Polyhydroxybutyrate (PHB) is a biodegradable thermoplastic that is produced by various microorganisms. Because of its potential to replace conventional plastics, it has been closely researched in the past few years. Methanotrophic bacteria, bacteria that consume methane, produce this bioplastic when it lacks certain nutrients. The utilization of methane to produce PHB shows much promise as methane is a cheap, plentiful gas. In this study, we observed the methanotroph, Methylosinus trichosporium OB3b , and its yield of PHB in the absence of nitrogen. The optical density of Methylosinus trichosporium OB3b was measured in order to observe cell growth and PHB production patterns over a 48 hour period.

Anaerobic methanotrophic communities thrive in deep submarine permafrost.

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Winkel, Matthias; Mitzscherling, Julia; Overduin, Pier P; Horn, Fabian; Winterfeld, Maria; Rijkers, Ruud; Grigoriev, Mikhail N; Knoblauch, Christian; Mangelsdorf, Kai; Wagner, Dirk; Liebner, Susanne

2018-01-22

Thawing submarine permafrost is a source of methane to the subsurface biosphere. Methane oxidation in submarine permafrost sediments has been proposed, but the responsible microorganisms remain uncharacterized. We analyzed archaeal communities and identified distinct anaerobic methanotrophic assemblages of marine and terrestrial origin (ANME-2a/b, ANME-2d) both in frozen and completely thawed submarine permafrost sediments. Besides archaea potentially involved in anaerobic oxidation of methane (AOM) we found a large diversity of archaea mainly belonging to Bathyarchaeota, Thaumarchaeota, and Euryarchaeota. Methane concentrations and δ 13 C-methane signatures distinguish horizons of potential AOM coupled either to sulfate reduction in a sulfate-methane transition zone (SMTZ) or to the reduction of other electron acceptors, such as iron, manganese or nitrate. Analysis of functional marker genes (mcrA) and fluorescence in situ hybridization (FISH) corroborate potential activity of AOM communities in submarine permafrost sediments at low temperatures. Modeled potential AOM consumes 72-100% of submarine permafrost methane and up to 1.2 Tg of carbon per year for the total expected area of submarine permafrost. This is comparable with AOM habitats such as cold seeps. We thus propose that AOM is active where submarine permafrost thaws, which should be included in global methane budgets.

[Comparative characterization of cultured methane-oxidizing bacteria by serological and molecular methods].

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Slobodova, N V; Kolganova, T V; Bulygina, E S; Kuznetsov, B B; Turova, T P; Kravchenko, I K

2006-01-01

Three stable methane-oxidizing enrichment cultures, SB26, SB31, and SB31A were analyzed by transmission electron microscopy and by serological and molecular techniques. Electron microscopy revealed the presence of both type I and type II methanotrophs in SB31 and SB31A enrichments; only type II methanotrophs were found in SB26 enrichment. Methylosinus trichosporium was detected in all three enrichments by the application of species-specific antibodies. Additionally, Methylocystis echinoides was found in SB26 culture; Methylococcus capsulatus, in SB31 and SB31A; and Methylomonas methanica, in SB31. The analysis with pmoA and nifH gene sequences as phylogenetic markers revealed the presence of Methylosinus/Methylocystis group in all communities. Moreover, the analysis of pmoA sequences revealed the presence of Methylomonas in SB31. Methylocella was detected in SB31 and SB31A enrichments only by nifH analysis. It was concluded that the simultaneous application of different approaches reveals more reliable information on the diversity of methanotrophs.

Molecular diversity of the methanotrophic bacteria communities associated with disused tin-mining ponds in Kampar, Perak, Malaysia.

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Sow, S L S; Khoo, G; Chong, L K; Smith, T J; Harrison, P L; Ong, H K A

2014-10-01

In a previous study, notable differences of several physicochemical properties, as well as the community structure of ammonia oxidizing bacteria as judged by 16S rRNA gene analysis, were observed among several disused tin-mining ponds located in the town of Kampar, Malaysia. These variations were associated with the presence of aquatic vegetation as well as past secondary activities that occurred at the ponds. Here, methane oxidizing bacteria (MOB), which are direct participants in the nutrient cycles of aquatic environments and biological indicators of environmental variations, have been characterised via analysis of pmoA functional genes in the same environments. The MOB communities associated with disused tin-mining ponds that were exposed to varying secondary activities were examined in comparison to those in ponds that were left to nature. Comparing the sequence and phylogenetic analysis of the pmoA clone libraries at the different ponds (idle, lotus-cultivated and post-aquaculture), we found pmoA genes indicating the presence of type I and type II MOB at all study sites, but type Ib sequences affiliated with the Methylococcus/Methylocaldum lineage were most ubiquitous (46.7 % of clones). Based on rarefaction analysis and diversity indices, the disused mining pond with lotus culture was observed to harbor the highest richness of MOB. However, varying secondary activity or sample type did not show a strong variation in community patterns as compared to the ammonia oxidizers in our previous study.

Anaerobic/oxic/anoxic granular sludge process as an effective nutrient removal process utilizing denitrifying polyphosphate-accumulating organisms.

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Kishida, Naohiro; Kim, Juhyun; Tsuneda, Satoshi; Sudo, Ryuichi

2006-07-01

In a biological nutrient removal (BNR) process, the utilization of denitrifying polyphosphate-accumulating organisms (DNPAOs) has many advantages such as effective use of organic carbon substrates and low sludge production. As a suitable process for the utilization of DNPAOs in BNR, an anaerobic/oxic/anoxic granular sludge (AOAGS) process was proposed in this study. In spite of performing aeration for nitrifying bacteria, the AOAGS process can create anaerobic/anoxic conditions suitable for the cultivation of DNPAOs because anoxic zones exist inside the granular sludge in the oxic phase. Thus, DNPAOs can coexist with nitrifying bacteria in a single reactor. In addition, the usability of DNPAOs in the reactor can be improved by adding the anoxic phase after the oxic phase. These characteristics enable the AOAGS process to attain effective removal of both nitrogen and phosphorus. When acetate-based synthetic wastewater (COD: 600 mg/L, NH4-N: 60 mg/L, PO(4)-P: 10 mg/L) was supplied to a laboratory-scale sequencing batch reactor under the operation of anaerobic/oxic/anoxic cycles, granular sludge with a diameter of 500 microm was successfully formed within 1 month. Although the removal of both nitrogen and phosphorus was almost complete at the end of the oxic phase, a short anoxic period subsequent to the oxic phase was necessary for further removal of nitrogen and phosphorus. As a result, effluent concentrations of NH(4)-N, NO(x)-N and PO(4)-P were always lower than 1 mg/L. It was found that penetration depth of oxygen inside the granular sludge was approximately 100 microm by microsensor measurements. In addition, from the microbiological analysis by fluorescence in situ hybridization, existence depth of polyphosphate-accumulating organisms was further than the maximum oxygen penetration depth. The water quality data, oxygen profiles and microbial community structure demonstrated that DNPAOs inside the granular sludge may be responsible for denitrification in the

Structure of nitrogen-converting communities induced by hydraulic retention time and COD/N ratio in constantly aerated granular sludge reactors treating digester supernatant.

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Cydzik-Kwiatkowska, Agnieszka; Rusanowska, Paulina; Zielińska, Magdalena; Bernat, Katarzyna; Wojnowska-Baryła, Irena

2014-02-01

This study investigated how hydraulic retention time (HRT) and COD/N ratio affect nitrogen-converting consortia in constantly aerated granules treating high-ammonium digester supernatant. Three HRTs (10, 13, 19 h) were tested at COD/N ratios of 4.5 and 2.3. Denaturing gradient gel electrophoresis and relative real-time PCR were used to characterize the microbial communities. When changes in HRT and COD/N increased nitrogen loading, the ratio of the relative abundance of aerobic to anaerobic ammonium-oxidizers decreased. The COD/N ratio determined the species composition of the denitrifiers; however, Thiobacillus denitrificans, Pseudomonas denitrificans and Azoarcus sp. showed a high tolerance to the environmental conditions and occurred in the granules from all reactors. Denitrifier genera that support granule formation were identified, such as Pseudomonas, Shinella, and Flavobacterium. In aerated granules, nirK-possessing bacteria were more diverse than nirS-possessing bacteria. At a low COD/N ratio, N2O-reducer diversity increased because of the presence of bacteria known as aerobic denitrifiers. Copyright © 2013 Elsevier Ltd. All rights reserved.

Removal of pharmaceutical and personal care products (PPCPs) under nitrifying and denitrifying conditions.

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Suarez, Sonia; Lema, Juan M; Omil, Francisco

2010-05-01

The contribution of volatilization, sorption and transformation to the removal of 16 Pharmaceutical and Personal Care Products (PPCPs) in two lab-scale conventional activated sludge reactors, working under nitrifying (aerobic) and denitrifying (anoxic) conditions for more than 1.5 years, have been assessed. Pseudo-first order biological degradation rate constants (k(biol)) were calculated for the selected compounds in both reactors. Faster degradation kinetics were measured in the nitrifying reactor compared to the denitrifying system for the majority of PPCPs. Compounds could be classified according to their k(biol) into very highly (k(biol)>5Lg(SS)(-1)d(-1)), highly (1fragrances (HHCB, AHTN and ADBI) were transformed to a large extent under aerobic (>75%) and anoxic (>65%) conditions, whereas naproxen (NPX), ethinylestradiol (EE2), roxithromycin (ROX) and erythromycin (ERY) were only significantly transformed in the aerobic reactor (>80%). The anti-depressant citalopram (CTL) was moderately biotransformed under both, aerobic and anoxic conditions (>60% and >40%, respectively). Some compounds, as carbamazepine (CBZ), diazepam (DZP), sulfamethoxazole (SMX) and trimethoprim (TMP), manifested high resistance to biological transformation. Solids Retention Time (SRT(aerobic) >50d and 20d and <20d) had a slightly positive effect on the removal of FLX, NPX, CTL, EE2 and natural estrogens (increase in removal efficiencies <10%). Removal of diclofenac (DCF) in the aerobic reactor was positively affected by the development of nitrifying biomass and increased from 0% up to 74%. Similarly, efficient anoxic transformation of ibuprofen (75%) was observed after an adaptation period of 340d. Temperature (16-26 degrees C) only had a slight effect on the removal of CTL which increased in 4%.

Versatile transformations of hydrocarbons in anaerobic bacteria: substrate ranges and regio- and stereo-chemistry of activation reactions†

Science.gov (United States)

Jarling, René; Kühner, Simon; Basílio Janke, Eline; Gruner, Andrea; Drozdowska, Marta; Golding, Bernard T.; Rabus, Ralf; Wilkes, Heinz

2015-01-01

Anaerobic metabolism of hydrocarbons proceeds either via addition to fumarate or by hydroxylation in various microorganisms, e.g., sulfate-reducing or denitrifying bacteria, which are specialized in utilizing n-alkanes or alkylbenzenes as growth substrates. General pathways for carbon assimilation and energy gain have been elucidated for a limited number of possible substrates. In this work the metabolic activity of 11 bacterial strains during anaerobic growth with crude oil was investigated and compared with the metabolite patterns appearing during anaerobic growth with more than 40 different hydrocarbons supplied as binary mixtures. We show that the range of co-metabolically formed alkyl- and arylalkyl-succinates is much broader in n-alkane than in alkylbenzene utilizers. The structures and stereochemistry of these products are resolved. Furthermore, we demonstrate that anaerobic hydroxylation of alkylbenzenes does not only occur in denitrifiers but also in sulfate reducers. We propose that these processes play a role in detoxification under conditions of solvent stress. The thermophilic sulfate-reducing strain TD3 is shown to produce n-alkylsuccinates, which are suggested not to derive from terminal activation of n-alkanes, but rather to represent intermediates of a metabolic pathway short-cutting fumarate regeneration by reverse action of succinate synthase. The outcomes of this study provide a basis for geochemically tracing such processes in natural habitats and contribute to an improved understanding of microbial activity in hydrocarbon-rich anoxic environments. PMID:26441848

Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

NARCIS (Netherlands)

Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

1998-01-01

The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative. denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing bacteria,

Nitrate removal from water using denitrifier-bacteria immobilized on activated carbon at fluidized-bed reactor

Directory of Open Access Journals (Sweden)

hatam Godini

2012-09-01

Results: The experimental results demonstrated that the over 94% of NO3-N was removed in the 2-3 h retention time, almost no NO2-N accumulated in treated water when the concentration of NO3-N was around 50-100 mg/L in influent. Under the experimental conditions, drinking water quality was achieved at a nitrogen loading lower than 2.4 kgNO3-N•m-3•day-1 with an influent COD/N ratio of 3 if ethanol was used as the carbon source. The denitrification rate increased up to a maximal value of 3.45 kgNO3-N•m-3•day-1 with increasing nitrogen loading rate (4.8 kgNO3-N•m-3•day-1. Conclusion: This study demonstrates that GAC offers favorable attachment sites for bacteria on the surfaces of GAC for Nitrate removal and denitrification of water was carried out in a fluidized bed reactor (FBR using Activated carbon-immobilized cell beads as support carriers.

Development of hydrogen oxidizing bacteria using hydrogen from radiolysis or metal corrosion

International Nuclear Information System (INIS)

Libert, M.F.; Sellier, R.; Marty, V.; Camaro, S.

2000-01-01

The effect of many parameters need to be studied to characterize the long term behavior of nuclear waste in a deep repository. These parameters concern the chemical effects, radiolytic effects, mechanical properties, water composition, and microbiological activity. To evaluate microbial activity in such an environment, work was focused on an inventory of key nutrients (C, H, 0, N, P, S) and energy sources required for bacterial growth. The production of hydrogen in the nuclear waste environment leads to the growth of hydrogen oxidizing bacteria, which modify the gas production balance. A deep repository containing bituminized waste drums implies several sources of hydrogen: - water radiolysis; -corrosion of metal containers; - radiolysis of the embedding matrix (bitumen). Two deep geological disposal conditions leading to H 2 production in a bituminized nuclear waste environment were simulated in the present study: - H 2 production by iron corrosion under anaerobic conditions was simulated by adding 10% of H 2 in the atmosphere; - H 2 production by radiolysis of bitumen matrix was approached by subjecting this material to external gamma irradiation with a dose rate near real conditions (6 Gy/h). The presence of dissolved H 2 in water allows the growth of hydrogen oxidizing bacteria leading to: - CO 2 and N 2 production; - H 2 consumption; - lower NO 3 - concentration caused by reduction to nitrogen. In the first case, hydrogen consumption is limited by the NO 3 - release rate from the bitumen matrix. In the second case, however, under gamma radiation at a low dose rate, hydrogen production is weak, and the hydrogen is completely consumed by microorganisms. Knowledge about these hydrogen oxidizing bacteria is just beginning to emerge. Heterotrophic denitrifying bacteria adapt well to hydrogen metabolism (autotrophic metabolism) by oxidizing H 2 instead of hydrocarbons. (authors)

Activity and diversity of methane-oxidizing bacteria along a Norwegian sub-Arctic glacier forefield.

Science.gov (United States)

Mateos-Rivera, Alejandro; Øvreås, Lise; Wilson, Bryan; Yde, Jacob C; Finster, Kai W

2018-05-01

Methane (CH4) is one of the most abundant greenhouse gases in the atmosphere and identification of its sources and sinks is crucial for the reliability of climate model outputs. Although CH4 production and consumption rates have been reported from a broad spectrum of environments, data obtained from glacier forefields are restricted to a few locations. We report the activities of methanotrophic communities and their diversity along a chronosequence in front of a sub-Arctic glacier using high-throughput sequencing and gas flux measurements. CH4 oxidation rates were measured in the field throughout the growing season during three sampling times at eight different sampling points in combination with laboratory incubation experiments. The overall results showed that the methanotrophic community had similar trends of increased CH4 consumption and increased abundance as a function of soil development and time of year. Sequencing results revealed that the methanotrophic community was dominated by a few OTUs and that a short-term increase in CH4 concentration, as performed in the field measurements, altered slightly the relative abundance of the OTUs.

Effects of long-term elevated CO2 on N2-fixing, denitrifying and nitrifying enzyme activities in forest soils under Pinus sylvestriformis in Changbai Mountain

Institute of Scientific and Technical Information of China (English)

ZHENG Jun-Qiang; HAN Shi-Jie; REN Fei-Rong; ZHOU Yu-Mei; ZHANG Yan

2008-01-01

A study was conducted to determine the effects of elevated CO2 on soil N process at Changbai Mountain in Jilin Province,northeastern China (42o24'N,128o06'E,and 738 m elevation).A randomized complete block design of ambient and elevated CO2 was established in an open-top chamber facility in the spring of 1999.Changpai Scotch pine (Pinus sylvestris var.sylvestriformis seeds were sowed in May,1999 and CO2 fumigation treatments began after seeds germination.In each year,the exposure started at the end of April and stopped at the end of October.Soil samples were collected in June and August 2006 and in June 2007,and soil nitrifying,denitrifying and N2-fixing enzyme activities were measured.Results show that soil nitrifying enzyme activities (NEA) in the 5-10 cm soil layer were significantly increased at elevated CO2 by 30.3% in June 2006,by 30.9% in August 2006 and by 11.3% in June 2007.Soil denitrifying enzyme activities (DEA) were significantly decreased by elevated CO2 treatment in June 2006 (P < 0.012) and August 2006 (P < 0.005) samplings in our study; no significant difference was detected in June 2007,and no significant changes in N2-fixing enzyme activity were found.This study suggests that elevated CO2 can alter soil nitrifying enzyme and denitrifying enzyme activities.

Anaerobic methane oxidation coupled to denitrification is the dominant methane sink in a deep lake

Science.gov (United States)

Deutzmann, Joerg S.; Stief, Peter; Brandes, Josephin; Schink, Bernhard

2014-01-01

Anaerobic methane oxidation coupled to denitrification, also known as “nitrate/nitrite-dependent anaerobic methane oxidation” (n-damo), was discovered in 2006. Since then, only a few studies have identified this process and the associated microorganisms in natural environments. In aquatic sediments, the close proximity of oxygen- and nitrate-consumption zones can mask n-damo as aerobic methane oxidation. We therefore investigated the vertical distribution and the abundance of denitrifying methanotrophs related to Candidatus Methylomirabilis oxyfera with cultivation-independent molecular techniques in the sediments of Lake Constance. Additionally, the vertical distribution of methane oxidation and nitrate consumption zones was inferred from high-resolution microsensor profiles in undisturbed sediment cores. M. oxyfera-like bacteria were virtually absent at shallow-water sites (littoral sediment) and were very abundant at deep-water sites (profundal sediment). In profundal sediment, the vertical distribution of M. oxyfera-like bacteria showed a distinct peak in anoxic layers that coincided with the zone of methane oxidation and nitrate consumption, a strong indication for n-damo carried out by M. oxyfera-like bacteria. Both potential n-damo rates calculated from cell densities (660–4,890 µmol CH4⋅m−2⋅d−1) and actual rates calculated from microsensor profiles (31–437 µmol CH4⋅m−2⋅d−1) were sufficiently high to prevent methane release from profundal sediment solely by this process. Additionally, when nitrate was added to sediment cores exposed to anoxic conditions, the n-damo zone reestablished well below the sediment surface, completely preventing methane release from the sediment. We conclude that the previously overlooked n-damo process can be the major methane sink in stable freshwater environments if nitrate is available in anoxic zones. PMID:25472842

Anaerobic methane oxidation coupled to denitrification is the dominant methane sink in a deep lake.

Science.gov (United States)

Deutzmann, Joerg S; Stief, Peter; Brandes, Josephin; Schink, Bernhard

2014-12-23

Anaerobic methane oxidation coupled to denitrification, also known as "nitrate/nitrite-dependent anaerobic methane oxidation" (n-damo), was discovered in 2006. Since then, only a few studies have identified this process and the associated microorganisms in natural environments. In aquatic sediments, the close proximity of oxygen- and nitrate-consumption zones can mask n-damo as aerobic methane oxidation. We therefore investigated the vertical distribution and the abundance of denitrifying methanotrophs related to Candidatus Methylomirabilis oxyfera with cultivation-independent molecular techniques in the sediments of Lake Constance. Additionally, the vertical distribution of methane oxidation and nitrate consumption zones was inferred from high-resolution microsensor profiles in undisturbed sediment cores. M. oxyfera-like bacteria were virtually absent at shallow-water sites (littoral sediment) and were very abundant at deep-water sites (profundal sediment). In profundal sediment, the vertical distribution of M. oxyfera-like bacteria showed a distinct peak in anoxic layers that coincided with the zone of methane oxidation and nitrate consumption, a strong indication for n-damo carried out by M. oxyfera-like bacteria. Both potential n-damo rates calculated from cell densities (660-4,890 µmol CH4⋅m(-2)⋅d(-1)) and actual rates calculated from microsensor profiles (31-437 µmol CH4⋅m(-2)⋅d(-1)) were sufficiently high to prevent methane release from profundal sediment solely by this process. Additionally, when nitrate was added to sediment cores exposed to anoxic conditions, the n-damo zone reestablished well below the sediment surface, completely preventing methane release from the sediment. We conclude that the previously overlooked n-damo process can be the major methane sink in stable freshwater environments if nitrate is available in anoxic zones.

Performance of denitrifying microbial fuel cell with biocathode over nitrite

Directory of Open Access Journals (Sweden)

Zhao eHuimin

2016-03-01

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

Microbial reduction of 99Tc in organic matter-rich soils

International Nuclear Information System (INIS)

Abdelouas, A.; Grambow, B.; Fattahi, M.; Andres, Y.; Leclerc-Cessac, E.

2005-01-01

For safety assessment purposes, it is necessary to study the mobility of long-lived radionuclides in the geosphere and the biosphere. Within this framework, we studied the behaviour of 99 Tc in biologically active organic matter-rich soils. To simulate the redox conditions in soils, we stimulated the growth of aerobic and facultative denitrifying and anaerobic sulphate-reducing bacteria (SRB). In the presence of either a pure culture of denitrifiers (Pseudomonas aeruginosa) or a consortium of soil denitrifiers, the solubility of TcO 4 - was not affected. The nonsorption of TcO 4 - onto bacteria was confirmed in biosorption experiments with washed cells of P. aeruginosa regardless of the pH. At the end of denitrification with indigenous denitrifiers in soil/water batch experiments, the redox potential (E H ) dropped and this was accompanied by an increase of Fe concentration in solution as a result of reduction of less soluble Fe(III) to Fe(II) from the soil particles. It is suggested that this is due to the growth of a consortium of anaerobic bacteria (e.g., Fe-reducing bacteria). The drop in E H was accompanied by a strong decrease in Tc concentration as a result of Tc(VII) reduction to Tc(IV). Thermodynamic calculations suggested the precipitation of TcO 2 . The stimulation of the growth of indigenous sulphate-reducing bacteria in soil/water systems led to even lower E H with final Tc concentration of 10 -8 M. Experiments with glass columns filled with soil reproduced the results obtained with batch cultures. Sequential chemical extraction of precipitated Tc in soils showed that this radionuclide is strongly immobilised within soil particles under anaerobic conditions. More than 90% of Tc is released together with organic matter (60-66%) and Fe-oxyhydroxides (23-31%). The present work shows that ubiquitous indigenous anaerobic bacteria in soils play a major role in Tc immobilisation. In addition, organic matter plays a key role in the stability of the reduced Tc

Nitrate Removal from Wastewater through Biological Denitrification with OGA 24 in a Batch Reactor

Directory of Open Access Journals (Sweden)

Federico Rossi

2014-12-01

Full Text Available Nitrates pollution of waters is a worldwide problem and its remediation is a big challenge from the technical and the scientific point of view. One of the most used and promising cleaning techniques is the biological treatment of wastewaters operated by denitrifying bacteria. In this paper we begin a thorough study of denitrifying performances of the bacterium Azospira sp. OGA 24, recently isolated from the highly polluted Sarno river in the south of Italy. Here, the kinetics of nitrates consumption operated by bacteria in a specifically devised batch bioreactor, in anoxic condition and with acetate as the organic substrate, has been characterized. Experimental data were then used in a simplified model of a real wastewater treatment plant to find that OGA 24 can clean water with efficiency up to 90%. The denitrifying performances of OGA 24 match the requirements of Italian laws and make the bacterium suitable for its employment in treatment plants.

Optimizing BTEX biodegradation under denitrifying conditions

International Nuclear Information System (INIS)

Hutchins, S.R.

1991-01-01

Leaking underground storage tanks are a major source of ground water contamination by petroleum hydrocarbons. Gasoline and other fuels contain benzene, toluene, ethylbenzene, and xylenes (collectively known as BTEX), which are hazardous compounds, regulated by the U.S. Environmental Protection Agency (EPA). Laboratory tests were conducted to determine optimum conditions for benzene, toluene, ethylbenzene, and xylene (collectively known as BTEX) biodegradation by aquifer microorganisms under denitrifying conditions. Microcosms, constructed with aquifer samples from Traverse City, Michigan, were amended with selected concentrations of nutrients and one or more hydrocarbons. Toluene, ethylbenzene, m-xylene, and p-xylene, were degraded to below 5 micrograms/L when present as sole source substrates; stoichiometric calculations indicated that nitrate removal was sufficient to account for 70 to 80% of the compounds being mineralized. o-Xylene was recalcitrant when present as a sole source substrate, but was slowly degraded in the presence of the other hydrocarbons. Benzene was not degraded within one year, regardless of whether it was available as a sole source substrate or in combination with toluene, phenol, or catechol. Pre-exposure to low levels of BTEX and nutrients had variable effects, as did the addition of different concentrations of ammonia and phosphate. Nitrate concentrations as high as 500 mg/L NO3-N were slightly inhibitory. These data indicate that nitrate-mediated biodegradation of BTEX at Traverse City can occur under a variety of environmental conditions with rates relatively independent of nutrient concentrations. However, the data reaffirm that benzene is recalcitrant under strictly anaerobic conditions in these samples

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

Directory of Open Access Journals (Sweden)

Z. Kobierzyńska-Gołąb

2015-06-01

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

Short exposure to acetylene to distinguish between nitrifier and denitrifier nitrous oxide production in soil and sediment samples

OpenAIRE

Kester, R.A.; Boer, W. de; Laanbroek, H.J.

1996-01-01

The contribution of nitrifiers and denitrifiers to the nitrous oxide production in slurries of calcareous silt loam and river bank sediment at different oxygen concentrations was determined using acetylene as nitrification inhibitor. The addition of 10 Pa acetylene resulted in inhibition of nitrous oxide production at oxic conditions, but strongly enhanced the nitrous oxide production at oxygen-poor and anoxic conditions. Inhibition of nitrification by short exposure (1 to 24 h) to high conce...

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

OpenAIRE

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

2014-01-01

Heterotrophic nitrifiers synthesize nitrogenous gasses when nitrifying ammonium ion. A Cupriavidus pauculus, previously thought an Alcaligenes sp. and noted as an active heterotrophic nitrifier-denitrifier, was examined for its ability to produce nitrogen gas (N2) and nitrous oxide (N2O) while heterotrophically nitrifying the organic substrate pyruvic oxime [CH3–C(NOH)–COOH]. Neither N2 nor N2O were produced. Nucleotide and phylogenetic analyses indicated that the organism is a member of a g...

Conformational control of the binding of diatomic gases to cytochrome c'.

Science.gov (United States)

Manole, Andreea; Kekilli, Demet; Svistunenko, Dimitri A; Wilson, Michael T; Dobbin, Paul S; Hough, Michael A

2015-06-01

The cytochromes c' (CYTcp) are found in denitrifying, methanotrophic and photosynthetic bacteria. These proteins are able to form stable adducts with CO and NO but not with O2. The binding of NO to CYTcp currently provides the best structural model for the NO activation mechanism of soluble guanylate cyclase. Ligand binding in CYTcps has been shown to be highly dependent on residues in both the proximal and distal heme pockets. Group 1 CYTcps typically have a phenylalanine residue positioned close to the distal face of heme, while for group 2, this residue is typically leucine. We have structurally, spectroscopically and kinetically characterised the CYTcp from Shewanella frigidimarina (SFCP), a protein that has a distal phenylalanine residue and a lysine in the proximal pocket in place of the more common arginine. Each monomer of the SFCP dimer folds as a 4-alpha-helical bundle in a similar manner to CYTcps previously characterised. SFCP exhibits biphasic binding kinetics for both NO and CO as a result of the high level of steric hindrance from the aromatic side chain of residue Phe 16. The binding of distal ligands is thus controlled by the conformation of the phenylalanine ring. Only a proximal 5-coordinate NO adduct, confirmed by structural data, is observed with no detectable hexacoordinate distal NO adduct.

Nitrite survival and nitrous oxide production of denitrifying phosphorus removal sludges in long-term nitrite/nitrate-fed sequencing batch reactors.

Science.gov (United States)

Wang, Yayi; Zhou, Shuai; Ye, Liu; Wang, Hong; Stephenson, Tom; Jiang, Xuxin

2014-12-15

Nitrite-based phosphorus (P) removal could be useful for innovative biological P removal systems where energy and carbon savings are a priority. However, using nitrite for denitrification may cause nitrous oxide (N2O) accumulation and emissions. A denitrifying nitrite-fed P removal system [Formula: see text] was successfully set up in a sequencing batch reactor (SBR) and was run for 210 days. The maximum pulse addition of nitrite to [Formula: see text] was 11 mg NO2(-)-N/L in the bulk, and a total of 34 mg NO2(-)-N/L of nitrite was added over three additions. Fluorescent in situ hybridization results indicated that the P-accumulating organisms (PAOs) abundance was 75 ± 1.1% in [Formula: see text] , approximately 13.6% higher than that in a parallel P removal SBR using nitrate [Formula: see text] . Type II Accumulibacter (PAOII) (unable to use nitrate as an electron acceptor) was the main PAOs species in [Formula: see text] , contributing 72% to total PAOs. Compared with [Formula: see text] , [Formula: see text] biomass had enhanced nitrite/free nitrous acid (FNA) endurance, as demonstrated by its higher nitrite denitrification and P uptake rates. N2O accumulated temporarily in [Formula: see text] after each pulse of nitrite. Peak N2O concentrations in the bulk for [Formula: see text] were generally 6-11 times higher than that in [Formula: see text] ; these accumulations were rapidly denitrified to nitrogen gases. N2O concentration increased rapidly in nitrate-cultivated biomass when 5 or 10 mg NO2(-)-N/L per pulse was added. Whereas, N2O accumulation did not occur in nitrite-cultivated biomass until up to 30 mg NO2(-)-N/L per pulse was added. Long-term acclimation to nitrite and pulse addition of nitrite in [Formula: see text] reduced the risk of nitrite accumulation, and mitigated N2O accumulation and emissions from denitrifying P removal by nitrite. Copyright © 2014 Elsevier Ltd. All rights reserved.

Bacterial community involved in the nitrogen cycle in a down-flow sponge-based trickling filter treating UASB effluent.

Science.gov (United States)

Mac Conell, E F A; Almeida, P G S; Martins, K E L; Araújo, J C; Chernicharo, C A L

2015-01-01

The bacterial community composition of a down-flow sponge-based trickling filter treating upflow anaerobic sludge blanket (UASB) effluent was investigated by pyrosequencing. Bacterial community composition considerably changed along the reactor and over the operational period. The dominant phyla detected were Proteobacteria, Verrucomicrobia, and Planctomycetes. The abundance of denitrifiers decreased from the top to the bottom and it was consistent with the organic matter concentration gradients. At lower loadings (organic and nitrogen loading rates), the abundance of anammox bacteria was higher than that of the ammonium-oxidizing bacteria in the upper portion of the reactor, suggesting that aerobic and anaerobic ammonium oxidation occurred. Nitrification occurred in all the compartments, while anammox bacteria prominently appeared even in the presence of high organic carbon to ammonia ratios (around 1.0-2.0 gCOD gN(-1)). The results suggest that denitrifiers, nitrifiers, and anammox bacteria coexisted in the reactor; thus, different metabolic pathways were involved in ammonium removal in the post-UASB reactor sponge-based.

Interactions between ammonia and nitrite oxidizing bacteria in co-cultures: Is there evidence for mutualism, commensalism, or competition?

Energy Technology Data Exchange (ETDEWEB)

Sayavedra-Soto, Luis [Oregon State Univ., Corvallis, OR (United States); Arp, Daniel [Oregon State Univ., Corvallis, OR (United States)

2017-08-01

Nitrification is a two-step environmental microbial process in the nitrogen cycle in which ammonia is oxidized to nitrate. Ammonia-oxidizing bacteria and archaea oxidize ammonia to nitrite and nitrite is oxidized to nitrate by nitrite-oxidizing bacteria. These microorganisms, which likely act in concert in a microbial community, play critical roles in the movement of inorganic N in soils, sediments and waters and are essential to the balance of the nitrogen cycle. Anthropogenic activity has altered the balance of the nitrogen cycle through agriculture practices and organic waste byproducts. Through their influence on available N for plant growth, nitrifying microorganisms influence plant productivity for food and fiber production and the associated carbon sequestration. N Fertilizer production, primarily as ammonia, requires large inputs of natural gas and hydrogen. In croplands fertilized with ammonia-based fertilizers, nitrifiers contribute to the mobilization of this N by producing nitrate (NO3-), wasting the energy used in the production and application of ammonia-based fertilizer. The resulting nitrate is readily leached from these soils, oxidized to gaseous N oxides (greenhouse gases), and denitrified to N2 (which is no longer available as a plant N source). Still, ammonia oxidizers are beneficial in the treatment of wastewater and they also show potential to contribute to microbial bioremediation strategies for clean up of environments contaminated with chlorinated hydrocarbons. Mitigation of the negative effects and exploitation of the beneficial effects of nitrifiers will be facilitated by a systems-level understanding of the interactions of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria with the environment and with each other.

Microbial diversity in Frenulata (Siboglinidae, Polychaeta) species from mud volcanoes in the Gulf of Cadiz (NE Atlantic).

Science.gov (United States)

Rodrigues, Clara F; Hilário, Ana; Cunha, Marina R; Weightman, Andrew J; Webster, Gordon

2011-06-01

Frenulates are a group of gutless marine annelids belonging to the Siboglinidae that are nutritionally dependent upon endosymbiotic bacteria. We have characterized the bacteria associated with several frenulate species from mud volcanoes in the Gulf of Cadiz by PCR-DGGE of bacterial 16S rRNA genes, coupled with analysis of 16S rRNA gene libraries. In addition to the primary symbiont, bacterial consortia (microflora) were found in all species analysed. Phylogenetic analyses indicate that the primary symbiont in most cases belongs to the Gammaproteobacteria and were related to thiotrophic and methanotrophic symbionts from other marine invertebrates, whereas members of the microflora were related to multiple bacterial phyla. This is the first molecular evidence of methanotrophic bacteria in at least one frenulate species. In addition, the occurrence of the same bacterial phylotype in different Frenulata species, from different depths and mud volcanoes suggests that there is no selection for specific symbionts and corroborates environmental acquisition as previously proposed for this group of siboglinids.

Complete Nutrient Removal Coupled to Nitrous Oxide Production as a Bioenergy Source by Denitrifying Polyphosphate-Accumulating Organisms.

Science.gov (United States)

Gao, Han; Liu, Miaomiao; Griffin, James S; Xu, Longcheng; Xiang, Da; Scherson, Yaniv D; Liu, Wen-Tso; Wells, George F

2017-04-18

Coupled aerobic-anoxic nitrous decomposition operation (CANDO) is a promising emerging bioprocess for wastewater treatment that enables direct energy recovery from nitrogen (N) in three steps: (1) ammonium oxidation to nitrite; (2) denitrification of nitrite to nitrous oxide (N 2 O); and (3) N 2 O conversion to N 2 with energy generation. However, CANDO does not currently target phosphorus (P) removal. Here, we demonstrate that denitrifying polyphosphate-accumulating organism (PAO) enrichment cultures are capable of catalyzing simultaneous biological N and P removal coupled to N 2 O generation in a second generation CANDO process, CANDO+P. Over 7 months (>300 cycles) of operation of a prototype lab-scale CANDO+P sequencing batch reactor treating synthetic municipal wastewater, we observed stable and near-complete N removal accompanied by sustained high-rate, high-yield N 2 O production with partial P removal. A substantial increase in abundance of the PAO Candidatus Accumulibacter phosphatis was observed, increasing from 5% of the total bacterial community in the inoculum to over 50% after 4 months. PAO enrichment was accompanied by a strong shift in the dominant Accumulibacter population from clade IIC to clade IA, based on qPCR monitoring of polyphosphate kinase 1 (ppk1) gene variants. Our work demonstrates the feasibility of combining high-rate, high-yield N 2 O production for bioenergy production with combined N and P removal from wastewater, and it further suggests a putative denitrifying PAO niche for Accumulibacter clade IA.

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

Directory of Open Access Journals (Sweden)

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

Complex community of nitrite-dependent anaerobic methane oxidation bacteria in coastal sediments of the Mai Po wetland by PCR amplification of both 16S rRNA and pmoA genes.

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Chen, Jing; Zhou, Zhichao; Gu, Ji-Dong

2015-02-01

In the present work, both 16S rRNA and pmoA gene-based PCR primers were employed successfully to study the diversity and distribution of n-damo bacteria in the surface and lower layer sediments at the coastal Mai Po wetland. The occurrence of n-damo bacteria in both the surface and subsurface sediments with high diversity was confirmed in this study. Unlike the two other known n-damo communities from coastal areas, the pmoA gene-amplified sequences in the present work clustered not only with some freshwater subclusters but also within three newly erected marine subclusters mostly, indicating the unique niche specificity of n-damo bacteria in this wetland. Results suggested vegetation affected the distribution and community structures of n-damo bacteria in the sediments and n-damo could coexist with sulfate-reducing methanotrophs in the coastal ecosystem. Community structures of the Mai Po n-damo bacteria based on 16S rRNA gene were different from those of either the freshwater or the marine. In contrast, structures of the Mai Po n-damo communities based on pmoA gene grouped with the marine ones and were clearly distinguished from the freshwater ones. The abundance of n-damo bacteria at this wetland was quantified using 16S rRNA gene PCR primers to be 2.65-6.71 × 10(5) copies/g dry sediment. Ammonium and nitrite strongly affected the community structures and distribution of n-damo bacteria in the coastal Mai Po wetland sediments.

Pseudomonads Isolated from Pristine Background Groundwater Proliferate More Effectively in Co-culture than in Monoculture Under Denitrifying Conditions

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Aaring, A. B.; Lancaster, A.; Novichkov, P.; Adams, M. W. W.; Deutschbauer, A. M.; Chakraborty, R.

2016-12-01

As part of the Ecosystems and Networks Integrated with Genes and Molecular Assemblies (ENIGMA) consortium, we study the microbial community at the U.S. Department of Energy's Field Research Center (FRC) in Oak Ridge. The groundwater at this site contains plumes of nitrate with concentrations up to 14,000mg/L among other contaminants, though molybdenum concentrations are low. Because molybdenum is essential to nitrate reduction, this can be inhibitory to growth. Several strains of Pseudomonas were isolated from the same background groundwater sample. These isolates utilized diverse carbon sources ranging from acetate to glucose while growing under denitrifying conditions. The strains were also screened for nitrate tolerance and a couple of them were shown to be tolerant to 300-400 mM nitrate under anaerobic conditions. In the field site the bacteria live in consortia rather than in isolation, therefore we hypothesized that growth of these strains will be more robust in co-culture, as the denitrification pathway was segmented between the species. Three of the isolates (Pseudomonas fluorescens strains N1B4, N2E2, N2E3) were selected for in-depth analysis based on growth in pairwise co-cultures relative to monocultures, and the availability of the relevant genetic tools, such as transposon mutant libraries. Full genome sequencing showed that strain N2E3 has a truncated dentrification pathway: it lacks nitrous oxide reductase. Our results show strain N2E2 grow to maximum cell density an average of 45 hours more quickly when grown with strain N2E3 than in monoculture. Utilizing RB-TnSeq libraries of our strains, it was also found that some genes involved in nitrate reduction, sulfate permeability, molybdenum utilization, and anaerobic reduction are important for growth under these conditions. In addition, a few unexpected genes were also shown to be positively correlated to growth, such as genes homologous to genes for DNA proofreading or antibiotic production. These

Heterotrophs are key contributors to nitrous oxide production in mixed liquor under low C-to-N ratios during nitrification - batch experiments and modelling

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Domingo Felez, Carlos; Pellicer i Nàcher, Carles; Petersen, Morten S.

2017-01-01

Nitrous oxide (N2O), a by-product of biological nitrogen removal during wastewater treatment, is produced by ammonia-oxidizing bacteria (AOB) and heterotrophic denitrifying bacteria (HB). Mathematical models are used to predict N2O emissions, often including AOB as the main N2O producer. Several...

Using natural biomass microorganisms for drinking water denitrification.

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Costa, Darleila Damasceno; Gomes, Anderson Albino; Fernandes, Mylena; Lopes da Costa Bortoluzzi, Roseli; Magalhães, Maria de Lourdes Borba; Skoronski, Everton

2018-07-01

Among the methods that are studied to eliminate nitrate from drinking water, biological denitrification is an attractive strategy. Although several studies report the use of denitrifying bacteria for nitrate removal, they usually involve the use of sewage sludge as biomass to obtain the microbiota. In the present study, denitrifying bacteria was isolated from bamboo, and variable parameters were controlled focusing on optimal bacterial performance followed by physicochemical analysis of water adequacy. In this way, bamboo was used as a source of denitrifying microorganisms, using either Immobilized Microorganisms (IM) or Suspended Microorganisms (SM) for nitrate removal. Denitrification parameters optimization was carried out by analysis of denitrification at different pH values, temperature, nitrate concentrations, carbon sources as well as different C/N ratios. In addition, operational stability and denitrification kinetics were evaluated. Microorganisms present in the biomass responsible for denitrification were identified as Proteus mirabilis. The denitrified water was submitted to physicochemical treatment such as coagulation and flocculation to adjust to the parameters of color and turbidity to drinking water standards. Denitrification using IM occurred with 73% efficiency in the absence of an external carbon source. The use of SM provided superior denitrification efficiency using ethanol (96.46%), glucose (98.58%) or glycerol (98.5%) as carbon source. The evaluation of the operational stability allowed 12 cycles of biomass reuse using the IM and 9 cycles using the SM. After physical-chemical treatment, only SM denitrified water remained within drinking water standards parameters of color and turbidity. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effects of Bubble-Mediated Processes on Nitrous Oxide Dynamics in Denitrifying Bioreactors

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McGuire, P. M.; Falk, L. M.; Reid, M. C.

2017-12-01

To mitigate groundwater and surface water impacts of reactive nitrogen (N), agricultural and stormwater management practices can employ denitrifying bioreactors (DNBs) as low-cost solutions for enhancing N removal. Due to the variable nature of hydrologic events, DNBs experience dynamic flows which can impact physical and biological processes within the reactors and affect performance. A particular concern is incomplete denitrification, which can release the potent greenhouse gas nitrous oxide (N2O) to the atmosphere. This study aims to provide insight into the effects of varying hydrologic conditions upon the operation of DNBs by disentangling abiotic and biotic controls on denitrification and N2O dynamics within a laboratory-scale bioreactor. We hypothesize that under transient hydrologic flows, rising water levels lead to air entrapment and bubble formation within the DNB porous media. Mass transfer of oxygen (O2) between trapped gas and liquid phases creates aerobic microenvironments that can inhibit N2O reductase (NosZ) enzymes and lead to N2O accumulation. These bubbles also retard N2O transport and make N2O unavailable for biological reduction, further enhancing atmospheric fluxes when water levels fall. The laboratory-scale DNB permits measurements of longitudinal and vertical profiles of dissolved constituents as well as trace gas concentrations in the reactor headspace. We describe a set of experiments quantifying denitrification pathway biokinetics under steady-state and transient hydrologic conditions and evaluate the role of bubble-mediated processes in enhancing N2O accumulation and fluxes. We use sulfur hexafluoride and helium as dissolved gas tracers to examine the impact of bubble entrapment upon retarded gas transport and enhanced trace gas fluxes. A planar optode sensor within the bioreactor provides near-continuous 2-D profiles of dissolved O2 within the bioreactor and allows for identification of aerobic microenvironments. We use qPCR to

Bacterial endosymbioses of gutless tube-dwelling worms in nonhydrothermal vent habitats.

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Naganuma, Takeshi; Elsaied, Hosam E; Hoshii, Daiki; Kimura, Hiroyuki

2005-01-01

Gutless tube-dwelling worms of pogonophorans (also known as frenulates) and vestimentiferans depend on primary production of endosymbiotic bacteria. The endosymbionts include thiotrophs that oxidize sulfur for autotrophic production and methanotrophs that oxidize and assimilate methane. Although most of the pogonophoran and vestimentiferan tube worms possess single thiotrophic 16S rRNA genes (16S rDNA) related to gamma-proteobacteria, some pogonohorans are known to bear single methanotroph species or even dual symbionts of thiotrophs and methanotrophs. The vestimentiferan Lamellibrachia sp. L1 shows symbiotic 16S rDNA sequences of alpha-, beta-, gamma-, and epsilon-proteobacteria, varying among specimens, with RuBisCO form II gene (cbbM) sequences related to beta-proteobacteria. An unidentified pogonophoran from the world's deepest cold seep, 7326-m deep in the Japan Trench, hosts a symbiotic thiotroph based on 16S rDNA with the RuBisCO form I gene (cbbL). In contrast, a shallow-water pogonophoran (Oligobrachia mashikoi) in coastal Japan Sea has a methanotrophic 16S rDNA and thiotrophic cbbL, which may suggest the feature of type X methanotrophs. These observations demonstrate that pogonophoran and vestimentiferan worms have higher plasticity in bacterial symbioses than previously suspected.

Nitrate Removal Rates in Denitrifying Bioreactors During Storm Flows

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Pluer, W.; Walter, T.

2017-12-01

Field denitrifying bioreactors are designed to reduce excess nitrate (NO3-) pollution in runoff from agricultural fields. Field bioreactors saturate organic matter to create conditions that facilitate microbial denitrification. Prior studies using steady flow in lab-scale bioreactors showed that a hydraulic retention time (HRT) between 4 and 10 hours was optimal for reducing NO3- loads. However, during storm-induced events, flow rate and actual HRT fluctuate. These fluctuations have the potential to disrupt the system in significant ways that are not captured by the idealized steady-flow HRT models. The goal of this study was to investigate removal rate during dynamic storm flows of variable rates and durations. Our results indicate that storm peak flow and duration were not significant controlling variables. Instead, we found high correlations (p=0.004) in average removal rates between bioreactors displaying a predominantly uniform flow pattern compared with bioreactors that exhibited preferential flow (24.4 and 21.4 g N m-3 d-1, respectively). This suggests that the internal flow patterns are a more significant driver of removal rate than external factors of the storm hydrograph. Designing for flow patterns in addition to theoretical HRT will facilitate complete mixing within the bioreactors. This will help maximize excess NO3- removal during large storm-induced runoff events.

Potential Role of Nitrite for Abiotic Fe(II) Oxidation and Cell Encrustation during Nitrate Reduction by Denitrifying Bacteria

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Klueglein, Nicole; Zeitvogel, Fabian; Stierhof, York-Dieter; Floetenmeyer, Matthias; Konhauser, Kurt O.; Obst, Martin

2014-01-01

Microorganisms have been observed to oxidize Fe(II) at neutral pH under anoxic and microoxic conditions. While most of the mixotrophic nitrate-reducing Fe(II)-oxidizing bacteria become encrusted with Fe(III)-rich minerals, photoautotrophic and microaerophilic Fe(II) oxidizers avoid cell encrustation. The Fe(II) oxidation mechanisms and the reasons for encrustation remain largely unresolved. Here we used cultivation-based methods and electron microscopy to compare two previously described nitrate-reducing Fe(II) oxidizers ( Acidovorax sp. strain BoFeN1 and Pseudogulbenkiania sp. strain 2002) and two heterotrophic nitrate reducers (Paracoccus denitrificans ATCC 19367 and P. denitrificans Pd 1222). All four strains oxidized ∼8 mM Fe(II) within 5 days in the presence of 5 mM acetate and accumulated nitrite (maximum concentrations of 0.8 to 1.0 mM) in the culture media. Iron(III) minerals, mainly goethite, formed and precipitated extracellularly in close proximity to the cell surface. Interestingly, mineral formation was also observed within the periplasm and cytoplasm; intracellular mineralization is expected to be physiologically disadvantageous, yet acetate consumption continued to be observed even at an advanced stage of Fe(II) oxidation. Extracellular polymeric substances (EPS) were detected by lectin staining with fluorescence microscopy, particularly in the presence of Fe(II), suggesting that EPS production is a response to Fe(II) toxicity or a strategy to decrease encrustation. Based on the data presented here, we propose a nitrite-driven, indirect mechanism of cell encrustation whereby nitrite forms during heterotrophic denitrification and abiotically oxidizes Fe(II). This work adds to the known assemblage of Fe(II)-oxidizing bacteria in nature and complicates our ability to delineate microbial Fe(II) oxidation in ancient microbes preserved as fossils in the geological record. PMID:24271182

Methylocystis bryophila sp. nov., a facultatively methanotrophic bacterium from acidic Sphagnum peat, and emended description of the genus Methylocystis (ex Whittenbury et al. 1970) Bowman et al. 1993.

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Belova, Svetlana E; Kulichevskaya, Irina S; Bodelier, Paul L E; Dedysh, Svetlana N

2013-03-01

A novel species is proposed for two facultatively methanotrophic representatives of the genus Methylocystis, strains H2s(T) and S284, which were isolated from an acidic (pH 4.3) Sphagnum peat-bog lake (Teufelssee, Germany) and an acidic (pH 3.8) peat bog (European North Russia), respectively. Cells of strains H2s(T) and S284 are aerobic, Gram-negative, non-motile, curved coccoids or short rods that contain an intracytoplasmic membrane system typical of type-II methanotrophs. They possess both a soluble and a particulate methane monooxygenase (MMO); the latter is represented by two isozymes, pMMO1 and pMMO2. The preferred growth substrates are methane and methanol. In the absence of C1 substrates, however, these methanotrophs are capable of slow growth on acetate. Atmospheric nitrogen is fixed by means of an aerotolerant nitrogenase. Strains H2s(T) and S284 grow between pH 4.2 and 7.6 (optimum pH 6.0-6.5) and at 8-37 °C (optimum 25-30 °C). The major fatty acids are C18 : 1ω8c, C18 : 1ω7c and C16 : 1ω7c; the major quinone is Q-8. The DNA G+C content is 62.0-62.3 mol%. Strains H2s(T) and S284 share identical 16S rRNA gene sequences, which displayed 96.6-97.3 % similarity to sequences of other taxonomically characterized members of the genus Methylocystis. Therefore, strains H2s(T) and S284 are classified as members of a novel species, for which the name Methylocystis bryophila sp. nov. is proposed; strain H2s(T) ( = DSM 21852(T)  = VKM B-2545(T)) is the type strain.

Community Composition of Nitrous Oxide Consuming Bacteria in the Oxygen Minimum Zone of the Eastern Tropical South Pacific

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Xin Sun

2017-06-01

Full Text Available The ozone-depleting and greenhouse gas, nitrous oxide (N2O, is mainly consumed by the microbially mediated anaerobic process, denitrification. N2O consumption is the last step in canonical denitrification, and is also the least O2 tolerant step. Community composition of total and active N2O consuming bacteria was analyzed based on total (DNA and transcriptionally active (RNA nitrous oxide reductase (nosZ genes using a functional gene microarray. The total and active nosZ communities were dominated by a limited number of nosZ archetypes, affiliated with bacteria from marine, soil and marsh environments. In addition to nosZ genes related to those of known marine denitrifiers, atypical nosZ genes, related to those of soil bacteria that do not possess a complete denitrification pathway, were also detected, especially in surface waters. The community composition of the total nosZ assemblage was significantly different from the active assemblage. The community composition of the total nosZ assemblage was significantly different between coastal and off-shore stations. The low oxygen assemblages from both stations were similar to each other, while the higher oxygen assemblages were more variable. Community composition of the active nosZ assemblage was also significantly different between stations, and varied with N2O concentration but not O2. Notably, nosZ assemblages were not only present but also active in oxygenated seawater: the abundance of total and active nosZ bacteria from oxygenated surface water (indicated by nosZ gene copy number was similar to or even larger than in anoxic waters, implying the potential for N2O consumption even in the oxygenated surface water.

Community Composition of Nitrous Oxide Consuming Bacteria in the Oxygen Minimum Zone of the Eastern Tropical South Pacific

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Sun, Xin; Jayakumar, Amal; Ward, Bess B.

2017-01-01

The ozone-depleting and greenhouse gas, nitrous oxide (N2O), is mainly consumed by the microbially mediated anaerobic process, denitrification. N2O consumption is the last step in canonical denitrification, and is also the least O2 tolerant step. Community composition of total and active N2O consuming bacteria was analyzed based on total (DNA) and transcriptionally active (RNA) nitrous oxide reductase (nosZ) genes using a functional gene microarray. The total and active nosZ communities were dominated by a limited number of nosZ archetypes, affiliated with bacteria from marine, soil and marsh environments. In addition to nosZ genes related to those of known marine denitrifiers, atypical nosZ genes, related to those of soil bacteria that do not possess a complete denitrification pathway, were also detected, especially in surface waters. The community composition of the total nosZ assemblage was significantly different from the active assemblage. The community composition of the total nosZ assemblage was significantly different between coastal and off-shore stations. The low oxygen assemblages from both stations were similar to each other, while the higher oxygen assemblages were more variable. Community composition of the active nosZ assemblage was also significantly different between stations, and varied with N2O concentration but not O2. Notably, nosZ assemblages were not only present but also active in oxygenated seawater: the abundance of total and active nosZ bacteria from oxygenated surface water (indicated by nosZ gene copy number) was similar to or even larger than in anoxic waters, implying the potential for N2O consumption even in the oxygenated surface water. PMID:28702012

Heterotrophic ammonium removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium, Providencia rettgeri YL

Institute of Scientific and Technical Information of China (English)

TAYLOR Shauna M; HE Yiliang; ZHAO Bin; HUANG Jue

2009-01-01

Bacterium Providencia rettgeri YL was found to exhibit an unusual ability to heterotrophically nitrify and aerobically denitrify various concentrations of ammonium (NH4+-N). In order to further analyze its removal ability, several experiments were conducted to identify the growth and ammonium removal response in different carbon to nitrogen (C/N) mass ratios, shaking speeds, temperatures, ammonium concentrations and to qualitatively verify the production of nitrogen gas using gas chromatography techniques. Results showed that under optimum conditions (C/N 10, 30℃, 120 r/min), YL can significantly remove low and high concentrations of ammonium within 12 to 48 h of growth. The nitrification products hydroxylamine (NH2OH), nitrite (NO2-) and nitrate (NO3-) as well as the denitrification product, nitrogen gas (N2), were detected under completely aerobic conditions.

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

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

2016-02-01

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

Fatty Acid and Carbon Isotopic Evidence for type I Methanotrophs in Microbial Mats from a Shallow Marine Gas Seep, Coal Oil Point, CA.

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Ding, H.; Valentine, D.

2005-12-01

To study the microbial community in a Southern California seep field, sediment and bacterial mat samples were collected from natural gas-bearing and gas-free surfaces at two distinct seeps in the Coal Oil Point seep field, offshore Santa Barbara. Fatty acids in these samples were extracted, analyzed and identified. Using gas chromatography (GC), more than 30 different fatty acids were separated. Generally, fatty acid concentrations in natural gas-bearing samples were about 5-fold higher compared to gas-free samples. Using gas chromatography mass sepctrometry (GC-MS), all separated fatty acids were identified in each sample. The major constituents included saturated 14:0, 16:0, 18:0, branched i-15, a-15 and unsaturated 16:1 and 18:1 series fatty acids. GC-IRMS (isotope ratio mass spectrometry) analysis provided the 13C of all major fatty acids and some 16:1 series fatty acids were found to be more depleted than -40% in samples associated with gas seepage. After treatment with dimethyl disufide (DMDS), the 16:1 series fatty acids were resolved into five distinct components, including common composition 16:1(7), bacterial specific i-16:1(7) and typical biomarkers of type I methnotrophs 16:1(8), 16(6) and 16:1(5), suggesting an important role for methnotrophs in seep sediments and microbial mats. These results provide evidence for the activity of type I methanotrophic bacteria in microbial mats and surficial sediments at the Coal Oil Point seep field, and have implications for methane cycling in this and other seep

Temperature-Induced Increase in Methane Release from Peat Bogs: A Mesocosm Experiment

NARCIS (Netherlands)

van Winden, J.F.; Reichart, G.J.; McNamara, N.P.; Benthien, A.; Sinninghe Damsté, J.S.

2012-01-01

Peat bogs are primarily situated at mid to high latitudes and future climatic change projections indicate that these areas may become increasingly wetter and warmer. Methane emissions from peat bogs are reduced by symbiotic methane oxidizing bacteria (methanotrophs). Higher temperatures and

Temperature-induced increase in methane release from peat bogs: A mesocosm experiment

NARCIS (Netherlands)

Winden, J.F. van; Reichart, G.-J.; McNamara, N.P.; Benthien, A.; Sinninghe Damsté, J.S.

2012-01-01

Peat bogs are primarily situated at mid to high latitudes and future climatic change projections indicate that these areas may become increasingly wetter and warmer. Methane emissions from peat bogs are reduced by symbiotic methane oxidizing bacteria (methanotrophs). Higher temperatures and

Mechanisms of Persistence of the Ammonia-Oxidizing Bacteria Nitrosomonas to the Biocide Free Nitrous Acid.

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Laloo, Andrew E; Wei, Justin; Wang, Dongbo; Narayanasamy, Shaman; Vanwonterghem, Inka; Waite, David; Steen, Jason; Kaysen, Anne; Heintz-Buschart, Anna; Wang, Qilin; Schulz, Benjamin; Nouwens, Amanda; Wilmes, Paul; Hugenholtz, Philip; Yuan, Zhiguo; Bond, Philip L

2018-05-01

Free nitrous acid (FNA) exerts a broad range of antimicrobial effects on bacteria, although susceptibility varies considerably among microorganisms. Among nitrifiers found in activated sludge of wastewater treatment processes (WWTPs), nitrite-oxidizing bacteria (NOB) are more susceptible to FNA compared to ammonia-oxidizing bacteria (AOB). This selective inhibition of NOB over AOB in WWTPs bypasses nitrate production and improves the efficiency and costs of the nitrogen removal process in both the activated sludge and anaerobic ammonium oxidation (Anammox) system. However, the molecular mechanisms governing this atypical tolerance of AOB to FNA have yet to be understood. Herein we investigate the varying effects of the antimicrobial FNA on activated sludge containing AOB and NOB using an integrated metagenomics and label-free quantitative sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) metaproteomic approach. The Nitrosomonas genus of AOB, on exposure to FNA, maintains internal homeostasis by upregulating a number of known oxidative stress enzymes, such as pteridine reductase and dihydrolipoyl dehydrogenase. Denitrifying enzymes were upregulated on exposure to FNA, suggesting the detoxification of nitrite to nitric oxide. Interestingly, proteins involved in stress response mechanisms, such as DNA and protein repair enzymes, phage prevention proteins, and iron transport proteins, were upregulated on exposure to FNA. In addition enzymes involved in energy generation were also upregulated on exposure to FNA. The total proteins specifically derived from the NOB genus Nitrobacter was low and, as such, did not allow for the elucidation of the response mechanism to FNA exposure. These findings give us an understanding of the adaptive mechanisms of tolerance within the AOB Nitrosomonas to the biocidal agent FNA.

Molecular identification of potential denitrifying bacteria and use of D-optimal mixture experimental design for the optimization of denitrification process.

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Ben Taheur, Fadia; Fdhila, Kais; Elabed, Hamouda; Bouguerra, Amel; Kouidhi, Bochra; Bakhrouf, Amina; Chaieb, Kamel

2016-04-01

Three bacterial strains (TE1, TD3 and FB2) were isolated from date palm (degla), pistachio and barley. The presence of nitrate reductase (narG) and nitrite reductase (nirS and nirK) genes in the selected strains was detected by PCR technique. Molecular identification based on 16S rDNA sequencing method was applied to identify positive strains. In addition, the D-optimal mixture experimental design was used to optimize the optimal formulation of probiotic bacteria for denitrification process. Strains harboring denitrification genes were identified as: TE1, Agrococcus sp LN828197; TD3, Cronobacter sakazakii LN828198 and FB2, Pedicoccus pentosaceus LN828199. PCR results revealed that all strains carried the nirS gene. However only C. sakazakii LN828198 and Agrococcus sp LN828197 harbored the nirK and the narG genes respectively. Moreover, the studied bacteria were able to form biofilm on abiotic surfaces with different degree. Process optimization showed that the most significant reduction of nitrate was 100% with 14.98% of COD consumption and 5.57 mg/l nitrite accumulation. Meanwhile, the response values were optimized and showed that the most optimal combination was 78.79% of C. sakazakii LN828198 (curve value), 21.21% of P. pentosaceus LN828199 (curve value) and absence (0%) of Agrococcus sp LN828197 (curve value). Copyright © 2016 Elsevier Ltd. All rights reserved.

A performance study of simultaneous microbial removal of no and SO2 in a biotrickling-filter under anaerobic condition

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Yaqiong Han

2011-06-01

Full Text Available The behaviors of simultaneous removal of NO and SO2 using the coculture of anaerobic denitrifying bacteria and sulfate reducing bacteria was investigated in a bench-scale biotrickling-filter. When the combined NO/SO2 removal biotrickling-filter was operated at an empty bed residence time of 76 s with NO and SO2 feed concentrations of 2 and 2 g/m3, respectively, the SO2 removal efficiency was always above 95%, while NO removal exhibited an evident periodicity of 5-6 days for the initial 60 days after the attachment phase. A steady-state NO-removal efficiency of around 90% was obtained after 130 days of continuous operation. Experimental results indicated that the coculture in the combined NO/SO2 removal biotrickling-filter showed a higher resistance to shock NO-loadings and a better tolerance to starvation than the single denitrifying bacteria in the NO removal biotrickling-filter.

Long-term operation of oxygen-limiting membrane bioreactor (MBR) for the development of simultaneous partial nitrification, anammox and denitrification (SNAD) process.

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Zhao, Chuanqi; Wang, Gang; Xu, Xiaochen; Yang, Yuesuo; Yang, Fenglin

2017-07-18

In this study, an oxygen-limiting membrane bioreactor (MBR) with recirculation of biogas for relieving membrane fouling was successfully operated to realize the simultaneous partial nitrification, anammox and denitrification (SNAD) process. The MBR operation was considered effective in the long-term test with total nitrogen (TN) and chemical oxygen demand (COD) removal efficiencies of 94.86% and 98.91%, respectively. Membrane fouling was significantly alleviated due to the recirculation of biogas and the membrane had been cleaned four times with a normal filtration period of 52 days. The co-existence of ammonia-oxidizing bacteria (AOB), anammox and denitrifying bacteria in MBR was confirmed by scanning electron microscopy (SEM) and fluorescence in situ hybridizations (FISH) analysis. Furthermore, AOB were found close to the granule surface, while denitrifying bacteria and anammox were in the deeper layer of granules. Potential in excellent TN and COD removal, operational stability and sustainability, as well as in alleviating membrane fouling is expected by using this oxygen-limiting MBR.

Microbiology of Lonar Lake and other soda lakes

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Paul Antony, Chakkiath; Kumaresan, Deepak; Hunger, Sindy; Drake, Harold L; Murrell, J Colin; Shouche, Yogesh S

2013-01-01

Soda lakes are saline and alkaline ecosystems that are believed to have existed throughout the geological record of Earth. They are widely distributed across the globe, but are highly abundant in terrestrial biomes such as deserts and steppes and in geologically interesting regions such as the East African Rift valley. The unusual geochemistry of these lakes supports the growth of an impressive array of microorganisms that are of ecological and economic importance. Haloalkaliphilic Bacteria and Archaea belonging to all major trophic groups have been described from many soda lakes, including lakes with exceptionally high levels of heavy metals. Lonar Lake is a soda lake that is centered at an unusual meteorite impact structure in the Deccan basalts in India and its key physicochemical and microbiological characteristics are highlighted in this article. The occurrence of diverse functional groups of microbes, such as methanogens, methanotrophs, phototrophs, denitrifiers, sulfur oxidizers, sulfate reducers and syntrophs in soda lakes, suggests that these habitats harbor complex microbial food webs that (a) interconnect various biological cycles via redox coupling and (b) impact on the production and consumption of greenhouse gases. Soda lake microorganisms harbor several biotechnologically relevant enzymes and biomolecules (for example, cellulases, amylases, ectoine) and there is the need to augment bioprospecting efforts in soda lake environments with new integrated approaches. Importantly, some saline and alkaline lake ecosystems around the world need to be protected from anthropogenic pressures that threaten their long-term existence. PMID:23178675

Ammonia-Oxidizing Archaea Are More Resistant Than Denitrifiers to Seasonal Precipitation Changes in an Acidic Subtropical Forest Soil

Directory of Open Access Journals (Sweden)

Jie Chen

2017-07-01

Full Text Available Seasonal precipitation changes are increasingly severe in subtropical areas. However, the responses of soil nitrogen (N cycle and its associated functional microorganisms to such precipitation changes remain unclear. In this study, two projected precipitation patterns were manipulated: intensifying the dry-season drought (DD and extending the dry-season duration (ED but increasing the wet-season storms following the DD and ED treatment period. The effects of these two contrasting precipitation patterns on soil net N transformation rates and functional gene abundances were quantitatively assessed through a resistance index. Results showed that the resistance index of functional microbial abundance (-0.03 ± 0.08 was much lower than that of the net N transformation rate (0.55 ± 0.02 throughout the experiment, indicating that microbial abundance was more responsive to precipitation changes compared with the N transformation rate. Spring drought under the ED treatment significantly increased the abundances of both nitrifying (amoA and denitrifying genes (nirK, nirS, and nosZ, while changes in these gene abundances overlapped largely with control treatment during droughts in the dry season. Interestingly, the resistance index of the ammonia-oxidizing archaea (AOA amoA abundance was significantly higher than that of the denitrifying gene abundances, suggesting that AOA were more resistant to the precipitation changes. This was attributed to the stronger environmental adaptability and higher resource utilization efficiency of the AOA community, as indicated by the lack of correlations between AOA gene abundance and environmental factors [i.e., soil water content, ammonium (NH4+ and dissolved organic carbon concentrations] during the experiment.

Acetate utilization as a survival strategy of peat-inhabiting Methylocystis spp

NARCIS (Netherlands)

Belova, S.E.; Baani, M.; Suzina, N.E.; Bodelier, P.L.E.; Liesack, W.; Dedysh, S.N.

2011-01-01

Representatives of the genus Methylocystis are traditionally considered to be obligately methanotrophic bacteria, which are incapable of growth on multicarbon substrates. Here, we describe a novel member of this genus, strain H2s, which represents a numerically abundant and ecologically important

Methane fluxes and the functional groups of methanotrophs and methanogens in a young Arctic landscape on Disko Island, West Greenland

DEFF Research Database (Denmark)

Christiansen, Jesper Riis; Barrera Romero, Alejandro Jose; Jørgensen, Niels O. G.

2015-01-01

and activity indicates that the age of an Arctic landscape is not important for the CH4 consumption but can be very important for CH4 production. Considering the prevalence of dry landscapes and contrasting ages of high Arctic soils, our results highlight that well-drained soils should not be overlooked......Arctic soils are known to be important methane (CH4) consumers and sources. This study integrates in situ fluxes of CH4 between upland and wetland soils with potential rates of CH4 oxidation and production as well as abundance and diversity of the methanotrophs and methanogens measured...... as an important component of Arctic net CH4 budget....

Day/night temperature differences (DNTD) trigger changes in nutrient removal and functional bacteria in membrane bioreactors.

Science.gov (United States)

Zhang, Shaoqing; Sheng, Binbin; Lin, Wenting; Meng, Fangang

2018-09-15

Temperature is a well-known environmental stress that influences both microbial metabolism and community structure in the biological wastewater treatment systems. In this study, responses of biological performance and sludge microbiota to the long-term day/night temperature differences (DNTD) were investigated in membrane bioreactors (MBRs). The results showed that the functional bacteria could sustained their ecological functions at low DNTD (20/30 °C), resulting in relatively stable performance with respect to nutrient removal. However, when the activated sludge was subjected to a high DNTD (17/33 °C), the effluent concentrations of COD, TN and TP were significantly higher in MBR-B than that in MBR-A. In addition, more severe membrane fouling occurred under the perturbation of high DNTD as revealed by the transmembrane pressure (TMP) profile, which was mainly attributed to the accumulation of extracellular polymeric substances (EPS). The results of 16S rRNA gene sequencing showed that DNTD showed negligible effect on the bacterial community structures. Nonetheless, the functional bacteria responded differently to DNTD, which were in accordance with the bioreactor performances. Specifically, Nitrospina (NOB) and Tetrasphaera (PAOs) appeared to be sensitive to both low and high DNTD. In contrast, a low DNTD showed marginal effects on the denitrifiers, while a high DNTD significantly decreased their abundances. More strikingly, filamentous bulking bacteria were found to be well-adapted to DNTD, indicating their tolerance to the daily temperature fluctuation. This study will advance our knowledge regarding the response of microbial ecology of activated sludge to daily temperature variations in full-scale MBRs. Copyright © 2018 Elsevier B.V. All rights reserved.

Development of a system for genetic manipulation of the facultative methanotroph Methylocella silvestris BL2.

Science.gov (United States)

Crombie, Andrew; Murrell, J Colin

2011-01-01

An understanding of the metabolism and metabolic regulation of the facultative methanotroph Methylocella silvestris BL2 is required to understand its role in methane oxidation in the environment, and methods for genetics manipulation are essential tools in these investigations. In addition, the ability to engineer the metabolic capabilities of M. silvestris may well have useful biotechnological applications. We describe a simple and effective method of genetic manipulation for this organism which relies on the electroporation of a linear DNA fragment to introduce chromosomal gene deletions. In a two-step procedure, the gene of interest is first replaced with an antibiotic-resistance cassette which is subsequently removed, resulting in an unmarked gene deletion. This method is illustrated by the deletion of isocitrate lyase, which abolished growth on one-carbon and severely disabled growth on two-carbon compounds. Subsequent complementation with the wild-type gene and promoter restored growth, demonstrating stable transcription from the broad-host-range plasmid employed. Copyright © 2011 Elsevier Inc. All rights reserved.

Nitrous Oxide Metabolism in Nitrate-Reducing Bacteria: Physiology and Regulatory Mechanisms.

Science.gov (United States)

Torres, M J; Simon, J; Rowley, G; Bedmar, E J; Richardson, D J; Gates, A J; Delgado, M J

2016-01-01

Nitrous oxide (N2O) is an important greenhouse gas (GHG) with substantial global warming potential and also contributes to ozone depletion through photochemical nitric oxide (NO) production in the stratosphere. The negative effects of N2O on climate and stratospheric ozone make N2O mitigation an international challenge. More than 60% of global N2O emissions are emitted from agricultural soils mainly due to the application of synthetic nitrogen-containing fertilizers. Thus, mitigation strategies must be developed which increase (or at least do not negatively impact) on agricultural efficiency whilst decrease the levels of N2O released. This aim is particularly important in the context of the ever expanding population and subsequent increased burden on the food chain. More than two-thirds of N2O emissions from soils can be attributed to bacterial and fungal denitrification and nitrification processes. In ammonia-oxidizing bacteria, N2O is formed through the oxidation of hydroxylamine to nitrite. In denitrifiers, nitrate is reduced to N2 via nitrite, NO and N2O production. In addition to denitrification, respiratory nitrate ammonification (also termed dissimilatory nitrate reduction to ammonium) is another important nitrate-reducing mechanism in soil, responsible for the loss of nitrate and production of N2O from reduction of NO that is formed as a by-product of the reduction process. This review will synthesize our current understanding of the environmental, regulatory and biochemical control of N2O emissions by nitrate-reducing bacteria and point to new solutions for agricultural GHG mitigation. © 2016 Elsevier Ltd. All rights reserved.

DRIVERS OF THE DYNAMICS OF DIAZOTROPHS AND DENITRIFIERS IN NORTH SEA BOTTOM WATERS AND SEDIMENTS

Directory of Open Access Journals (Sweden)

Lucas eStal

2015-07-01

Full Text Available The fixation of dinitrogen (N2 and denitrification are two opposite processes in the nitrogen cycle. The former transfers atmospheric dinitrogen gas into bound nitrogen in the biosphere, while the latter returns this bound nitrogen back to atmospheric dinitrogen. It is unclear whether or not these processes are intimately connected in any microbial ecosystem or that they are spatially and/or temporally separated. Here, we measured seafloor nitrogen fixation and denitrification as well as pelagic nitrogen fixation by using the stable isotope technique. Alongside, we measured the diversity, abundance, and activity of nitrogen-fixing and denitrifying microorganisms at three stations in the southern North Sea. Nitrogen fixation ranged from undetectable to 2.4 nmol N L-1 d-1 and from undetectable to 8.2 nmol N g-1 d-1 in the water column and seafloor, respectively. The highest rates were measured in August at Doggersbank, both for the water column and for the seafloor. Denitrification ranged from 1.7 to 208.8 µmol m-2 d-1 and the highest rates were measured in May at the Oyster Grounds. DNA sequence analysis showed sequences of nifH, a structural gene for nitrogenase, related to sequences from anaerobic sulfur/iron reducers and sulfate reducers. Sequences of the structural gene for nitrite reductase, nirS, were related to environmental clones from marine sediments. Quantitative polymerase chain reaction (qPCR data revealed the highest abundance of nifH and nirS genes at the Oyster Grounds. Quantitative reverse transcription polymerase chain reaction (qRT-PCR data revealed the highest nifH expression at Doggersbank and the highest nirS expression at the Oyster Grounds. The distribution of the diazotrophic and denitrifying communities seems to be subject to different selecting factors, leading to spatial and temporal separation of nitrogen fixation and denitrification. These selecting factors include temperature, organic matter availability, and

Storage and growth of denitrifiers in aerobic granules: part I. model development.

Science.gov (United States)

Ni, Bing-Jie; Yu, Han-Qing

2008-02-01

A mathematical model, based on the Activated Sludge Model No.3 (ASM3), is developed to describe the storage and growth activities of denitrifiers in aerobic granules under anoxic conditions. In this model, mass transfer, hydrolysis, simultaneous anoxic storage and growth, anoxic maintenance, and endogenous decay are all taken into account. The model established is implemented in the well-established AQUASIM simulation software. A combination of completely mixed reactor and biofilm reactor compartments provided by AQUASIM is used to simulate the mass transport and conversion processes occurring in both bulk liquid and granules. The modeling results explicitly show that the external substrate is immediately utilized for storage and growth at feast phase. More external substrates are diverted to storage process than the primary biomass production process. The model simulation indicates that the nitrate utilization rate (NUR) of granules-based denitrification process includes four linear phases of nitrate reduction. Furthermore, the methodology for determining the most important parameter in this model, that is, anoxic reduction factor, is established. (c) 2007 Wiley Periodicals, Inc.

Methylocella tundrae sp. nov., a novel methanotrophic bacterium from acidic tundra peatlands.

Science.gov (United States)

Dedysh, Svetlana N; Berestovskaya, Yulia Y; Vasylieva, Lina V; Belova, Svetlana E; Khmelenina, Valentina N; Suzina, Natalia E; Trotsenko, Yuri A; Liesack, Werner; Zavarzin, George A

2004-01-01

A novel species, Methylocella tundrae, is proposed for three methanotrophic strains (T4T, TCh1 and TY1) isolated from acidic Sphagnum tundra peatlands. These strains are aerobic, Gram-negative, non-motile, dinitrogen-fixing rods that possess a soluble methane monooxygenase and utilize the serine pathway for carbon assimilation. Strains T4T, TCh1 and TY1 are moderately acidophilic organisms capable of growth between pH 4.2 and 7.5 (optimum 5.5-6.0) and between 5 and 30 degrees C (optimum 15 degrees C). The major phospholipid fatty acid is 18:1omega7c. The DNA G+C content of strain T4T is 63.3 mol%. The three strains possess almost identical 16S rRNA gene sequences and are most closely related to two previously identified species of Methylocella, Methylocella palustris (97% similarity) and Methylocella silvestris (97.5% similarity). DNA-DNA hybridization values of strain T4T with Methylocella palustris KT and Methylocella silvestris BL2T were respectively 27 and 36%. Thus, the tundra strains represent a novel species, for which the name Methylocella tundrae sp. nov. is proposed. Strain T4T (=DSM 15673T=NCIMB 13949T) is the type strain.

Sodium lauryl ether sulfate (SLES) degradation by nitrate-reducing bacteria.

Science.gov (United States)

Paulo, Ana M S; Aydin, Rozelin; Dimitrov, Mauricio R; Vreeling, Harm; Cavaleiro, Ana J; García-Encina, Pedro A; Stams, Alfons J M; Plugge, Caroline M

2017-06-01

The surfactant sodium lauryl ether sulfate (SLES) is widely used in the composition of detergents and frequently ends up in wastewater treatment plants (WWTPs). While aerobic SLES degradation is well studied, little is known about the fate of this compound in anoxic environments, such as denitrification tanks of WWTPs, nor about the bacteria involved in the anoxic biodegradation. Here, we used SLES as sole carbon and energy source, at concentrations ranging from 50 to 1000 mg L -1 , to enrich and isolate nitrate-reducing bacteria from activated sludge of a WWTP with the anaerobic-anoxic-oxic (A 2 /O) concept. In the 50 mg L -1 enrichment, Comamonas (50%), Pseudomonas (24%), and Alicycliphilus (12%) were present at higher relative abundance, while Pseudomonas (53%) became dominant in the 1000 mg L -1 enrichment. Aeromonas hydrophila strain S7, Pseudomonas stutzeri strain S8, and Pseudomonas nitroreducens strain S11 were isolated from the enriched cultures. Under denitrifying conditions, strains S8 and S11 degraded 500 mg L -1 SLES in less than 1 day, while strain S7 required more than 6 days. Strains S8 and S11 also showed a remarkable resistance to SLES, being able to grow and reduce nitrate with SLES concentrations up to 40 g L -1 . Strain S11 turned out to be the best anoxic SLES degrader, degrading up to 41% of 500 mg L -1 . The comparison between SLES anoxic and oxic degradation by strain S11 revealed differences in SLES cleavage, degradation, and sulfate accumulation; both ester and ether cleavage were probably employed in SLES anoxic degradation by strain S11.

Nitrification inhibition as measured by RNA- and DNA-based function-specific assays and microbial community structure analyses

Science.gov (United States)

Abstract: The biological removal of ammonia in conventional wastewater treatment plants (WWTPs) is performed by promoting nitrification, which transforms ammonia into nitrate, which in turn is converted into nitrogen gas by denitrifying bacteria. The first step in nitrification, ...

Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

NARCIS (Netherlands)

Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

1998-01-01

The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative, denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing

Activity and diversity of methane-oxidizing bacteria along a Norwegian sub-Arctic glacier forefield

DEFF Research Database (Denmark)

Mateos-Rivera, Alejandro; Øvreås, Lise; Wilson, Bryan

2018-01-01

throughout the growing season during three sampling times at eight different sampling points in combination with laboratory incubation experiments. The overall results showed that the methanotrophic community had similar trends of increased CH4 consumption and increased abundance as a function of soil...

[Effects of Different Reclaimed Scenarios on Soil Microbe and Enzyme Activities in Mining Areas].

Science.gov (United States)

Li, Jun-jian; Liu, Feng; Zhou, Xiao-mei

2015-05-01

Abstract: Ecological degradation in the mining areas is greatly aggravated in recent several decades, and ecological restoration has become the primary measure for the sustainable development. Soil microbe and enzyme activity are sensitive indices to evaluate soil quality. Ecological reconstruction was initiated in Antaibao mining area, and we tested soil physicochemical properties, microbial populations of azotobacteria, nitrifying-bacteria and denitrifying-bacteria, and enzyme activities (including sucrose, polyphenol oxidase, dehydrogenase and urease) under different regeneration scenarios. Regeneration scenarios had significant effects on soil physicochemical properties, microbial population and enzyme activities. Total nitrogen was strongly correlated with azotobacteria and nitrifying-bacteria, however, total nitrogen was not correlated with denitrifying-bacteria. Phenol oxidase activity was negatively correlated with soil organic carbon and total nitrogen, but other enzyme activities were positively correlated with soil organic carbon and total nitrogen. Principal Component Analysis ( PCA) was applied to analyze the integrated fertility index (IFI). The highest and lowest IFIs were in Robinia pseudoacacia-Pinus tabuliformis mixed forests and un-reclaimed area, respectively. R. pseudoacacia-P. tabuliformis mixed forests were feasible for reclaimed mining areas in semi-arid region Northwest Shanxi.

Do freshwater macrophytes influence the community structure of ammonia-oxidizing and denitrifying bacteria in the rhizospere?

DEFF Research Database (Denmark)

Herrmann, Martina; Schramm, Andreas

2006-01-01

to unvegetated sediment, especially with respect to the availability of oxygen, organic carbon, and inorganic nitrogen. We hypothesize that macrophyte species create specific niches for ammonia oxidizing and nitrate-reducing bacteria in their rhizosphere, leading to plant-dependant differences in abundance...... dortmanna have been shown to release oxygen from their roots and to stimulate nitrification and coupled nitrification-denitrification in the rhizosphere. Together with the excretion of root exudates, this effect leads to strongly modified microenvironments at the root surface and in the rhizosphere compared......-denitrification using the 15N isotope pairing technique. Ammonia-oxidizing and nitrate-reducing populations are analyzed based on the ammonia monooxygenase gene (amoA) and the nitrate reductase gene (narG) as functional markers. Preliminary data indicate that there in fact exist differences in the community composition...

Insights into the effect of soil pH on N.sub.2./sub.O and N.sub.2./sub. emissions and denitrifier community size and activity

Czech Academy of Sciences Publication Activity Database

Čuhel, Jiří; Šimek, Miloslav; Laughlin, R.J.; Bru, D.; Chéneby, D.; Watson, C.J.; Philippot, L.

2010-01-01

Roč. 76, č. 6 (2010), s. 1870-1878 ISSN 0099-2240 R&D Projects: GA MŠk MEB020726; GA MŠk LC06066; GA AV ČR IAA600660605 Institutional research plan: CEZ:AV0Z60660521 Keywords : soil pH * N 2 O and N 2 emissions * denitrifier community Subject RIV: EH - Ecology, Behaviour Impact factor: 3.778, year: 2010

Identification and Characterization of EctR1, a New Transcriptional Regulator of the Ectoine Biosynthesis Genes in the Halotolerant Methanotroph Methylomicrobium alcaliphilum 20Z▿ †

OpenAIRE

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

2009-01-01

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, ectAp1 and ectAp2 (ectAp1p2), similar to the σ70-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 ...

Methane and carbon dioxide flux in the profile of wood ant (Formica aquilonia) nests and the surrounding forest floor during a laboratory incubation

Czech Academy of Sciences Publication Activity Database

Jílková, Veronika; Picek, T.; Šestauberová, Martina; Krištůfek, Václav; Cajthaml, Tomáš; Frouz, Jan

2016-01-01

Roč. 92, č. 10 (2016), č. článku fiw141. ISSN 0168-6496 Institutional support: RVO:60077344 ; RVO:61388971 Keywords : aboveground parts * available nutrients * bacteria * fungi * methanotrophs * temperate forest Subject RIV: EH - Ecology, Behaviour; EE - Microbiology, Virology (MBU-M) Impact factor: 3.720, year: 2016

Elementary sulfur in effluent from denitrifying sulfide removal process as adsorbent for zinc(II).

Science.gov (United States)

Chen, Chuan; Zhou, Xu; Wang, Aijie; Wu, Dong-hai; Liu, Li-hong; Ren, Nanqi; Lee, Duu-Jong

2012-10-01

The denitrifying sulfide removal (DSR) process can simultaneously convert sulfide, nitrate and organic compounds into elementary sulfur (S(0)), di-nitrogen gas and carbon dioxide, respectively. However, the S(0) formed in the DSR process are micro-sized colloids with negatively charged surface, making isolation of S(0) colloids from other biological cells and metabolites difficult. This study proposed the use of S(0) in DSR effluent as a novel adsorbent for zinc removal from wastewaters. Batch and continuous tests were conducted for efficient zinc removal with S(0)-containing DSR effluent. At pHremoval rates of zinc(II) were increased with increasing pH. The formed S(0) colloids carried negative charge onto which zinc(II) ions could be adsorbed via electrostatic interactions. The zinc(II) adsorbed S(0) colloids further enhanced coagulation-sedimentation efficiency of suspended solids in DSR effluents. The DSR effluent presents a promising coagulant for zinc(II) containing wastewaters. Copyright © 2012 Elsevier Ltd. All rights reserved.

Sulfate-reducing bacteria from the Arabian Sea - their distribution in relation to thiosulfate-oxidising and heterotrophic bacteria

Digital Repository Service at National Institute of Oceanography (India)

LokaBharathi, P.A.; Chandramohan, D.

able to oxidize sulfate but also denitrify (Loka Bharathi et aI., 1988). Positive correlations were found between SRB and TLOan and SRB and AnB when the whole set ofsamples were examined together and for the region in the case ofmud bank samples (Table... at Delaware Inlet, New Zealand. The infor mation available about these forms in Indian waters has been restricted to es tuarine ecosystems (Loka Bharathi and Chandramohan, 1985; Saxena et aI., 1988). We describe here their distribution in marine sediments...

Heat Production by the Denitrifying Bacterium Pseudomonas fluorescens and the Dissimilatory Ammonium-Producing Bacterium Pseudomonas putrefaciens during Anaerobic Growth with Nitrate as the Electron Acceptor

OpenAIRE

Samuelsson, M.-O.; Cadez, P.; Gustafsson, L.

1988-01-01

The heat production rate and the simultaneous nitrate consumption and production and consumption of nitrite and nitrous oxide were monitored during the anaerobic growth of two types of dissimilatory nitrate reducers. Pseudomonas fluorescens, a denitrifier, consumed nitrate and accumulated small amounts of nitrite or nitrous oxide. The heat production rate increased steadily during the course of nitrate consumption and decreased rapidly concomitant with the depletion of the electron acceptors....

From the Gut of an Insect to the Global Climate: Denitrification and Nitrous Oxide Production inside Lake Chironomidae

DEFF Research Database (Denmark)

Stief, Peter; Nielsen, Lars Peter; Revsbech, Niels Peter

2006-01-01

FROM THE GUT OF AN INSECT TO THE GLOBAL CLIMATE: DENITRIFICATION AND NITROUS OXIDE PRODUCTION INSIDE LAKE CHIRONOMIDAE P. Stief, L.P. Nielsen, N.P. Revsbech, A. Schramm Department of Biological Sciences, Microbiology, University of Aarhus, Denmark Denitrifying bacteria in lake sediments drive...

Bioremediation of toxic and hazardous wastes by denitrifying bacteria

International Nuclear Information System (INIS)

Barraquio, Wilfredo L.

2005-01-01

This papers discusses the wastes coming rom domestic, industrial and agricultural sources are polluting the forests, rivers lakes, groundwater, and air and there are some measures like the physicochemical and biological measures are being utilized to remedy the destruction of resources; and of the measures, bioremediation offers great potential in cleaning up the environment of pollutants which is a cost-effective and environment-friendly technology that uses microorganisms to degrade hazardous substances into less toxic

Descriptions of Roseiarcus fermentans gen. nov., sp. nov., a bacteriochlorophyll a-containing fermentative bacterium related phylogenetically to alphaproteobacterial methanotrophs, and of the family Roseiarcaceae fam. nov.

Science.gov (United States)

Kulichevskaya, Irina S; Danilova, Olga V; Tereshina, Vera M; Kevbrin, Vadim V; Dedysh, Svetlana N

2014-08-01

A light-pink-pigmented, microaerophilic bacterium was obtained from a methanotrophic consortium enriched from acidic Sphagnum peat and designated strain Pf56(T). Cells of this bacterium were Gram-negative, non-motile, thick curved rods that contained a vesicular intracytoplasmic membrane system characteristic of some purple non-sulfur alphaproteobacteria. The absorption spectrum of acetone/methanol extracts of cells grown in the light showed maxima at 363, 475, 505, 601 and 770 nm; the peaks at 363 and 770 nm are characteristic of bacteriochlorophyll a. However, in contrast to purple non-sulfur bacteria, strain Pf56(T) was unable to grow phototrophically under anoxic conditions in the light. Best growth occurred on some sugars and organic acids under micro-oxic conditions by means of fermentation. The fermentation products were propionate, acetate and hydrogen. Slow chemo-organotrophic growth was also observed under fully oxic conditions. Light stimulated growth. C1 substrates were not utilized. Strain Pf56(T) grew at pH 4.0-7.0 (optimum pH 5.5-6.5) and at 15-30 °C (optimum 22-28 °C). The major cellular fatty acids were 19 : 0 cyclo ω8c and 18 : 1ω7c; quinones were represented by ubiquinone Q-10. The G+C content of the DNA was 70.0 mol%. Strain Pf56 displays 93.6-94.7 and 92.7-93.7% 16S rRNA gene sequence similarity to members of the families Methylocystaceae and Beijerinckiaceae, respectively, and belongs to a large cluster of environmental sequences retrieved from various wetlands and forest soils in cultivation-independent studies. Phenotypic, genotypic and chemotaxonomic characteristics of strain Pf56(T) suggest that it represents a novel genus and species of bacteriochlorophyll a-containing fermentative bacteria, for which the name Roseiarcus fermentans gen. nov., sp. nov. is proposed. Strain Pf56(T) ( = DSM 24875(T) = VKM B-2876(T)) is the type strain of Roseiarcus fermentans, and is also the first characterized member of a novel family

Use of vegetable oil in a pilot-scale denitrifying barrier

Science.gov (United States)

Hunter, William J.

2001-12-01

Nitrate in drinking water is a hazard to both humans and animals. Contaminated water can cause methemoglobinemia and may pose a cancer risk. Permeable barriers containing innocuous oils, which stimulate denitrification, can remove nitrate from flowing groundwater. For this study, a sand tank (1.1×2.0×0.085 m in size) containing sand was used as a one-dimensional open-top scale model of an aquifer. A meter-long area near the center of the tank contained sand coated with soybean oil. This region served as a permeable denitrifying barrier. Water containing 20 mg l -1 nitrate-N was pumped through the barrier at a high flow rate, 1112 l week -1, for 30 weeks. During the 30-week study, the barrier removed 39% of the total nitrate-N present in the water. The barrier was most efficient during the first 10 weeks of the study when almost all of the nitrate and nitrogen was removed. Efficiency declined with time so that by week 30 almost no nitrate was removed by the system. Nitrite levels in the effluent water remained low throughout the study. Barriers could be used to protect groundwater from nitrate contamination or for the in situ treatment of contaminated water. At the low flow rates that exist in most aquifers, such barriers should be effective at removing nitrate from groundwater for a much longer period of time.

Production of Nitrous Oxide from Nitrite in Stable Type II Methanotrophic Enrichments.

Science.gov (United States)

Myung, Jaewook; Wang, Zhiyue; Yuan, Tong; Zhang, Ping; Van Nostrand, Joy D; Zhou, Jizhong; Criddle, Craig S

2015-09-15

The coupled aerobic-anoxic nitrous decomposition operation is a new process for wastewater treatment that removes nitrogen from wastewater and recovers energy from the nitrogen in three steps: (1) NH4(+) oxidation to NO2(-), (2) NO2(-) reduction to N2O, and (3) N2O conversion to N2 with energy production. Here, we demonstrate that type II methanotrophic enrichments can mediate step two by coupling oxidation of poly(3-hydroxybutyrate) (P3HB) to NO2(-) reduction. Enrichments grown with NH4(+) and NO2(-) were subject to alternating 48-h aerobic and anoxic periods, in which CH4 and NO2(-) were added together in a "coupled" mode of operation or separately in a "decoupled mode". Community structure was stable in both modes and dominated by Methylocystis. In the coupled mode, production of P3HB and N2O was low. In the decoupled mode, significant P3HB was produced, and oxidation of P3HB drove reduction of NO2(-) to N2O with ∼ 70% conversion for >30 cycles (120 d). In batch tests of wasted cells from the decoupled mode, N2O production rates increased at low O2 or high NO2(-) levels. The results are significant for the development of engineered processes that remove nitrogen from wastewater and for understanding of conditions that favor environmental production of N2O.

Benzene and ethylbenzene removal by denitrifying culture in a horizontal fixed bed anaerobic reactor

Energy Technology Data Exchange (ETDEWEB)

Gusmao, V.R.; Chinalia, F.A.; Sakamoto, I.K.; Varesche [Univ. de Sao Paulo (Brazil). Dept. de Hidraulica e Saneamento; Thiemann, O.H. [Univ. de Sao Paulo (Brazil). Inst. de Fisica de Sao Carlos

2004-07-01

Benzene, ethylbenzene, toluene, and xylene are toxic and are important constituents of gasoline and other petroleum fuels. These compounds are potential health hazards because of their high solubility and hence their ability to contaminate groundwater. Anaerobic immobilized biomass is a way of treating wastewater contaminated with the above compounds. The performance of a specially adapted biofilm is critical in the viability of this idea. In this investigation, an especially adapted biofilm was obtained using a denitrifying bacterial strain isolated from a slaughterhouse wastewater treatment plant. The strain was cultured in a liquid medium with added ethanol, nitrate, ethylbenzene, and benzene. To assess the viability of the strain for the purposes of degradation of ethylbenzene, and benzene two separate horizontal reactors were prepared with polyurethane foam in order to immobilize the biomass. Various concentrations of the two compounds were admitted. At high concentrations chemical oxygen demand decreased dramatically and benzene and ethylbenzene removal almost 100 per cent. DNA sequencing of the biofilm showed that Paracoccus versutus was the dominant species in the ethylbenzene reactor. 7 refs., 6 figs.

Microbial activity in subsurface samples before and during nitrate-enhanced bioremediation

International Nuclear Information System (INIS)

Thomas, J.M.; Gordy, V.R.; Bruce, C.L.; Ward, C.H.; Hutchins, S.R.; Sinclair, J.L.

1995-01-01

A study was conducted to determine the microbial activity at a site contaminated with JP-4 jet fuel before and during nitrate-enhanced bioremediation. Samples at three depths from six different locations were collected aseptically under anaerobic conditions before and during treatment. Cores were located in or close to the source of contamination, downgradient of the source, or outside the zone of contamination. Parameters for microbial characterization included (1) viable counts of aerobic heterotrophic, JP-4 degrading, and oligotrophic bacteria; (2) the most probable number (MPN) of aerobic and anaerobic protozoa; (3) the MPN of total denitrifiers; and (4) the MPN of denitrifiers in hydrocarbon-amended microcosms. The results indicate that the total number of denitrifiers increased by an order of magnitude during nitrate-enhanced bioremediation in most samples. The number of total heterotrophs and JP-4-degrading microorganisms growing aerobically also increased. In addition, the first anaerobic protozoa associated with hydrocarbon-contaminated subsurface materials were detected

Effects of porous carrier size on biofilm development, microbial distribution and nitrogen removal in microaerobic bioreactors

KAUST Repository

Ahmad, Muhammad; Liu, Sitong; Mahmood, Nasir; Mahmood, Asif; Ali, Muhammad; Zheng, Maosheng; Ni, Jinren

2017-01-01

In this study, effects of porous carrier’s size (polyurethane-based) on microbial characteristics were systematically investigated in addition to nitrogen removal performance in six microaerobic bioreactors. Among different sized carriers (50, 30, 20, 15,10, 5 mm), 15 mm carrier showed highest nitrogen removal (98%) due to optimal micro-environments created for aerobic nitrifiers in outer layer (0∼7 mm), nitrifiers and denitrifiers in middle layer (7∼10 mm) and anaerobic denitrifiers in inner layer (10∼15 mm). Candidatus brocadia, a dominant anammox bacteria, was solely concentrated close to centroid (0∼70 μm) and strongly co-aggregated with other bacterial communities in the middle layer of the carrier. Contrarily, carriers with a smaller (<15 mm) or larger size (>15 mm) either destroy the effective zone for anaerobic denitrifiers or damage the microaerobic environments due to poor mass transfer. This study is of particular use for optimal design of carriers in enhancing simultaneous nitrification-denitrification in microaerobic wastewater treatment processes.

Effects of porous carrier size on biofilm development, microbial distribution and nitrogen removal in microaerobic bioreactors

KAUST Repository

Ahmad, Muhammad

2017-03-15

In this study, effects of porous carrier’s size (polyurethane-based) on microbial characteristics were systematically investigated in addition to nitrogen removal performance in six microaerobic bioreactors. Among different sized carriers (50, 30, 20, 15,10, 5 mm), 15 mm carrier showed highest nitrogen removal (98%) due to optimal micro-environments created for aerobic nitrifiers in outer layer (0∼7 mm), nitrifiers and denitrifiers in middle layer (7∼10 mm) and anaerobic denitrifiers in inner layer (10∼15 mm). Candidatus brocadia, a dominant anammox bacteria, was solely concentrated close to centroid (0∼70 μm) and strongly co-aggregated with other bacterial communities in the middle layer of the carrier. Contrarily, carriers with a smaller (<15 mm) or larger size (>15 mm) either destroy the effective zone for anaerobic denitrifiers or damage the microaerobic environments due to poor mass transfer. This study is of particular use for optimal design of carriers in enhancing simultaneous nitrification-denitrification in microaerobic wastewater treatment processes.

Diversity of Metabolically Active Bacteria in Water-Flooded High-Temperature Heavy Oil Reservoir

Directory of Open Access Journals (Sweden)

Tamara N. Nazina

2017-04-01

Full Text Available The goal of this work was to study the overall genomic diversity of microorganisms of the Dagang high-temperature oilfield (PRC and to characterize the metabolically active fraction of these populations. At this water-flooded oilfield, the microbial community of formation water from the near-bottom zone of an injection well where the most active microbial processes of oil degradation occur was investigated using molecular, cultural, radiotracer, and physicochemical techniques. The samples of microbial DNA and RNA from back-flushed water were used to obtain the clone libraries for the 16S rRNA gene and cDNA of 16S rRNA, respectively. The DNA-derived clone libraries were found to contain bacterial and archaeal 16S rRNA genes and the alkB genes encoding alkane monooxygenases similar to those encoded by alkB-geo1 and alkB-geo6 of geobacilli. The 16S rRNA genes of methanogens (Methanomethylovorans, Methanoculleus, Methanolinea, Methanothrix, and Methanocalculus were predominant in the DNA-derived library of Archaea cloned sequences; among the bacterial sequences, the 16S rRNA genes of members of the genus Geobacillus were the most numerous. The RNA-derived library contained only bacterial cDNA of the 16S rRNA sequences belonging to metabolically active aerobic organotrophic bacteria (Tepidimonas, Pseudomonas, Acinetobacter, as well as of denitrifying (Azoarcus, Tepidiphilus, Calditerrivibrio, fermenting (Bellilinea, iron-reducing (Geobacter, and sulfate- and sulfur-reducing bacteria (Desulfomicrobium, Desulfuromonas. The presence of the microorganisms of the main functional groups revealed by molecular techniques was confirmed by the results of cultural, radioisotope, and geochemical research. Functioning of the mesophilic and thermophilic branches was shown for the microbial food chain of the near-bottom zone of the injection well, which included the microorganisms of the carbon, sulfur, iron, and nitrogen cycles.

Potential rates of ammonium oxidation, nitrite oxidation, nitrate reduction and denitrification in the young barley rhizosphere

DEFF Research Database (Denmark)

Højberg, Ole; Binnerup, S. J.; Sørensen, Jan

1996-01-01

Potential activities (enzyme contents) of ammonium (NH4+) oxidizing, nitrite (NO2-) oxidizing, nitrate (NO3-) reducing and denitrifying bacteria were measured in bulk and rhizosphere soil obtained from young barley plants in the field. The activities as well as pools of inorganic N (NH4+, NO2...

Impact Assessment and Bioremediation of oil Contaminated Soil: A ...

African Journals Online (AJOL)

MICHAEL HORSFALL

remediation process was descriptive using denitrifying bacteria for 27 days at 7 days intervals. The results ... environmental degradation has also been witnessed in the Niger ... in which toxic waste was deposited in pretence of ... wrapped in a well labeled foil paper to avoid ... organometallic compounds that migrate with.

Performance of Denitrifying Bioreactors at Reducing  Agricultural Nitrogen Pollution in a Humid  Subtropical Coastal Plain Climate

Directory of Open Access Journals (Sweden)

Timothy Rosen

2017-02-01

Full Text Available Denitrifying bioreactors are an agricultural best management practice developed in the  midwestern United States to treat agricultural drainage water enriched with nitrate‐nitrogen (NO3N. The practice is spreading rapidly to agricultural regions with poor water quality due to nutrient  enrichment. This makes it imperative to track bioreactor NO3‐N reduction efficiency as this practice  gets deployed to new regions. This study evaluated the application and performance of denitrifying  bioreactors in the humid subtropical coastal plain environment of the Chesapeake Bay catchment to  provide data about regionally specific NO3‐N reduction efficiencies. NO3‐N samples were taken  before  and  after  treatment  at  three  denitrifying  bioreactors,  in  addition  to  other  nutrients  (orthophosphate‐phosphorus,  PO4‐P;  ammonium‐nitrogen,  NH4‐N;  total  nitrogen,  TN;  total  phosphorus,  TP  and  water  quality  parameters  (dissolved  oxygen,  DO;  oxidation  reduction  potential,  ORP;  pH;  specific  conductance,  SPC.  Total  removal  ranged  drastically  between  bioreactors from 10 to 133 kg N, with removal efficiencies of 9.0% to 62% and N removal rates of  0.21 to 5.36 g N removed per m3 of bioreactor per day. As the first bioreactor study in the humid  subtropical coastal plain, this data provides positive proof of concept that denitrifying bioreactor is  another tool for reducing N loads in agricultural tile drainage in this region.

Increasing tetracycline concentrations on the performance and communities of mixed microalgae-bacteria photo-bioreactors

KAUST Repository

Xiong, Yanghui

2017-12-11

This study investigated the impact of varying concentrations of tetracycline on the performance of mixed microalgae-bacteria photo-bioreactors. Photo-bioreactors were assessed for their ability to remove carbon dioxide (CO2) from the biogas of anaerobic membrane bioreactor (anMBR), and nutrients from the anaerobic effluent. The varying concentrations of tetracycline had no impact on the removal of CO2 from biogas. 29% v/v of CO2 was completely removed to generate >20% v/v of oxygen (O2) in all reactors. Removal of nutrients and biomass was not affected at low concentrations of tetracycline (≤150μg/L), but 20mg/L of tetracycline lowered the biomass generation and removal efficiencies of phosphate. Conversely, high chlorophyll a and b content was observed at 20mg/L of tetracycline. High tetracycline level had no impact on the diversity of 18S rRNA gene-based microalgal communities but adversely affected the 16S rRNA gene-based microbial communities. Specifically, both Proteobacteria and Bacteroidetes phyla decreased in relative abundance but not phylum Chloroplast. Additionally, both nitrogen-fixing (e.g. Flavobacterium, unclassified Burkholderiales and unclassified Rhizobiaceae) and denitrifying groups (e.g. Hydrogenophaga spp.) were significantly reduced in relative abundance at high tetracycline concentration. Phosphate-accumulating microorganisms, Acinetobacter spp. and Pseudomonas spp. were similarly reduced upon exposure to high tetracycline concentration. Unclassified Comamonadaceae, however, increased in relative abundance, which correlated with an increase in the abundance of tetracycline resistance genes associated with efflux pump mechanism. Overall, the findings demonstrate that antibiotic concentrations in municipal wastewaters will not significantly affect the removal of nutrients by the mixed microalgae-bacteria photo-bioreactors. However, utilizing such photo-bioreactors as a polishing step for anMBRs that treat wastewaters with high tetracycline

Microbial populations and activities of mangrove, restinga and Atlantic forest soils from Cardoso Island, Brazil.

Science.gov (United States)

Pupin, B; Nahas, E

2014-04-01

Mangroves provide a distinctive ecological environment that differentiates them from other ecosystems. This study deal to evaluate the frequency of microbial groups and the metabolic activities of bacteria and fungi isolated from mangrove, restinga and Atlantic forest soils. Soil samples were collected during the summer and winter at depths of 0-2, 2-5 and 5-10 cm. Except for fungi, the counts of the total, sporulating, Gram-negative, actinomycetes, nitrifying and denitrifying bacteria decreased significantly in the following order: Atlantic forest >mangrove > restinga. The counts of micro-organisms decreased by 11 and 21% from the surface to the 2-5 and 5-10 cm layers, but denitrifying bacteria increased by 44 and 166%, respectively. A larger growth of micro-organisms was verified in the summer compared with the winter, except for actinomycetes and fungi. The average frequency of bacteria isolated from mangrove, restinga and Atlantic forest soils was 95, 77 and 78%, and 93, 90 and 95% for fungi, respectively. Bacteria were amylolytic (33%), producers of acid phosphatase (79%) and solubilizers (18%) of inorganic phosphate. The proportions of fungi were 19, 90 and 27%. The mangrove soil studied had higher chemical characteristics than the Atlantic forest, but the high salinity may have restricted the growth of microbial populations. Estimates of the microbial counts and activities were important to elucidate the differences of mangrove ecosystem from restinga and Atlantic forest. © 2013 The Society for Applied Microbiology.

Mathematical modeling of simultaneous carbon-nitrogen-sulfur removal from industrial wastewater.

Science.gov (United States)

Xu, Xi-Jun; Chen, Chuan; Wang, Ai-Jie; Ni, Bing-Jie; Guo, Wan-Qian; Yuan, Ye; Huang, Cong; Zhou, Xu; Wu, Dong-Hai; Lee, Duu-Jong; Ren, Nan-Qi

2017-01-05

A mathematical model of carbon, nitrogen and sulfur removal (C-N-S) from industrial wastewater was constructed considering the interactions of sulfate-reducing bacteria (SRB), sulfide-oxidizing bacteria (SOB), nitrate-reducing bacteria (NRB), facultative bacteria (FB), and methane producing archaea (MPA). For the kinetic network, the bioconversion of C-N by heterotrophic denitrifiers (NO 3 - →NO 2 - →N 2 ), and that of C-S by SRB (SO 4 2- →S 2- ) and SOB (S 2- →S 0 ) was proposed and calibrated based on batch experimental data. The model closely predicted the profiles of nitrate, nitrite, sulfate, sulfide, lactate, acetate, methane and oxygen under both anaerobic and micro-aerobic conditions. The best-fit kinetic parameters had small 95% confidence regions with mean values approximately at the center. The model was further validated using independent data sets generated under different operating conditions. This work was the first successful mathematical modeling of simultaneous C-N-S removal from industrial wastewater and more importantly, the proposed model was proven feasible to simulate other relevant processes, such as sulfate-reducing, sulfide-oxidizing process (SR-SO) and denitrifying sulfide removal (DSR) process. The model developed is expected to enhance our ability to predict the treatment of carbon-nitrogen-sulfur contaminated industrial wastewater. Copyright © 2016 Elsevier B.V. All rights reserved.

Development of hydrogen oxidizing bacteria using hydrogen from radiolysis or metal corrosion; Developpement de populations microbiennes oxydant l'hydrogene produit par radiolyse ou par corrosion des metaux

Energy Technology Data Exchange (ETDEWEB)

Libert, M F; Sellier, R; Marty, V; Camaro, S [CEA Cadarache, Dept. d' Entreposage et de Stockage des Dechets (DCC/DESD/SEP), 13 - Saint-Paul-lez-Durance (France)

2000-07-01

The effect of many parameters need to be studied to characterize the long term behavior of nuclear waste in a deep repository. These parameters concern the chemical effects, radiolytic effects, mechanical properties, water composition, and microbiological activity. To evaluate microbial activity in such an environment, work was focused on an inventory of key nutrients (C, H, 0, N, P, S) and energy sources required for bacterial growth. The production of hydrogen in the nuclear waste environment leads to the growth of hydrogen oxidizing bacteria, which modify the gas production balance. A deep repository containing bituminized waste drums implies several sources of hydrogen: - water radiolysis; -corrosion of metal containers; - radiolysis of the embedding matrix (bitumen). Two deep geological disposal conditions leading to H{sub 2} production in a bituminized nuclear waste environment were simulated in the present study: - H{sub 2} production by iron corrosion under anaerobic conditions was simulated by adding 10% of H{sub 2} in the atmosphere; - H{sub 2} production by radiolysis of bitumen matrix was approached by subjecting this material to external gamma irradiation with a dose rate near real conditions (6 Gy/h). The presence of dissolved H{sub 2} in water allows the growth of hydrogen oxidizing bacteria leading to: - CO{sub 2} and N{sub 2} production; - H{sub 2} consumption; - lower NO{sub 3}{sup -} concentration caused by reduction to nitrogen. In the first case, hydrogen consumption is limited by the NO{sub 3}{sup -} release rate from the bitumen matrix. In the second case, however, under gamma radiation at a low dose rate, hydrogen production is weak, and the hydrogen is completely consumed by microorganisms. Knowledge about these hydrogen oxidizing bacteria is just beginning to emerge. Heterotrophic denitrifying bacteria adapt well to hydrogen metabolism (autotrophic metabolism) by oxidizing H{sub 2} instead of hydrocarbons. (authors)

Anaerobic bacteria

Science.gov (United States)

Anaerobic bacteria are bacteria that do not live or grow when oxygen is present. In humans, these bacteria ... Brook I. Diseases caused by non-spore-forming anaerobic bacteria. In: Goldman L, Schafer AI, eds. Goldman-Cecil ...

Constraints on mechanisms and rates of anaerobic oxidation of methane by microbial consortia: process-based modeling of ANME-2 archaea and sulfate reducing bacteria interactions

Directory of Open Access Journals (Sweden)

B. Orcutt

2008-11-01

Full Text Available Anaerobic oxidation of methane (AOM is the main process responsible for the removal of methane generated in Earth's marine subsurface environments. However, the biochemical mechanism of AOM remains elusive. By explicitly resolving the observed spatial arrangement of methanotrophic archaea and sulfate reducing bacteria found in consortia mediating AOM, potential intermediates involved in the electron transfer between the methane oxidizing and sulfate reducing partners were investigated via a consortium-scale reaction transport model that integrates the effect of diffusional transport with thermodynamic and kinetic controls on microbial activity. Model simulations were used to assess the impact of poorly constrained microbial characteristics such as minimum energy requirements to sustain metabolism and cell specific rates. The role of environmental conditions such as the influence of methane levels on the feasibility of H₂, formate and acetate as intermediate species, and the impact of the abundance of intermediate species on pathway reversal were examined. The results show that higher production rates of intermediates via AOM lead to increased diffusive fluxes from the methane oxidizing archaea to sulfate reducing bacteria, but the build-up of the exchangeable species can cause the energy yield of AOM to drop below that required for ATP production. Comparison to data from laboratory experiments shows that under the experimental conditions of Nauhaus et al. (2007, none of the potential intermediates considered here is able to support metabolic activity matching the measured rates.

Denitrification in groundwater at uranium mill tailings sites

International Nuclear Information System (INIS)

Goering, Timothy J.; Groffman, Armando; Thomson, Bruce

1992-01-01

Nitrates are a major contaminant in groundwater at many Uranium Mill Tailings Remedial Action (UMTRA) sites. Microbial denitrification, the transformation of nitrate to nitrogen gas, may be occurring in groundwater at several UMTRA sites. Denitrification is a biologically mediated process whereby facultative anaerobes use nitrate for respiration under anaerobic conditions. Denitrifying bacteria are ubiquitous in soils, sediments, and water. Denitrification requires nitrate, organic carbon, oxygen-limiting conditions, and trace nutrients, especially phosphorus. The lack of organic carbon is the most common limiting factor for denitrification. Denitrification occurs under a limited range of temperature and pH. The uranium milling processes used at UMTRA sites provided a readily available source of carbon and nitrates for denitrifying bacteria. At the Maybell, Colorado, site, the denitrifying organisms Pseudomonas, Flavobacterium and Acinetobacter were identified in core samples of materials from beneath the tailings. In addition, microcosm experiments simulating aquifer conditions beneath the tailings pile showed an average 40 percent decrease in nitrate concentrations over 13 days. At the New Rifle, Colorado, site, aquifer conditions appear favorable for denitrification. Nitrate and organic carbon are readily available in the groundwater, and redox conditions beneath and downgradient of the tailings pile are relatively anoxic. Downgradient from the tailings, total nitrogen is being removed from the groundwater system at a greater rate than the geochemically conservative anion, chloride. This removal may be due to denitrification and adsorption of ammonium onto clay and silt particles. (author)

Denitrification in groundwater at uranium mill tailings sites

Energy Technology Data Exchange (ETDEWEB)

Goering, Timothy J [Jacobs Engineering Group, Inc., Albuquerque, NM (United States); Groffman, Armando [Roy F. Weston, Inc., Albuquerque, NM (United States); Thomson, Bruce [University of New Mexico, Albuquerque, NM (United States)

1992-07-01

Nitrates are a major contaminant in groundwater at many Uranium Mill Tailings Remedial Action (UMTRA) sites. Microbial denitrification, the transformation of nitrate to nitrogen gas, may be occurring in groundwater at several UMTRA sites. Denitrification is a biologically mediated process whereby facultative anaerobes use nitrate for respiration under anaerobic conditions. Denitrifying bacteria are ubiquitous in soils, sediments, and water. Denitrification requires nitrate, organic carbon, oxygen-limiting conditions, and trace nutrients, especially phosphorus. The lack of organic carbon is the most common limiting factor for denitrification. Denitrification occurs under a limited range of temperature and pH. The uranium milling processes used at UMTRA sites provided a readily available source of carbon and nitrates for denitrifying bacteria. At the Maybell, Colorado, site, the denitrifying organisms Pseudomonas, Flavobacterium and Acinetobacter were identified in core samples of materials from beneath the tailings. In addition, microcosm experiments simulating aquifer conditions beneath the tailings pile showed an average 40 percent decrease in nitrate concentrations over 13 days. At the New Rifle, Colorado, site, aquifer conditions appear favorable for denitrification. Nitrate and organic carbon are readily available in the groundwater, and redox conditions beneath and downgradient of the tailings pile are relatively anoxic. Downgradient from the tailings, total nitrogen is being removed from the groundwater system at a greater rate than the geochemically conservative anion, chloride. This removal may be due to denitrification and adsorption of ammonium onto clay and silt particles. (author)

Effect of bio-column composed of aged refuse on methane abatement--a novel configuration of biological oxidation in refuse landfill.

Science.gov (United States)

Han, Dan; Zhao, Youcai; Xue, Binjie; Chai, Xiaoli

2010-01-01

An experimental bio-column composed of aged refuse was installed around the exhaust pipe as a new way to mitigate methane in refuse landfill. One of the objectives of this work was to assess the effect of aged refuse thickness in bio-column on reducing CH4 emissions. Over the study period, methane oxidation was observed at various thicknesses, 5 cm (small size), 10 cm (middle size) and 15 cm (large size), representing one to three times of pipeline diameters. The middle and large size both showed over 90% methane conversion, and the highest methane conversion rate of above 95% occurred in the middle-size column cell. Michaelis-Menten equation addressed the methanotrophs diffusion in different layers of the bio-columns. Maximum methanotrophic activity (Vmax) measured at the three thicknesses ranged from 6.4 x 10(-3) to 15.6 x 10(-3) units, and the half-saturation value (K(M)) ranged from 0.85% to 1.67%. Both the highest Vmax and K(M) were observed at the middle-size of the bio-column, as well as the largest methanotrophs population, suggesting a significant efficiency of methane mitigation happened in the optimum zone with greatest affinity and methanotrophic bacteria activities. Therefore, bio-column is a potential style for methane abatement in landfill, and the aged refuse both naturally formed and artificially placed in the column plays a critical role in CH4 emission.

Crenothrix are major methane consumers in stratified lakes.

Science.gov (United States)

Oswald, Kirsten; Graf, Jon S; Littmann, Sten; Tienken, Daniela; Brand, Andreas; Wehrli, Bernhard; Albertsen, Mads; Daims, Holger; Wagner, Michael; Kuypers, Marcel Mm; Schubert, Carsten J; Milucka, Jana

2017-09-01

Methane-oxidizing bacteria represent a major biological sink for methane and are thus Earth's natural protection against this potent greenhouse gas. Here we show that in two stratified freshwater lakes a substantial part of upward-diffusing methane was oxidized by filamentous gamma-proteobacteria related to Crenothrix polyspora. These filamentous bacteria have been known as contaminants of drinking water supplies since 1870, but their role in the environmental methane removal has remained unclear. While oxidizing methane, these organisms were assigned an 'unusual' methane monooxygenase (MMO), which was only distantly related to 'classical' MMO of gamma-proteobacterial methanotrophs. We now correct this assignment and show that Crenothrix encode a typical gamma-proteobacterial PmoA. Stable isotope labeling in combination swith single-cell imaging mass spectrometry revealed methane-dependent growth of the lacustrine Crenothrix with oxygen as well as under oxygen-deficient conditions. Crenothrix genomes encoded pathways for the respiration of oxygen as well as for the reduction of nitrate to N 2 O. The observed abundance and planktonic growth of Crenothrix suggest that these methanotrophs can act as a relevant biological sink for methane in stratified lakes and should be considered in the context of environmental removal of methane.

Regulation of nitrous oxide emission associated with benthic invertebrates

DEFF Research Database (Denmark)

Stief, Peter; Schramm, Andreas

2010-01-01

1. A number of freshwater invertebrate species emit N2O, a greenhouse gas that is produced in their gut by denitrifying bacteria (direct N2O emission). Additionally, benthic invertebrate species may contribute to N2O emission from sediments by stimulating denitrification because of their bioirrig......1. A number of freshwater invertebrate species emit N2O, a greenhouse gas that is produced in their gut by denitrifying bacteria (direct N2O emission). Additionally, benthic invertebrate species may contribute to N2O emission from sediments by stimulating denitrification because...... of their bioirrigation behaviour (indirect N2O emission). 2. Two benthic invertebrate species were studied to determine (i) the dependence of direct N2O emission on the preferred diet of the animals, (ii) the regulation of direct N2O emission by seasonally changing factors, such as body size, temperature and NO3...... emitted by benthic invertebrates can be partially consumed in the sediment (E. danica), non-emitting species can still indirectly contribute to total N2O emission from sediment (S. lutaria)....

Antibiotics and Manure Effects on Microbial Communities Responsible for Nitrous Oxide Emissions from Grasslands

Science.gov (United States)

Semedo, M.; Song, B.; Sparrer, T.; Crozier, C.; Tobias, C. R.; Phillips, R. L.

2015-12-01

Agroecosystems are major contributors of nitrous oxide (N2O) emissions. Denitrification and nitrification are the primary pathways of N2O emission in soils. However, there is uncertainty regarding the organisms responsible for N2O production. Bacteria were previously considered the only microbial N2O source, however, current studies indicate that fungi also produce N2O by denitrification. Denitrifying bacteria can be a source or sink of N2O depending on the presence and expression of nitrous oxide reductase genes (nosZ), encoding for the enzyme converting N2O to N2. Fungal denitrification may produce only N2O as an end product due to missing the nosZ gene. Animal manures applied to agricultural fields can transfer antibiotics to soils as a result of antibiotic use in the livestock industry. These antibiotics target mostly bacteria and may promote fungal growth. The growth inhibition of denitrifying bacteria may favor fungal denitrifiers potentially enhancing N2O emissions. Our objective is to examine the effects of antibiotic exposure and manure fertilization on the microbial communities responsible for N2 and N2O production in grasslands. Soil slurry incubations were conducted with tetracycline at different concentrations. A mesocosm experiment was also performed with soil cores exposed to tetracycline and cow manure. Production of N2O and N2 was measured using gas chromatography with electron capture detector (GC-ECD) and isotope ratio mass spectrometry (IRMS), respectively. Antibiotic inhibition of soil N2 production was found to be dose dependent, reaching up to 80% inhibition with 1g Kg-1 of tetracycline treatment, while N2O production was enhanced up to 8 times. These results suggest higher fungal denitrification with a concomitant decrease in bacterial denitrification after antibiotic exposure. We also found higher N2O fluxes in the soil mesocosms treated with manure plus tetracycline. Quantitative PCR (qPCR) will be conducted to examine the changes in

Biopotentiality of High Efficient Aerobic Denitrifier Bacillus megaterium S379 for Intensive Aquaculture Water Quality Management.

Science.gov (United States)

Gao, Junqian; Gao, Dan; Liu, Hao; Cai, Jiajai; Zhang, Junqi; Qi, Zhengliang

2018-05-24

Excessive nitrite accumulation is a very tough issue for intensive aquaculture. A high efficient aerobic denitrifier Bacillus megaterium S379 with 91.71±0.17% of NO 2 - -N (65 mg L -1 ) removal was successfully isolated for solving the problem. Denitrification of S379 showed excellent environment adaptation that it kept high nitrite removal ratio (more than 85%) when temperature ranged from 25°C to 40°C and pH varied between 7.0 and 9.0, and could endure as high as 560 mg L -1 of NO 2 - -N. Immobilization of S379 could enhance denitrification even when NO 2 - -N adding amount got to 340 mg L -1 . Immobilized cells also showed well pollutants removal performance in aquaculture wastewater treatment. Moreover, S379 possessed positive hydrolase activities for starch, casein, cellulose and fat and bore more than 60 ppt of salinity. Totally, all the results revealed significant potentiality of immobilized S379 applied in aquaculture water quality management. Copyright © 2018 Elsevier Ltd. All rights reserved.

Methane-Oxidizing Enzymes: An Upstream Problem in Biological Gas-to-Liquids Conversion

OpenAIRE

Lawton, Thomas J.; Rosenzweig, Amy C.

2016-01-01

Biological conversion of natural gas to liquids (Bio-GTL) represents an immense economic opportunity. In nature, aerobic methanotrophic bacteria and anaerobic archaea are able to selectively oxidize methane using methane monooxygenase (MMO) and methyl coenzyme M reductase (MCR) enzymes. Although significant progress has been made toward genetically manipulating these organisms for biotechnological applications, the enzymes themselves are slow, complex, and not recombinantly tractable in tradi...

2012 Molecular Basis of Microbial One-Carbon Metabolism Gordon Research Conferences and Gordon Research Seminar, August 4-10,2012

Energy Technology Data Exchange (ETDEWEB)

Hanson, Thomas

2012-08-10

The 2012 Gordon Conference will present and discuss cutting-edge research in the field of microbial metabolism of C1 compounds. The conference will feature the roles and application of C1 metabolism in natural and synthetic systems at scales from molecules to ecosystems. The conference will stress molecular aspects of the unique metabolism exhibited by autotrophic bacteria, methanogens, methylotrophs, aerobic and anaerobic methanotrophs, and acetogens.

A novel methanotroph in the genus Methylomonas that contains a distinct clade of soluble methane monooxygenase.

Science.gov (United States)

Nguyen, Ngoc-Loi; Yu, Woon-Jong; Yang, Hye-Young; Kim, Jong-Geol; Jung, Man-Young; Park, Soo-Je; Roh, Seong-Woon; Rhee, Sung-Keun

2017-10-01

Aerobic methane oxidation is a key process in the global carbon cycle that acts as a major sink of methane. In this study, we describe a novel methanotroph designated EMGL16-1 that was isolated from a freshwater lake using the floating filter culture technique. Based on a phylogenetic analysis of 16S rRNA gene sequences, the isolate was found to be closely related to the genus Methylomonas in the family Methylococcaceae of the class Gammaproteobacteria with 94.2-97.4% 16S rRNA gene similarity to Methylomonas type strains. Comparison of chemotaxonomic and physiological properties further suggested that strain EMGL16-1 was taxonomically distinct from other species in the genus Methylomonas. The isolate was versatile in utilizing nitrogen sources such as molecular nitrogen, nitrate, nitrite, urea, and ammonium. The genes coding for subunit of the particulate form methane monooxygenase (pmoA), soluble methane monooxygenase (mmoX), and methanol dehydrogenase (mxaF) were detected in strain EMGL16-1. Phylogenetic analysis of mmoX indicated that mmoX of strain EMGL16-1 is distinct from those of other strains in the genus Methylomonas. This isolate probably represents a novel species in the genus. Our study provides new insights into the diversity of species in the genus Methylomonas and their environmental adaptations.

[Biosynthesis of the bioprotectant ectoin by aerobic methylotrophic bacteria from methanol].

Science.gov (United States)

Doronina, N V; Ezhov, V A; Beschastnyĭ, A P; Trotsenko, Iu A

2010-01-01

It is shown that neutrophilic methylobacteria Methylophaga thalassica and M. marina have higher rates of growth and ectoin accumulation compared to the haloalkaliphilic species M. alcalica and M. natronia and methanotrophs Methylomicrobium alcaliphilum and M. kenyense. The conditions of M. thalassica cultivation in methanol-containing medium were optimized. The yield of this process reached 60 g/l of absolutely dry biomass containing 15-19% (9-11 g/l) ectoine. The scheme of ectoin isolation from the biomass by extraction and subsequent purification, which allowed obtaining preparations of different degree of purity, was developed.

[Biotechnological potential of methylotrophic bacteria: a review of current status and future prospects].

Science.gov (United States)

Trotsenko, Iu A; Doronina, N V; Khmelenina, V N

2005-01-01

Major results of the authors' findings on the implementation of biotechnological potential of aerobic methylobacteria and methanotrophs for obtaining forage proteins, biopolymers (polybutyrate and polysaccharides), enzymes (oxidoreductases), and bioprotectors (ectoin), as well as for degrading toxic C1 and Cn compounds have been reviewed. Unique features of the structural and functional organization of the metabolism of extremophilic (tolerant) methylotrophs are discussed, with a view for their prospective use in various fields of modern biotechnology, including biocatalysis and nanotechnology.

Activity of type i methanotrophs dominates under high methane concentration: Methanotrophic activity in slurry surface crusts as influenced by methane, oxygen, and inorganic nitrogen

DEFF Research Database (Denmark)

Duan, Yun Feng; Reinsch, Sabine; Ambus, Per

2017-01-01

Livestock slurry is a major source of atmospheric methane (CH4), but surface crusts harboring methane-oxidizing bacteria (MOB) could mediate against CH4 emissions. This study examined conditions for CH4 oxidation by in situ measurements of oxygen (O2) and nitrous oxide (N2O), as a proxy for inorg......Livestock slurry is a major source of atmospheric methane (CH4), but surface crusts harboring methane-oxidizing bacteria (MOB) could mediate against CH4 emissions. This study examined conditions for CH4 oxidation by in situ measurements of oxygen (O2) and nitrous oxide (N2O), as a proxy...... for inorganic N transformations, in intact crusts using microsensors. This was combined with laboratory incubations of crust material to investigate the effects of O2, CH4, and inorganic N on CH4 oxidation, using 13CH4 to trace C incorporation into lipids of MOB. Oxygen penetration into the crust was 2 to 14 mm...

Nitrogen removal in the bioreactor landfill system with intermittent aeration at the top of landfilled waste

International Nuclear Information System (INIS)

He Ruo; Shen Dongsheng

2006-01-01

High ammonia concentration of recycled landfill leachate makes it very difficult to treat. In this work, a vertical aerobic/anoxic/anaerobic lab-scale bioreactor landfill system, which was constructed by intermittent aeration at the top of landfilled waste, as a bioreactor for in situ nitrogen removal was investigated during waste stabilization. Intermittent aeration at the top of landfilled waste might stimulate the growth of nitrifying bacteria and denitrifying bacteria in the top and middle layers of waste. The nitrifying bacteria population for the landfill bioreactor with intermittent aeration system reached between10 6 and 10 8 cells/dry g waste, although it decreased 2 orders of magnitude on day 30, due to the inhibitory effect of the acid environment and high organic matter in the landfilled waste. The denitrifying bacteria population increased by between 4 and 13 orders of magnitude compared with conventional anaerobic landfilled waste layers. Leachate NO 3 - -N concentration was very low in both two experimental landfill reactors. After 105 days operation, leachate NH 4 + -N and TN concentrations for the landfill reactor with intermittent aeration system dropped to 186 and 289 mg/l, respectively, while they were still kept above 1000 mg/l for the landfill reactor without intermittent aerobic system. In addition, there is an increase in the rate of waste stabilization as well as an increase of 12% in the total waste settlement for the landfill reactor with intermittent aeration system

Technology summary of the in situ bioremediation demonstration (methane biostimulation) via horizontal wells at the Savannah River Site Integrated Demonstration Project

International Nuclear Information System (INIS)

Hazen, T.C.; Looney, B.B.; Fliermans, C.B.; Eddy-Dilek, C.A.; Lombard, K.H.; Enzien, M.V.; Dougherty, J.M.; Wear, J.

1994-01-01

The US Department of Energy, Office of Technology Development, has been sponsoring full-scale environmental restoration technology demonstrations for the past 4 years. The Savannah River Site Integrated Demonstration focuses on ''Clean-up of Soils ad Groundwater Contaminated with Chlorinated VOCs.'' Several laboratories including our own had demonstrated the ability of methanotrophic bacteria to completely degrade or mineralize chlorinated solvents, and these bacteria were naturally found in soil and aquifer material. Thus the test consisted of injection of methane mixed with air into the contaminated aquifer via a horizontal well and extraction from the vadose zone via a parallel horizontal well

The effect of poly-β-hydroxyalkanoates degradation rate on nitrous oxide production in a denitrifying phosphorus removal system.

Science.gov (United States)

Wei, Yan; Wang, Shuying; Ma, Bin; Li, Xiyao; Yuan, Zhiguo; He, Yuelan; Peng, Yongzhen

2014-10-01

Poly-β-hydroxyalkanoates (PHAs) and free nitrous acid (FNA) have been revealed as significant factors causing nitrous oxide (N2O) production in denitrifying phosphorus removal systems. In this study, the effect of PHA degradation rate on N2O production was studied at low FNA levels. N2O production always maintained at approximately 40% of the amount of nitrite reduced independent of the PHA degradation rate. The electrons distributed to nitrite reduction were 1.6 times that to N2O reduction. This indicated that electron competition between these two steps was not affected by the PHA degradation rate. Continuous feed of nitrate was proposed, and demonstrated to reduce N2O accumulation by 75%. While being kept low, a possible compounding effect of a low-level FNA could not be ruled out. The sludge used likely contained both polyphosphate- and glycogen-accumulating organisms, and the results could not be simply attributed to either group of organisms. Copyright © 2014 Elsevier Ltd. All rights reserved.

Composition of methane-oxidizing bacterial communities as a function of nutrient loading in the Florida everglades.

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Chauhan, Ashvini; Pathak, Ashish; Ogram, Andrew

2012-10-01

Agricultural runoff of phosphorus (P) in the northern Florida Everglades has resulted in several ecosystem level changes, including shifts in the microbial ecology of carbon cycling, with significantly higher methane being produced in the nutrient-enriched soils. Little is, however, known of the structure and activities of methane-oxidizing bacteria (MOB) in these environments. To address this, 0 to 10 cm plant-associated soil cores were collected from nutrient-impacted (F1), transition (F4), and unimpacted (U3) areas, sectioned in 2-cm increments, and methane oxidation rates were measured. F1 soils consumed approximately two-fold higher methane than U3 soils; additionally, most probable numbers of methanotrophs were 4-log higher in F1 than U3 soils. Metabolically active MOB containing pmoA sequences were characterized by stable-isotope probing using 10 % (v/v) (13)CH(4). pmoA sequences, encoding the alpha subunit of methane monooxygenase and related to type I methanotrophs, were identified from both impacted and unimpacted soils. Additionally, impacted soils also harbored type II methanotrophs, which have been shown to exhibit preferences for high methane concentrations. Additionally, across all soils, novel pmoA-type sequences were also detected, indicating presence of MOB specific to the Everglades. Multivariate statistical analyses confirmed that eutrophic soils consisted of metabolically distinct MOB community that is likely driven by nutrient enrichment. This study enhances our understanding on the biological fate of methane being produced in productive wetland soils of the Florida Everglades and how nutrient-enrichment affects the composition of methanotroph bacterial communities.

Inference of coastal submergence from the study of beach rock off Visakhapatnam

Digital Repository Service at National Institute of Oceanography (India)

Rajamanickam, G.V.; Rao, K.M.; Rao, T.C.S.

. and the respective weight percentages were used tocalculat~the size parameters following Folk (I974). The composition of the cementing material w.as examined by directly staining with alizarin red-S on a weB poJished and Hel etched rock surface and by X... Bacteria and on the action of denitrifying bacteria in tropical and temperate seas, pap. Totuga Lab Carnegie Inst., Wash Publ., p. 182. Emery, K.O. and Cox, D.C., (I 956) Beach rock in the Jawaliah Islands, Pacific Science, V. 10, pp', 383. Folk, R.L, (1974...

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.

Nitrous Oxide Emission and Denitrifier Abundance in Two Agricultural Soils Amended with Crop Residues and Urea in the North China Plain.

Directory of Open Access Journals (Sweden)

Jianmin Gao

Full Text Available The application of crop residues combined with Nitrogen (N fertilizer has been broadly adopted in China. Crop residue amendments can provide readily available C and N, as well as other nutrients to agricultural soils, but also intensify the N fixation, further affecting N2O emissions. N2O pulses are obviously driven by rainfall, irrigation and fertilization. Fertilization before rainfall or followed by flooding irrigation is a general management practice for a wheat-maize rotation in the North China Plain. Yet, little is known on the impacts of crop residues combined with N fertilizer application on N2O emission under high soil moisture content. A laboratory incubation experiment was conducted to investigate the effects of two crop residue amendments (maize and wheat, individually or in combination with N fertilizer, on N2O emissions and denitrifier abundance in two main agricultural soils (one is an alluvial soil, pH 8.55, belongs to Ochri-Aquic Cambosols, OAC, the other is a lime concretion black soil, pH 6.61, belongs to Hapli-Aquic Vertosols, HAV under 80% WFPS (the water filled pore space in the North China Plain. Each type soil contains seven treatments: a control with no N fertilizer application (CK, N0, 200 kg N ha-1 (N200, 250 kg N ha-1 (N250, maize residue plus N200 (MN200, maize residue plus N250 (MN250, wheat residue plus N200 (WN200 and wheat residue plus N250 (WN250. Results showed that, in the HAV soil, MN250 and WN250 increased the cumulative N2O emissions by 60% and 30% compared with N250 treatment, respectively, but MN200 and WN200 decreased the cumulative N2O emissions by 20% and 50% compared with N200. In the OAC soil, compared with N200 or N250, WN200 and WN250 increased the cumulative N2O emission by 40%-50%, but MN200 and MN250 decreased the cumulative N2O emission by 10%-20%. Compared with CK, addition of crop residue or N fertilizer resulted in significant increases in N2O emissions in both soils. The cumulative N2O

Isotopes and their possible use as bio markers of microbial products

International Nuclear Information System (INIS)

Zyakun, A.M.

1992-01-01

The purpose of this presentation is to determine the range of possible variations in the distribution of carbon isotopes ( 12 C and 13 C) in the metabolic products of the basic biological systems (production of organic matter by photosynthetic bacteria, its consumption by heterotrophic organisms, biological production of methane, its utilization by methanotrophic organisms, biological production of carbon monoxide and its bacterial oxidation). 32 refs., 11 figs., 3 tabs

Anaerobic degradation of benzene by marine sulfate-reducing bacteria

Science.gov (United States)

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

2010-05-01

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

Mitigation of nitrous oxide (N2O) emissions from denitrifying fluidized bed bioreactors (DFBBRs) using calcium.

Science.gov (United States)

Eldyasti, Ahmed; Nakhla, George; Zhu, Jesse

2014-12-01

Nitrous oxide (N2O) is a significant anthropogenic greenhouse gases (AnGHGs) emitted from biological nutrient removal (BNR) processes. In this study, N2O production from denitrifying fluidized bed bioreactors (DFBBR) was reduced using calcium (Ca2+) dosage. The DFBBRs were operated on a synthetic municipal wastewater at four different calcium concentrations ranging from the typical municipal wastewater Ca2+ concentration (60 mg Ca2+/L) to 240 mg Ca2+/L at two different COD/N ratios. N2O emission rates, extracellular polymeric substances (EPS), water quality parameters, and microscopic images were monitored regularly in both phases. Calcium concentrations played a significant role in biofilm morphology with the detachment rates for R120Ca, R180Ca, and R240Ca 75% lower than for R60Ca, respectively. The N2O conversion rate at the typical municipal wastewater Ca2+ concentration (R60Ca) was about 0.53% of the influent nitrogen loading as compared with 0.34%, 0.42%, and 0.41% for R120Ca, R180Ca, and R240Ca, respectively corresponding to 21-36% reduction. Copyright © 2014 Elsevier Ltd. All rights reserved.

Continuous measurements of nitrous oxide isotopomers during incubation experiments

Science.gov (United States)

Winther, Malte; Balslev-Harder, David; Christensen, Søren; Priemé, Anders; Elberling, Bo; Crosson, Eric; Blunier, Thomas

2018-02-01

Nitrous oxide (N2O) is an important and strong greenhouse gas in the atmosphere. It is produced by microbes during nitrification and denitrification in terrestrial and aquatic ecosystems. The main sinks for N2O are turnover by denitrification and photolysis and photo-oxidation in the stratosphere. In the linear N = N = O molecule 15N substitution is possible in two distinct positions: central and terminal. The respective molecules, 14N15N16O and 15N14N16O, are called isotopomers. It has been demonstrated that N2O produced by nitrifying or denitrifying microbes exhibits a different relative abundance of the isotopomers. Therefore, measurements of the site preference (difference in the abundance of the two isotopomers) in N2O can be used to determine the source of N2O, i.e., nitrification or denitrification. Recent instrument development allows for continuous position-dependent δ15N measurements at N2O concentrations relevant for studies of atmospheric chemistry. We present results from continuous incubation experiments with denitrifying bacteria, Pseudomonas fluorescens (producing and reducing N2O) and Pseudomonas chlororaphis (only producing N2O). The continuous measurements of N2O isotopomers reveals the transient isotope exchange among KNO3, N2O, and N2. We find bulk isotopic fractionation of -5.01 ‰ ± 1.20 for P. chlororaphis, in line with previous results for production from denitrification. For P. fluorescens, the bulk isotopic fractionation during production of N2O is -52.21 ‰ ± 9.28 and 8.77 ‰ ± 4.49 during N2O reduction.The site preference (SP) isotopic fractionation for P. chlororaphis is -3.42 ‰ ± 1.69. For P. fluorescens, the calculations result in SP isotopic fractionation values of 5.73 ‰ ± 5.26 during production of N2O and 2.41 ‰ ± 3.04 during reduction of N2O. In summary, we implemented continuous measurements of N2O isotopomers during incubation of denitrifying bacteria and believe that similar experiments will lead to a better

Effect of aging on the concentrations of nitrous oxide in exhaled air

Energy Technology Data Exchange (ETDEWEB)

Mitsui, T.; Shimaoka, K.; Miyamura, M. [Research Center of Health, Physical Fitness and Sports, Nagoya University, Nagoya (Japan); Kato, N. [Chukyo Women`s University, Obu (Japan)

1997-12-03

Trace gases in exhaled air have been used as a simple means of assessing metabolic reactions. The investigations of trace gases derived from bacteria in human exhalation are usually hydrogen (H{sub 2}) or methane (CH{sub 4}). On the other hand, nitrous oxide (N{sub 2}O) is also derived from microorganisms, especially denitrifying bacteria. Although many kinds of denitrifying bacteria have been isolated on and in the human body, there has been few concerning N{sub 2}O. We studied 222 healthy people from the age of 5 to 85 years. The analysis of N{sub 2}O in exhaled air was carried out by a infrared-photoacoustic (IR-PAS) analyzer. It was found that N{sub 2}0 ranged from 0 to 1670 ppbv in exhaled air and that 59% (131) of the subjects were producers N{sub 2}O. A highly significant relationship was observed between age and concentrations of N{sub 2}O (r=0.40, P<0.01). The rate of production in young children and in the aged was significantly higher than that in adults aged 20-39 years (P<0.01), and less than 30% were producers during puberty. The change of normal microflora and in human body with aging may have caused the significant relationship between age and emissions of N{sub 2}O

Efficient simultaneous partial nitrification, anammox and denitrification (SNAD) system equipped with a real-time dissolved oxygen (DO) intelligent control system and microbial community shifts of different substrate concentrations.

Science.gov (United States)

Wen, Xin; Gong, Benzhou; Zhou, Jian; He, Qiang; Qing, Xiaoxia

2017-08-01

Simultaneous partial nitrification, anammox and denitrification (SNAD) process was studied in a sequencing batch biofilm reactor (SBBR) fed with synthetic wastewater in a range of 2200 mgN/L ∼ 50 mgN/L. Important was an external real-time precision dissolved oxygen (DO) intelligent control system that consisted of feed forward control system and feedback control system. This DO control system permitted close control of oxygen supply according to influent concentration, effluent quality and other environmental factors in the reactor. In this study the operation was divided into six phases according to influent nitrogen applied. SNAD system was successfully set up after adding COD into a CANON system. And the presence of COD enabled the survival of denitrifiers, and made Thauera and Pseudomonas predominant as functional denitrifiers in this system. Denaturing gradient gel electrophoresis (DGGE), fluorescence in situ hybridization (FISH) and 16S rRNA amplicon pyrosequencing were used to analyze the microbial variations of different substrate concentrations. Results indicated that the relative population of ammonia oxidizing bacteria (AOB) members decreased when influent ammonia concentration decreased from 2200 mg/L to 50 mg/L, while no dramatic drop of the percent of anammox bacteria was seen. And Nitrosomonas europaea was the predominant AOB in SNAD system treating sewage, while Candidatus Brocadia was the dominant anammox bacteria. Copyright © 2017 Elsevier Ltd. All rights reserved.

DECREASING OF SODIUM NITRITE CONTENT IN COOKED SAUSAGES USING DENITRIFYING MICROORGANISMS

Directory of Open Access Journals (Sweden)

Bal-Prylypko L. V.

2015-08-01

Full Text Available The purpose of this work was to study reduction of sodium nitrite in cooked sausages by adding of the optimized amount of denitrifying microorganisms to the bacterial preparation maintaining quality characteristics of the product. To develop biotechnology of boiled sausages «Naturel» we selected bacterial preparation based on nitrite-reducing strains of Staphylococcus carnosus and S. carnosus ssp.utilis. It was used generally accepted and special methods. The content of total pigments and nitrozopigments was determined by a method based on the extraction of meat pigments by aqueous acetone; color stability of final products was evaluated as the difference in optical density of nitroso pigment extracts before and after exposure (40 min of the sample under the light source; analytical processing of the experimental data was carried out using modern software; quantitative evaluation of color characteristics was performed in the RGB using a multifunctional device Epson Stylus TX400. Mathematical modeling was carried out on the basis of full factorial experiment such as 22, the optimization was performed by Box–Wilson. According to the study, using of the bacterial preparation based on nitrite-reducing strains of Staphylococcus carnosus and S. carnosus ssp. utilis in biotechnology of boiled sausages «Naturel» has a positive effect on the formation of the complex of required color characteristics of final products (for prototypes of sausages the index redness was 1. 61 times higher compared to the control. Degradation of sodium nitrite and formation of nitroso pigments were intensified that improved the stability of color during the storage (the index of color fastness of experimental cooked sausages was higher by 19%. The results of performed investigations illustrate the possibility of production of cooked sausages with a minimized content of synthetic food additives and ingredients.

Life on N2O: deciphering the ecophysiology of N2O respiring bacterial communities in a continuous culture.

Science.gov (United States)

Conthe, Monica; Wittorf, Lea; Kuenen, J Gijs; Kleerebezem, Robbert; van Loosdrecht, Mark C M; Hallin, Sara

2018-04-01

Reduction of the greenhouse gas N 2 O to N 2 is a trait among denitrifying and non-denitrifying microorganisms having an N 2 O reductase, encoded by nosZ. The nosZ phylogeny has two major clades, I and II, and physiological differences among organisms within the clades may affect N 2 O emissions from ecosystems. To increase our understanding of the ecophysiology of N 2 O reducers, we determined the thermodynamic growth efficiency of N 2 O reduction and the selection of N 2 O reducers under N 2 O- or acetate-limiting conditions in a continuous culture enriched from a natural community with N 2 O as electron acceptor and acetate as electron donor. The biomass yields were higher during N 2 O limitation, irrespective of dilution rate and community composition. The former was corroborated in a continuous culture of Pseudomonas stutzeri and was potentially due to cytotoxic effects of surplus N 2 O. Denitrifiers were favored over non-denitrifying N 2 O reducers under all conditions and Proteobacteria harboring clade I nosZ dominated. The abundance of nosZ clade II increased when allowing for lower growth rates, but bacteria with nosZ clade I had a higher affinity for N 2 O, as defined by μ max /K s . Thus, the specific growth rate is likely a key factor determining the composition of communities living on N 2 O respiration under growth-limited conditions.

Fatty Acid Biosynthesis Pathways in Methylomicrobium buryatense 5G(B1).

Science.gov (United States)

Demidenko, Aleksandr; Akberdin, Ilya R; Allemann, Marco; Allen, Eric E; Kalyuzhnaya, Marina G

2016-01-01

Methane utilization by methanotrophic bacteria is an attractive application for biotechnological conversion of natural or biogas into high-added-value products. Haloalcaliphilic methanotrophic bacteria belonging to the genus Methylomicrobium are among the most promising strains for methane-based biotechnology, providing easy and inexpensive cultivation, rapid growth, and the availability of established genetic tools. A number of methane bioconversions using these microbial cultures have been discussed, including the derivation of biodiesel, alkanes, and OMEGA-3 supplements. These compounds are derived from bacterial fatty acid pools. Here, we investigate fatty acid biosynthesis in Methylomicrobium buryatense 5G(B1) . Most of the genes homologous to typical Type II fatty acid biosynthesis pathways could be annotated by bioinformatics analyses, with the exception of fatty acid transport and regulatory elements. Different approaches for improving fatty acid accumulation were investigated. These studies indicated that both fatty acid degradation and acetyl- and malonyl-CoA levels are bottlenecks for higher level fatty acid production. The best strain generated in this study synthesizes 111 ± 2 mg/gDCW of extractable fatty acids, which is ~20% more than the original strain. A candidate gene for fatty acid biosynthesis regulation, farE , was identified and studied. Its deletion resulted in drastic changes to the fatty acid profile, leading to an increased pool of C18-fatty acid methyl ester. The FarE-regulon was further investigated by RNA-seq analysis of gene expression in farE -knockout mutants and farE -overexpressing strains. These gene profiles highlighted a novel set of enzymes and regulators involved in fatty acid biosynthesis. The gene expression and fatty acid profiles of the different farE -strains support the hypothesis that metabolic fluxes upstream of fatty acid biosynthesis restrict fatty acid production in the methanotroph.

Nitrate remediation in a novel upflow bio-electrochemical reactor (UBER) using palm shell activated carbon as cathode material

International Nuclear Information System (INIS)

Ghafari, Shahin; Hasan, Masitah; Aroua, Mohamed Kheireddine

2009-01-01

This study investigated the biological denitrification method which is a treatment method able to reduce inorganic nitrate compounds to harmless nitrogen gas. Autohydrogenotrophic denitrifying bacteria were used in this study to prevent any problematic outcomes associated with heterotrophic microorganisms. An upflow bio-electrochemical reactor (UBER) was used to accommodate hydrogenotrophic denitrifying bacteria employing palm shell granular activated carbon (GAC) as the biocarrier and cathode material. Bicarbonate as the external inorganic carbon source was fed to the reactor and hydrogen as the electron donor was generated in situ through electrolysis of water. Central composite design (CCD) and response surface methodology (RSM) were applied to investigate the effects of two operating parameters, namely electric current (I) and hydraulic retention time (HRT), on performance of the UBER. Electric current range of 0-20 mA and HRT range of 6-36 h were examined and results showed that nitrate can be entirely reduced within application of a wide operational range of electric current (10-16 mA) as well as HRT (13.5-30 h). However, increase of pH at cathode zone up to 10.5 inhibited nitrite reduction, and it was not reduced to the satisfactory level.

The importance of methane and thiosulfate in the metabolism of the bacterial symbionts of two deep-sea mussels

Science.gov (United States)

Fisher, C.R.; Childress, J.J.; Oremland, R.S.; Bidigare, R.R.

1987-01-01

Undescribed hydrocarbon-seep mussels were collected from the Louisiana Slope, Gulf of Mexico, during March 1986, and the ultrastructure of their gills was examined and compared to Bathymodiolus thermophilus, a mussel collected from the deep-sea hydrothermal vents on the Gala??pagos Rift in March 1985. These closely related mytilids both contain abundant symbiotic bacteria in their gills. However, the bacteria from the two species are distinctly different in both morphology and biochemistry, and are housed differently within the gills of the two mussels. The symbionts from the seep mussel are larger than the symbionts from B. thermophilus and, unlike the latter, contain stacked intracytoplasmic membranes. In the seep mussel three or fewer symbionts appear to be contained in each host-cell vacuole, while in B. thermophilus there are often more than twenty bacteria visible in a single section through a vacuole. The methanotrophic nature of the seep-mussel symbionts was confirmed in 14C-methane uptake experiments by the appearance of label in both CO2 and acid-stable, non-volatile, organic compounds after a 3 h incubation of isolated gill tissue. Furthermore, methane consumption was correlated with methanol dehydrogenase activity in isolated gill tissue. Activity of ribulose-1,5-biphosphate (RuBP) carboxylase and 14CO2 assimilation studies indicate the presence of either a second type of symbiont or contaminating bacteria on the gills of freshly captured seep mussels. A reevaluation of the nutrition of the symbionts in B. thermophilus indicates that while the major symbiont is not a methanotroph, its status as a sulfur-oxidizing chemoautotroph, as has been suggested previously, is far from proven. ?? 1987 Springer-Verlag.

Application of the differential soil bioreactor to in-situ biodegradation of trichloroethylene at the Savannah River Site

International Nuclear Information System (INIS)

Andrews, G.F.; Hansen, S.G.

1994-01-01

The differential soil bioreactor is a continuous-flow, laboratory treatability-study device in which groundwater, supplemented with nutrients, is recirculated through a disc of aquifer material at a rate that simulates actual groundwater flow. A high recycle ratio ensures that all bacteria in the disc are exposed to the same physiochemical environment, so rate and yield parameters needed for modeling in-situ bioremediation can, in principle, be derived directly from measurements of inlet and outlet concentrations of contaminants, nutrients and cells. Results are shown for the biodegradation of trichloroethylene by methanotrophic bacteria in sediments from the Savannah River site. The limitations of the technique for slow-flowing aquifers are discussed

Using Pure Cultures to Define the Site Preference of Nitrous Oxide Produced by Microbial Nitrification and Denitrification

Science.gov (United States)

Sutka, R. L.; Breznak, J. A.; Ostrom, N. E.; Ostrom, P. H.; Gandhi, H.

2004-12-01

Defining the site preference of nitrous oxide (N2O) produced in pure culture studies is crucial to interpreting field data. We have previously demonstrated that the intramolecular distribution of nitrogen isotopes (isotopomers) can be used to differentiate N2O produced by nitrifier denitrification and nitrification in cultures of Nitrosomonas europaea. Here, we have expanded on our initial results and evaluated the isotopomeric composition of N2O produced during nitrification and nitrifier denitrification with cultures of Nitrosospira multiformis. In addition, we have analyzed N2O produced during methanotrophic nitrification, denitrification, and fungal denitrification. To evaluate N2O production during nitrification and nitrifier denitrification, we compared the site preference of N2O formed as a result of nitrite reduction and hydroxylamine oxidation with Nitrosomonas europaea and Nitrosospira multiformis. The average site preference of N2O produced by hydroxylamine oxidation was similar for Nitrosomonas europaea (33.0 ± 3.5 ‰ ) and Nitrosospira multiformis (33.1 ± 4.2 ‰ ). Nitrous oxide produced by nitrifier-denitrification by Nitrosomonas europaea and Nitrosospira multiformis had a similar site preference of - 1.4 ± 4.4 ‰ and - 1.1 ± 2.6 ‰ respectively. The results indicate that it is possible to differentiate between N2O produced by nitrite reduction and hydroxylamine oxidation by ammonia oxidizing bacteria. Methanotrophic nitrification was evaluated by analyzing the N2O produced during hydroxylamine oxidation in concentrated cell suspensions of two methane oxidizing bacteria. The site preference of N2O produced by the two methane oxidizers, Methylococcus capsulatus Bath and Methylosinus trichosporium was 31.8 ± 4.7 ‰ and 33.0 ± 4.5 ‰ respectively. The results indicate that a site preference of 33 ‰ is applicable for nitrification regardless of whether a methane oxidizer or ammonia oxidizer is involved in the reaction. To determine the site

Reducing Open Cell Landfill Methane Emissions with a Bioactive Alternative Daily

Energy Technology Data Exchange (ETDEWEB)

Helene Hilger; James Oliver; Jean Bogner; David Jones

2009-03-31

Methane and carbon dioxide are formed in landfills as wastes degrade. Molecule-for-molecule, methane is about 20 times more potent than carbon dioxide at trapping heat in the earth's atmosphere, and thus, it is the methane emissions from landfills that are scrutinized. For example, if emissions composed of 60% methane and 40% carbon dioxide were changed to a mix that was 40% methane and 60% carbon dioxide, a 30% reduction in the landfill's global warming potential would result. A 10% methane, 90% carbon dioxide ratio will result in a 75% reduction in global warming potential compared to the baseline. Gas collection from a closed landfill can reduce emissions, and it is sometimes combined with a biocover, an engineered system where methane oxidizing bacteria living in a medium such as compost, convert landfill methane to carbon dioxide and water. Although methane oxidizing bacteria merely convert one greenhouse gas (methane) to another (carbon dioxide), this conversion can offer significant reductions in the overall greenhouse gas contribution, or global warming potential, associated with the landfill. What has not been addressed to date is the fact that methane can also escape from a landfill when the active cell is being filled with waste. Federal regulations require that newly deposited solid waste to be covered daily with a 6 in layer of soil or an alternative daily cover (ADC), such as a canvas tarp. The aim of this study was to assess the feasibility of immobilizing methane oxidizing bacteria into a tarp-like matrix that could be used for alternative daily cover at open landfill cells to prevent methane emissions. A unique method of isolating methanotrophs from landfill cover soil was used to create a liquid culture of mixed methanotrophs. A variety of prospective immobilization techniques were used to affix the bacteria in a tarp-like matrix. Both gel encapsulation of methanotrophs and gels with liquid cores containing methanotrophs were readily

The friendly bacteria within us Commensal bacteria of the intestine ...

Indian Academy of Sciences (India)

Balance of bacterial species in the gut · Immunosensory detection of intestinal bacteria · Pathogenic bacteria release interleukin-8 from HT-29 cells · Lactobacillus GG prevents the IL-8 release in response to pathogens · Effect of probiotic bacteria on chemokine response of epithelia to pathogens · PCR array studies in colon ...

Diversity and activity of benthic microbial communities at the North Alex mud volcano, Eastern Mediterranean

Science.gov (United States)

Makarow, Dorothee; Feseker, Tomas; Schmitz, Ruth; Treude, Tina

2010-05-01

North Alex mud volcano, located on the upper slope of the western Nile deep-sea fan, is characterized by an active seepage center transporting pore fluids, hydrocarbons and gases from deep subsurface sources to the sediment-water interface. Surface sediments feature steep temperature gradient of 8.5°C m-1. We sampled the top 40 cm of the sediments at different locations between the center and rim of the mud volcano to study the diversity, activity, and physiological characteristics of benthic microorganisms. The sediments revealed the activity of anaerobic oxidation of methane coupled to sulfate reduction with a mesophilic temperature optimum. Organisms involved in the process include consortia of methanotrophic archaea (ANME-2 group) and an unknown bacterial partner. Besides methanotrophic organisms the sediments harbored a variety of other bacterial and archaeal groups - including potentially thermophilic bacteria that could be involved in sulfur cycling. This poster presentation will provide an overview of microbial activities and community compositions of North Alex mud volcano sediments.

Microbial production of poly(hydroxybutyrate) from C₁ carbon sources.

Science.gov (United States)

Khosravi-Darani, Kianoush; Mokhtari, Zahra-Beigom; Amai, Tomohito; Tanaka, Kenji

2013-02-01

Polyhydroxybutyrate (PHB) is an attractive substitute for petrochemical plastic due to its similar properties, biocompatibility, and biodegradability. The cost of scaled-up PHB production inhibits its widespread usage. Intensive researches are growing to reduce costs and improve thermomechanical, physical, and processing properties of this green biopolymer. Among cheap substrates which are used for reducing total cost of PHB production, some C₁ carbon sources, e.g., methane, methanol, and CO₂ have received a great deal of attention due to their serious role in greenhouse problem. This article reviews the fundamentals of strategies for reducing PHA production and moves on to the applications of several cheap substrates with a special emphasis on methane, methanol, and CO₂. Also, some explanation for involved microorganisms including the hydrogen-oxidizing bacteria and methanotrophs, their history, culture condition, and nutritional requirements are given. After description of some important strains among the hydrogen-oxidizing and methanotrophic producers of PHB, the article is focused on limitations, threats, and opportunities for application and their future trends.

Systems Level Dissection of Anaerobic Methane Cycling: Quantitative Measurements of Single Cell Ecophysiology, Genetic Mechanisms, and Microbial Interactions

Energy Technology Data Exchange (ETDEWEB)

Orphan, Victoria [California Inst. of Technology (CalTech), Pasadena, CA (United States); Tyson, Gene [University of Queensland, Brisbane Australia; Meile, Christof [University of Georgia, Athens, Georgia; McGlynn, Shawn [California Inst. of Technology (CalTech), Pasadena, CA (United States); Yu, Hang [California Inst. of Technology (CalTech), Pasadena, CA (United States); Chadwick, Grayson [California Inst. of Technology (CalTech), Pasadena, CA (United States); Marlow, Jeffrey [California Inst. of Technology (CalTech), Pasadena, CA (United States); Trembath-Reichert, Elizabeth [California Inst. of Technology (CalTech), Pasadena, CA (United States); Dekas, Anne [California Inst. of Technology (CalTech), Pasadena, CA (United States); Hettich, Robert [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Pan, Chongle [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Ellisman, Mark [University of California San Diego; Hatzenpichler, Roland [California Inst. of Technology (CalTech), Pasadena, CA (United States); Skennerton, Connor [California Inst. of Technology (CalTech), Pasadena, CA (United States); Scheller, Silvan [California Inst. of Technology (CalTech), Pasadena, CA (United States)

2017-12-25

The global biological CH4 cycle is largely controlled through coordinated and often intimate microbial interactions between archaea and bacteria, the majority of which are still unknown or have been only cursorily identified. Members of the methanotrophic archaea, aka ‘ANME’, are believed to play a major role in the cycling of methane in anoxic environments coupled to sulfate, nitrate, and possibly iron and manganese oxides, frequently forming diverse physical and metabolic partnerships with a range of bacteria. The thermodynamic challenges overcome by the ANME and their bacterial partners and corresponding slow rates of growth are common characteristics in anaerobic ecosystems, and, in stark contrast to most cultured microorganisms, this type of energy and resource limited microbial lifestyle is likely the norm in the environment. While we have gained an in-depth systems level understanding of fast-growing, energy-replete microorganisms, comparatively little is known about the dynamics of cell respiration, growth, protein turnover, gene expression, and energy storage in the slow-growing microbial majority. These fundamental properties, combined with the observed metabolic and symbiotic versatility of methanotrophic ANME, make these cooperative microbial systems a relevant (albeit challenging) system to study and for which to develop and optimize culture-independent methodologies, which enable a systems-level understanding of microbial interactions and metabolic networks. We used an integrative systems biology approach to study anaerobic sediment microcosms and methane-oxidizing bioreactors and expanded our understanding of the methanotrophic ANME archaea, their interactions with physically-associated bacteria, ecophysiological characteristics, and underlying genetic basis for cooperative microbial methane-oxidation linked with different terminal electron acceptors. Our approach is inherently multi-disciplinary and multi-scaled, combining transcriptional and

Isolation and Characterization of Aerobic Denitrifiers from Agricultural Soil

OpenAIRE

ÇELEN, Ebru; KILIÇ, Mehmet Akif

2004-01-01

Denitrification is generally considered an anaerobic process. However, in recent years it has been shown that bacteria can also reduce nitrate to nitrite under aerobic conditions. The characterization of biologically available nitrogen forms and their biological cycling mechanisms is important for ecological and agricultural implications. In this study, aerobic nitrate reducers were isolated from greenhouse soil. Using a nitrate reduction assay, it was found that 39 out of 60 isolates can red...

Strategies to Optimize Microbially-Mediated Mitigation of Greenhouse Gas Emissions from Landfill Cover Soils

Energy Technology Data Exchange (ETDEWEB)

Jeremy Semrau; Sung-Woo Lee; Jeongdae Im; Sukhwan Yoon; Michael Barcelona

2010-09-30

The overall objective of this project, 'Strategies to Optimize Microbially-Mediated Mitigation of Greenhouse Gas Emissions from Landfill Cover Soils' was to develop effective, efficient, and economic methodologies by which microbial production of nitrous oxide can be minimized while also maximizing microbial consumption of methane in landfill cover soils. A combination of laboratory and field site experiments found that the addition of nitrogen and phenylacetylene stimulated in situ methane oxidation while minimizing nitrous oxide production. Molecular analyses also indicated that methane-oxidizing bacteria may play a significant role in not only removing methane, but in nitrous oxide production as well, although the contribution of ammonia-oxidizing archaea to nitrous oxide production can not be excluded at this time. Future efforts to control both methane and nitrous oxide emissions from landfills as well as from other environments (e.g., agricultural soils) should consider these issues. Finally, a methanotrophic biofiltration system was designed and modeled for the promotion of methanotrophic activity in local methane 'hotspots' such as landfills. Model results as well as economic analyses of these biofilters indicate that the use of methanotrophic biofilters for controlling methane emissions is technically feasible, and provided 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.

Big bacteria

DEFF Research Database (Denmark)

Schulz, HN; Jørgensen, BB

2001-01-01

A small number of prokaryotic species have a unique physiology or ecology related to their development of unusually large size. The biomass of bacteria varies over more than 10 orders of magnitude, from the 0.2 mum wide nanobacteria to the largest cells of the colorless sulfur bacteria......, Thiomargarita namibiensis, with a diameter of 750 mum. All bacteria, including those that swim around in the environment, obtain their food molecules by molecular diffusion. Only the fastest and largest swimmers known, Thiovulum majus, are able to significantly increase their food supply by motility...... and by actively creating an advective flow through the entire population. Diffusion limitation generally restricts the maximal size of prokaryotic cells and provides a selective advantage for mum-sized cells at the normally low substrate concentrations in the environment. The largest heterotrophic bacteria...

Conventional methanotrophs are responsible for atmospheric methane oxidation in paddy soils

NARCIS (Netherlands)

Cai, Yuanfeng; Yan, Zheng; Bodelier, P.L.E.; Conrad, R.; Jia, Zhongjun

2016-01-01

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

A pilot study of bacterial regrowth in water pipelines.

Science.gov (United States)

Mancini, Laura; Marcheggiani, Stefania; Grasso, Cinzia; Romanelli, Cristina; Mistretta, Antonio; Marranzano, Marina

2016-01-01

Bacterial regrowth in water distribution systems results in deterioration of bacteriological quality of drinking water, as well as accelerated corrosion of the pipelines. The aim of the present study was to analyze the phenomena of colonization and bacterial regrowth in source water and in the water distribution systems of three distribution networks in the province of Catania (Sicily, Italy). The BART™ (Biological Activity Reaction Test) method was used, which is also capable of determining the potential aggressiveness of microbial species in water. We searched for sulfate reducing bacteria, iron-related bacteria, nitrifying and denitrifying bacteria, heterotrophic bacteria, slime-forming bacteria and fluorescent Pseudomonads. A high concentration of heterotrophic bacteria was found in almost every water sample analyzed. Sulfate reducing bacteria and iron-related bacteria were found in all three distribution networks, while non-fluorescing Pseudomonas were detected in source water of only one of the distribution networks. The BART™ method was found to be a practical and easy to use tool to detect the different bacteria groups involved in regrowth phenomena.

A broad-spectrum analysis of the effects of teflubenzuron exposure on the biochemical activities and microbial community structure of soil.

Science.gov (United States)

Cycoń, Mariusz; Wójcik, Marcin; Borymski, Sławomir; Piotrowska-Seget, Zofia

2012-10-15

We evaluated the response of soil bacteria to applications of the insecticide teflubenzuron at the field rate dosage (FR; 0.15 mg/kg of soil) and at a higher dosage (10*FR; 1.5 mg/kg of soil). When applied at the FR dosage, teflubenzuron had no effect on several biochemical parameters of the soil, including substrate-induced respiration (SIR), dehydrogenase (DHA) and phosphatase activities (PHOS), and N-NO(3)(-) and N-NH(4)(+) concentrations. Additionally, no differences were observed in the culturable fraction of the soil bacteria (the number of heterotrophic, nitrifying, denitrifying and N(2)-fixing bacteria; the growth strategy; the ecophysiological and colony development indices; and the physiological state). In contrast, treatment with the 10*FR dosage of the insecticide significantly increased SIR, DHA, PHOS and N-NH(4)(+) levels and the number of heterotrophic and denitrifying bacteria. Decreases in urease activity (URE) and the number of nitrifying and N(2)-fixing bacteria were also observed. A phospholipid fatty acid (PLFA) method-based analysis of the entire soil microorganism population revealed that teflubenzuron treatment affected the total fatty acid level as well as those considered to be of Gram-positive bacteria, Gram-negative bacteria and fungi. This effect was observed on days 1 and 14 post-treatment. A principal component analysis (PCA) of the PLFAs showed that teflubenzuron treatment significantly shifted the microbial community structure; however, all of the observed effects were transient. Studies on the degradation of teflubenzuron revealed that this process is characterised by a short lag phase and a rate constant (k) of 0.020/day. This degradation rate follows first-order kinetics, and the DT50 was 33.5 days. This is the first study that thoroughly examines the functional and structural status of both the culturable and non-culturable fractions of the soil microbial community after teflubenzuron application. Copyright © 2012 Elsevier Ltd

Big bacteria

DEFF Research Database (Denmark)

Schulz, HN; Jørgensen, BB

2001-01-01

A small number of prokaryotic species have a unique physiology or ecology related to their development of unusually large size. The biomass of bacteria varies over more than 10 orders of magnitude, from the 0.2 mum wide nanobacteria to the largest cells of the colorless sulfur bacteria...... and by actively creating an advective flow through the entire population. Diffusion limitation generally restricts the maximal size of prokaryotic cells and provides a selective advantage for mum-sized cells at the normally low substrate concentrations in the environment. The largest heterotrophic bacteria......, the 80 x 600 mum large Epulopiscium sp. from the gut of tropical fish, are presumably living in a very nutrient-rich medium. Many large bacteria contain numerous inclusions in the cells that reduce the volume of active cytoplasm. The most striking examples of competitive advantage from large cell size...

Methylocella silvestris sp. nov., a novel methanotroph isolated from an acidic forest cambisol.

Science.gov (United States)

Dunfield, Peter F; Khmelenina, Valentina N; Suzina, Natalia E; Trotsenko, Yuri A; Dedysh, Svetlana N

2003-09-01

Two strains of Gram-negative, aerobic, non-pigmented, non-motile, rod-shaped, methane-oxidizing bacteria were isolated from an acidic forest cambisol near Marburg, Germany, and were designated as strains BL2(T) and A1. These bacteria were morphologically and phenotypically similar to Methylocella palustris K(T). The cells possess a highly specific bipolar appearance. They lack the intracytoplasmic membranes common to all methane-oxidizing bacteria except Methylocella, but contain a vesicular membrane system connected to the cytoplasmic membrane. A soluble methane monooxygenase was present, but no particulate methane monooxygenase could be detected. These bacteria utilize the serine pathway for carbon assimilation. Strains BL2(T) and A1 are moderately acidophilic, mesophilic organisms capable of growth at pH values between 4.5 and 7 (with an optimum at pH 5.5) and at temperatures between 4 and 30 degrees C. Compared with Methylocella palustris K(T), these strains have greater tolerance of cold temperatures, dissolved salts and methanol. On the basis of 16S rRNA gene sequence identity, of species with validly published names, strain BL2(T) is most closely related to Methylocella palustris K(T) (97.3 % identity), Beijerinckia indica subsp. indica ATCC 9039(T) (97.1 %) and Methylocapsa acidiphila B2(T) (96.2 %). The DNA G+C content is 60 mol% and the major phospholipid fatty acid is 18 : 1omega7. Strain BL2(T) showed only 21-22 % DNA-DNA hybridization with Methylocella palustris K(T). The data therefore suggest that strains BL2(T) and A1 represent a novel species of Methylocella; the name Methylocella silvestris sp. nov. is proposed, with strain BL2(T) (=DSM 15510(T)=NCIMB 13906(T)) as the type strain.

Microbial uptake of uranium, cesium, and radium

Energy Technology Data Exchange (ETDEWEB)

Strandberg, G.W.; Shumate, S.E. II; Parrott, J.R. Jr.; McWhirter, D.A.

1980-01-01

The ability of diverse microbial species to concentrate uranium, cesium, and radium was examined. Saccharomyces cerevisiae, Pseudomonas aeruginosa, and a mixed culture of denitrifying bacteria accumulated uranium to 10 to 15% of the dry cell weight. Only a fraction of the cells in a given population had visible uranium deposits in electron micrographs. While metabolism was not required for uranium uptake, mechanistic differences in the metal uptake process were indicated. Uranium accumulated slowly (hours) on the surface of S. cerevisiae and was subject to environmental factors (i.e., temperature, pH, interfering cations and anions). In contrast, P. aeruginosa and the mixed culture of denitrifying bacteria accumulated uranium rapidly (minutes) as dense, apparently random, intracellular deposits. This very rapid accumulation has prevented us from determining whether the uptake rate during the transient between the initial and equilibrium distribution of uranium is affected by environmental conditions. However, the final equilibrium distributions are not affected by those conditions which affect uptake by S. cerevisiae. Cesium and radium were concentrated to a considerably lesser extent than uranium by the several microbial species tested. The potential utility of microorganisms for the removal and concentration of these metals from nuclear processing wastes and several bioreactor designs for contacting microorganisms with contaminated waste streams will be discussed.

The metatranscriptome of a deep-sea hydrothermal plume is dominated by water column methanotrophs and lithotrophs.

Science.gov (United States)

Lesniewski, Ryan A; Jain, Sunit; Anantharaman, Karthik; Schloss, Patrick D; Dick, Gregory J

2012-12-01

Microorganisms mediate geochemical processes in deep-sea hydrothermal vent plumes, which are a conduit for transfer of elements and energy from the subsurface to the oceans. Despite this important microbial influence on marine geochemistry, the ecology and activity of microbial communities in hydrothermal plumes is largely unexplored. Here, we use a coordinated metagenomic and metatranscriptomic approach to compare microbial communities in Guaymas Basin hydrothermal plumes to background waters above the plume and in the adjacent Carmen Basin. Despite marked increases in plume total RNA concentrations (3-4 times) and microbially mediated manganese oxidation rates (15-125 times), plume and background metatranscriptomes were dominated by the same groups of methanotrophs and chemolithoautotrophs. Abundant community members of Guaymas Basin seafloor environments (hydrothermal sediments and chimneys) were not prevalent in the plume metatranscriptome. De novo metagenomic assembly was used to reconstruct genomes of abundant populations, including Marine Group I archaea, Methylococcaceae, SAR324 Deltaproteobacteria and SUP05 Gammaproteobacteria. Mapping transcripts to these genomes revealed abundant expression of genes involved in the chemolithotrophic oxidation of ammonia (amo), methane (pmo) and sulfur (sox). Whereas amo and pmo gene transcripts were abundant in both plume and background, transcripts of sox genes for sulfur oxidation from SUP05 groups displayed a 10-20-fold increase in plumes. We conclude that the biogeochemistry of Guaymas Basin hydrothermal plumes is mediated by microorganisms that are derived from seawater rather than from seafloor hydrothermal environments such as chimneys or sediments, and that hydrothermal inputs serve as important electron donors for primary production in the deep Gulf of California.

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

Science.gov (United States)

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

2014-01-01

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

Anaerobic degradation of anionic surfactants by indigenous microorganisms from sediments of a tropical polluted river in Brazil

Directory of Open Access Journals (Sweden)

Iolanda Cristina Silveira Duarte

2015-03-01

Full Text Available Linear alkylbenzene sulfonate (LAS is widely used in the formulation of domestic and industrial cleaning products, the most synthetic surfactants used worldwide. These products can reach water bodies through the discharge of untreated sewage or non-effective treatments. This study evaluates the ability of the microorganisms found in the Tietê river sediment to degrade this synthetic surfactant. The experiment was conducted in a bioreactor, operated in batch sequences under denitrifying conditions, with cycles of 24 hours and stirring at 150rpm, using 430mL of sediments and 1 070mL of a synthetic substrate consisting of yeast extract, soluble starch, sodium bicarbonate and sucrose. LAS was added at different concentrations of 15mg/L and 30mg/L. The reactor operation was divided into the biomass adaptation to the synthetic substrate without LAS and three experimental conditions: a addition of 15mg/L of LAS; b 50% reduction the co-substrate concentration and 15mg/L of LAS, and c addition of 30mg/L of LAS and 100% co-substrate concentration. The results showed that the degradation efficiency of LAS was directly related to the addition of co-substrates and the population of denitrifying bacteria. The removal of LAS and nitrate can be achieved simultaneously in wastewater with low organic loads. The reduction in the co-substrates concentration was directly influenced by the number of denitrifying bacteria (2.2x10(13 to 1.0x10(8MPN/gTVS, and consequently, LAS degradation (60.1 to 55.4%. The sediment microorganisms in the Tietê river can be used as an alternative inoculum in the treatment of wastewater with nitrate and LAS contamination.

Myriophyllum aquaticum Constructed Wetland Effectively Removes Nitrogen in Swine Wastewater

Directory of Open Access Journals (Sweden)

Haishu Sun

2017-10-01

Full Text Available Removal of nitrogen (N is a critical aspect in the functioning of constructed wetlands (CWs, and the N treatment in CWs depends largely on the presence and activity of macrophytes and microorganisms. However, the effects of plants on microorganisms responsible for N removal are poorly understood. In this study, a three-stage surface flow CW was constructed in a pilot-scale within monospecies stands of Myriophyllum aquaticum to treat swine wastewater. Steady-state conditions were achieved throughout the 600-day operating period, and a high (98.3% average ammonia removal efficiency under a N loading rate of 9 kg ha-1 d-1 was observed. To determine whether this high efficiency was associated with the performance of active microbes, the abundance, structure, and interactions of microbial community were compared in the unvegetated and vegetated samples. Real-time quantitative polymerase chain reactions showed the abundances of nitrifying genes (archaeal and bacterial amoA and denitrifying genes (nirS, nirK, and nosZ were increased significantly by M. aquaticum in the sediments, and the strongest effects were observed for the archaeal amoA (218-fold and nirS genes (4620-fold. High-throughput sequencing of microbial 16S rRNA gene amplicons showed that M. aquaticum greatly changed the microbial community, and ammonium oxidizers (Nitrosospira and Nitrososphaera, nitrite-oxidizing bacteria (Nitrospira, and abundant denitrifiers including Rhodoplanes, Bradyrhizobium, and Hyphomicrobium, were enriched significantly in the sediments. The results of a canonical correspondence analysis and Mantle tests indicated that M. aquaticum may shift the sediment microbial community by changing the sediment chemical properties. The enriched nitrifiers and denitrifiers were distributed widely in the vegetated sediments, showing positive ecological associations among themselves and other bacteria based on phylogenetic molecular ecological networks.

Bacteria and lignin degradation

Institute of Scientific and Technical Information of China (English)

Jing LI; Hongli YUAN; Jinshui YANG

2009-01-01

Lignin is both the most abundant aromatic (phenolic) polymer and the second most abundant raw material.It is degraded and modified by bacteria in the natural world,and bacteria seem to play a leading role in decomposing lignin in aquatic ecosystems.Lignin-degrading bacteria approach the polymer by mechanisms such as tunneling,erosion,and cavitation.With the advantages of immense environmental adaptability and biochemical versatility,bacteria deserve to be studied for their ligninolytic potential.

[Distribution Characteristics of Nitrifiers and Denitrifiers in the River Sediments of Tongling City].

Science.gov (United States)

Cheng, Jian-hua; Dou, Zhi-yong; Sun, Qing-ye

2016-04-15

Rivers in mining areas were influenced by contaminants such as nitrogen, phosphorus and organic matter due to domestic and agricultural wastewater discharge in addition to pollutants caused by mining activities. In this study, surface sediment samples of rivers in Tongling city were collected to address the effect of season and pollution type on the abundance of nitrifiers and denitrifiers using quantitative polymerase chain reaction (QPCR) technique targeting at the ammonia monooxygenase (amoA) and nitrite reductase (nir) genes. The results showed that the average ahundance of ammonia oxidizing archaea (AGA) (ranging from 1.74 x 10⁵ to 1.45 x 10⁸ copies · g⁻¹) was 4.39 times that of ammonia oxidizing hacteria (AGH) (ranging from 1.39 x 10⁵ to 3.39 x 10⁷ copies · g⁻¹); and the average abundance of nirK gene (ranging from 4.45 x 10⁶ to 1.51 x 10⁸ copies · g) was almost a thirtieth part of nirS gene (ranging from 1.69 x 10⁷ to 8.55 x 10⁹ copies · g⁻¹). The abundance of AOA was higher in spring and autumn, and lower in summer and winter. And sediment AOB abundance was higher in spring and winter than in summer and autumn. Meanwhile, the abundance of nir genes was in the order of spring (nirS )/autumn (nirK) > summer > winter > autumn (nirS )/spring (nirK). Moreover, the abundance of bacterial and archaeal arnoA and nirS genes in sediments influenced by mine pollution was generally higher than that in sediments influenced by agricultural non-point pollution, whereas the abundance of nirK gene showed an opposite trend.

Production of poly-β-hydroxybutyrate (PHB) by Methylobacterium organophilum isolated from a methanotrophic consortium in a two-phase partition bioreactor

International Nuclear Information System (INIS)

Zuniga, C.; Morales, M.; Le Borgne, S.; Revah, S.

2011-01-01

The biodegradation of methane, a greenhouse gas, and the accumulation of poly-β-hydroxybutyrate (PHB) were studied using a methanotrophic consortium and an isolated strain thereof. The specific rates for methane consumption were 100 and 17mg CH 4 g x -1 h -1 for the isolate and the consortium, respectively. Also the effect of including 10% (v v -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 CH 4 g x -1 h -1 for the isolated strain. The accumulated PHB in the reactor was 34% and 38% (w w -1 ) for the consortium and the isolate, respectively. The highest productivity was obtained in the TPPB and was 1.61mg PHB g x -1 h -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 -1 ) of PHB under nitrogen limitation in microcosm experiments.

Enumeration and characterization of microorganisms associated with the uranium ore deposit at Cigar Lake, Canada. Informal report

International Nuclear Information System (INIS)

Francis, A.J.; Joshi-Tope, G.; Gillow, J.B.; Dodge, C.J.

1994-03-01

The high-grade uranium deposit at Cigar Lake, Canada, is being investigated as a natural analog for the disposal of nuclear fuel waste. Geochemical aspects of the site have been studied in detail, but the microbial ecology has not been fully investigated. Microbial populations in an ore sample and in groundwater samples from the vicinity of the ore zone were examined to determine their effect on uranium mobility. Counts of the total number of bacteria and of respiring bacteria were obtained by direct microscopy, and the viable aerobic and anaerobic bacteria were assessed as colony forming units (CFUs) by the dilution plating technique. In addition, the population distribution of denitrifiers, fermenters, iron- and sulfur-oxidizers, iron- and sulfate-reducers, and methanogens was determined by the most probable number (MPN) technique

Microbial reduction of SO{sub 2} and NO{sub x} as a means of by-product recovery/disposal from regenerable processes for the desulfurization of flue gas. Final report

Energy Technology Data Exchange (ETDEWEB)

Sublette, K.L.

1994-03-01

The main objective of this research was to investigate microorganisms capable of fossil fuel flue gas desulfurization and denitrification. The study used municipal sewage sludge as a carbon and energy source for SO{sub 2}-reducing cultures. The individual tasks developed a consortium of sulfate-reducing bacteria, investigated the design parameters for a continuous process, preformed a cost analysis, and screened sulfate-reducing bacteria. In the investigation of microbial reduction of NO{sub x} to nitrogen, tasks included screening denitrifying bacteria for NO and NO{sub 2} activity, developing optimum NO-reducing cultures, and investigating design parameters for a continuous system. This final report reviews the work previous to the current project, describes project objectives and the specific work plan, and reports results from the work completed during the previous reporting periods.

Site properties have a stronger influence than fire severity on ectomycorrhizal fungi and associated N-cycling bacteria in regenerating post-beetle-killed lodgepole pine forests.

Science.gov (United States)

Kennedy, Nabla M; Robertson, Susan J; Green, D Scott; Scholefield, Scott R; Arocena, Joselito M; Tackaberry, Linda E; Massicotte, Hugues B; Egger, Keith N

2015-09-01

Following a pine beetle epidemic in British Columbia, Canada, we investigated the effect of fire severity on rhizosphere soil chemistry and ectomycorrhizal fungi (ECM) and associated denitrifying and nitrogen (N)-fixing bacteria in the root systems of regenerating lodgepole pine seedlings at two site types (wet and dry) and three fire severities (low, moderate, and high). The site type was found to have a much larger impact on all measurements than fire severity. Wet and dry sites differed significantly for almost all soil properties measured, with higher values identified from wet types, except for pH and percent sand that were greater on dry sites. Fire severity caused few changes in soil chemical status. Generally, bacterial communities differed little, whereas ECM morphotype analysis revealed ectomycorrhizal diversity was lower on dry sites, with a corresponding division in community structure between wet and dry sites. Molecular profiling of the fungal ITS region confirmed these results, with a clear difference in community structure seen between wet and dry sites. The ability of ECM fungi to colonize seedlings growing in both wet and dry soils may positively contribute to subsequent regeneration. We conclude that despite consecutive landscape disturbances (mountain pine beetle infestation followed by wildfire), the "signature" of moisture on chemistry and ECM community structure remained pronounced.

Biotic manganese oxidation coupled with methane oxidation using a continuous-flow bioreactor system under marine conditions.

Science.gov (United States)

Kato, Shingo; Miyazaki, Masayuki; Kikuchi, Sakiko; Kashiwabara, Teruhiko; Saito, Yumi; Tasumi, Eiji; Suzuki, Katsuhiko; Takai, Ken; Cao, Linh Thi Thuy; Ohashi, Akiyoshi; Imachi, Hiroyuki

2017-10-01

Biogenic manganese oxides (BioMnOx) can be applied for the effective removal and recovery of trace metals from wastewater because of their high adsorption capacity. Although a freshwater continuous-flow system for a nitrifier-based Mn-oxidizing microbial community for producing BioMnOx has been developed so far, a seawater continuous-flow bioreactor system for BioMnOx production has not been established. Here, we report BioMnOx production by a methanotroph-based microbial community by using a continuous-flow bioreactor system. The bioreactor system was operated using a deep-sea sediment sample as the inoculum with methane as the energy source for over 2 years. The BioMnOx production became evident after 370 days of reactor operation. The maximum Mn oxidation rate was 11.4 mg L -1 day -1 . An X-ray diffraction analysis showed that the accumulated BioMnOx was birnessite. 16S rRNA gene-based clone analyses indicated that methanotrophic bacterial members were relatively abundant in the system; however, none of the known Mn-oxidizing bacteria were detected. A continuous-flow bioreactor system coupled with nitrification was also run in parallel for 636 days, but no BioMnOx production was observed in this bioreactor system. The comparative experiments indicated that the methanotroph-based microbial community, rather than the nitrifier-based community, was effective for BioMnOx production under the marine environmental conditions.

The fecal bacteria

Science.gov (United States)

Sadowsky, Michael J.; Whitman, Richard L.

2011-01-01

The Fecal Bacteria offers a balanced, integrated discussion of fecal bacteria and their presence and ecology in the intestinal tract of mammals, in the environment, and in the food supply. This volume covers their use in examining and assessing water quality in order to offer protection from illnesses related to swimming in or ingesting contaminated water, in addition to discussing their use in engineering considerations of water quality, modeling, monitoring, and regulations. Fecal bacteria are additionally used as indicators of contamination of ready-to-eat foods and fresh produce. The intestinal environment, the microbial community structure of the gut microbiota, and the physiology and genomics of this broad group of microorganisms are explored in the book. With contributions from an internationally recognized group of experts, the book integrates medicine, public health, environmental, and microbiological topics in order to provide a unique, holistic understanding of fecal bacteria. Moreover, it shows how the latest basic science and applied research findings are helping to solve problems and develop effective management strategies. For example, readers will discover how the latest tools and molecular approaches have led to our current understanding of fecal bacteria and enabled us to improve human health and water quality. The Fecal Bacteria is recommended for microbiologists, clinicians, animal scientists, engineers, environmental scientists, food safety experts, water quality managers, and students. It will help them better understand fecal bacteria and use their knowledge to protect human and environmental health. They can also apply many of the techniques and molecular tools discussed in this book to the study of a broad range of microorganisms in a variety of habitats.

The nitrogen cycle in anaerobic methanotrophic mats of the Black Sea is linked to sulfate reduction and biomass decomposition.

Science.gov (United States)

Siegert, Michael; Taubert, Martin; Seifert, Jana; von Bergen-Tomm, Martin; Basen, Mirko; Bastida, Felipe; Gehre, Matthias; Richnow, Hans-Hermann; Krüger, Martin

2013-11-01

Anaerobic methanotrophic (ANME) mats host methane-oxidizing archaea and sulfate-reducing prokaryotes. Little is known about the nitrogen cycle in these communities. Here, we link the anaerobic oxidation of methane (AOM) to the nitrogen cycle in microbial mats of the Black Sea by using stable isotope probing. We used four different (15)N-labeled sources of nitrogen: dinitrogen, nitrate, nitrite and ammonium. We estimated the nitrogen incorporation rates into the total biomass and the methyl coenzyme M reductase (MCR). Dinitrogen played an insignificant role as nitrogen source. Assimilatory and dissimilatory nitrate reduction occurred. High rates of nitrate reduction to dinitrogen were stimulated by methane and sulfate, suggesting that oxidation of reduced sulfur compounds such as sulfides was necessary for AOM with nitrate as electron acceptor. Nitrate reduction to dinitrogen occurred also in the absence of methane as electron donor but at six times slower rates. Dissimilatory nitrate reduction to ammonium was independent of AOM. Ammonium was used for biomass synthesis under all conditions. The pivotal enzyme in AOM coupled to sulfate reduction, MCR, was synthesized from nitrate and ammonium. Results show that AOM coupled to sulfate reduction along with biomass decomposition drive the nitrogen cycle in the ANME mats of the Black Sea and that MCR enzymes are involved in this process. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Bleach vs. Bacteria

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... Articles | Inside Life Science Home Page Bleach vs. Bacteria By Sharon Reynolds Posted April 2, 2014 Your ... hypochlorous acid to help kill invading microbes, including bacteria. Researchers funded by the National Institutes of Health ...

Potential role of bacteria packaging by protozoa in the persistence and transmission of pathogenic bacteria

OpenAIRE

Denoncourt, Alix M.; Paquet, Valérie E.; Charette, Steve J.

2014-01-01

Many pathogenic bacteria live in close association with protozoa. These unicellular eukaryotic microorganisms are ubiquitous in various environments. A number of protozoa such as amoebae and ciliates ingest pathogenic bacteria, package them usually in membrane structures, and then release them into the environment. Packaged bacteria are more resistant to various stresses and are more apt to survive than free bacteria. New evidence indicates that protozoa and not bacteria control the packaging...

Performance and microbial community of anammox in presence of micro-molecule carbon source.

Science.gov (United States)

He, Shilong; Yang, Wan; Qin, Meng; Mao, Zhen; Niu, Qigui; Han, Ming

2018-08-01

Because ammonium (NH 4 + -N) coexists with organic matter in some wastewaters, the possible adverse influences of organic matter become a major concern in the applications of anaerobic ammonium oxidation (anammox). In this study, the effects of acetate, as a representative of micro-molecule organic matter, on anammox were investigated. Efficient nitrogen removal was realized because denitrifying bacteria and anammox bacteria (AnAOB) had a better synergistic effect under the condition of chemical oxygen demand (COD) concentrations lower than 251 ± 7 mg L -1 . Furthermore, the nitrogen removal efficiency (NRE) decreased to 82.02 ± 3.14% when COD was increased to 730 ± 9 mg L -1 , and effluent free ammonia (FA) reached 21.93 ± 4.71 mg L -1 might be one of factors leading to inhibition. However, the nitrogen-removal contribution rate of anammox remained steady at 61.97 ± 2.84% at COD of 730 ± 9 mg L -1 , which indicated that anammox was still dominant in the system. AnAOB, such as Ca. Kuenenia and Ca. Jettenia, and denitrifying bacteria, such as Denitratisoma and Thauera, were found to coexist in the reactor. Interestingly, Ca. Kuenenia presented in the trend of first decreased then increased with the increasing of organic matter concentration, which might be one of reasons that anammox played an important role in nitrogen removal at high COD concentration. Copyright © 2018 Elsevier Ltd. All rights reserved.

Bacteria-Mineral Interactions on the Surfaces of Metal-Resistant Bacteria

International Nuclear Information System (INIS)

Malkin, A.J.

2010-01-01

The extraordinary ability of indigenous microorganisms, like metal-resistant bacteria, for biotransformation of toxic compounds is of considerable interest for the emerging area of environmental bioremediation. However, the underlying mechanisms by which metal-resistant bacteria transform toxic compounds are currently unknown and await elucidation. The project's objective was to study stress-induced responses of metal-resistant bacteria to environmental changes and chemical stimulants. This project involved a multi-institutional collaboration of our LLNL group with the group of Dr. H.-Y. Holman (Lawrence Berkeley National Laboratory). In this project, we have utilized metal-resistant bacteria Arthrobacter oxydans as a model bacterial system. We have utilized atomic force microscopy (AFM) to visualize for the first time at the nanometer scale formation of stress-induced structures on bacterial surfaces in response to Cr (VI) exposure. We have demonstrated that structure, assembly, and composition of these stress-induced structures are dependent on Cr (VI) concentrations. Our AFM observations of the appearance and development of stress-induced layers on the surfaces of Arthrobacter oxydans bacteria exposed to Cr (VI) were confirmed by Dr. Holman's biochemical, electron microscopy, and synchrotron infrared spectromicroscopy studies. In general, in vitro imaging of live microbial and cellular systems represents one of the most challenging issues in application of AFM. Various approaches for immobilization of bacteria on the substrate for in vitro imaging were tested in this project. Imaging of live bacteria was achieved, however further optimization of experimental methods are needed for high-resolution visualization of the cellular environmental structural dynamics by AFM. This project enhanced the current insight into molecular architecture, structural and environmental variability of bacterial systems. The project partially funded research for two book chapters (1

iDynoMiCS: next‐generation individual‐based modelling of biofilms

DEFF Research Database (Denmark)

Lardon, Laurent; Merkey, Brian; Martins, Sónia

2011-01-01

how environmentally fluctuating oxygen availability affects the diversity and composition of a community of denitrifying bacteria that induce the denitrification pathway under anoxic or low oxygen conditions. We tested the hypothesis that the existence of these diverse strategies of denitrification...... and higher at intermediate frequency of change. The highly modular nature of the new computational model made this case study straightforward to implement, and reflects the sort of novel studies that can easily be executed with the new model....

[Darwin and bacteria].

Science.gov (United States)

Ledermann D, Walter

2009-02-01

As in 2009 the scientific world celebrates two hundreds years from the birthday of Charles Darwin and one hundred and fifty from the publication of The Origin of Species, an analysis of his complete work is performed, looking for any mention of bacteria. But it seems that the great naturahst never took knowledge about its existence, something rather improbable in a time when the discovery of bacteria shook the medical world, or he deliberately ignored them, not finding a place for such microscopic beings into his theory of evolution. But the bacteria badly affected his familiar life, killing scarlet fever one of his children and worsening to death the evolution of tuberculosis of his favourite Annie. Darwin himself could suffer the sickness of Chagas, whose etiological agent has a similar level to bacteria in the scale of evolution.

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

Directory of Open Access Journals (Sweden)

Miguel Ramirez

2014-01-01

Full Text Available Heterotrophic nitrifiers synthesize nitrogenous gasses when nitrifying ammonium ion. A Cupriavidus pauculus, previously thought an Alcaligenes sp. and noted as an active heterotrophic nitrifier-denitrifier, was examined for its ability to produce nitrogen gas (N2 and nitrous oxide (N2O while heterotrophically nitrifying the organic substrate pyruvic oxime [CH3–C(NOH–COOH]. Neither N2 nor N2O were produced. Nucleotide and phylogenetic analyses indicated that the organism is a member of a genus (Cupriavidus known for its resistance to metals and its metabolism of xenobiotics. The microbe (a Cupriavidus pauculus designated as C. pauculus strain UM1 was examined for its ability to perform heterotrophic nitrification in the presence of Cu2+ and Ni2+ and to metabolize the xenobiotic phenol. The bacterium heterotrophically nitrified well when either 1 mM Cu2+ or 0.5 mM Ni2+ was present in either enriched or minimal medium. The organism also used phenol as a sole carbon source in either the presence or absence of 1 mM Cu2+ or 0.5 mM Ni2+. The ability of this isolate to perform a number of different metabolisms, its noteworthy resistance to copper and nickel, and its potential use as a bioremediation agent are discussed.

The Experiment Study of Anaerobic Ammonia Oxidation Start-up by Using the Upflow Double Layer Anaerobic Filter

Directory of Open Access Journals (Sweden)

YAO Li

2018-02-01

Full Text Available Anammox is an efficient nitrogen removal process, but it is difficult to start-up and operate, and ananammox reactor is the efficient way to resolve this problem. The start-up of anammox reactor by upflow anaerobic filter was studied. Denitrifying sludge, anaerobic sludge, and mixed sludge was inoculated on the packing materials, respectively and an autotrophic denitrification condition was provided by the simulated wastewater influent. Along with the gradual increase of matrix concentration and hydraulic load, the microflora was converted to the anaerobic ammonium oxidation(anammoxreaction. The results showed that the anammox reaction could be started by all the three sludge, and the time of start-up of denitrifying sludge, anaerobic sludge, mixed sludge was 42, 54 days and 45 days, respectively. The best result was that inoculated with denitrifying sludge with 82.2% of the total nitrogen removal rate, which started-up quickly and nitrogen was removed efficiently. Double packing effectively improved the stability of anammox process in the reactor, in which the suitable influent concentration loading for the anammox bacteria was 270 mg·L-1 and 360 mg·L-1 for ammonia nitrogen and nitrite nitrogen, respectively, and the COD concentration could not be more than 150 mg· L-1. Furthermore, there was a coexist-effect for anaerobic ammonia oxidation and methanation in this reactor system.

Nitrate reducing bacterial activity in concrete cells of nuclear waste disposal

Science.gov (United States)

Alquier, M.; Kassim, C.; Bertron, A.; Rafrafi, Y.; Sablayrolles, C.; Albrecht, A.; Erable, B.

2013-07-01

Leaching experiments of solid matrices (bitumen and cement pastes) have been first implemented to define the physicochemical conditions that microorganisms are likely to meet at the bitumen-concrete interface (see the paper of Bertron et al.). Of course, as might be suspected, the cement matrix imposes highly alkaline pH conditions (10 denitrifying bacterial strains led us to select Halomonas desiderata as a model bacterium capable of catalyzing the reaction of nitrate reduction in these extreme conditions of pH. The denitrifying activity of Halomonas desiderata was quantified in batch bioreactor in the presence of solid matrices and / or leachate from bitumen and cement matrices. Denitrification was relatively fast in the presence of cement matrix (<100 hours) and 2 to 3 times slower in the presence of bituminous matrix. Overall, the presence of solid cement promoted the kinetics of denitrification. The observation of solid surfaces at the end of the experiment revealed the presence of a biofilm of Halomonas desiderata on the cement paste surface. These attached bacteria showed a denitrifying activity comparable to planktonic bacterial culture. On the other side, no colonization of bitumen could be highlighted as either by SEM or epifluorescence microscopy. Now, we are currently developing a continuous experimental bioreactor which should allow us a more rational understanding of the bitumen-cement-microbe interactions.

Nitrate reducing bacterial activity in concrete cells of nuclear waste disposal

Directory of Open Access Journals (Sweden)

Albrecht A.

2013-07-01

Full Text Available Leaching experiments of solid matrices (bitumen and cement pastes have been first implemented to define the physicochemical conditions that microorganisms are likely to meet at the bitumen-concrete interface (see the paper of Bertron et al.. Of course, as might be suspected, the cement matrix imposes highly alkaline pH conditions (10 denitrifying bacterial strains led us to select Halomonas desiderata as a model bacterium capable of catalyzing the reaction of nitrate reduction in these extreme conditions of pH. The denitrifying activity of Halomonas desiderata was quantified in batch bioreactor in the presence of solid matrices and / or leachate from bitumen and cement matrices. Denitrification was relatively fast in the presence of cement matrix (<100 hours and 2 to 3 times slower in the presence of bituminous matrix. Overall, the presence of solid cement promoted the kinetics of denitrification. The observation of solid surfaces at the end of the experiment revealed the presence of a biofilm of Halomonas desiderata on the cement paste surface. These attached bacteria showed a denitrifying activity comparable to planktonic bacterial culture. On the other side, no colonization of bitumen could be highlighted as either by SEM or epifluorescence microscopy. Now, we are currently developing a continuous experimental bioreactor which should allow us a more rational understanding of the bitumen-cement-microbe interactions.

Microbial Diversity and Characteristics in Anaerobic Environments in KURT Groundwater

International Nuclear Information System (INIS)

Roh, Yul; Oh, Jong Min; Rhee, Sung Keun; Yong, Jong Joong

2008-03-01

The Underground Research Tunnel (URT) located in Korea Atomic Energy Research Institute (KAERI), Daejeon, South Korea was recently constructed as an experimental site to study radionuclide transport, biogeochemistry, radionuclide-mineral interactions for the geological disposal of high level nuclear waste. Groundwater sampled from URT was used to examine microbial diversity and to enrich metal reducing bacteria for studying microbe-metal interactions. Genomic analysis indicated that the groundwater contained diverse microorganisms such as metal reducers, metal oxidizers, anaerobic denitrifying bacteria, and bacteria for reductive dechlorination. Metal-reducing bacteria enriched from the groundwater was used to study metal reduction and biomineralization. The metal-reducing bacteria enriched with acetate or lactate as the electron donors showed the bacteria reduced Fe(III)-citrate, Fe(III) oxyhydroxide, Mn(IV) oxide, and Cr(VI) as the electron acceptors. Preliminary study indicated that the enriched bacteria were able to use glucose, lactate, acetate, and hydrogen as electron donors while reducing Fe(III)-citrate or Fe(III) oxyhydroxide as the electron acceptor. The bacteria exhibited diverse mineral precipitation capabilities including the formation of magnetite, siderite, and rhodochrosite. The results indicated that Fe(III)- and metal-reducing communities are present in URT at the KAERI

Microbial Diversity in KURT Groundwater and Biomineralization Characteristics

International Nuclear Information System (INIS)

Roh, Yul; Rhee, Sung Keun; Oh, Jong Min; Park, Byung Jun

2009-03-01

The Underground Research Tunnel (URT) located in Korea Atomic Energy Research Institute (KAERI), Daejeon, South Korea was recently constructed as an experimental site to study radionuclide transport, biogeochemistry, radionuclide-mineral interactions for the geological disposal of high level nuclear waste. Groundwater sampled from URT was used to examine microbial diversity and to enrich metal reducing bacteria for studying microbe-metal interactions. Genomic analysis indicated that the groundwater contained diverse microorganisms such as metal reducers, metal oxidizers, anaerobic denitrifying bacteria, and bacteria for reductive dechlorination. Metal-reducing bacteria enriched from the groundwater was used to study metal reduction and biomineralization. The metal-reducing bacteria enriched with acetate or lactate as the electron donors showed the bacteria reduced Fe(III)-citrate, Fe(III) oxyhydroxides, Mn(IV) oxide, and Cr(VI) as the electron acceptors. Preliminary study indicated that the enriched bacteria were able to use glucose, lactate, acetate, and hydrogen as electron donors while reducing Fe(III)-citrate or Fe(III) oxyhydroxide as the electron acceptor. The bacteria exhibited diverse mineral precipitation capabilities including the formation of magnetite, siderite, and rhodochrosite. The results indicated that Fe(III)- and metal-reducing communities are present in URT at the KAERI

Sequentially aerated membrane biofilm reactors for autotrophic nitrogen removal: microbial community composition and dynamics

DEFF Research Database (Denmark)

Pellicer i Nàcher, Carles; Franck, Stephanie; Gülay, Arda

2014-01-01

Membrane-aerated biofilm reactors performing autotrophic nitrogen removal can be successfully applied to treat concentrated nitrogen streams. However, their process performance is seriously hampered by the growth of nitrite oxidizing bacteria (NOB). In this work we document how sequential aeration...... (rich in oxygen) and AnAOB in regions neighbouring the liquid phase. Both communities were separated by a transition region potentially populated by denitrifying heterotrophic bacteria. AOB and AnAOB bacterial groups were more abundant and diverse than NOB, and dominated by the r......-strategists Nitrosomonas europaea and Ca. Brocadia anammoxidans, respectively. Taken together, the present work presents tools to better engineer, monitor and control the microbial communities that support robust, sustainable and efficient nitrogen removal....

Potential role of bacteria packaging by protozoa in the persistence and transmission of pathogenic bacteria

Directory of Open Access Journals (Sweden)

Alix M Denoncourt

2014-05-01

Full Text Available Many pathogenic bacteria live in close association with protozoa. These unicellular eukaryotic microorganisms are ubiquitous in various environments. A number of protozoa such as amoebae and ciliates ingest pathogenic bacteria, package them usually in membrane structures, and then release them into the environment. Packaged bacteria are more resistant to various stresses and are more apt to survive than free bacteria. New evidence indicates that protozoa and not bacteria control the packaging process. It is possible that packaging is more common than suspected and may play a major role in the persistence and transmission of pathogenic bacteria. To confirm the role of packaging in the propagation of infections, it is vital that the molecular mechanisms governing the packaging of bacteria by protozoa be identified as well as elements related to the ecology of this process in order to determine whether packaging acts as a Trojan Horse.