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Sample records for 15n enriched ammonium

  1. Ammonium and nitrate uptake lengths in a small forested stream determined by {sup 15}N tracer and short-term nutrient enrichment experiments

    SciTech Connect

    Mulholland, P.J.; Tank, J.L.; Sanzone, D.M.; Webster, J.R.; Wollheim, W.; Peterson, B.J.; Meyer, J.L.

    1998-11-01

    Nutrient cycling is an important characteristic of all ecosystems, including streams. Nutrients often limit the growth rates of stream algae and heterotrophic microbes and the decomposition rate of allochthonous organic matter. Nutrient uptake (S{sub W}), defined as the mean distance traveled by a nutrient atom dissolved in stream water before uptake by biota is often used as an index of nutrient cycling in streams. It is often overlooked, however, that S{sub W} is not a measure of nutrient uptake rate per se, but rather a measure of the efficiency with which a stream utilizes the available nutrient supply. The ideal method for measuring S{sub W} involves short-term addition of a nutrient tracer. Regulatory constraints often preclude use of nutrient radiotracers in field studies and methodological difficulties and high analytical costs have previously hindered the use of stable isotope nutrient tracers (e.g., {sup 15}N). Short-term nutrient enrichments are an alternative to nutrient tracer additions for measuring S{sub W}.

  2. Stepwise enrichment of 15N along food chains: Further evidence and the relation between δ 15N and animal age

    NASA Astrophysics Data System (ADS)

    Minagawa, Masao; Wada, Eitaro

    1984-05-01

    The isotopic composition of nitrogen was measured in marine and fresh-water animals from the East China Sea, The Bering Sea, Lake Ashinoko and Usujiri intertidal zone. Primary producers, showed average δ15Nversus atmospheric nitrogen of +5.0%. (+3.4 to +7.5) in the Bering Sea and Lake Ashinoko, and +6.8%. (+6.0 to +7.6) in Usujiri intertidal zone. Blue green algae from the East China Sea show an average -0.55%. (-0.8 to +1.2). All consumers, Zooplankton, fish and bird exhibited Stepwise enrichment of 15N with increasing trophic level. The 15N enrichment at a single feeding process ranged from +1.3 to +5.3 averaging +3.4 ± 1.1%.. This isotopic fractionation seems to be independent of habitat. The effect of age in animals was obtained by analyzing two marine mussels. The soft tissue nitrogen showed +2.0%. enrichment relative to that of primary producers, and the magnitude was almost constant with shell ages ranging from 0 to 8 years. A similar 15N enrichment occurs in all Molluscs, Crustaceans, Insecta, Amphibia, Fish, Ave and Mammal species regardless of the difference in the form of excreted nitrogen and in laboratory cultured fish, brine shrimp and mice (+2.9 to +4.9%.). The excreted ammonia from guppy was sufficiently light to balance the concentration of 15N to animal body.

  3. Creating 13C- and 15N-enriched tree leaf litter for decomposition experiments

    NASA Astrophysics Data System (ADS)

    Szlavecz, K. A.; Pitz, S.; Chang, C.; Bernard, M.

    2013-12-01

    Labeling plant material with heavy isotopes of carbon and nitrogen can produce a traceable nutrient signal that can be followed into the different trophic levels and decomposer food web. We treated 60 tree saplings with 13C-enriched CO2 gas and 15N-enriched ammonium nitrate over a three-month period to create dually-labeled plant material for future decomposition experiments. The trees included both early (Red maple, Sweetgum, Tulip poplar) and late (American beech, White oak) successional deciduous tree species, and a conifer, White pine. We constructed a 2.4 m × 2.4 m × 2.4 m environmental chamber that was climate-controlled using an air conditioning system. An Arduino microcontroller interfaced with a Vaisala GMP343 CO2 probe maintained a CO2 concentration between 500-520 ppm by controlling a solenoid valve on the CO2 tank regulator. The trees were placed into the chamber in August 2012 and remained until senescence unless they were lost to death or disease. Ammonium nitrate was added twice, in September and October. Leaf samples were collected prior to the start of the experiment and after senescence, whereas root samples were collected only in December. Samples were dried, ground and analyzed using an isotope ratio mass spectrometer. American beech and White oak had 40% mortality, and 34% of tulip poplar trees were removed because of powdery mildew overgrowth or death. Most tulip poplar trees exhibited a second leaf out following senescence in late September. Nearly 1 kg of litter was produced with tulip poplar representing over half of the total mass. Levels of enrichment varied greatly by species. Beech (-14.2‰) and White oak (-4.8‰) had low levels of enrichment in comparison to early successional species such as Sweetgum (41.7‰) and Tulip poplar (30.7‰ [first leaf fall] and 238.0‰ [second leaf fall]). Leaf enrichment with 15N followed a similar pattern, though it was achieved at a higher level with δ15N values varying from 271.6‰ to 1354.2

  4. Accessible NMR Experiments Studying the Hydrodynamics of [subscript 15]N-Enriched Ubiquitin at Low Fields

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

    We have recently developed and implemented two experiments in biomolecular NMR for an undergraduate-level biophysical chemistry laboratory with commercially available [subscript 15]N-enriched human ubiquitin. These experiments take advantage of [subscript 15]N direct detection of the NMR signal. The first experiment develops skills in acquiring…

  5. Accessible NMR Experiments Studying the Hydrodynamics of [superscript 15]N-Enriched Ubiquitin at Low Fields

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

    We have recently developed and implemented two experiments in biomolecular NMR for an undergraduate-level biophysical chemistry laboratory with commercially available [superscript 15]N-enriched human ubiquitin. These experiments take advantage of [superscript 15]N direct detection of the NMR signal. The first experiment develops skills in…

  6. Increase of Natural 15N Enrichment of Soybean Nodules with Mean Nodule Mass 1

    PubMed Central

    Shearer, Georgia; Bryan, Barbara A.; Kohl, Daniel H.

    1984-01-01

    The 15N abundance of soybean (Glycine max L. Merrill var Harosoy) nodules is usually greater than it is for other tissues or for atmospheric N2. Results of experiments in which nodules were separated by size show that the magnitude of the 15N enrichment is correlated with nodule mass. The results support the hypothesis that 15N enrichment of nodules results from differential N isotopic fractionation for synthesis of nodule tissue versus synthesis of compounds for export from the nodule. The physiological significance of this hypothesis is that it requires that a substantial fraction of the N for nodule tissue synthesis in 15N-enriched nodules be N recently fixed within the same nodule. PMID:16663917

  7. Preparation and characterization of 15N-enriched, size-defined heparan sulfate precursor oligosaccharides

    PubMed Central

    Sigulinsky, Crystal; Babu, Ponnusamy; Victor, Xylophone V.; Kuberan, Balagurunathan

    2009-01-01

    We report the preparation of size-defined [15N]N-acetylheparosan oligosaccharides from Escherichia coli-derived 15N-enriched N-acetylheparosan. Optimized growth conditions of E. coli in minimal media containing 15NH4Cl yielded [15N]N-acetylheparosan on a preparative scale. Depolymerization of [15N]N-acetylheparosan by heparitinase I yielded resolvable, even-numbered oligosaccharides ranging from disaccharide to icosaccharide. Anion-exchange chromatography-assisted fractionation afforded size-defined [15N]N-acetylheparosan oligosaccharides identifiable by ESI-TOFMS. These isotopically labeled oligosaccharides will prove to be valuable research tools for the chemoenzymatic synthesis of heparin and heparan sulfate oligosaccharides and for the study of their structural biology. PMID:19945695

  8. Highly 15N-Enriched Chondritic Clasts in the Isheyevo Meteorite

    SciTech Connect

    Bonal, L; Huss, G R; Krot, A N; Nagashima, K; Ishii, H A; Bradley, J P; Hutcheon, I D

    2009-01-14

    The metal-rich carbonaceous chondrites (CB and CH) have the highest whole-rock {sup 15}N enrichment ({delta}{sup 15}N up to +1500{per_thousand}), similar to {delta}{sup 15}N values reported in micron-sized regions (hotspots) of Interplanetary Dust Particles (IDPs) of possibly cometary origin and fine-grained matrices of unmetamorphosed chondrites. These {sup 15}N-rich hotspots are commonly attributed to low-temperature ion-molecule reactions in the protosolar molecular cloud or in the outer part of the protoplanetary disk. The nature of the whole-rock {sup 15}N enrichment of the metal-rich chondrites is not understood. We report a discovery of a unique type of primitive chondritic clasts in the CH/CB-like meteorite Isheyevo, which provides important constraints on the origin of {sup 15}N anomaly in metal-rich chondrites and nitrogen-isotope fractionation in the Solar System. These clasts contain tiny chondrules and refractory inclusions (5-15 {micro}m in size), and abundant ferromagnesian chondrule fragments (1-50 {micro}m in size) embedded in the partly hydrated, fine-grained matrix material composed of olivines, pyroxenes, poorly-organized aromatic organics, phyllosilicates and other hydrous phases. The mineralogy and oxygen isotope compositions of chondrules and refractory inclusions in the clasts are similar to those in the Isheyevo host, suggesting formation at similar heliocentric distances. In contrast to the previously known extraterrestrial samples, the fine-grained material in the clasts is highly and rather uniformly enriched in {sup 15}N, with bulk {delta}{sup 15}N values ranging between +1000 and +1300{per_thousand}; the {delta}{sup 15}N values in rare hotspots range from +1400 to +4000{per_thousand}. Since fine-grained matrices in the lithic clasts are the only component containing thermally unprocessed (during CAI and chondrule formation or during impact melting) materials that accreted into the metal rich chondrite parent body(ies), the {sup 15}N-enriched

  9. Soil Microbial 15N-Natural Abundance is Enriched Relative to Other Soil N Pools and Indicates Microbial C-Limitation

    NASA Astrophysics Data System (ADS)

    Dijkstra, P.; Doucett, R.; Hart, S. C.; Boring, L. L.; Schwartz, E.; Hungate, B. A.

    2004-12-01

    Soil microbial biomass is responsible for many of the nitrogen (N) transformations that occur between different soil organic matter pools, plants and atmosphere. For this reason, it is important to learn more about the 15N natural abundance of these organisms. The microbial biomass takes up organic carbon (C) and N and inorganic N for assimilation and respiration. Under C-limited conditions, organic N compounds are mainly utilized as C-source, and excess N leaves the cell (mineralization). The processes of N assimilation, N dissimilation, and export discriminate against the heavier 15N isotope. This leaves the cells 15N enriched compared to their supposed substrates, or total soil N. Selective uptake of enriched ammonium (as a result of nitrification) may contribute to the higher \\delta15N of the micro-organisms. We measured 15N natural abundances of the microbial biomass, using the chloroform-fumigation-extraction method, in grassland soils along an elevation gradient, a fire-disturbance gradient in Florida, a ponderosa forest restoration study, and a dung-gradient near a water source in desert grassland and found 0-12\\permil difference between the soil extractable N and the microbial biomass. We also found that the enrichment is quantitatively dependent on C availability. We speculate that, in addition to an indicator for C availability to the microbes, microbial transformations may explain the 15N-enrichment of soil organic matter with depth.

  10. In Vivo Fluxes in the Ammonium-Assimilatory Pathways in Corynebacterium glutamicum Studied by 15N Nuclear Magnetic Resonance

    PubMed Central

    Tesch, M.; de Graaf, A. A.; Sahm, H.

    1999-01-01

    Glutamate dehydrogenase (GDH) and glutamine synthetase (GS)–glutamine 2-oxoglutarate-aminotransferase (GOGAT) represent the two main pathways of ammonium assimilation in Corynebacterium glutamicum. In this study, the ammonium assimilating fluxes in vivo in the wild-type ATCC 13032 strain and its GDH mutant were quantitated in continuous cultures. To do this, the incorporation of 15N label from [15N]ammonium in glutamate and glutamine was monitored with a time resolution of about 10 min with in vivo 15N nuclear magnetic resonance (NMR) used in combination with a recently developed high-cell-density membrane-cyclone NMR bioreactor system. The data were used to tune a standard differential equation model of ammonium assimilation that comprised ammonia transmembrane diffusion, GDH, GS, GOGAT, and glutamine amidotransferases, as well as the anabolic incorporation of glutamate and glutamine into biomass. The results provided a detailed picture of the fluxes involved in ammonium assimilation in the two different C. glutamicum strains in vivo. In both strains, transmembrane equilibration of 100 mM [15N]ammonium took less than 2 min. In the wild type, an unexpectedly high fraction of 28% of the NH4+ was assimilated via the GS reaction in glutamine, while 72% were assimilated by the reversible GDH reaction via glutamate. GOGAT was inactive. The analysis identified glutamine as an important nitrogen donor in amidotransferase reactions. The experimentally determined amount of 28% of nitrogen assimilated via glutamine is close to a theoretical 21% calculated from the high peptidoglycan content of C. glutamicum. In the GDH mutant, glutamate was exclusively synthesized over the GS/GOGAT pathway. Its level was threefold reduced compared to the wild type. PMID:10049869

  11. Why is Mineral-Associated Organic Matter Enriched in 15N? Evidence from Grazed Pasture Soil

    NASA Astrophysics Data System (ADS)

    Baisden, W. T.; Wells, N. S.; Mudge, P. L.; Clough, T. J.; Schipper, L. A.; Ghani, A.; Stevenson, B.

    2014-12-01

    Throughout the scientific literature, measurements across soil depth and density fractions suggest that, with few exceptions, mineral-associated organic matter (OM) has higher δ15N than non-mineral-associated OM. This implies that the δ15N difference between N inputs and mineral-stabilized OM may characterize the microbial processes involved in stabilization and mineral association. Yet current understanding of observed N isotope fractionation in terrestrial ecosystems suggests the large isotope effects are expressed during inorganic N transformations from NH4 to gaseous loss pathways of NH3 volatilization and denitrification. How can the relative importance of N isotope fractionation during OM stabilization versus loss pathways be resolved? We recently examined N isofluxes when a temporary nitrogen excess is created by urine deposition in a New Zealand dairy pasture. We found that the N isotopic composition of volatilized NH3, and NO3 available for leaching or denitrification could not be linked back to the added N using Rayleigh distillation models. Instead, the results imply that the added N was immobilized, and the N available for losses was increasingly derived from mineralization of organic matter during the course of the experiment. These results are consistent with recent evidence of enhanced OM mineralization in urine patches, understanding of N isotope mass balances and long-standing evidence that gross mineralization and immobilization fluxes greatly exceed net mineralization and nitrification, except at very high N saturation. These results suggest that where 15N enrichment occurs due to fractionating loss pathways, the isotope effects are primarily transmitted to immobilized N, forming 15N enriched stabilized OM. This further explains earlier findings that the δ15N of soil OM represents an integrated indicator of losses, reflecting the intensity and duration of pastoral agriculture. We suggest that development of an indicator based on δ15N in

  12. Food webs of two intermittently open estuaries receiving 15N-enriched sewage effluent

    NASA Astrophysics Data System (ADS)

    Hadwen, Wade L.; Arthington, Angela H.

    2007-01-01

    Carbon and nitrogen stable isotope signatures were used to assess the response of food webs to sewage effluent discharged into two small intermittently open estuaries in northern New South Wales, Australia. One of these systems, Tallows Creek, has a history of direct sewage inputs, whilst the other, Belongil Creek, receives wastewater via an extensive wetland treatment system. The food webs of both systems were driven by algal sources of carbon, reflecting high autotrophic productivity in response to the nutrients entering the system from sewage effluent. All aquatic biota collected from Tallows Creek had significantly enriched δ15N signatures relative to their conspecifics from Belongil Creek, indicating that sewage nitrogen had been assimilated and transferred throughout the Tallows Creek food web. These δ15N values were higher than those reported from studies in permanently open estuaries receiving sewage effluent. We suggest that these enriched signatures and the transfer of nitrogen throughout the entire food web reflect differences in hydrology and associated nitrogen cycling processes between permanently open and intermittently open estuaries. Although all organisms in Tallows Creek were generally 15N-enriched, isotopically light (less 15N-enriched) individuals of estuary perchlet ( Ambassis marianus) and sea mullet ( Mugil cephalus) were also collected. These individuals were most likely recent immigrants into Tallows Creek, as this system had only recently been opened to the ocean. This isotopic discrimination between resident (enriched) and immigrant (significantly less enriched) individuals can provide information on fish movement patterns and the role of heavily polluted intermittently open estuaries in supporting commercially and recreationally valuable estuarine species.

  13. Plant community change mediates the response of foliar δ(15)N to CO 2 enrichment in mesic grasslands.

    PubMed

    Polley, H Wayne; Derner, Justin D; Jackson, Robert B; Gill, Richard A; Procter, Andrew C; Fay, Philip A

    2015-06-01

    Rising atmospheric CO2 concentration may change the isotopic signature of plant N by altering plant and microbial processes involved in the N cycle. CO2 may increase leaf δ(15)N by increasing plant community productivity, C input to soil, and, ultimately, microbial mineralization of old, (15)N-enriched organic matter. We predicted that CO2 would increase aboveground productivity (ANPP; g biomass m(-2)) and foliar δ(15)N values of two grassland communities in Texas, USA: (1) a pasture dominated by a C4 exotic grass, and (2) assemblages of tallgrass prairie species, the latter grown on clay, sandy loam, and silty clay soils. Grasslands were exposed in separate experiments to a pre-industrial to elevated CO2 gradient for 4 years. CO2 stimulated ANPP of pasture and of prairie assemblages on each of the three soils, but increased leaf δ(15)N only for prairie plants on a silty clay. δ(15)N increased linearly as mineral-associated soil C declined on the silty clay. Mineral-associated C declined as ANPP increased. Structural equation modeling indicted that CO2 increased ANPP partly by favoring a tallgrass (Sorghastrum nutans) over a mid-grass species (Bouteloua curtipendula). CO2 may have increased foliar δ(15)N on the silty clay by reducing fractionation during N uptake and assimilation. However, we interpret the soil-specific, δ(15)N-CO2 response as resulting from increased ANPP that stimulated mineralization from recalcitrant organic matter. By contrast, CO2 favored a forb species (Solanum dimidiatum) with higher δ(15)N than the dominant grass (Bothriochloa ischaemum) in pasture. CO2 enrichment changed grassland δ(15)N by shifting species relative abundances. PMID:25604918

  14. Plant community change mediates the response of foliar δ(15)N to CO 2 enrichment in mesic grasslands.

    PubMed

    Polley, H Wayne; Derner, Justin D; Jackson, Robert B; Gill, Richard A; Procter, Andrew C; Fay, Philip A

    2015-06-01

    Rising atmospheric CO2 concentration may change the isotopic signature of plant N by altering plant and microbial processes involved in the N cycle. CO2 may increase leaf δ(15)N by increasing plant community productivity, C input to soil, and, ultimately, microbial mineralization of old, (15)N-enriched organic matter. We predicted that CO2 would increase aboveground productivity (ANPP; g biomass m(-2)) and foliar δ(15)N values of two grassland communities in Texas, USA: (1) a pasture dominated by a C4 exotic grass, and (2) assemblages of tallgrass prairie species, the latter grown on clay, sandy loam, and silty clay soils. Grasslands were exposed in separate experiments to a pre-industrial to elevated CO2 gradient for 4 years. CO2 stimulated ANPP of pasture and of prairie assemblages on each of the three soils, but increased leaf δ(15)N only for prairie plants on a silty clay. δ(15)N increased linearly as mineral-associated soil C declined on the silty clay. Mineral-associated C declined as ANPP increased. Structural equation modeling indicted that CO2 increased ANPP partly by favoring a tallgrass (Sorghastrum nutans) over a mid-grass species (Bouteloua curtipendula). CO2 may have increased foliar δ(15)N on the silty clay by reducing fractionation during N uptake and assimilation. However, we interpret the soil-specific, δ(15)N-CO2 response as resulting from increased ANPP that stimulated mineralization from recalcitrant organic matter. By contrast, CO2 favored a forb species (Solanum dimidiatum) with higher δ(15)N than the dominant grass (Bothriochloa ischaemum) in pasture. CO2 enrichment changed grassland δ(15)N by shifting species relative abundances.

  15. Determination of the delta(15N/14N)of Ammonium (NH4+) in Water: RSIL Lab Code 2898

    USGS Publications Warehouse

    Hannon, Janet E.; Böhlke, John Karl

    2008-01-01

    The purpose of the technique described by Reston Stable Isotope Laboratory (RSIL) lab code 2898 is to determine the N isotopic composition, delta(15N/14N), abbreviated as d15N, of ammonium (NH4+) in water (freshwater and saline water). The procedure involves converting dissolved NH4+ into NH3 gas by raising the pH of the sample to above 9 with MgO and subsequently trapping the gas quantitatively as (NH4)2SO4 on a glass fiber (GF) filter. The GF filter is saturated with NaHSO4 and pressure sealed between two gas-permeable polypropylene filters. The GF filter 'sandwich' floats on the surface of the water sample in a closed bottle. NH3 diffuses from the water through the polypropylene filter and reacts with NaHSO4, forming (NH4)2SO4 on the GF filter. The GF filter containing (NH4)2SO4 is dried and then combusted with a Carlo Erba NC 2500 elemental analyzer (EA), which is used to convert total nitrogen in a solid sample into N2 gas. The EA is connected to a continuous-flow isotope-ratio mass spectrometer (CF-IRMS), which determines the relative difference in ratios of the amounts of the stable isotopes of nitrogen (15N and 14N) of the product N2 gas and a reference N2 gas. The filters containing the samples are compressed in tin capsules and loaded into a Costech Zero-Blank Autosampler on the EA. Under computer control, samples then are dropped into a heated reaction tube that contains an oxidant, where combustion takes place in a He atmosphere containing an excess of O2 gas. To remove S-O gases produced from the NaHSO4, a plug of Ag-coated Cu wool is inserted at the bottom of the reaction tube. Combustion products are transported by a He carrier through a reduction furnace to remove excess O2, toconvert all nitrogen oxides to N2, and to remove any remaining S-O gases. The gases then pass through a drying tube to remove water. The gas-phase products, mainly N2 and a small amount of background CO2, are separated by a gas chromatograph (GC). The gas is then introduced

  16. A suite of sensitive chemical methods to determine the δ15N of ammonium, nitrate and total dissolved N in soil extracts.

    PubMed

    Lachouani, Petra; Frank, Alexander H; Wanek, Wolfgang

    2010-12-30

    Natural (15) N abundances (δ(15) N values) of different soil nitrogen pools deliver crucial information on the soil N cycle for the analysis of biogeochemical processes. Here we report on a complete suite of methods for sensitive δ(15) N analysis in soil extracts. A combined chemical reaction of vanadium(III) chloride (VCl(3) ) and sodium azide under acidic conditions is used to convert nitrate into N(2) O, which is subsequently analyzed by purge-and-trap isotope ratio mass spectrometry (PTIRMS) with a cryo-focusing unit. Coupled with preparation steps (microdiffusion for collection of ammonium, alkaline persulfate oxidation to convert total dissolved N (TDN) or ammonium into nitrate) this allows the determination of the δ(15) N values of nitrate, ammonium and total dissolved N (dissolved organic N, microbial biomass N) in soil extracts with the same basic protocol. The limits of quantification for δ(15) N analysis with a precision of 0.5‰ were 12.4 µM for ammonium, 23.7 µM for TDN, 16.5 µM for nitrate and 22.7 µM for nitrite.

  17. Litterfall 15N abundance indicates declining soil nitrogen availability in a free-air CO2 enrichment experiment.

    PubMed

    Garten, Charles T; Iversen, Colleen M; Norby, Richard J

    2011-01-01

    Forest productivity increases in response to carbon dioxide (CO2) enrichment of the atmosphere. However, in nitrogen-limited ecosystems, increased productivity may cause a decline in soil nitrogen (N) availability and induce a negative feedback on further enhancement of forest production. In a free-air CO2 enrichment (FACE) experiment, the response of sweetgum (Liquidambar styraciflua L.) productivity to elevated CO2 concentrations [CO2] has declined over time, but documenting an associated change in soil N availability has been difficult. Here we assess the time history of soil N availability through analysis of natural 15N abundance in archived samples of freshly fallen leaf litterfall. Litterfall delta15N declined from 1998 to 2005, and the rate of decline was significantly faster in elevated [CO2]. Declining leaf litterfall delta15N is indicative of a tighter ecosystem N cycle and more limited soil N availability. By integrating N availability over time and throughout the soil profile, temporal dynamics in leaf litterfall delta15N provide a powerful tool for documenting changes in N availability and the critical feedbacks between C and N cycles that will control forest response to elevated atmospheric CO2 concentrations.

  18. Solving the woolly mammoth conundrum: amino acid 15N-enrichment suggests a distinct forage or habitat

    PubMed Central

    Schwartz-Narbonne, Rachel; Longstaffe, Fred J.; Metcalfe, Jessica Z.; Zazula, Grant

    2015-01-01

    Understanding woolly mammoth ecology is key to understanding Pleistocene community dynamics and evaluating the roles of human hunting and climate change in late Quaternary megafaunal extinctions. Previous isotopic studies of mammoths’ diet and physiology have been hampered by the ‘mammoth conundrum’: woolly mammoths have anomalously high collagen δ15N values, which are more similar to coeval carnivores than herbivores, and which could imply a distinct diet and (or) habitat, or a physiological adaptation. We analyzed individual amino acids from collagen of adult woolly mammoths and coeval species, and discovered greater  15N enrichment in source amino acids of woolly mammoths than in most other herbivores or carnivores. Woolly mammoths consumed an isotopically distinct food source, reflective of extreme aridity, dung fertilization, and (or) plant selection. This dietary signal suggests that woolly mammoths occupied a distinct habitat or forage niche relative to other Pleistocene herbivores. PMID:26056037

  19. Solving the woolly mammoth conundrum: amino acid 15N-enrichment suggests a distinct forage or habitat

    NASA Astrophysics Data System (ADS)

    Schwartz-Narbonne, Rachel; Longstaffe, Fred J.; Metcalfe, Jessica Z.; Zazula, Grant

    2015-06-01

    Understanding woolly mammoth ecology is key to understanding Pleistocene community dynamics and evaluating the roles of human hunting and climate change in late Quaternary megafaunal extinctions. Previous isotopic studies of mammoths’ diet and physiology have been hampered by the ‘mammoth conundrum’: woolly mammoths have anomalously high collagen δ15N values, which are more similar to coeval carnivores than herbivores, and which could imply a distinct diet and (or) habitat, or a physiological adaptation. We analyzed individual amino acids from collagen of adult woolly mammoths and coeval species, and discovered greater  15N enrichment in source amino acids of woolly mammoths than in most other herbivores or carnivores. Woolly mammoths consumed an isotopically distinct food source, reflective of extreme aridity, dung fertilization, and (or) plant selection. This dietary signal suggests that woolly mammoths occupied a distinct habitat or forage niche relative to other Pleistocene herbivores.

  20. Simple approach for the preparation of (15-15)N2-enriched water for nitrogen fixation assessments: evaluation, application and recommendations.

    PubMed

    Klawonn, Isabell; Lavik, Gaute; Böning, Philipp; Marchant, Hannah K; Dekaezemacker, Julien; Mohr, Wiebke; Ploug, Helle

    2015-01-01

    Recent findings revealed that the commonly used (15)N2 tracer assay for the determination of dinitrogen (N2) fixation can underestimate the activity of aquatic N2-fixing organisms. Therefore, a modification to the method using pre-prepared (15-15)N2-enriched water was proposed. Here, we present a rigorous assessment and outline a simple procedure for the preparation of (15-15)N2-enriched water. We recommend to fill sterile-filtered water into serum bottles and to add (15-15)N2 gas to the water in amounts exceeding the standard N2 solubility, followed by vigorous agitation (vortex mixing ≥ 5 min). Optionally, water can be degassed at low-pressure (≥950 mbar) for 10 min prior to the (15-15)N2 gas addition to indirectly enhance the (15-15)N2 concentration. This preparation of (15-15)N2-enriched water can be done within 1 h using standard laboratory equipment. The final (15)N-atom% excess was 5% after replacing 2-5% of the incubation volume with (15-15)N2-enriched water. Notably, the addition of (15-15)N2-enriched water can alter levels of trace elements in the incubation water due to the contact of (15-15)N2-enriched water with glass, plastic and rubber ware. In our tests, levels of trace elements (Fe, P, Mn, Mo, Cu, Zn) increased by up to 0.1 nmol L(-1) in the final incubation volume, which may bias rate measurements in regions where N2 fixation is limited by trace elements. For these regions, we tested an alternative way to enrich water with (15-15)N2. The (15-15)N2 was injected as a bubble directly to the incubation water, followed by gentle shaking. Immediately thereafter, the bubble was replaced with water to stop the (15-15)N2 equilibration. This approach achieved a (15)N-atom% excess of 6.6 ± 1.7% when adding 2 mL (15-15)N2 per liter of incubation water. The herein presented methodological tests offer guidelines for the (15)N2 tracer assay and thus, are crucial to circumvent methodological draw-backs for future N2 fixation assessments. PMID:26300853

  1. Simple approach for the preparation of 15−15N2-enriched water for nitrogen fixation assessments: evaluation, application and recommendations

    PubMed Central

    Klawonn, Isabell; Lavik, Gaute; Böning, Philipp; Marchant, Hannah K.; Dekaezemacker, Julien; Mohr, Wiebke; Ploug, Helle

    2015-01-01

    Recent findings revealed that the commonly used 15N2 tracer assay for the determination of dinitrogen (N2) fixation can underestimate the activity of aquatic N2-fixing organisms. Therefore, a modification to the method using pre-prepared 15−15N2-enriched water was proposed. Here, we present a rigorous assessment and outline a simple procedure for the preparation of 15−15N2-enriched water. We recommend to fill sterile-filtered water into serum bottles and to add 15−15N2 gas to the water in amounts exceeding the standard N2 solubility, followed by vigorous agitation (vortex mixing ≥ 5 min). Optionally, water can be degassed at low-pressure (≥950 mbar) for 10 min prior to the 15−15N2 gas addition to indirectly enhance the 15−15N2 concentration. This preparation of 15−15N2-enriched water can be done within 1 h using standard laboratory equipment. The final 15N-atom% excess was 5% after replacing 2–5% of the incubation volume with 15−15N2-enriched water. Notably, the addition of 15−15N2-enriched water can alter levels of trace elements in the incubation water due to the contact of 15−15N2-enriched water with glass, plastic and rubber ware. In our tests, levels of trace elements (Fe, P, Mn, Mo, Cu, Zn) increased by up to 0.1 nmol L−1 in the final incubation volume, which may bias rate measurements in regions where N2 fixation is limited by trace elements. For these regions, we tested an alternative way to enrich water with 15−15N2. The 15−15N2 was injected as a bubble directly to the incubation water, followed by gentle shaking. Immediately thereafter, the bubble was replaced with water to stop the 15−15N2 equilibration. This approach achieved a 15N-atom% excess of 6.6 ± 1.7% when adding 2 mL 15−15N2 per liter of incubation water. The herein presented methodological tests offer guidelines for the 15N2 tracer assay and thus, are crucial to circumvent methodological draw-backs for future N2 fixation assessments. PMID:26300853

  2. Measurement of (15)N enrichment of glutamine and urea cycle amino acids derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate using liquid chromatography-tandem quadrupole mass spectrometry.

    PubMed

    Nakamura, Hidehiro; Karakawa, Sachise; Watanabe, Akiko; Kawamata, Yasuko; Kuwahara, Tomomi; Shimbo, Kazutaka; Sakai, Ryosei

    2015-05-01

    6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is an amino acid-specific derivatizing reagent that has been used for sensitive amino acid quantification by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). In this study, we aimed to evaluate the ability of this method to measure the isotopic enrichment of amino acids and to determine the positional (15)N enrichment of urea cycle amino acids (i.e., arginine, ornithine, and citrulline) and glutamine. The distribution of the M and M+1 isotopomers of each natural AQC-amino acid was nearly identical to the theoretical distribution. The standard deviation of the (M+1)/M ratio for each amino acid in repeated measurements was approximately 0.1%, and the ratios were stable regardless of the injected amounts. Linearity in the measurements of (15)N enrichment was confirmed by measuring a series of (15)N-labeled arginine standards. The positional (15)N enrichment of urea cycle amino acids and glutamine was estimated from the isotopic distribution of unique fragment ions generated at different collision energies. This method was able to identify their positional (15)N enrichment in the plasma of rats fed (15)N-labeled glutamine. These results suggest the utility of LC-MS/MS detection of AQC-amino acids for the measurement of isotopic enrichment in (15)N-labeled amino acids and indicate that this method is useful for the study of nitrogen metabolism in living organisms.

  3. Fossil Fuel Combustion-Related Emissions Dominate Atmospheric Ammonia Sources during Severe Haze Episodes: Evidence from (15)N-Stable Isotope in Size-Resolved Aerosol Ammonium.

    PubMed

    Pan, Yuepeng; Tian, Shili; Liu, Dongwei; Fang, Yunting; Zhu, Xiaying; Zhang, Qiang; Zheng, Bo; Michalski, Greg; Wang, Yuesi

    2016-08-01

    The reduction of ammonia (NH3) emissions is urgently needed due to its role in aerosol nucleation and growth causing haze formation during its conversion into ammonium (NH4(+)). However, the relative contributions of individual NH3 sources are unclear, and debate remains over whether agricultural emissions dominate atmospheric NH3 in urban areas. Based on the chemical and isotopic measurements of size-resolved aerosols in urban Beijing, China, we find that the natural abundance of (15)N (expressed using δ(15)N values) of NH4(+) in fine particles varies with the development of haze episodes, ranging from -37.1‰ to -21.7‰ during clean/dusty days (relative humidity: ∼ 40%), to -13.1‰ to +5.8‰ during hazy days (relative humidity: 70-90%). After accounting for the isotope exchange between NH3 gas and aerosol NH4(+), the δ(15)N value of the initial NH3 during hazy days is found to be -14.5‰ to -1.6‰, which indicates fossil fuel-based emissions. These emissions contribute 90% of the total NH3 during hazy days in urban Beijing. This work demonstrates the analysis of δ(15)N values of aerosol NH4(+) to be a promising new tool for partitioning atmospheric NH3 sources, providing policy makers with insights into NH3 emissions and secondary aerosols for regulation in urban environments. PMID:27359161

  4. Fossil Fuel Combustion-Related Emissions Dominate Atmospheric Ammonia Sources during Severe Haze Episodes: Evidence from (15)N-Stable Isotope in Size-Resolved Aerosol Ammonium.

    PubMed

    Pan, Yuepeng; Tian, Shili; Liu, Dongwei; Fang, Yunting; Zhu, Xiaying; Zhang, Qiang; Zheng, Bo; Michalski, Greg; Wang, Yuesi

    2016-08-01

    The reduction of ammonia (NH3) emissions is urgently needed due to its role in aerosol nucleation and growth causing haze formation during its conversion into ammonium (NH4(+)). However, the relative contributions of individual NH3 sources are unclear, and debate remains over whether agricultural emissions dominate atmospheric NH3 in urban areas. Based on the chemical and isotopic measurements of size-resolved aerosols in urban Beijing, China, we find that the natural abundance of (15)N (expressed using δ(15)N values) of NH4(+) in fine particles varies with the development of haze episodes, ranging from -37.1‰ to -21.7‰ during clean/dusty days (relative humidity: ∼ 40%), to -13.1‰ to +5.8‰ during hazy days (relative humidity: 70-90%). After accounting for the isotope exchange between NH3 gas and aerosol NH4(+), the δ(15)N value of the initial NH3 during hazy days is found to be -14.5‰ to -1.6‰, which indicates fossil fuel-based emissions. These emissions contribute 90% of the total NH3 during hazy days in urban Beijing. This work demonstrates the analysis of δ(15)N values of aerosol NH4(+) to be a promising new tool for partitioning atmospheric NH3 sources, providing policy makers with insights into NH3 emissions and secondary aerosols for regulation in urban environments.

  5. Salt marsh ecosystem biogeochemical responses to nutrient enrichment: a paired 15N tracer study.

    PubMed

    Drake, D C; Peterson, Bruce J; Galván, Kari A; Deegan, Linda A; Hopkinson, Charles; Johnson, J Michael; Koop-Jakobsen, K; Lemay, Lynsey E; Picard, Christian

    2009-09-01

    We compared processing and fate of dissolved NO3- in two New England salt marsh ecosystems, one receiving natural flood tide concentrations of approximately 1-4 micromol NO3-/ L and the other receiving experimentally fertilized flood tides containing approximately 70-100 micromol NO3-/ L. We conducted simultaneous 15NO3- (isotope) tracer additions from 23 to 28 July 2005 in the reference (8.4 ha) and fertilized (12.4 ha) systems to compare N dynamics and fate. Two full tidal cycles were intensively studied during the paired tracer additions. Resulting mass balances showed that essentially 100% (0.48-0.61 mol NO3-N.ha(-1).h(-1)) of incoming NO3- was assimilated, dissimilated, sorbed, or sedimented (processed) within a few hours in the reference system when NO3- concentrations were 1.3-1.8 micromol/L. In contrast, only 50-60% of incoming NO3- was processed in the fertilized system when NO3- concentrations were 84-96 micromol/L; the remainder was exported in ebb tidewater. Gross NO3- processing was approximately 40 times higher in the fertilized system at 19.34-24.67 mol NO3-N.ha(-1).h(-1). Dissimilatory nitrate reduction to ammonium was evident in both systems during the first 48 h of the tracer additions but <1% of incoming 15NO3- was exported as 15NH4+. Nitrification rates calculated by 15NO3- dilution were 6.05 and 4.46 mol.ha(-1).h(-1) in the fertilized system but could not be accurately calculated in the reference system due to rapid (<4 h) NO3- turnover. Over the five-day paired tracer addition, sediments sequestered a small fraction of incoming NO3-, although the efficiency of sequestration was 3.8% in the reference system and 0.7% in the fertilized system. Gross sediment N sequestration rates were similar at 13.5 and 12.6 mol.ha(-1).d(-1), respectively. Macrophyte NO3- uptake efficiency, based on tracer incorporation in aboveground tissues, was considerably higher in the reference system (16.8%) than the fertilized system (2.6%), although bulk uptake of NO3

  6. CO2 enrichment modulates ammonium nutrition in tomato adjusting carbon and nitrogen metabolism to stomatal conductance.

    PubMed

    Vega-Mas, Izargi; Marino, Daniel; Sánchez-Zabala, Joseba; González-Murua, Carmen; Estavillo, Jose María; González-Moro, María Begoña

    2015-12-01

    Ammonium (NH4(+)) toxicity typically occurs in plants exposed to high environmental NH4(+) concentration. NH4(+) assimilating capacity may act as a biochemical mechanism avoiding its toxic accumulation but requires a fine tuning between nitrogen assimilating enzymes and carbon anaplerotic routes. In this work, we hypothesized that extra C supply, exposing tomato plants cv. Agora Hybrid F1 to elevated atmospheric CO2, could improve photosynthetic process and thus ameliorate NH4(+) assimilation and tolerance. Plants were grown under nitrate (NO3(-)) or NH4(+) as N source (5-15mM), under two atmospheric CO2 levels, 400 and 800ppm. Growth and gas exchange parameters, (15)N isotopic signature, C and N metabolites and enzymatic activities were determined. Plants under 7.5mM N equally grew independently of the N source, while higher ammonium supply resulted toxic for growth. However, specific stomatal closure occurred in 7.5mM NH4(+)-fed plants under elevated CO2 improving water use efficiency (WUE) but compromising plant N status. Elevated CO2 annulled the induction of TCA anaplerotic enzymes observed at non-toxic NH4(+) nutrition under ambient CO2. Finally, CO2 enrichment benefited tomato growth under both nutritions, and although it did not alleviate tomato NH4(+) tolerance it did differentially regulate plant metabolism in N-source and -dose dependent manner. PMID:26706056

  7. CO2 enrichment modulates ammonium nutrition in tomato adjusting carbon and nitrogen metabolism to stomatal conductance.

    PubMed

    Vega-Mas, Izargi; Marino, Daniel; Sánchez-Zabala, Joseba; González-Murua, Carmen; Estavillo, Jose María; González-Moro, María Begoña

    2015-12-01

    Ammonium (NH4(+)) toxicity typically occurs in plants exposed to high environmental NH4(+) concentration. NH4(+) assimilating capacity may act as a biochemical mechanism avoiding its toxic accumulation but requires a fine tuning between nitrogen assimilating enzymes and carbon anaplerotic routes. In this work, we hypothesized that extra C supply, exposing tomato plants cv. Agora Hybrid F1 to elevated atmospheric CO2, could improve photosynthetic process and thus ameliorate NH4(+) assimilation and tolerance. Plants were grown under nitrate (NO3(-)) or NH4(+) as N source (5-15mM), under two atmospheric CO2 levels, 400 and 800ppm. Growth and gas exchange parameters, (15)N isotopic signature, C and N metabolites and enzymatic activities were determined. Plants under 7.5mM N equally grew independently of the N source, while higher ammonium supply resulted toxic for growth. However, specific stomatal closure occurred in 7.5mM NH4(+)-fed plants under elevated CO2 improving water use efficiency (WUE) but compromising plant N status. Elevated CO2 annulled the induction of TCA anaplerotic enzymes observed at non-toxic NH4(+) nutrition under ambient CO2. Finally, CO2 enrichment benefited tomato growth under both nutritions, and although it did not alleviate tomato NH4(+) tolerance it did differentially regulate plant metabolism in N-source and -dose dependent manner.

  8. Macroalgal bioindicators (growth, tissue N, delta(15)N) detect nutrient enrichment from shrimp farm effluent entering Opunohu Bay, Moorea, French Polynesia.

    PubMed

    Lin, David T; Fong, Peggy

    2008-02-01

    Nutrient enrichment from shrimp aquaculture poses an increasing environmental threat due to the industry's projected rapid growth and unsustainable management practices. Traditional methods to monitor impacts emphasize water quality sampling; however, there are many advantages to bioindicators, especially in developing countries. We investigated the usefulness of three bioindicators -- growth, tissue nitrogen content and nitrogen stable isotope signature (delta(15)N) -- in the tropical red macroalga Acanthophora spicifera. Algae were collected, cultured, and deployed in a spatial array around the outflow from a shrimp farm in Moorea, French Polynesia, to detect nitrogenous wastes. All three parameters were highest adjacent to the shrimp farm indicating nutrient enrichment, and delta(15)N values confirmed the shrimp farm as the dominant nutrient source (5.63-5.96 per thousand). Isotope ratios proved the most sensitive indicator, as delta(15)N signatures were detected at the most distant sites tested, confirming their usefulness in tracing nutrients and mapping the spatial extent of enrichment.

  9. Precision and sensitivity of the measurement of 15N enrichment in D-alanine from bacterial cell walls using positive/negative ion mass spectrometry

    NASA Technical Reports Server (NTRS)

    Tunlid, A.; Odham, G.; Findlay, R. H.; White, D. C.

    1985-01-01

    Sensitive detection of cellular components from specific groups of microbes can be utilized as 'signatures' in the examination of microbial consortia from soils, sediments or biofilms. Utilizing capillary gas chromatography/mass spectrometry and stereospecific derivatizing agents, D-alanine, a component localized in the prokaryotic (bacterial) cell wall, can be detected reproducibly. Enrichments of D-[15N]alanine determined in E. coli grown with [15N]ammonia can be determined with precision at 1.0 atom%. Chemical ionization with methane gas and the detection of negative ions (M - HF)- and (M - F or M + H - HF)- formed from the heptafluorobutyryl D-2 butanol ester of D-alanine allowed as little as 8 pg (90 fmol) to be detected reproducibly. This method can be utilized to define the metabolic activity in terms of 15N incorporation at the level of 10(3)-10(4) cells, as a function of the 15N-14N ratio.

  10. Plant community change mediates the response of foliar delta15N to CO2 enrichment in mesic grasslands

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rising atmospheric CO2 concentration may change the isotopic signature of plant N by altering plant and microbial processes involved in the N cycle. Isotope fractionation theory and limited experimental evidence indicate that CO2 may increase leaf delta15N by increasing plant community productivity,...

  11. N,N-Di- n-octyl- N,N-dimethyl and N,N-di- n-nonyl- N,N-dimethyl ammonium cholates: 13C and 15N CPMAS NMR, powder X-ray diffraction and thermoanalytical characterization

    NASA Astrophysics Data System (ADS)

    Kolehmainen, Erkki; Lahtinen, Manu; Valkonen, Arto; Behera, Babita; Kauppinen, Reijo

    2009-07-01

    N,N-Di- n-octyl- N,N-dimethyl cholate ( 1) and N,N-di- n-nonyl- N,N-dimethyl ammonium cholate ( 2) have been prepared by crystallization from equimolar mixtures of sodium cholate and quaternary N,N-di- n-alkyl- N,N-dimethyl ( n-octyl or n-nonyl) ammonium bromides. The formed crystalline materials have been structurally characterized by 13C and 15N cross polarization magic angle spinning (CPMAS) NMR, powder X-ray diffraction (PXRD) and thermoanalytical (TGA/DTA and DSC) methods and compared with each other. Powder X-ray diffraction patterns of 1 and 2 reveal clear similarities. Combined with the thermoanalytical data of these structures an existence of two hydrated polymorphs (most probably mono- and dihydrates) can be proposed. This presumption is further supported by 13C CPMAS NMR showing clearly double resonances for the carboxylic and majority of other carbons in these quaternary ammonium cholates. Owing to the endogenous character of cholate anion these ionic structures possess great potential in many pharmaceutical applications such as controlled drug delivery.

  12. Extreme 15N-enrichments in 2.72-Gyr-old sediments: evidence for a turning point in the nitrogen cycle.

    PubMed

    Thomazo, C; Ader, M; Philippot, P

    2011-03-01

    Although nitrogen is a key element in organic molecules such as nucleic acids and proteins, the timing of the emergence of its modern biogeochemical cycle is poorly known. Recent studies on the antiquity of the nitrogen cycle and its interaction with free oxygen suggests the establishment of a complete aerobic N biogeochemical cycle with nitrification, denitrification, and nitrogen fixation at about 2.68 Gyr. Here, we report new bulk nitrogen isotope data for the 2.72 billion-year-old sedimentary succession of the Tumbiana Formation (Pilbara Craton, Western Australia). The nitrogen isotopic compositions vary widely from +8.6‰ up to +50.4‰ and are inversely correlated with the very low δ(13)C values of associated organic matter defining the Fortescue excursion (down to about -56‰). We propose that this (15)N-enrichment records the onset of nitrification coupled to the continuous removal of its derivatives (nitrite and nitrate) by denitrification. This finding implies an increase in the availability of electron acceptors and probably oxygen in the Tumbiana depositional environment, 300 million years before the oxygenation of the Earth's atmosphere.

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

    PubMed

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

    2013-03-01

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

  14. Stable isotope probing with 15N achieved by disentangling the effects of genome G+C content and isotope enrichment on DNA density.

    PubMed

    Buckley, Daniel H; Huangyutitham, Varisa; Hsu, Shi-Fang; Nelson, Tyrrell A

    2007-05-01

    Stable isotope probing (SIP) of nucleic acids is a powerful tool that can identify the functional capabilities of noncultivated microorganisms as they occur in microbial communities. While it has been suggested previously that nucleic acid SIP can be performed with 15N, nearly all applications of this technique to date have used 13C. Successful application of SIP using 15N-DNA (15N-DNA-SIP) has been limited, because the maximum shift in buoyant density that can be achieved in CsCl gradients is approximately 0.016 g ml-1 for 15N-labeled DNA, relative to 0.036 g ml-1 for 13C-labeled DNA. In contrast, variation in genome G+C content between microorganisms can result in DNA samples that vary in buoyant density by as much as 0.05 g ml-1. Thus, natural variation in genome G+C content in complex communities prevents the effective separation of 15N-labeled DNA from unlabeled DNA. We describe a method which disentangles the effects of isotope incorporation and genome G+C content on DNA buoyant density and makes it possible to isolate 15N-labeled DNA from heterogeneous mixtures of DNA. This method relies on recovery of "heavy" DNA from primary CsCl density gradients followed by purification of 15N-labeled DNA from unlabeled high-G+C-content DNA in secondary CsCl density gradients containing bis-benzimide. This technique, by providing a means to enhance separation of isotopically labeled DNA from unlabeled DNA, makes it possible to use 15N-labeled compounds effectively in DNA-SIP experiments and also will be effective for removing unlabeled DNA from isotopically labeled DNA in 13C-DNA-SIP applications.

  15. Stable Isotope Probing with 15N Achieved by Disentangling the Effects of Genome G+C Content and Isotope Enrichment on DNA Density▿ †

    PubMed Central

    Buckley, Daniel H.; Huangyutitham, Varisa; Hsu, Shi-Fang; Nelson, Tyrrell A.

    2007-01-01

    Stable isotope probing (SIP) of nucleic acids is a powerful tool that can identify the functional capabilities of noncultivated microorganisms as they occur in microbial communities. While it has been suggested previously that nucleic acid SIP can be performed with 15N, nearly all applications of this technique to date have used 13C. Successful application of SIP using 15N-DNA (15N-DNA-SIP) has been limited, because the maximum shift in buoyant density that can be achieved in CsCl gradients is approximately 0.016 g ml−1 for 15N-labeled DNA, relative to 0.036 g ml−1 for 13C-labeled DNA. In contrast, variation in genome G+C content between microorganisms can result in DNA samples that vary in buoyant density by as much as 0.05 g ml−1. Thus, natural variation in genome G+C content in complex communities prevents the effective separation of 15N-labeled DNA from unlabeled DNA. We describe a method which disentangles the effects of isotope incorporation and genome G+C content on DNA buoyant density and makes it possible to isolate 15N-labeled DNA from heterogeneous mixtures of DNA. This method relies on recovery of “heavy” DNA from primary CsCl density gradients followed by purification of 15N-labeled DNA from unlabeled high-G+C-content DNA in secondary CsCl density gradients containing bis-benzimide. This technique, by providing a means to enhance separation of isotopically labeled DNA from unlabeled DNA, makes it possible to use 15N-labeled compounds effectively in DNA-SIP experiments and also will be effective for removing unlabeled DNA from isotopically labeled DNA in 13C-DNA-SIP applications. PMID:17369331

  16. Differential growth of the fungus Absidia cylindrospora on 13C/15N-labelled media.

    PubMed

    Crotty, F V; Blackshaw, R P; Murray, P J

    2011-06-15

    Many studies utilise enrichment of stable isotopes as tracers to follow the interactions occurring within soil food webs and methods have been developed to enrich bacteria, soil fauna and plant litter, Here for the first time we attempt to enrich a soil fungus to 99 atom% with (13)C and (15)N stable isotopes. In this study our objectives were to (a) assess whether the saprotrophic zygomycete fungus Absidia cylindrospora could grow on a medium enriched to 99 atom% with (13)C-glucose and (15)N-ammonium chloride, (b) to determine the level of enrichment obtained, and (c) to examine the change in growth rate of this fungus while it was growing on the dually enriched medium. To achieve this, the fungus was grown on agar enriched with (13)C and (15)N to 99 atom% and its growth rate monitored. The results showed that A. cylindrospora would grow on the highly labelled growth medium, but that its rate of growth was affected compared with the rate on either natural abundance media or media highly enriched with a single isotope ((13)C or (15)N). The implications of these results is that although the fungus is able to utilise these heavier isotopes, the biochemical processes involved in growth are affected, and consideration should be given to these differences when using stable isotope tracers in, for example, soil food web studies.

  17. Biosynthetic uniform 13C,15N-labelling of zervamicin IIB. Complete 13C and 15N NMR assignment.

    PubMed

    Ovchinnikova, Tatyana V; Shenkarev, Zakhar O; Yakimenko, Zoya A; Svishcheva, Natalia V; Tagaev, Andrey A; Skladnev, Dmitry A; Arseniev, Alexander S

    2003-01-01

    Zervamicin IIB is a member of the alpha-aminoisobutyric acid containing peptaibol antibiotics. A new procedure for the biosynthetic preparation of the uniformly 13C- and 15N-enriched peptaibol is described This compound was isolated from the biomass of the fungus-producer Emericellopsis salmosynnemata strain 336 IMI 58330 obtained upon cultivation in the totally 13C, 15N-labelled complete medium. To prepare such a medium the autolysed biomass and the exopolysaccharides of the obligate methylotrophic bacterium Methylobacillus flagellatus KT were used. This microorganism was grown in totally 13C, 15N-labelled minimal medium containing 13C-methanol and 15N-ammonium chloride as the only carbon and nitrogen sources. Preliminary NMR spectroscopic analysis indicated a high extent of isotope incorporation (> 90%) and led to the complete 13C- and 15N-NMR assignment including the stereospecific assignment of Aib residues methyl groups. The observed pattern of the structurally important secondary chemical shifts of 1H(alpha), 13C=O and 13C(alpha) agrees well with the previously determined structure of zervamicin IIB in methanol solution. PMID:14658801

  18. Coupling between anammox and autotrophic denitrification for simultaneous removal of ammonium and sulfide by enriched marine sediments.

    PubMed

    Rios-Del Toro, E Emilia; Cervantes, Francisco J

    2016-06-01

    In the present study, the capacity of enrichments derived from marine sediments collected from different sites of the Mexican littoral to perform anaerobic ammonium oxidation (anammox) coupled to sulfide-dependent denitrification for simultaneous removal of ammonium and sulfide linked to nitrite reduction was evaluated. Sulfide-dependent denitrification out-competed anammox during the simultaneous oxidation of sulfide and ammonium. Significant accumulation of elemental sulfur (ca. 14-30 % of added sulfide) occurred during the coupling between the two respiratory processes, while ammonium was partly oxidized (31-47 %) due to nitrite limitation imposed in sediment incubations. Nevertheless, mass balances revealed up to 38 % more oxidation of the electron donors available (ammonium and sulfide) than that expected from stoichiometry. Recycling of nitrite, from nitrate produced through anammox, is proposed to contribute to extra oxidation of sulfide, while additional ammonium oxidation is suggested by sulfate-reducing anammox (SR-anammox). The complex interaction between nitrogenous and sulfurous compounds occurring through the concomitant presence of autotrophic denitrification, conventional anammox and SR-anammox may significantly drive the nitrogen and sulfur fluxes in marine environments. PMID:26994921

  19. Isotopically enriched ammonium shows high nitrogen transformation in the pile top zone of dairy manure compost

    NASA Astrophysics Data System (ADS)

    Maeda, Koki; Toyoda, Sakae; Yano, Midori; Hattori, Shohei; Fukasawa, Makoto; Nakajima, Keiichi; Yoshida, Naohiro

    2016-03-01

    Nitrogen isotope ratios (δ15N) of NH4+ in dairy manure compost piles with and without bulking agent (10 % w/w) were compared to understand the effects of the use of bulking agent on nitrogen conversion during manure composting. The δ15N-NH4+ values in each of three pile zones (top, side and core) were also compared. At the end of the process, piles with bulking agent showed significantly higher δ15N values (17.7 ± 1.3 ‰) than piles without bulking agent (11.8 ± 0.9 ‰), reflecting the significantly higher nitrogen conversion and NH3 loss in the former. The samples from the top zone, especially in the piles with bulking agent, showed very high NH4+ concentrations with significantly high 15N15N: 12.7-29.8 ‰) values, indicating that extremely high nitrogen conversion, nitrification-denitrification activity of the microbes and NH3 volatilization occurred in this zone.

  20. Biochar alleviates combined stress of ammonium and acids by firstly enriching Methanosaeta and then Methanosarcina.

    PubMed

    Lü, Fan; Luo, Chenghao; Shao, Liming; He, Pinjing

    2016-03-01

    This investigation evaluated the effectiveness of biochar of different particle sizes in alleviating ammonium (NH4(+)) inhibition (up to 7 g-N/L) during anaerobic digestion of 6 g/L glucose. Compared to the control treatment without biochar addition, treatments that included biochar particles 2-5 mm, 0.5-1 mm and 75-150 μm in size reduced the methanization lag phase by 23.9%, 23.8% and 5.9%, respectively, and increased the maximum methane production rate by 47.1%, 23.5% and 44.1%, respectively. These results confirmed that biochar accelerated the initiation of methanization during anaerobic digestion under double inhibition risk from both ammonium and acids. Furthermore, fine biochar significantly promoted the production of volatile fatty acids (VFAs). Comparative analysis on the archaeal and bacterial diversity at the early and later stages of digestion, and in the suspended, biochar loosely bound, and biochar tightly bound fractions suggested that, in suspended fractions, hydrogenotrophic Methanobacterium was actively resistant to ammonium. However, acetoclastic Methanosaeta can survive at VFAs concentrations up to 60-80 mmol-C/L by improved affinity to conductive biochar, resulting in the accelerated initiation of acetate degradation. Improved methanogenesis was followed by the colonization of the biochar tightly bound fractions by Methanosarcina. The selection of appropriate biochar particles sizes was important in facilitating the initial colonization of microbial cells.

  1. The 15N isotope to evaluate fertilizer nitrogen absorption efficiency by the coffee plant.

    PubMed

    Fenilli, Tatiele A B; Reichart, Klaus; Bacchi, Osny O S; Trivelin, Paulo C O; Dourado-Neto, Durval

    2007-12-01

    The use of the 15N label for agronomic research involving nitrogen (N) cycling and the fate of fertilizer-N is well established, however, in the case of long term experimentation with perennial crops like citrus, coffee and rubber tree, there are still shortcomings mainly due to large plant size, sampling procedures, detection levels and interferences on the system. This report tries to contribute methodologically to the design and development of 15N labeled fertilizer experiments, using as an example a coffee crop fertilized with 15N labeled ammonium sulfate, which was followed for two years. The N of the plant derived from the fertilizer was studied in the different parts of the coffee plant in order to evaluate its distribution within the plant and the agronomic efficiency of the fertilizer application practice. An enrichment of the fertilizer-N of the order of 2% 15N abundance was sufficient to study N absorption rates and to establish fertilizer-N balances after one and two years of coffee cropping. The main source of errors in the estimated values lies in the inherent variability among field replicates and not in the measurements of N contents and 15N enrichments of plant material by mass-spectrometry. PMID:18066442

  2. The 15N isotope to evaluate fertilizer nitrogen absorption efficiency by the coffee plant.

    PubMed

    Fenilli, Tatiele A B; Reichart, Klaus; Bacchi, Osny O S; Trivelin, Paulo C O; Dourado-Neto, Durval

    2007-12-01

    The use of the 15N label for agronomic research involving nitrogen (N) cycling and the fate of fertilizer-N is well established, however, in the case of long term experimentation with perennial crops like citrus, coffee and rubber tree, there are still shortcomings mainly due to large plant size, sampling procedures, detection levels and interferences on the system. This report tries to contribute methodologically to the design and development of 15N labeled fertilizer experiments, using as an example a coffee crop fertilized with 15N labeled ammonium sulfate, which was followed for two years. The N of the plant derived from the fertilizer was studied in the different parts of the coffee plant in order to evaluate its distribution within the plant and the agronomic efficiency of the fertilizer application practice. An enrichment of the fertilizer-N of the order of 2% 15N abundance was sufficient to study N absorption rates and to establish fertilizer-N balances after one and two years of coffee cropping. The main source of errors in the estimated values lies in the inherent variability among field replicates and not in the measurements of N contents and 15N enrichments of plant material by mass-spectrometry.

  3. Nitrification-driven forms of nitrogen metabolism in microbial mat communities thriving along an ammonium-enriched subsurface geothermal stream

    NASA Astrophysics Data System (ADS)

    Nishizawa, Manabu; Koba, Keisuke; Makabe, Akiko; Yoshida, Naohiro; Kaneko, Masanori; Hirao, Shingo; Ishibashi, Jun-ichiro; Yamanaka, Toshiro; Shibuya, Takazo; Kikuchi, Tohru; Hirai, Miho; Miyazaki, Junichi; Nunoura, Takuro; Takai, Ken

    2013-07-01

    ) was nearly constant (250 μM) throughout the stream. Based on the level of detectable dissolved molecular oxygen (O2) of the stream water (⩾38 μM) along with metabolic measurements, it was predicted that nitrification by thermophilic AOA and NOB components in the microbial mats that were exposed to the stream water would constrain the concentrations and isotopic ratios of ΣNH3, NO2- and NO3- of the stream water. The δ15N value of ΣNH3 increased from 0‰ to 7‰ with decreasing concentration, which was consistent with the previously reported isotopic fractionation for microbial ΣNH3 oxidation. In contrast, the δ15N value of NO2- was 22‰ lighter than that of NO3- in the steam water at the same site, indicating an inverse isotopic fractionation for microbial NO2- oxidation. The variation in concentrations and δ15N values of ΣNH3, NO2- and NO3- was largely explained using a two-step nitrification model, and the apparent nitrogen isotopic fractionations of ΣNH3 oxidation and NO2- oxidation were estimated to be 0.986 and 1.020, respectively. In the interstitial water within the microbial mats, the compositional and isotopic properties of TIN at the downstream site indicated potential denitrification by the anaerobic microbial components. The geochemically deduced transition of microbial nitrogen metabolism was substantiated through cultivation-independent microbiological analyses.

  4. Ammonium and phosphate enrichment across the dry-wet transition and their ecological relevance in a subtropical reservoir, China.

    PubMed

    Mo, Qiongli; Chen, Nengwang; Zhou, Xingpeng; Chen, Jixin; Duan, Shuiwang

    2016-07-13

    Small river reservoirs are widespread and can be ecologically sensitive across the dry-wet transition under monsoon climate with respect to nutrient loading and phenology. Monthly sampling and high-frequency in situ measurements were conducted for a river reservoir (southeast China) in 2013-2014 to examine the seasonal pattern of nutrients and phytoplankton. We found that nutrient concentrations were runoff-mediated and determined by watershed inputs and, in some cases, by internal cycling depending on hydrology and temperature. Ammonium and phosphate were relatively enriched in February-March (a transitional period from dry/cold to wet/hot climate), which can be ascribed to initial flushing runoff from human/animal waste and spring fertilizer use. A phytoplankton bloom (mainly Chlorophyta) occurred during April after a surge of water temperature, probably due to the higher availability of inorganic nutrients and sunlight and suitable hydraulic residence time (medium flow) in the transitional period. The concentration of phytoplankton was low during May-June (wet-hot climate) when the concentrations of total suspended matter (TSM) were highest, likely owing to the "shading" effect of TSM and turbulence of high flow conditions. Nutrient-algae shifts across the dry-wet season and vertical profiles suggested that algal blooms seem to be fueled primarily by phosphate and ammonium rather than nitrate. Current findings of a strong temporal pattern and the relationship between physical parameters, nutrient and biota would improve our understanding of drivers of change in water quality and ecosystem functions with dam construction. PMID:27329744

  5. Ammonium and phosphate enrichment across the dry-wet transition and their ecological relevance in a subtropical reservoir, China.

    PubMed

    Mo, Qiongli; Chen, Nengwang; Zhou, Xingpeng; Chen, Jixin; Duan, Shuiwang

    2016-07-13

    Small river reservoirs are widespread and can be ecologically sensitive across the dry-wet transition under monsoon climate with respect to nutrient loading and phenology. Monthly sampling and high-frequency in situ measurements were conducted for a river reservoir (southeast China) in 2013-2014 to examine the seasonal pattern of nutrients and phytoplankton. We found that nutrient concentrations were runoff-mediated and determined by watershed inputs and, in some cases, by internal cycling depending on hydrology and temperature. Ammonium and phosphate were relatively enriched in February-March (a transitional period from dry/cold to wet/hot climate), which can be ascribed to initial flushing runoff from human/animal waste and spring fertilizer use. A phytoplankton bloom (mainly Chlorophyta) occurred during April after a surge of water temperature, probably due to the higher availability of inorganic nutrients and sunlight and suitable hydraulic residence time (medium flow) in the transitional period. The concentration of phytoplankton was low during May-June (wet-hot climate) when the concentrations of total suspended matter (TSM) were highest, likely owing to the "shading" effect of TSM and turbulence of high flow conditions. Nutrient-algae shifts across the dry-wet season and vertical profiles suggested that algal blooms seem to be fueled primarily by phosphate and ammonium rather than nitrate. Current findings of a strong temporal pattern and the relationship between physical parameters, nutrient and biota would improve our understanding of drivers of change in water quality and ecosystem functions with dam construction.

  6. Nitrogen concentration and δ 15N of altered oceanic crust obtained on ODP Legs 129 and 185: Insights into alteration-related nitrogen enrichment and the nitrogen subduction budget

    NASA Astrophysics Data System (ADS)

    Li, Long; Bebout, Gray E.; Idleman, Bruce D.

    2007-05-01

    Knowledge of the subduction input flux of nitrogen (N) in altered oceanic crust (AOC) is critical in any attempt to mass-balance N across arc-trench systems on a global or individual-margin basis. We have employed sealed-tube, carrier-gas-based methods to examine the N concentrations and isotopic compositions of AOC. Analyses of 53 AOC samples recovered on DSDP/ODP legs from the North and South Pacific, the North Atlantic, and the Antarctic oceans (with larger numbers of samples from Site 801 outboard of the Mariana trench and Site 1149 outboard of the Izu trench), and 14 composites for the AOC sections at Site 801, give N concentrations of 1.3 to 18.2 ppm and δ 15N Air of -11.6‰ to +8.3‰, indicating significant N enrichment probably during the early stages of hydrothermal alteration of the oceanic basalts. The N-δ 15N modeling for samples from Sites 801 and 1149 ( n = 39) shows that the secondary N may come from (1) the sedimentary N in the intercalated sediments and possibly overlying sediments via fluid-sediment/rock interaction, and (2) degassed mantle N 2 in seawater via alteration-related abiotic reduction processes. For all Site 801 samples, weak correlation of N and K 2O contents indicates that the siting of N in potassic alteration phases strongly depends on N availability and is possibly influenced by highly heterogeneous temperature and redox conditions during hydrothermal alteration. The upper 470-m AOC recovered by ODP Legs 129 and 185 delivers approximately 8 × 10 5 g/km N annually into the Mariana margin. If the remaining less-altered oceanic crust (assuming 6.5 km, mostly dikes and gabbros) has MORB-like N of 1.5 ppm, the entire oceanic crust transfers 5.1 × 10 6 g/km N annually into that trench. This N input flux is twice as large as the annual N input of 2.5 × 10 6 g/km in seafloor sediments subducting into the same margin, demonstrating that the N input in oceanic crust, and its isotopic consequences, must be considered in any assessment

  7. The response of nitrifying microbial assemblages to ammonium (NH4+) enrichment from salmon farm activities in a northern Chilean Fjord

    NASA Astrophysics Data System (ADS)

    Elizondo-Patrone, Claudia; Hernández, Klaudia; Yannicelli, Beatriz; Olsen, Lasse Mork; Molina, Verónica

    2015-12-01

    The consequences of aquaculture include alterations in nitrogen cycling in aquatic environments that may lead to ecosystem degradation. Herein salmon aquaculture release of ammonium (NH4+) to the water column and its effects on natural archaea and bacteria ammonia-oxidizers (AOA and AOB) and nitrite-oxidizing bacteria (NOB) community structure were studied in the Comau fjord using molecular approaches, such as: cloning (AOA and AOB richness), qPCR for C. Nitrosopumilus maritimus (AOA) and Nitrospina sp. (NOB) abundance (DNA) and RT-qPCR only for Nitrospina sp activity (RNA). Sampling was carried out in brackish (0.7-25 salinity, <5 m depth) and marine (>30 salinity, 25 m depth) waters during contrasting salmon production periods: rest (winter 2012), growth and harvest (summer and winter 2013). During the rest period, the highest NH4+ concentration was observed at Vodudahue River, whereas during productive periods NH4+ accumulated in the brackish layer inside salmon cages and in the vicinty (up to 700 m distance from the cages). The nitrifier community from the fjord reference station (Stn-C) was characterized by C. N. maritimus (AOA) and Nitrosomonas sp. (AOB) sequences affiliated with cosmopolitan ecotypes (e.g., marine, freshwater, hydrothermal), maxima abundances of C. N. maritimus (AOA) and Nitrospina sp. and extreme ranges of Nitrospina sp. activity occurred in the brackish layer. During productive periods, abundances of C. N. maritimus were co-varied with NH4+ concentrations inside salmon cages (summer) and the adjacent areas (winter). Productive periods were characterized by lower abundances but more homogeneity between brackish and marine areas than for the Stn-C nitrifiers. The physiological state of Nitrospina sp. estimated from cDNA:DNA ratios indicated higher growth during winter 2013 associated with NH4+ enrichment derived from production and river input. Our results suggest that in Comau Fjord, NH4+ enrichment events occur during salmon production and

  8. Characterization of incubation experiments and development of an enrichment culture capable of ammonium oxidation under iron-reducing conditions

    NASA Astrophysics Data System (ADS)

    Huang, S.; Jaffé, P. R.

    2015-02-01

    Incubation experiments were conducted using soil samples from a forested riparian wetland where we have previously observed anaerobic ammonium oxidation coupled to iron reduction. Production of both nitrite and ferrous iron was measured repeatedly during incubations when the soil slurry was supplied with either ferrihydrite or goethite and ammonium chloride. Significant changes in the microbial community were observed after 180 days of incubation as well as in a continuous flow membrane reactor, using 16S rRNA gene PCR-denaturing gradient gel electrophoresis, 454 pyrosequencing, and real-time quantitative PCR analysis. We be Acidimicrobiaceae bacterium A6), belonging to the Acidimicrobiaceae family, whose closest cultivated relative is Ferrimicrobium acidiphilum (with 92% identity) and Acidimicrobium ferrooxidans (with 90% identity), might play a key role in this anaerobic biological process that uses ferric iron as an electron acceptor while oxidizing ammonium to nitrite. After ammonium was oxidized to nitrite, nitrogen loss proceeded via denitrification and/or anammox.

  9. Ammonium assmilation in spruce ectomycorrhizas

    SciTech Connect

    Chalot, M.; Brun, A.; Botton, B. ); Stewart, G. )

    1990-05-01

    Assimilation of labelled NH{sub 4}{sup +} into amino acids has been followed in ectomycorrhizal roots of spruce. Over an 18 h period of NH{sub 4}{sup +} feeding, Gln, Glu and Ala became the most abundant amino acids. Gln was also the most highly labelled amino acid during the experiment, followed by Glu and Ala. This result indicates that Gln synthesis is an important ammonium utilization reaction in spruce mycorrhizas. Addition of MSX to NH{sub 4}{sup +} fed mycorrhizas caused an inhibition of Gln accumulation with a corresponding increase in Glu, Ala and Asn levels. The supply of MSX induced a sharp diminution of {sup 15}N enrichment in both amino and amido groups of glutamine. In contrast, the {sup 15}N incorporation into Glu and derivatives (Ala and Asp) remained very high. This study demonstrates that the fungal glutamate dehydrogenase is quite operative in spruce ectomycorrhizas since it is able to sustain ammonium assimilation when glutamine synthetase is inhibited.

  10. Characterization of incubation experiments and development of an enrichment culture capable of ammonium oxidation under iron reducing conditions

    NASA Astrophysics Data System (ADS)

    Huang, S.; Jaffé, P. R.

    2014-08-01

    Incubation experiments were conducted using soil samples from a forested riparian wetland where we have previously observed anaerobic ammonium oxidation coupled to iron reduction. Production of both nitrite and ferrous iron were measured repeatedly during incubations when the soil slurry was supplied with either ferrihydrite or goethite and ammonium chloride. Significant changes in the microbial community were observed after 180 days of incubation as well as in a continuous flow membrane reactor, using 16S rRNA gene PCR-denaturing gradient gel electrophoresis, 454-pyrosequencing, and real-time quantitative PCR analysis. We believe that one of the dominant microbial species in our system (an uncultured Acidimicrobiaceae bacterium A6), belonging to the Acidimicrobiaceae family, whose closest cultivated relative is Ferrimicrobium acidiphilum (with 92% identity) and Acidimicrobium ferrooxidans (with 90% identity), might play a key role in this anaerobic biological process that uses ferric iron as an electron acceptor while oxidizing ammonium to nitrite. After ammonium was oxidized to nitrite, nitrogen loss proceeded via denitrification and/or anammox.

  11. Use of a 15N tracer to determine linkages between a mangrove and an upland freshwater swamp

    NASA Astrophysics Data System (ADS)

    MacKenzie, R. A.; Cormier, N.

    2005-05-01

    Mangrove forests and adjacent upland freshwater swamps are important components of subsistence-based economies of Pacific islands. Mangroves provide valuable firewood (Rhizophora apiculata) and mangrove crabs (Scylla serrata); intact freshwater swamps are often used for agroforestry (e.g., taro cultivation). While these two systems are connected hydrologically via groundwater and surface flows, little information is available on how they may be biogeochemically or ecologically linked. For example, mangrove leaf litter was once thought to be an important food source for resident and transient nekton and invertebrates, but this value may have been overestimated. Instead, nutrients or allochthonous material (e.g., phytoplankton, detritus) delivered via groundwater or surface water from upland freshwater swamps may play a larger role in mangrove food webs. Understanding the linkages between these two ecologically and culturally important ecosystems will help us to understand the potential impacts of hydrological alterations that occur when roads or bridges are constructed through them. We conducted a 15N tracer study in the Yela watershed on the island of Kosrae, Federated States of Micronesia. K15NO3 was continually added at trace levels for 4 weeks to the Yela River in an upland freshwater swamp adjacent to a mangrove forest. Nitrate and ammonium pools, major primary producers, macroinvertebrates, and fish were sampled from stations 5 m upstream (freshwater swamp) and 138, 188, 213, and 313 m downstream (mangrove) from the tracer addition. Samples were collected once a week prior to, during, and after the 15N addition for a total of 6 weeks. Preliminary results revealed no significant enrichment (< 1 ‰) in the 15N isotope composition of either resident shrimp (Macrobrachium sp.) or mudskipper fish (Periophthalmus sp.). However, the 15N signature of ammonium pools was enriched 10-60 ‰ by the end of the third week. These results suggest that the tracer was present

  12. Nitrogen isotope fractionation during archaeal ammonia oxidation: Coupled estimates from isotopic measurements of ammonium and nitrite

    NASA Astrophysics Data System (ADS)

    Mooshammer, Maria; Stieglmeier, Michaela; Bayer, Barbara; Jochum, Lara; Melcher, Michael; Wanek, Wolfgang

    2014-05-01

    Ammonia-oxidizing archaea (AOA) are ubiquitous in marine and terrestrial environments and knowledge about the nitrogen (N) isotope effect associated with their ammonia oxidation activity will allow a better understanding of natural abundance isotope ratios, and therefore N transformation processes, in the environment. Here we examine the kinetic isotope effect for ammonia oxidation in a pure soil AOA culture (Ca. Nitrososphaera viennensis) and a marine AOA enrichment culture. We estimated the isotope effect from both isotopic signatures of ammonium and nitrite over the course of ammonia oxidation. Estimates of the isotope effect based on the change in the isotopic signature of ammonium give valuable insight, because these estimates are not subject to the same concerns (e.g., accumulation of an intermediate) as estimates based on isotopic measurements of nitrite. Our results show that both the pure soil AOA culture and a marine AOA enrichment culture have similar but substantial isotope effect during ammonia consumption (31-34 per mill; based on ammonium) and nitrite production (43-45 per mill; based on nitrite). The 15N fractionation factors of both cultures tested fell in the upper range of the reported isotope effects for archaeal and bacterial ammonia oxidation (10-41 per mill) or were even higher than those. The isotope fractionation for nitrite production was significantly larger than for ammonium consumption, indicating that (1) some intermediate (e.g., hydroxylamine) of ammonia oxidation accumulates, allowing for a second 15N fractionation step to be expressed, (2) a fraction of ammonia oxidized is lost via gaseous N forms (e.g., NO or N2O), which is 15N-enriched or (3) a fraction of ammonium is assimilated into AOA biomass, biomass becoming 15N-enriched. The significance of these mechanisms will be explored in more detail for the soil AOA culture, based on isotope modeling and isotopic measurements of biomass and N2O.

  13. Comparison of nitrogen removal rates and nitrous oxide production from enriched anaerobic ammonium oxidizing bacteria in suspended and attached growth reactors.

    PubMed

    Panwivia, Supaporn; Sirvithayapakorn, Sanya; Wantawin, Chalermraj; Noophan, Pongsak Lek; Munakata-Marr, Junko

    2014-01-01

    Attached growth-systems for the anaerobic ammonium oxidation (anammox) process have been postulated for implementation in the field. However, information about the anammox process in attached growth-systems is limited. This study compared nitrogen removal rates and nitrous oxide (N2O) production of enriched anammox cultures in both suspended and attached growth sequencing batch reactors (SBRs). Suspended growth reactors (SBR-S) and attached growth reactors using polystyrene sponge as a medium (SBR-A) were used in these experiments. After inoculation with an enriched anammox culture, significant nitrogen removals of ammonium (NH4 (+)) and nitrite (NO2 (-)) were observed under NH4 (+):NO2 (-) ratios ranging from 1:1 to 1:2 in both types of SBRs. The specific rates of total nitrogen removal in SBR-S and SBR-A were 0.52 mg N/mg VSS-d and 0.44 mg N/mg VSS-d, respectively, at an NH4 (+):NO2 (-) ratio of 1:2. N2O production by the enriched anammox culture in both SBR-S and SBR-A was significantly higher at NH4 (+):NO2 (-) ratio of 1:2 than at NH4 (+):NO2 (-) ratios of 1:1 and 1:1.32. In addition, N2O production was higher at a pH of 6.8 than at pH 7.3, 7.8, and 8.3 in both SBR-S and SBR-A. The results of this investigation demonstrate that the anammox process may avoid N2O emission by maintaining an NH4 (+):NO2 (-) ratio of less than 1:2 and pH higher than 6.8.

  14. Nitrogen input 15N-signatures are reflected in plant 15N natural abundances of N-rich tropical forest in China

    NASA Astrophysics Data System (ADS)

    Abdisa Gurmesa, Geshere; Lu, Xiankai; Gundersen, Per; Yunting, Fang; Mo, Jiangming

    2016-04-01

    In this study, we tested the measurement of natural abundance of 15N15N) for its ability to assess changes in N cycling due to increased N deposition in two forest types; namely, an old-growth broadleaved forest and a pine forest, in southern China. We measured δ15N values of inorganic N in input and output fluxes under ambient N deposition, and N concentration and δ15N of major ecosystem compartments under ambient and increased N deposition. Our results showed that N deposition to the forests was 15N-depleted, and was dominated by NH4-N. Plants were 15N-depleted due to imprint from the 15N-depleted atmospheric N deposition. The old-growth forest had larger N concentration and was more 15N-enriched than the pine forest. Nitrogen addition did not significantly affect N concentration, but it significantly increased δ15N values of plants, and slightly more so in the pine forest, toward the 15N signature of the added N in both forests. The result indicates that the pine forest may rely more on the 15N-depleted deposition N. Soil δ15N values were slightly decreased by the N addition. Our result suggests that ecosystem δ15N is more sensitive to the changes in ecosystem N status and N cycling than N concentration in N-saturated sub-tropical forests.

  15. Robust method for investigating nitrogen metabolism of 15N labeled amino acids using AccQ•Tag ultra performance liquid chromatography-photodiode array-electrospray ionization-mass spectrometry: application to a parasitic plant-plant interaction.

    PubMed

    Gaudin, Zachary; Cerveau, Delphine; Marnet, Nathalie; Bouchereau, Alain; Delavault, Philippe; Simier, Philippe; Pouvreau, Jean-Bernard

    2014-01-21

    An AccQ•Tag ultra performance liquid chromatography-photodiode array-electrospray ionization-mass spectrometry (AccQ•Tag-UPLC-PDA-ESI-MS) method is presented here for the fast, robust, and sensitive quantification of (15)N isotopologue enrichment of amino acids in biological samples, as for example in the special biotic interaction between the cultivated specie Brassica napus (rapeseed) and the parasitic weed Phelipanche ramosa (broomrape). This method was developed and validated using amino acid standard solutions containing (15)N amino acid isotopologues and/or biological unlabeled extracts. Apparatus optimization, limits of detection and quantification, quantification reproducibility, and calculation method of (15)N isotopologue enrichment are presented. Using this method, we could demonstrate that young parasite tubercles assimilate inorganic nitrogen as (15)N-ammonium when supplied directly through batch incubation but not when supplied by translocation from host root phloem, contrary to (15)N2-glutamine. (15)N2-glutamine mobility from host roots to parasite tubercles followed by its low metabolism in tubercles suggests that the host-derived glutamine acts as an important nitrogen containing storage compound in the young tubercle of Phelipanche ramosa. PMID:24359440

  16. Production of 15N-Labelled Liquid Organic Fertilisers Based on Manure and Crop Residue for Use in Fertigation Studies.

    PubMed

    Martínez-Alcántara, Belén; Martínez-Cuenca, Mary-Rus; Fernández, Carlos; Legaz, Francisco; Quiñones, Ana

    2016-01-01

    Large quantities of crop residue and animal manure from agricultural and livestock activities are annually produced worldwide. With proper management, these residues are potentially valuable sources of plant nutrients, mainly N. Recycling such subproducts in sustainably-based agricultural systems can minimise the use of mineral fertilisers, and hence reduce the potential risk of surface and groundwater pollution. Therefore, the purpose of this study was to obtain (small scale) two liquid labelled-organic fertilisers, an animal- and a vegetal-based organic (AO and VO, respectively) fertiliser, to be used as organic N sources in subsequent fertigation studies. Forage maize (Zea mays L.) grown under 15N-labelled fertiliser supply was used as raw material for VO fertiliser production, and also as 15N-labelled sheep feed to obtain 15N-labelled manure. The labelled faeces fraction was used as raw material for the AO fertiliser. The VO fertiliser was obtained after an acidic and an enzyme-driven hydrolysis. The AO fertiliser was obtained after acidic hydrolysis. The VO liquid fertiliser presented an N concentration of 330 mg·L-1, 85% of total N was organic, while ammonium and nitrate N accounted for 55% and 45% of the mineral nitrogen fraction, respectively. This fertiliser also exhibited high K, Ca and S concentrations and notable values for the remaining macro- and micronutrients. The AO liquid fertiliser had a similar total N concentration (496 mg·L-1, 82% of total N in an organic form) to that of VO, but its mineral N fraction significantly differed, which came in a predominantly (95%) ammonia form. It also had a high content of N, P, K and other macronutrients, and sufficient Fe, Zn, Mn, Cu and B levels, which suggests its suitability as a potential fertiliser. The percentage of 15N enrichment in both VO and AO liquid fertilisers exceeded 2% 15N atom excess, which enabled their use in subsequent assays run to assess nitrogen uptake efficiency.

  17. Phytoplankton-Specific Response to Enrichment of Phosphorus-Rich Surface Waters with Ammonium, Nitrate, and Urea

    PubMed Central

    Donald, Derek B.; Bogard, Matthew J.; Finlay, Kerri; Bunting, Lynda; Leavitt, Peter R.

    2013-01-01

    Supply of anthropogenic nitrogen (N) to the biosphere has tripled since 1960; however, little is known of how in situ response to N fertilisation differs among phytoplankton, whether species response varies with the chemical form of N, or how interpretation of N effects is influenced by the method of analysis (microscopy, pigment biomarkers). To address these issues, we conducted two 21-day in situ mesocosm (3140 L) experiments to quantify the species- and genus-specific responses of phytoplankton to fertilisation of P-rich lake waters with ammonium (NH4+), nitrate (NO3−), and urea ([NH2]2CO). Phytoplankton abundance was estimated using both microscopic enumeration of cell densities and high performance liquid chromatographic (HPLC) analysis of algal pigments. We found that total algal biomass increased 200% and 350% following fertilisation with NO3− and chemically-reduced N (NH4+, urea), respectively, although 144 individual taxa exhibited distinctive responses to N, including compound-specific stimulation (Planktothrix agardhii and NH4+), increased biomass with chemically-reduced N alone (Scenedesmus spp., Coelastrum astroideum) and no response (Aphanizomenon flos-aquae, Ceratium hirundinella). Principle components analyses (PCA) captured 53.2–69.9% of variation in experimental assemblages irrespective of the degree of taxonomic resolution of analysis. PCA of species-level data revealed that congeneric taxa exhibited common responses to fertilisation regimes (e.g., Microcystis aeruginosa, M. flos-aquae, M. botrys), whereas genera within the same division had widely divergent responses to added N (e.g., Anabaena, Planktothrix, Microcystis). Least-squares regression analysis demonstrated that changes in phytoplankton biomass determined by microscopy were correlated significantly (p<0.005) with variations in HPLC-derived concentrations of biomarker pigments (r2 = 0.13–0.64) from all major algal groups, although HPLC tended to underestimate the relative

  18. Marking Drosophila suzukii (Diptera: Drosophilidae) With Rubidium or 15N.

    PubMed

    Klick, J; Yang, W Q; Bruck, D J

    2015-06-01

    Drosophila suzukii Matsumura (Diptera: Drosophilidae) has caused significant economic damage to berry and stone fruit production regions. Markers that are systemic in plants and easily transferred to target organisms are needed to track D. suzukii exploitation of host resources and trophic interactions. High and low concentrations of the trace element, rubidium (Rb), and the stable isotope, 15N, were tested to mark D. suzukii larvae feeding on fruits of enriched strawberry plants grown in containers under greenhouse conditions. Fly marker content and proportion of flies marked 1, 7, and 14 d after emergence from enriched fruits and fly dry mass were analyzed. Nearly 100% of the flies analyzed 14 d after emerging from 15N-enriched plants were marked, whereas only 30-75% and 0-3% were marked 14 d after emerging from high and low Rb concentration plants, respectively. Rapid Rb decay, strong 15N persistence, and the economics of using these markers in the field to elucidate D. suzukii pest ecology are discussed. PMID:26470275

  19. Ntrace a 15N tracing model to analyse gross N transformations and sources of gaseous N emissions

    NASA Astrophysics Data System (ADS)

    Muller, Christoph

    2013-04-01

    Anthropogenically generated reactive nitrogen (N) cascades throughout the global environment ...(Galloway and Cowling, 2002). This reactive N may be lost from ecosystems via leaching, as nitrate (NO3-), or in gaseous forms such as ammonia and nitrous oxide (N2O) and the loss is governed by the N dynamics of the system. Thus, to be rendered environmentally benign NO3- must be reduced to a non-reactive form, dinitrogen (N2) which requires the evaluation of three major biological pathways of NO3- reduction: i) assimilatory NO3- reduction into biomass, ii) dissimilatory NO3- reduction to NH4+ (DNRA) and iii) dissimilatory NO3- reduction to N2 (denitrification) ...(Burger and Jackson, 2004). Advanced techniques based on 15N tracing in combination with suitable model analyses are the method of choice to analyse complex N interactions and simultaneous N transformation process. Techniques are based on dilution - enrichment principles and usually rely on the simultaneous labelling of various N pools. The data sets are then analysed by suitable 15N tracing models which allow the individual N transformation rates to be calculated based on realistic kinetic settings. The 15N tracing model Ntrace has been developed to analyse the simultaneously occurring N transformation rates in soil-plant systems and includes submodels for the evaluation of the processes associated with gaseous N emissions. The 15N tracing model and some typical model results will be presented. Literature cited .Burger, M., and Jackson, L. E. (2004). Plant and microbial use and turnover: rapid conversion of nitrate to ammonium in soil with roots. Plant and Soil 266, 289-301. Galloway, J. N., and Cowling, E. B. (2002). Reactive nitrogen and the world: 200 years of change. Ambio 31, 64-71. .

  20. Refining cotton-wick method for 15N plant labelling.

    NASA Astrophysics Data System (ADS)

    Fustec, Joëlle; Mahieu, Stéphanie

    2010-05-01

    The symbiosis Fabaceae/Rhizobiaceae plays a critical role in the nitrogen cycle. It gives the plant the ability to fix high amounts of atmospheric N. A part of this N can be transferred to the soil via rhizodeposition. The contribution of Fabaceae to the soil N pool is difficult to measure, since it is necessary for assessing N benefits for other crops, for soil biological activity, and for reducing water pollution in sustainable agriculture (Fustec, 2009). The aim of this study was to test and improve the reliability of the 15N cotton-wick method for measuring the soil N derived from plant rhizodeposition (Mahieu et al., 2007). The effects of the concentration of the 15N-urea labelling solution and of the feeding frequency (continuous or pulses) on the assessment of nitrogen rhizodeposition were studied in two greenhouse experiments using the field pea (Pisum sativum L.) and the non-nodulating isoline P2. The plant parts and the soil were prepared for 15N:14N measurements for assessing N rhizodeposition (Mahieu et al., 2009). The fraction of plants' belowground nitrogen allocated to rhizodeposition in both Frisson pea and P2 was 20 to more than 50% higher when plants were labelled continuously than when they were labelled using fortnightly pulses. Our results suggested that when 15N root enrichment was high, nitrogen rhizodeposition was underestimated only for plants that were 15N-fed by fortnightly pulses, and not in plants 15N-fed continuously. This phenomenon was especially observed for plants relying on symbiotic N fixation for N acquisition; it may be linked to the concentration of the labelling solution. In conclusion, N rhizodeposition assessment was strongly influenced by the 15N-feeding frequency and the concentration of the labelling solution. The estimation of N rhizodeposition was more reliable when plants were labelled continuously with a dilute solution of 15N urea. Fustec et al. 2009. Agron. Sustain. Dev., DOI 10.1051/agro/2009003, in press. Mahieu

  1. The effect of manuring on cereal and pulse amino acid δ(15)N values.

    PubMed

    Styring, Amy K; Fraser, Rebecca A; Bogaard, Amy; Evershed, Richard P

    2014-06-01

    Amino acid δ(15)N values of barley (Hordeum vulgare) and bread wheat (Triticum aestivum) grains and rachis and broad bean (Vicia faba) and pea (Pisum sativum) seeds, grown in manured and unmanured soil at the experimental farm stations of Rothamsted, UK and Bad Lauchstädt, Germany, were determined by GC-C-IRMS. Manuring was found to result in a consistent (15)N-enrichment of cereal grain amino acid δ(15)N values, indicating that manuring did not affect the metabolic routing of nitrogen (N) into cereal grain amino acids. The increase in cereal grain δ(15)N values with manuring is therefore due to a (15)N-enrichment in the δ(15)N value of assimilated inorganic-N. Greater variation was observed in the (15)N-enrichment of rachis amino acids with manuring, possibly due to enhanced sensitivity to changes in growing conditions and higher turnover of N in rachis cells compared to cereal grains. Total amino acid δ(15)N values of manured and unmanured broad beans and peas were very similar, indicating that the legumes assimilated N2 from the atmosphere rather than N from the soil, since there was no evidence for routing of (15)N-enriched manure N into any of the pulse amino acids. Crop amino acid δ(15)N values thus provide insights into the sources of N assimilated by non N2-fixing and N2-fixing crops grown on manured and unmanured soils, and reveal an effect of manure on N metabolism in different crop species and plant parts.

  2. Food web structure in two counter-rotating eddies based on δ15N and δ13C isotopic analyses

    NASA Astrophysics Data System (ADS)

    Waite, A. M.; Muhling, B. A.; Holl, C. M.; Beckley, L. E.; Montoya, J. P.; Strzelecki, J.; Thompson, P. A.; Pesant, S.

    2007-04-01

    We measured the natural inventories of nitrogen and carbon stable isotopes within various ecosystem fractions of two counter-rotating eddies associated with the poleward Leeuwin Current (LC), off Western Australia. Isotopic signatures ( δ15N and δ13C) were used as proxies for trophic transformation of inorganic and organic matter and are the basis for our discussion on food web functions in the two eddies. We present the first measurements of dissolved inorganic nitrogen (DIN) isotopic composition for the eastern Indian Ocean. We show that the large autotrophs (sampled within the >5-μm and >20-μm fractions of particulate organic matter (POM)), including a distinctive diatom population in the warm-core (WC) eddy, are likely to have taken up sources of DIN which were primarily nitrate, while the picoplankton are likely to have assimilated a large fraction of recycled ammonium. We show that phytoplankton in the cold-core (CC) eddy had distinctly more enriched δ15N signatures than in the WC eddy, probably due to the higher vertical fluxes of nitrate into the CC eddy. A clear negative correlation between mixed-layer depth and δ15N in POM across both eddies also supports the role of vertical nitrate fluxes in determining the primary δ15N signature of the autotrophs. Within the WC eddy, there was a significant δ13C-enrichment in comparison to the CC eddy across all size fractions of the mesozooplankton community, which, in combination with a low C:N molar ratio the >200- and >500-μm mesozooplankton size fractions, suggests a healthier mesozooplankton community, with greater lipid storage, in the WC eddy. This is consistent with the greater productivity and biomass of large diatoms in the WC eddy. Larval fish from the WC eddy also had an enriched δ13C signature compared to those from the CC eddy. The WC eddy had higher production rates than the CC eddy, and harboured a physiologically healthier population of zooplankton. Paradoxically, this seemed to occur

  3. Fungal Functioning In A Pine Forest: Evidence From A 15N-Labeled Global Change Experiment

    NASA Astrophysics Data System (ADS)

    Hobbie, E. A.; Hofmockel, K.; van Diepen, L.

    2012-12-01

    In this study, we used tracer 15N labeling and a concurrent six-year nitrogen fertilization experiment in a Pinus taeda Free Air CO2 Enrichment (FACE) experiment to investigate the functioning of saprotrophic and ectomycorrhizal fungi in nitrogen cycling. Ectomycorrhizal fungi with hydrophobic ectomycorrhizae acquired nitrogen from deeper in the soil profile than taxa with hydrophilic ectomycorrhizae, whereas saprotrophic fungi acquired nitrogen primarily from relatively recent surface litter. By combining natural abundance and tracer measurements, we estimated the 15N enrichment of fungi relative to source nitrogen at between 4‰ and 9‰, depending on genus, with some genera sampling pools more 15N-enriched in the labeling experiment than could be measured by bulk analyses. Thus, biologically relevant nitrogen fractions may correspond poorly to what can be extracted chemically. The multiple 15N labels in this FACE experiment proved useful for tracking nitrogen into different fungal taxa from potential nitrogen sources.

  4. An analytical method for hydrogeochemical surveys: Inductively coupled plasma-atomic emission spectrometry after using enrichment coprecipitation with cobalt and ammonium pyrrolidine dithiocarbamate

    USGS Publications Warehouse

    Hopkins, D.M.

    1991-01-01

    Trace metals that are commonly associated with mineralization were concentrated and separated from natural water by coprecipitation with ammonium pyrollidine dithiocarbamate (APDC) and cobalt and determined by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The method is useful in hydrogeochemical surveys because it permits preconcentration near the sample sites, and selected metals are preserved shortly after the samples are collected. The procedure is relatively simple: (1) a liter of water is filtered; (2) the pH is adjusted; (3) Co chloride and APDC are added to coprecipitate the trace metals; and (4) later, the precipitate is filtered, dissolved, and diluted to 10 ml for a 100-fold concentration enrichment of the separated metals. Sb(III), As(III), Cd, Cr, Cu, Fe, Pb, Mo, Ni, Ag, V, and Zn can then be determined simultaneously by ICP-AES. In an experiment designed to measure the coprecipitation efficiency, Sb(III), Cd and Ag were recovered at 70 to 75% of their original concentration. The remaining metals were recovered at 85 to 100% of their original concentrations, however. The range for the lower limits of determination for the metals after preconcentration is 0.1 to 3.0 ??g/l. The precision of the method was evaluated by replicate analyses of a Colorado creek water and two simulated water samples. The accuracy of the method was estimated using a water reference standard (SRM 1643a) certified by the U.S. National Bureau of Standards. In addition, the method was evaluated by analyzing groundwater samples collected near a porphyry copper deposit in Arizona and by analyzing meltwater from glacier-covered areas favorable for mineralization in south-central Alaska. The results for the ICP-AES analyses compared favorably with those obtained using the sequential technique of GFAAS on the acidified but unconcentrated water samples. ICP-AES analysis of trace-metal preconcentrates for hydrogeochemical surveys is more efficient than GFAAS because a

  5. Measuring denitrification after grassland renewal and grassland conversion to cropland by using the 15N gas-flux method

    NASA Astrophysics Data System (ADS)

    Buchen, Caroline; Eschenbach, Wolfram; Flessa, Heinz; Giesemann, Anette; Lewicka-Szczebak, Dominika; Well, Reinhard

    2015-04-01

    Denitrification, the reduction of oxidized forms of inorganic N to N2O and N2 is an important pathway of gaseous nitrogen losses. Measuring denitrification, especially the reduction of N2O to N2, expressed in the product ratio (N2O/(N2O + N2)), is rather difficult and hence rarely performed under field conditions. But using the 15N gas-flux method allows determining N transformation processes in their natural environment. In order to develop effective climate mitigation strategies understanding the N2O source is essential. We used the 15N gas-flux method to determine N2O and N2 emissions following grassland renewal and conversion techniques. Therefore we selected three different treatments: control (C), mechanical grassland renovation (GR) (autumn 2013) and grassland conversion to maize (GM) (spring 2014) from field plot trials on two different sites (Histic Gleysoil and Plaggic Anthrosol) near Oldenburg, Lower Saxony, Germany. We applied 15N labeled KNO3- (60 atom. % 15N) at a rate equivalent to common farming practices (150 kg N*ha-1) using needle injection of fertilizer solution in three different depths (10 cm, 15 cm, 20 cm) for homogeneous soil labeling up to 30 cm in microplots. During the first 10 days after application (May 2014) gas flux measurements from closed chambers were performed every second day and then weekly following a period of 8 weeks. Gas samples were analyzed for δ15N of N2 and N2O by IRMS according to Lewicka-Szczebak et al. (2013). Concentration and 15N enrichment of NO3- in soil water was determined on weekly samples using the SPIN-MAS technique (Stange et al. 2007). Fluxes of N2 and N2O evolved from the 15N labeled soil nitrogen pool were calculated using the equations of Spott et al. (2006). Peak events of N2 and N2O emissions occurred during the first 10 days of measurement, showing differences in soil types, as well as treatment variations. N2 fluxes up to 178 g*ha-1*day-1 and N2O fluxes up to 280 g*ha-1*day-1 were measured on the

  6. 15N-15N Proton Assisted Recoupling in Magic Angle Spinning NMR

    PubMed Central

    Lewandowski, Józef R.; De Paëpe, Gaël; Eddy, Matthew T.; Griffin, Robert G.

    2009-01-01

    We describe a new magic angle spinning (MAS) NMR experiment for obtaining 15N-15N correlation spectra. The approach yields direct information about the secondary and tertiary structure of proteins, including identification of α-helical stretches and inter-strand connectivity in antiparallel β-sheets, which are of major interest for structural studies of membrane proteins and amyloid fibrils. The method, 15N-15N proton assisted recoupling (PAR), relies on a second order mechanism, third spin assisted recoupling (TSAR), used previously in the context of 15N-13C and 13C-13C polarization transfer schemes. In comparison to 15N-15N proton driven spin diffusion experiments, the PAR technique accelerates polarization transfer between 15N’s by a factor of ~102−103, and is furthermore applicable over the entire range of currently available MAS frequencies (10–70 kHz). PMID:19334788

  7. Aerosol isotopic ammonium signatures over the remote Atlantic Ocean

    NASA Astrophysics Data System (ADS)

    Lin, C. T.; Jickells, T. D.; Baker, A. R.; Marca, A.; Johnson, M. T.

    2016-05-01

    We report aerosol ammonium 15N signatures for samples collected from research cruises on the South Atlantic and Caribbean using a new high sensitivity method. We confirm a pattern of isotopic signals from generally light (δ15N -5 to -10‰), for aerosols with very low (<2 nmol m-3) ammonium concentrations from the remote high latitude ocean, to generally heavier values (δ15N +5 to +10‰), for aerosols collected in temperate and tropical latitudes and with higher ammonium concentrations (>2 nmol m-3). We discuss whether this reflects a mixing of aerosols from two end-members (polluted continental and remote marine emissions), or isotopic fractionation during aerosol transport.

  8. Production of 15N-Labelled Liquid Organic Fertilisers Based on Manure and Crop Residue for Use in Fertigation Studies.

    PubMed

    Martínez-Alcántara, Belén; Martínez-Cuenca, Mary-Rus; Fernández, Carlos; Legaz, Francisco; Quiñones, Ana

    2016-01-01

    Large quantities of crop residue and animal manure from agricultural and livestock activities are annually produced worldwide. With proper management, these residues are potentially valuable sources of plant nutrients, mainly N. Recycling such subproducts in sustainably-based agricultural systems can minimise the use of mineral fertilisers, and hence reduce the potential risk of surface and groundwater pollution. Therefore, the purpose of this study was to obtain (small scale) two liquid labelled-organic fertilisers, an animal- and a vegetal-based organic (AO and VO, respectively) fertiliser, to be used as organic N sources in subsequent fertigation studies. Forage maize (Zea mays L.) grown under 15N-labelled fertiliser supply was used as raw material for VO fertiliser production, and also as 15N-labelled sheep feed to obtain 15N-labelled manure. The labelled faeces fraction was used as raw material for the AO fertiliser. The VO fertiliser was obtained after an acidic and an enzyme-driven hydrolysis. The AO fertiliser was obtained after acidic hydrolysis. The VO liquid fertiliser presented an N concentration of 330 mg·L-1, 85% of total N was organic, while ammonium and nitrate N accounted for 55% and 45% of the mineral nitrogen fraction, respectively. This fertiliser also exhibited high K, Ca and S concentrations and notable values for the remaining macro- and micronutrients. The AO liquid fertiliser had a similar total N concentration (496 mg·L-1, 82% of total N in an organic form) to that of VO, but its mineral N fraction significantly differed, which came in a predominantly (95%) ammonia form. It also had a high content of N, P, K and other macronutrients, and sufficient Fe, Zn, Mn, Cu and B levels, which suggests its suitability as a potential fertiliser. The percentage of 15N enrichment in both VO and AO liquid fertilisers exceeded 2% 15N atom excess, which enabled their use in subsequent assays run to assess nitrogen uptake efficiency. PMID:26982183

  9. Production of 15N-Labelled Liquid Organic Fertilisers Based on Manure and Crop Residue for Use in Fertigation Studies

    PubMed Central

    Martínez-Alcántara, Belén; Martínez-Cuenca, Mary-Rus; Fernández, Carlos; Legaz, Francisco; Quiñones, Ana

    2016-01-01

    Large quantities of crop residue and animal manure from agricultural and livestock activities are annually produced worldwide. With proper management, these residues are potentially valuable sources of plant nutrients, mainly N. Recycling such subproducts in sustainably-based agricultural systems can minimise the use of mineral fertilisers, and hence reduce the potential risk of surface and groundwater pollution. Therefore, the purpose of this study was to obtain (small scale) two liquid labelled-organic fertilisers, an animal- and a vegetal-based organic (AO and VO, respectively) fertiliser, to be used as organic N sources in subsequent fertigation studies. Forage maize (Zea mays L.) grown under 15N-labelled fertiliser supply was used as raw material for VO fertiliser production, and also as 15N-labelled sheep feed to obtain 15N-labelled manure. The labelled faeces fraction was used as raw material for the AO fertiliser. The VO fertiliser was obtained after an acidic and an enzyme-driven hydrolysis. The AO fertiliser was obtained after acidic hydrolysis. The VO liquid fertiliser presented an N concentration of 330 mg·L-1, 85% of total N was organic, while ammonium and nitrate N accounted for 55% and 45% of the mineral nitrogen fraction, respectively. This fertiliser also exhibited high K, Ca and S concentrations and notable values for the remaining macro- and micronutrients. The AO liquid fertiliser had a similar total N concentration (496 mg·L-1, 82% of total N in an organic form) to that of VO, but its mineral N fraction significantly differed, which came in a predominantly (95%) ammonia form. It also had a high content of N, P, K and other macronutrients, and sufficient Fe, Zn, Mn, Cu and B levels, which suggests its suitability as a potential fertiliser. The percentage of 15N enrichment in both VO and AO liquid fertilisers exceeded 2% 15N atom excess, which enabled their use in subsequent assays run to assess nitrogen uptake efficiency. PMID:26982183

  10. Symbiotic nitrogen fixation in an arid ecosystem measured by sup 15 N natural abundance

    SciTech Connect

    Johnson, G.V. )

    1990-05-01

    Plants dependent on nitrogen fixation have an {sup 15}N abundance similar to the atmosphere, while non-nitrogen fixing plants usually are enriched in {sup 15}N and are similar to soil nitrogen values. The natural abundance of {sup 15}N in leaf tissues and soils was determined to evaluate symbiotic nitrogen fixation by several legumes and actinorhizal species in the Sevilleta Long-term Ecological Research area in central New Mexico. Comparison of {delta}{sup 15}N values for the legume Prosopis glandulosa (mesquite) to adjacent Atriplex canascens (fourwing saltbush) indicated that P. glandulosa obtained 66% of its nitrogen by fixation. The legume Hoffmanseggia jamesii was found to be utilizing soil nitrogen. The {delta}{sup 15}N values for the actinorhizal plants, Elaeagnus angustifolia and Cercocarpus montanus, while below values for soil nitrogen, did not differ from associated non-fixing plants.

  11. Unusually negative nitrogen isotopic compositions (δ15N) of mangroves and lichens in an oligotrophic, microbially-influenced ecosystem

    NASA Astrophysics Data System (ADS)

    Fogel, M. L.; Wooller, M. J.; Cheeseman, J.; Smallwood, B. J.; Roberts, Q.; Romero, I.; Jacobsen Meyers, M.

    2008-02-01

    Extremes in (δ15N values in mangrove tissues and lichens (range = +4 to -22‰) were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, atmospheric ammonia, mangrove leaves, roots, stems, and wood, and lichens, were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Porewater ammonium concentrations had little to no relationship to N isotopic fractionation in mangrove tissues. The δ15N of fine and coarse roots was 9‰ more positive than leaf tissue from the same tree. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year to a &delta:15N closer to the &delta:15N of porewater ammonium15N=+4‰). Isotopically negative ammonia in the atmosphere (δ15N=-18‰) and in rainwater (δ15N=-9‰) were found on Twin Cays and may be sources of available N for isotopically depleted mangrove trees and lichens. In highly stressed, severely P limited trees, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

  12. A new organic reference material, L-glutamic acid, USGS41a, for δ13C and δ15N measurements − a replacement for USGS41

    USGS Publications Warehouse

    Qi, Haiping; Coplen, Tyler B.; Mroczkowski, Stanley J.; Brand, Willi A.; Brandes, Lauren; Geilmann, Heike; Schimmelmann, Arndt

    2016-01-01

    RationaleThe widely used l-glutamic acid isotopic reference material USGS41, enriched in both 13C and 15N, is nearly exhausted. A new material, USGS41a, has been prepared as a replacement for USGS41.MethodsUSGS41a was prepared by dissolving analytical grade l-glutamic acid enriched in 13C and 15N together with l-glutamic acid of normal isotopic composition. The δ13C and δ15N values of USGS41a were directly or indirectly normalized with the international reference materials NBS 19 calcium carbonate (δ13CVPDB = +1.95 mUr, where milliurey = 0.001 = 1 ‰), LSVEC lithium carbonate (δ13CVPDB = −46.6 mUr), and IAEA-N-1 ammonium sulfate (δ15NAir = +0.43 mUr) and USGS32 potassium nitrate (δ15N = +180 mUr exactly) by on-line combustion, continuous-flow isotope-ratio mass spectrometry, and off-line dual-inlet isotope-ratio mass spectrometry.ResultsUSGS41a is isotopically homogeneous; the reproducibility of δ13C and δ15N is better than 0.07 mUr and 0.09 mUr, respectively, in 200-μg amounts. It has a δ13C value of +36.55 mUr relative to VPDB and a δ15N value of +47.55 mUr relative to N2 in air. USGS41 was found to be hydroscopic, probably due to the presence of pyroglutamic acid. Experimental results indicate that the chemical purity of USGS41a is substantially better than that of USGS41.ConclusionsThe new isotopic reference material USGS41a can be used with USGS40 (having a δ13CVPDB value of −26.39 mUr and a δ15NAir value of −4.52 mUr) for (i) analyzing local laboratory isotopic reference materials, and (ii) quantifying drift with time, mass-dependent isotopic fractionation, and isotope-ratio-scale contraction for isotopic analysis of biological and organic materials. Published in 2016. This article is a U.S. Government work and is in the public domain in the USA.

  13. Plant and Soil Natural Abundance delta-15N: Indicators of Nitrogen Cycling in the Catskill Mountains, New York, USA

    NASA Astrophysics Data System (ADS)

    Templer, P. H.; Lovett, G. M.; Weathers, K.; Arthur, M. A.

    2002-12-01

    We examined the potential use of natural abundance 15N of plants and soils as an indicator of forest nitrogen (N) cycling rates within the Catskill Mountains, NY. These watersheds receive among the highest rates of N deposition in the northeastern United States and are beginning to show signs of N saturation. Many studies have shown a link between increased N cycling rates and 15N enrichment of soil and plant pools. Faster rates of N cycling processes, especially nitrification, lead to fractionation of 14/15N, creating N products that are relatively depleted in 15N. This can lead to enrichment of soil pools, as lighter 14N is lost from the system via leaching or denitrification. Plant N pools can become increasingly enriched as they take up 15N-enriched soil N. Despite similar amounts of N deposition across the Catskill Mountains, forests dominated by different tree species appear to vary in the amount of N retained or lost to nearby streams. To determine if plant and soil 15N could be used as indicators of N cycling rates, we collected foliage, wood, litterfall, organic and mineral soil, and fine roots from single species stands of American beech (Fagus grandifolia), eastern hemlock (Tsuga canadensis), red oak (Quercus rubra), and sugar maple (Acer saccharum). Fine roots and soil 15N were highest within sugar maple stands (p<0.05). Sugar maple soils also had the highest rates of net nitrification and N leaching. Therefore, soil 15N appears to correlate with forest N retention and loss. However, 15N enrichment was highest within foliage, litterfall and wood of beech trees (p<0.05). The decoupling between foliage 15N and N cycling, as well as between 15N of foliage and fine roots, illustrates that it may not be possible to use a single plant pool as an indicator of N cycling rates.

  14. Human dietary δ(15)N intake: representative data for principle food items.

    PubMed

    Huelsemann, F; Koehler, K; Braun, H; Schaenzer, W; Flenker, U

    2013-09-01

    Dietary analysis using δ(15)N values of human remains such as bone and hair is usually based on general principles and limited data sets. Even for modern humans, the direct ascertainment of dietary δ(15)N is difficult and laborious, due to the complexity of metabolism and nitrogen fractionation, differing dietary habits and variation of δ(15)N values of food items. The objective of this study was to summarize contemporary regional experimental and global literature data to ascertain mean representative δ(15)N values for distinct food categories. A comprehensive data set of more than 12,000 analyzed food samples was summarized from the literature. Data originated from studies dealing with (1) authenticity tracing or origin control of food items, and (2) effects of fertilization or nutrition on δ(15)N values of plants or animals. Regional German food δ(15)N values revealed no major differences compared with the mean global values derived from the literature. We found that, in contrast to other food categories, historical faunal remains of pig and poultry are significantly enriched in (15)N compared to modern samples. This difference may be due to modern industrialized breeding practices. In some food categories variations in agricultural and feeding regimens cause significant differences in δ(15)N values that may lead to misinterpretations when only limited information is available.

  15. Regulation of [15N]urea synthesis from [5-15N]glutamine. Role of pH, hormones, and pyruvate.

    PubMed

    Nissim, I; Yudkoff, M; Brosnan, J T

    1996-12-01

    We have utilized both [5-15N]glutamine and [3-13C] pyruvate as metabolic tracers in order to: (i) examine the effect of pH, glucagon (GLU), or insulin on the precursor-product relationship between 15NH3, [15N]citrulline, and, thereby, [15N]urea synthesis and (ii) elucidate the mechanism(s) by which pyruvate stimulates [15N] urea synthesis. Hepatocytes isolated from rat were incubated at pH 6.8, 7.4, or 7.6 with 1 mM [5-15N]glutamine and 0.1 mM 14NH4Cl in the presence or the absence of [3-13C] pyruvate (2 mM). A separate series of experiments was performed at pH 7.4 in the presence of insulin or GLU. 15NH3 enrichment exceeded or was equal to that of [15N]citrulline under all conditions except for pH 7.6, when the 15N enrichment in citrulline exceeded that in ammonia. The formation of [15N]citrulline (atom % excess) was increased with higher pH. Flux through phosphate-dependent glutaminase (PDG) and [15N]urea synthesis were stimulated (p < 0.05) at pH 7.6 or with GLU and decreased (p < 0.05) at pH 6.8. Insulin had no significant effect on flux through PDG or on [15N]urea synthesis. Decreased [15N]urea production at pH 6.8 was associated with depleted aspartate and glutamate levels. Pyruvate attenuated this decrease in the aspartate and glutamate pools and stimulated [15N]urea synthesis. Production of Asp from pyruvate was increased with increasing medium pH. Approximately 80% of Asp was derived from [3-13C]pyruvate regardless of incubation pH or addition of hormone. Furthermore, approximately 20, 40, and 50% of the mitochondrial N-acetylglutamate (NAG) pool was derived from [3-13C]pyruvate at pH 6.8, 7.4, and 7.6, respectively. Both the concentration and formation of [13C]NAG from [3-13C]pyruvate were increased (p < 0.05) with glucagon and decreased (p < 0.05) with insulin or at pH 6.8. The data suggest a correlation between changes in [15N]urea synthesis and alterations in the level and synthesis of [13C]NAG from pyruvate. The current observations suggest that the

  16. Unusually negative nitrogen isotopic compositions (δ15N) of mangroves and lichens in an oligotrophic, microbially-influenced ecosystem

    NASA Astrophysics Data System (ADS)

    Fogel, M. L.; Wooller, M. J.; Cheeseman, J.; Smallwood, B. J.; Roberts, Q.; Romero, I.; Meyers, M. J.

    2008-12-01

    Extremes in δ15N values in mangrove tissues and lichens (range =+4 to -22‰) were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, and atmospheric ammonia, and the δ15N of lichens, mangrove leaves, roots, stems, and wood were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Dwarfed Rhizophora mangle trees had the most negative δ15N, whereas fringing Rhizophora trees, the most positive δ15N values. Porewater ammonium concentrations had little relationship to N isotopic fractionation in mangrove tissues. In dwarfed mangroves, the δ15N of fine and coarse roots were 6-9‰ more positive than leaf tissue from the same tree, indicating different sources of N for root and leaf tissues. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year from -12‰ to -2‰, approaching the δ15N of porewater ammonium15N=+4‰). Isotopically depleted ammonia in the atmosphere (δ15N=-19‰) and in rainwater (δ15N=-10‰) were found on Twin Cays. We propose that foliar uptake of these atmospheric sources by P-stressed, dwarfed mangrove trees and lichens can explain their very negative δ15N values. In environments where P is limiting for growth, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

  17. Light-mediated 15N fractionation in Caribbean gorgonian octocorals: implications for pollution monitoring

    NASA Astrophysics Data System (ADS)

    Baker, D. M.; Kim, K.; Andras, J. P.; Sparks, J. P.

    2011-09-01

    The stable nitrogen isotope ratio ( δ 15N) of coral tissue is a useful recorder of anthropogenic pollution in tropical marine ecosystems. However, little is known of the natural environmentally induced fractionations that affect our interpretation of coral δ 15N values. In symbiotic scleractinians, light affects metabolic fractionation of N during photosynthesis, which may confound the identification of N pollution between sites of varied depth or turbidity. Given the superiority of octocorals for δ 15N studies, our goal was to quantify the effect of light on gorgonian δ 15N in the context of monitoring N pollution sources. Using field collections, we show that δ 15N declined by 1.4‰ over 20 m depth in two species of gorgonians, the common sea fan, Gorgonia ventalina, and the slimy sea plume, Pseudopterogorgia americana. An 8-week laboratory experiment with P. americana showed that light, not temperature causes this variation, whereby the lowest fractionation of the N source was observed in the highest light treatment. Finally, we used a yearlong reciprocal depth transplant experiment to quantify the time frame over which δ 15N changes in G. ventalina as a function of light regime . Over the year, δ 15N was unchanged and increased slightly in the deep control colonies and shallow colonies transplanted to the deep site, respectively. Within 6 months, colonies transplanted from deep to shallow became enriched by 0.8‰, mirroring the enrichment observed in the shallow controls, which was likely due to the combined effect of an increase in the source δ 15N and reduced fractionation. We conclude that light affects gorgonian δ 15N fractionation and should be considered in sampling designs for N pollution monitoring. However, these fractionations are small relative to differences observed between natural and anthropogenic N sources.

  18. Fate and metabolism of [15N]2,4,6-trinitrotoluene in soil.

    PubMed

    Weiss, Martin; Geyer, Roland; Russow, Rolf; Richnow, Hans H; Kästner, Matthias

    2004-08-01

    The fates of the labels from [14C] and [15N] trinitrotoluene were analyzed in bioreactors under aerobic conditions in soil treated by a fungal bioremediation process with Stropharia rugosoannulata and in control soil. Up to 17.5% of the 15N label had a different fate than the 14C label. Three N-mineralization processes were identified in detailed experiments with [15N]TNT. About 2% of the 15N label was found as NO3- and NH4+, showing simultaneous processes of direct TNT denitration (I) and reduction with cleavage of the amino groups (II). The enrichment of NO2-/NO3- (up to 7.5 atom% 15N abundance) indicates the formation of Meisenheimer complexes with a denitration of [15N]TNT. A 1.4% of the label was found distributed between N2O and N2. However, the 15N enrichment of the N2O (up to 38 atom%) demonstrated that both N atoms were generated from the labeled TNT and clearly indicates a novel formation process (III). We propose, as an explanation, the generation of N2O by cleavage from condensed azoxy metabolites. In addition, 1.7% of the 15N label was detected as biogenic amino acids in the wheat straw containing the fungus. Overall, 60 to 85% of the applied [15N]TNT was degraded and 52 to 64% was found as nonextractable residues in the soil matrix. Three percent was detected as 2-amino-4,6-dinitrotoluene and 4-amino-2,6-dinitrotoluene.

  19. Use of /sup 15/N to measure nitrogen uptake in eutrophic oceans; experimental considerations

    SciTech Connect

    Not Available

    1986-07-01

    The use of /sup 15/N to measure the flux of nitrogen compounds has become increasingly popular as the techniques and instrumentation for stable isotope analysis have become more widely available. Questions concerning equations for calculating uptake, effect of isotope dilution (in the case of ammonium), duration of incubation, and relationship between disappearance of a nitrogen compound and the /sup 15/N uptake measurement have arisen, especially for the research conducted in oligotrophic regions. Fewer problems seem to have occurred ineutrophic areas. However, sufficient literature now exists to allow some generally accepted experimental procedures for /sup 15/N studies in eutrophic regions to be laid down. Incubation periods of 2-6 h appear to avoid problems related to isotope dilution and to overcome the bias introduced in some cases by initial high rate or surge uptake. During such incubation periods, assimilation is measured rather than uptake or transport into the cell. Incorporation of /sup 15/N into the particulate fraction is usually linear with time over the periods currently used. The /sup 15/N method provides a better estimate of incorporation into phytoplankton than /sup 14/N disappearance, but a small fraction appears to be lost. Although most workers suggest the loss to be a result of dissolved organic nitrogen production, direct evidence is lacking. If the considerations discussed here are applied with the /sup 15/N techniques currently available, reliable estimates of phytoplankton nitrogen flux in eutrophic areas can be obtained.

  20. Production of 15N-depleted biomass during cyanobacterial N2-fixation at high Fe concentrations

    NASA Astrophysics Data System (ADS)

    Zerkle, Aubrey L.; Junium, Christopher K.; Canfield, Donald E.; House, Christopher H.

    2008-09-01

    In this study we examine the effects of varying Fe, Mo, and P concentrations on δ15N fractionation during N2 fixation in the cyanobacterium Anabaena variabilis. We show that when grown in Fe-enriched media ([Fe] ≥ 50 nM), this organism produces biomass up to 3‰ lower in δ15N than when grown in Fe-limited media ([Fe] < 50 nM). A compilation of our data with previous measurements of δ15N in N2-fixing cyanobacteria reveals a general trend toward the production of more 15N-depleted biomass at higher Fe concentrations. We discuss our results in the context of negative δ15N values preserved in Archean and some Phanerozoic sediments, generally attributed to the production of marine organic matter with low δ15N by N2 fixation (and potentially NH4+ regeneration) during periods of fluctuating nutrient dynamics. We suggest that enhanced Fe availability during periods of widespread ocean anoxia can further stimulate the production of 15N-depleted biomass by N2-fixing organisms, contributing to the isotopic record.

  1. Hydrogen doppler spectroscopy using 15N ions

    NASA Astrophysics Data System (ADS)

    Borucki, L.; Becker, H. W.; Gorris, F.; Kubsky, S.; Schulte, W. H.; Rolfs, C.

    The energy spread of atomic and molecular ion beams from the 4 MV Dynamitron tandem accelerator at the Ruhr-Universität Bochum has been studied and in part minimized. Using the ER= 6.40 MeV narrow resonance in 1H(15N,αγ)12C with an 15N energy spread of 4.55 keV, the Doppler broadening for several hydrogen-bearing gases was found to be in good agreement with expectation: e.g. for NH3 gas a rotational-vibrational Doppler width of 10.41 +/- 0.25 keV was observed (theory = 10.4 keV). Studies of the vibrational Doppler widths of H-bonds on a Si <100> surface were performed using a 4πγ-ray detection system together with UHV-chambers for sample preparation, transport, and analysis. The results showed that further improvements in the experimental set-ups are needed for such investigations.

  2. Two new organic reference materials for δ13C and δ15N measurements and a new value for the δ13C of NBS 22 oil

    USGS Publications Warehouse

    Qi, Haiping; Coplen, Tyler B.; Geilmann, Heike; Brand, Willi A.; Bohlke, John Karl

    2003-01-01

    Analytical grade L-glutamic acid is chemically stable and has a C/N mole ratio of 5, which is close to that of many of natural biological materials, such as blood and animal tissue. Two L-glutamic acid reference materials with substantially different 13C and 15N abundances have been prepared for use as organic reference materials for C and N isotopic measurements. USGS40 is analytical grade L-glutamic acid and has a δ13C value of −26.24‰ relative to VPDB and a δ15N value of −4.52‰ relative to N2 in air. USGS41 was prepared by dissolving analytical grade L-glutamic acid with L-glutamic acid enriched in 13C and 15N. USGS41 has a δ13C value of +37.76‰ and a δ15N value of +47.57‰. The δ13C and δ15N values of both materials were measured against the international reference materials NBS 19 calcium carbonate (δ13C = +1.95‰), L-SVEC lithium carbonate (δ13C = −46.48‰), IAEA-N-1 ammonium sulfate (δ15N = 0.43‰), and USGS32 potassium nitrate (δ15N = 180‰) by on-line combustion continuous-flow and off-line dual-inlet isotope-ratio mass spectrometry. Both USGS40 and USGS41 are isotopically homogeneous; reproducibility of δ13C is better than 0.13‰, and that of δ15N is better than 0.13‰ in 100-μg amounts. These two isotopic reference materials can be used for (i) calibrating local laboratory reference materials, and (ii) quantifying drift with time, mass-dependent fractionations, and isotope-ratio-scale contraction in the isotopic analysis of various biological materials. Isotopic results presented in this paper yield a δ13C value for NBS 22 oil of −29.91‰, in contrast to the commonly accepted value of −29.78‰ for which off-line blank corrections probably have not been quantified satisfactorily.

  3. Ammonium methacrylate

    Integrated Risk Information System (IRIS)

    Ammonium methacrylate ; CASRN 16325 - 47 - 6 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcin

  4. Ammonium sulfamate

    Integrated Risk Information System (IRIS)

    Ammonium sulfamate ; CASRN 7773 - 06 - 0 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogen

  5. Ammonium acetate

    Integrated Risk Information System (IRIS)

    Ammonium acetate ; CASRN 631 - 61 - 8 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic

  6. Stem injection of 15N-NH4NO3 into mature Sitka spruce (Picea sitchensis).

    PubMed

    Nair, Richard; Weatherall, Andrew; Perks, Mike; Mencuccini, Maurizio

    2014-10-01

    Stem injection techniques can be used to introduce (15)N into trees to overcome a low variation in natural abundance and label biomass with a distinct (15)N signature, but have tended to target small and young trees, of a variety of species, with little replication. We injected 98 atom% (15)N ammonium nitrate (NH4NO3) solution into 13 mature, 9- to 13-m tall edge-profile Sitka spruce trees in order to produce a large quantity of labelled litter, examining the distribution of the isotope throughout the canopy after felling in terms of both total abundance of (15)N and relative distribution of the isotope throughout individual trees. Using a simple mass balance of the canopy alone, based on observed total needle biomass and modelled branch biomass, all of the isotope injected was accounted for, evenly split between needles and branches, but with a high degree of variability both within individual trees, and among trees. Both (15)N abundance and relative within-canopy distribution were biased towards the upper and middle crown in foliage. Recovery of the label in branches was much more variable than in needles, possibly due to differences in nitrogen allocation for both growth and storage, which differ seasonally between foliage and woody biomass.

  7. Compound-Specific δ15N Amino Acid Measurements in Littoral Mussels in the California Upwelling Ecosystem: A New Approach to Generating Baseline δ15N Isoscapes for Coastal Ecosystems

    PubMed Central

    Vokhshoori, Natasha L.; McCarthy, Matthew D.

    2014-01-01

    We explored δ15N compound-specific amino acid isotope data (CSI-AA) in filter-feeding intertidal mussels (Mytilus californianus) as a new approach to construct integrated isoscapes of coastal primary production. We examined spatial δ15N gradients in the California Upwelling Ecosystem (CUE), determining bulk δ15N values of mussel tissue from 28 sites between Port Orford, Oregon and La Jolla, California, and applying CSI-AA at selected sites to decouple trophic effects from isotopic values at the base of the food web. Bulk δ15N values showed a strong linear trend with latitude, increasing from North to South (from ∼7‰ to ∼12‰, R2 = 0.759). In contrast, CSI-AA trophic position estimates showed no correlation with latitude. The δ15N trend is therefore most consistent with a baseline δ15N gradient, likely due to the mixing of two source waters: low δ15N nitrate from the southward flowing surface California Current, and the northward transport of the California Undercurrent (CUC), with15N-enriched nitrate. This interpretation is strongly supported by a similar linear gradient in δ15N values of phenylalanine (δ15NPhe), the best AA proxy for baseline δ15N values. We hypothesize δ15NPhe values in intertidal mussels can approximate annual integrated δ15N values of coastal phytoplankton primary production. We therefore used δ15NPhe values to generate the first compound-specific nitrogen isoscape for the coastal Northeast Pacific, which indicates a remarkably linear gradient in coastal primary production δ15N values. We propose that δ15NPhe isoscapes derived from filter feeders can directly characterize baseline δ15N values across major biochemical provinces, with potential applications for understanding migratory and feeding patterns of top predators, monitoring effects of climate change, and study of paleo- archives. PMID:24887109

  8. Determination of nitrogen-15 enrichment of nitrate and nitrite using thermospray liquid chromatography-mass spectrometry

    SciTech Connect

    Hogge, L.R.; Hynes, R.K.; Nelson, L.M.; Vestal, M.L.

    1986-11-01

    A new method has been developed for the determination of percent /sup 15/N enrichment in NO/sub 2//sup -/ and NO/sub 3//sup -/ that is both rapid and sensitive. NO/sub 2//sup -/ and NO/sub 3//sup -/ separation was done by use of reverse-phase liquid chromatography with a mobile phase consisting of ammonium acetate with tetrabutylammonium acetate as the ion pair reagent. The % /sup 15/N in isotopically enriched anions was determined by using thermospray liquid chromatography-mass spectrometry (LC-MS). Enrichment values were obtained on as little as 20 ng of NO/sub 2//sup -/-N/20 ..mu..L and 4 ng of NO/sub 3//sup -/-N/20 ..mu..L. The linear regression curve was plotted for the observed enrichment vs. the actual enrichment over the range from the natural abundance to 25% enrichment for each anion and the equation for the linear regression calculated (y = 1.16x + 0.7577 for NO/sub 2//sup -/ and y = 0.951x + 0.447 for NO/sub 3//sup -/). The coefficient of determination for the NO/sub 2//sup -/ and NO/sub 3//sup -/ curves was 0.988 and 0.987, respectively. The dynamic range of the method was found to be suitable for the range of NO/sub 2//sup -/ and NO/sub 3//sup -/ expected in plant xylem sap. Xylem sap samples obtained from NO/sub 3//sup -/ fed pea, Pisum sativum cv. Homesteader (3.9 ..mu..g of NO/sub 3//sup -/-N/mL, 24.7% /sup 15/N), contained a /sup 15/N enrichment of 25.4% +/- 0.7%.

  9. Compound-specific 15N analysis of amino acids in 15N tracer experiments provide an estimate of newly synthesised soil protein from inorganic and organic substrates

    NASA Astrophysics Data System (ADS)

    Charteris, Alice; Michaelides, Katerina; Evershed, Richard

    2015-04-01

    unlike previous works analyses for amino acids (representing organic products) rather than ammonium (NH4+) and nitrate (NO3-). Amino acids are commonly referred to as 'the building blocks of life' as they form the proteins which regulate life's essential biochemical reactions. Proteinaceous matter generally comprises 20-40% of total soil N and is ubiquitous in living organisms, so is a likely 'organic product' of microbial activity/assimilation. Hence, we consider it likely that amino acids represent the major organic nitrogenous products and a reasonable 'proxy' for/measure of the assimilation of an applied 15N substrate by the soil microbial biomass and an estimate of the newly synthesized soil protein. Brookes, P. C. et al. Soil Biol Biochem. 1985, 17, 837-842. Jenkinson, D. S. et al. Soil Biol Biochem. 2004, 36, 5-7. Nannipieri, P. et al. Plant Soil. 1999, 208, 43-56. Pilbeam, C. J. et al. J Agr Sci. 1997, 128, 415-424. Sebilo, M. et al. PNAS. 2013, 110, 18185-18189.

  10. Nitrogen Isotopic Ratios in Cometary NH2: Implication for 15N-fractionation in Ammonia

    NASA Astrophysics Data System (ADS)

    Shinnaka, Yoshiharu; Kawakita, Hideyo; Jehin, Emmanuël; Decock, Alice; Hutsemékers, Damien; Manfroid, Jean; Arai, Akira

    2015-11-01

    Isotopic ratios in cometary molecules are diagnostic for the physico-chemical conditions where molecules formed and are processed, from the interstellar medium to the solar nebula. Usually temperatures at the molecular formation control the fractionation of the heavier element in molecular species, e.g., D-fractionation in water.In cometary volatiles, the 14N/15N ratios in CN have been well observed (Manfroid et al. 2009, A&A, 503, 613, and reference therein) and is consistent with the ratio in HCN (a most probable parent of CN) measured in few comets (Bockelée-Morvan et al. 2008, ApJ, 679, L49). Those ratios are enriched compared to the proto-solar value by a factor of ~3. In contrast to those Nitriles, there are only few reports on 14N/15N ratios in Ammonia (as Amine) (Rousselot et al. 2014, ApJ, 780, L17; Shinnaka et al. 2014, ApJ, 782, L16). Ammonia (NH3) is usually the most abundant and HCN is the second most abundant N-bearing volatiles in cometary ice. Especially, recent observations of 15NH2 revealed the 14N/15N ratios in NH3 are comparable to those of CN. However, from the viewpoint of theoretical work, the enrichment of 15N in cometary NH3 cannot be reproduced by current chemical network models. Information about the diversity of the 14N/15N ratios in NH3 of individual comets is needed to understand the formation mechanisms/environments of NH3 in the early solar system.To clarify the diversity of the 14N/15N ratios in cometary NH3, we determine the 14N/15N ratios in NH3 for more than ten comets individually which include not only Oort cloud comets but also short period comets by using the high-resolution optical spectra of NH2. These spectra were obtained with both the UVES mounted on the VLT in Chile and the HDS on the Subaru Telescope in Hawaii.The derived 14N/15N ratios in NH3 for more than ten comets show high 15N-enrichment compared with the elemental abundances of nitrogen in the Sun by about factor of ~3 and has no large diversity depending on

  11. 15N Abundance of Nodules as an Indicator of N Metabolism in N2-Fixing Plants 1

    PubMed Central

    Shearer, Georgia; Feldman, Lori; Bryan, Barbara A.; Skeeters, Jerri L.; Kohl, Daniel H.; Amarger, Nöelle; Mariotti, Françoise; Mariotti, André

    1982-01-01

    This paper expands upon previous reports of 15N elevation in nodules (compared to other tissues) of N2-fixing plants. N2-Fixing nodules of Glycine max (soybeans), Vigna unguiculata (cowpea), Phaseolus vulgaris (common bean), Phaseolus coccineus (scarlet runner bean), Prosopis glandulosa (mesquite), and Olneya tesota (desert ironwood) were enriched in 15N. Nodules of Vicia faba (fava beans), Arachis hypogaea (peanut), Trifolium pratense (red clover), Pisum sativum (pea), Lathyrus sativus (grass pea), Medicago sativa (alfalfa), and Lupinus mutabilis (South American lupine) were not; nor were the nodules of nine species of N2-fixing nonlegumes. The nitrogen of ineffective nodules of soybeans and cowpeas was not enriched in 15N. Thus, 15N elevation in nodules of these plants depends on active N2-fixation. Results obtained so far on the generality of 15N enrichment in N2-fixing nodules suggest that only the nodules of plants which actively fix N2 and which transport allantoin or allantoic acid exhibit 15N enrichment. PMID:16662517

  12. The First in Vivo Observation of 13C- 15N Coupling in Mammalian Brain

    NASA Astrophysics Data System (ADS)

    Kanamori, Keiko; Ross, Brian D.

    2001-12-01

    [5-13C,15N]Glutamine, with 1J(13C-15N) of 16 Hz, was observed in vivo in the brain of spontaneously breathing rats by 13C MRS at 4.7 T. The brain [5-13C]glutamine peak consisted of the doublet from [5-13C,15N]glutamine and the center [5-13C,14N]glutamine peak, resulting in an apparent triplet with a separation of 8 Hz. The time course of formation of brain [5-13C,15N]glutamine was monitored in vivo with a time resolution of 20-35 min. This [5-13C,15N]glutamine was formed by glial uptake of released neurotransmitter [5-13C]glutamate and its reaction with 15NH3 catalyzed by the glia-specific glutamine synthetase. The neurotransmitter glutamate C5 was selectively13C-enriched by intravenous [2,5-13C]glucose infusion to 13C-label whole-brain glutamate C5, followed by [12C]glucose infusion to chase 13C from the small and rapidly turning-over glial glutamate pool, leaving 13C mainly in the neurotransmitter [5-13C]glutamate pool, which is sequestered in vesicles until release. Hence, the observed [5-13C,15N]glutamine arises from a coupling between 13C of neuronal origin and 15N of glial origin. Measurement of the rate of brain [5-13C,15N]glutamine formation provides a novel noninvasive method of studying the kinetics of neurotransmitter uptake into glia in vivo, a process that is crucial for protecting the brain from glutamate excitotoxicity.

  13. Effects of four different restoration treatments on the natural abundance of (15)n stable isotopes in plants.

    PubMed

    Temperton, Vicky M; Märtin, Lea L A; Röder, Daniela; Lücke, Andreas; Kiehl, Kathrin

    2012-01-01

    δ(15)N signals in plant and soil material integrate over a number of biogeochemical processes related to nitrogen (N) and therefore provide information on net effects of multiple processes on N dynamics. In general little is known in many grassland restoration projects on soil-plant N dynamics in relation to the restoration treatments. In particular, δ(15)N signals may be a useful tool to assess whether abiotic restoration treatments have produced the desired result. In this study we used the range of abiotic and biotic conditions provided by a restoration experiment to assess to whether the restoration treatments and/or plant functional identity and legume neighborhood affected plant δ(15)N signals. The restoration treatments consisted of hay transfer and topsoil removal, thus representing increasing restoration effort, from no restoration measures, through biotic manipulation to major abiotic manipulation. We measured δ(15)N and %N in six different plant species (two non-legumes and four legumes) across the restoration treatments. We found that restoration treatments were clearly reflected in δ(15)N of the non-legume species, with very depleted δ(15)N associated with low soil N, and our results suggest this may be linked to uptake of ammonium (rather than nitrate). The two non-legume species differed considerably in their δ(15)N signals, which may be related to the two species forming different kinds of mycorrhizal symbioses. Plant δ(15)N signals could clearly separate legumes from non-legumes, but our results did not allow for an assessment of legume neighborhood effects on non-legume δ(15)N signals. We discuss our results in the light of what the δ(15)N signals may be telling us about plant-soil N dynamics and their potential value as an indicator for N dynamics in restoration.

  14. Effects of Four Different Restoration Treatments on the Natural Abundance of 15N Stable Isotopes in Plants

    PubMed Central

    Temperton, Vicky M.; Märtin, Lea L. A.; Röder, Daniela; Lücke, Andreas; Kiehl, Kathrin

    2012-01-01

    δ15N signals in plant and soil material integrate over a number of biogeochemical processes related to nitrogen (N) and therefore provide information on net effects of multiple processes on N dynamics. In general little is known in many grassland restoration projects on soil–plant N dynamics in relation to the restoration treatments. In particular, δ15N signals may be a useful tool to assess whether abiotic restoration treatments have produced the desired result. In this study we used the range of abiotic and biotic conditions provided by a restoration experiment to assess to whether the restoration treatments and/or plant functional identity and legume neighborhood affected plant δ15N signals. The restoration treatments consisted of hay transfer and topsoil removal, thus representing increasing restoration effort, from no restoration measures, through biotic manipulation to major abiotic manipulation. We measured δ15N and %N in six different plant species (two non-legumes and four legumes) across the restoration treatments. We found that restoration treatments were clearly reflected in δ15N of the non-legume species, with very depleted δ15N associated with low soil N, and our results suggest this may be linked to uptake of ammonium (rather than nitrate). The two non-legume species differed considerably in their δ15N signals, which may be related to the two species forming different kinds of mycorrhizal symbioses. Plant δ15N signals could clearly separate legumes from non-legumes, but our results did not allow for an assessment of legume neighborhood effects on non-legume δ15N signals. We discuss our results in the light of what the δ15N signals may be telling us about plant–soil N dynamics and their potential value as an indicator for N dynamics in restoration. PMID:22645597

  15. (15)N- and (2)H proteomic stable isotope probing links nitrogen flow to archaeal heterotrophic activity.

    PubMed

    Justice, Nicholas B; Li, Zhou; Wang, Yingfeng; Spaudling, Susan E; Mosier, Annika C; Hettich, Robert L; Pan, Chongle; Banfield, Jillian F

    2014-10-01

    Understanding how individual species contribute to nutrient transformations in a microbial community is critical to prediction of overall ecosystem function. We conducted microcosm experiments in which floating acid mine drainage (AMD) microbial biofilms were submerged - recapitulating the final stage in a natural biofilm life cycle. Biofilms were amended with either (15)NH4(+) or deuterium oxide ((2)H2O) and proteomic stable isotope probing (SIP) was used to track the extent to which different members of the community used these molecules in protein synthesis across anaerobic iron-reducing, aerobic iron-reducing and aerobic iron-oxidizing environments. Sulfobacillus spp. synthesized (15)N-enriched protein almost exclusively under iron-reducing conditions whereas the Leptospirillum spp. synthesized (15)N-enriched protein in all conditions. There were relatively few (15)N-enriched archaeal proteins, and all showed low atom% enrichment, consistent with Archaea synthesizing protein using the predominantly (14)N biomass derived from recycled biomolecules. In parallel experiments using (2)H2O, extensive archaeal protein synthesis was detected in all conditions. In contrast, the bacterial species showed little protein synthesis using (2)H2O. The nearly exclusive ability of Archaea to synthesize proteins using (2)H2O may be due to archaeal heterotrophy, whereby Archaea offset deleterious effects of (2)H by accessing (1)H generated by respiration of organic compounds.

  16. Food webs in Mongolian grasslands: the analysis of 13C and 15N natural abundances.

    PubMed

    Kohzu, Ayato; Iwata, T; Kato, M; Nishikawa, J; Wada, Eitaro; Amartuvshin, N; Namkhaidorj, B; Fujita, N

    2009-09-01

    Overgrazing often lowers species richness and productivity of grassland communities. For Mongolian grassland ecosystems, a lack of detailed information about food-web structures makes it difficult to predict the effects of overgrazing on species diversity and community composition. We analysed the delta13C and delta15N signatures of herbaceous plants, arthropods (grouped by feeding habit), wild and domestic mammals, and humans in central Mongolia to understand the predominant food-web pathways in this grassland ecosystem. The delta13C and delta15N values of mammals showed little variation within species, but varied considerably with slope position for arthropods. The apparent isotopic discrimination between body tissue and hair of mammals was estimated as 2.0 per thousand for delta13C and 2.1 per thousand for delta15N, which was large enough to cause overestimation of the trophic level of mammals if not taken into account when using hair samples to measure isotopic enrichment.

  17. Vertical δ13C and δ15N changes during pedogenesis

    NASA Astrophysics Data System (ADS)

    Brunn, Melanie; Spielvogel, Sandra; Wells, Andrew; Condron, Leo; Oelmann, Yvonne

    2015-04-01

    The natural abundance of soil organic matter (SOM) stable C and N isotope ratios are subjected to vertical changes throughout the soil profile. This vertical distribution is a widely reported phenomenon across varieties of ecosystems and constitutes important insights of soil carbon cycling. In most ecosystems, SOM becomes enriched in heavy isotopes by several per mill in the first few centimeters of the topsoil. The enrichment of 13C in SOM with soil depth is attributed to biological and physical-chemical processes in soil e.g., plant physiological impacts, microbial decomposition, sorption and transport processes. Such vertical trends in 13C and 15N abundance have rarely been related to SOM composition during pedogenesis. The aims of our study were to investigate short and long-term δ13C and δ15N depth changes and their interrelations under progressing pedogenesis and ecosystem development. We sampled soils across the well studied fordune progradation Haast-chronosequence, a dune ridge system under super-humid climate at the West Coast of New Zealand's South Island (43° 53' S, 169° 3' E). Soils from 11 sites with five replicates each covered a time span of around 2870 yr of soil development (from Arenosol to Podzol). Vertical changes of δ13C and δ15N values of SOM were investigated in the organic layers and in 1-cm depth intervals of the upper 10 cm of the mineral soil. With increasing soil depth SOM became enriched in δ13C by 1.9 ± SE 0.1 o and in δ15N by 6.0 ± 0.4 ‰˙Litter δ13C values slightly decreased with increasing soil age (r = -0.61; p = 0.00) likely due to less efficient assimilation linked to nutrient limitations. Fractionation processes during mycorrhizal transfer appeared to affect δ15N values in the litter. We found a strong decrease of δ15N in the early succession stages ≤ 300 yr B.P. (r = -0.95; p = 0.00). Positive relations of vertical 13C and 15N enrichment with soil age might be related to decomposition and appeared to be

  18. Macroalgae δ15N values in well-mixed estuaries: Indicator of anthropogenic nitrogen input or macroalgae metabolism?

    NASA Astrophysics Data System (ADS)

    Raimonet, Mélanie; Guillou, Gaël; Mornet, Françoise; Richard, Pierre

    2013-03-01

    Although nitrogen stable isotope ratio (δ15N) in macroalgae is widely used as a bioindicator of anthropogenic nitrogen inputs to the coastal zone, recent studies suggest the possible role of macroalgae metabolism in δ15N variability. Simultaneous determinations of δ15N of dissolved inorganic nitrogen (DIN) along the land-sea continuum, inter-species variability of δ15N and its sensitivity to environmental factors are necessary to confirm the efficiency of macroalgae δ15N in monitoring nitrogen origin in mixed-use watersheds. In this study, δ15N of annual and perennial macroalgae (Ulva sp., Enteromorpha sp., Fucus vesiculosus and Fucus serratus) are compared to δ15N-DIN along the Charente Estuary, after characterizing δ15N of the three main DIN sources (i.e. cultivated area, pasture, sewage treatment plant outlet). During late winter and spring, when human activities produce high DIN inputs, DIN sources exhibit distinct δ15N signals in nitrate (NO) and ammonium (NH): cultivated area (+6.5 ± 0.6‰ and +9.0 ± 11.0‰), pasture (+9.2 ± 1.8‰ and +12.4‰) and sewage treatment plant discharge (+16.9 ± 8.7‰ and +25.4 ± 5.9‰). While sources show distinct δN- in this multiple source catchment, the overall mixture of NO sources - generally >95% DIN - leads to low variations of δN-NO at the mouth of the estuary (+7.7 to +8.4‰). Even if estuarine δN-NO values are not significantly different from pristine continental and oceanic site (+7.3‰ and +7.4‰), macroalgae δ15N values are generally higher at the mouth of the estuary. This highlights high anthropogenic DIN inputs in the estuary, and enhanced contribution of 15N-depleted NH in oceanic waters. Although seasonal variations in δN-NO are low, the same temporal trends in macroalgae δ15N values at estuarine and oceanic sites, and inter-species differences in δ15N values, suggest that macroalgae δ15N values might be modified by the metabolic response of macroalgae to environmental parameters (e

  19. Afforestation impacts microbial biomass and its natural (13)C and (15)N abundance in soil aggregates in central China.

    PubMed

    Wu, Junjun; Zhang, Qian; Yang, Fan; Lei, Yao; Zhang, Quanfa; Cheng, Xiaoli

    2016-10-15

    We investigated soil microbial biomass and its natural abundance of δ(13)C and δ(15)N in aggregates (>2000μm, 250-2000μm, 53-250μm and <53μm) of afforested (implementing woodland and shrubland plantations) soils, adjacent croplands and open area (i.e., control) in the Danjiangkou Reservoir area of central China. The afforested soils averaged higher microbial biomass carbon (MBC) and nitrogen (MBN) levels in all aggregates than in open area and cropland, with higher microbial biomass in micro-aggregates (<250μm) than in macro-aggregates (>2000μm). The δ(13)C of soil microbial biomass was more enriched in woodland soils than in other land use types, while δ(15)N of soil microbial biomass was more enriched compared with that of organic soil in all land use types. The δ(13)C and δ(15)N of microbial biomass were positively correlated with the δ(13)C and δ(15)N of organic soil across aggregates and land use types, whereas the (13)C and (15)N enrichment of microbial biomass exhibited linear decreases with the corresponding C:N ratio of organic soil. Our results suggest that shifts in the natural (13)C and (15)N abundance of microbial biomass reflect changes in the stabilization and turnover of soil organic matter (SOM) and thereby imply that afforestation can greatly impact SOM accumulation over the long-term.

  20. Insights into Nitrogen Isotopic Fractionation During Algal Assimilation of Nitrate and Ammonium

    NASA Astrophysics Data System (ADS)

    Evans, S. L.; Swart, P. K.; Capo, T. R.

    2008-12-01

    Nitrogen availability is an important factor controlling algal growth in marine environments, representing a limiting nutrient throughout much of the global ocean. Anthropogenic inputs to the coastal zone may shift the nutrient regime, leading to questions regarding the extent of anthropogenic nutrient impacts in near-shore environments. A large body of work has been completed relating the δ15N of algae, seagrasses, and other benthic organisms to anthropogenic nutrient sources. However, previous work by our research group characterizing the δ15N of organic material associated with waste water discharge points, and in reef and embayment environments of the south Florida coastal zone, has suggested that δ15N values alone do not provide unequivocal evidence of anthropogenic nitrogen loading. Greater understanding of nitrogen processing and isotopic fractionation in coastal benthic organisms is necessary before blanket assumptions regarding nutrient uptake and source association can be universally accepted. Closed system mesocosm incubations examining fractionation associated with assimilation of nitrate and ammonium in cultured red algae, Gracilaria sp. and Agardhiella sp., were completed under varied nitrate and ammonium concentrations from 10 to 500 μM with initial nitrogen isotopic compositions of 2.7-3 ‰. Following 8-day incubations, the isotopic composition of new algal growth ranged between +2.43 and -5.77 ‰, with more depleted values coincident with higher N-availability. Rayleigh fractionation calculations yield fractionation factors of 4-9 ‰ (α values of 1.0045 to 1.008), which represent significantly larger values than those previously reported in the literature for macroalgae. 15N-tracer experiments (initial δ15N = 1000 ‰) were also conducted to assess nutrient preferences in the cultured algae. Isotopic composition of new algal growth varied from -1.3 to +495.0 ‰ with only Agardhiella exhibiting an obvious preference for ammonium

  1. Sewage derive [sup 15]N in the Baltic traced in fucus

    SciTech Connect

    Hobbie, J.E.; Fry, B. ); Larsson, U.; Elmgren, R. )

    1990-01-09

    Himmerfjarden, a fjord-like bay on the eastern shore of the Baltic, receives treated sewage from 250,000 inhabitants. Because the inorganic N in the effluent is enriched in [sup 15]N through denitrification, nitrification, and ammonia volatilization, an analysis of the distribution of [sup 15]N in the Bay tells how far from the source the sewage nitrogen moves. The attached macroalga Fucus vesiculosus was collected in early May from rocky shore at 0-0.5 m depth and the [sup 15]N content of the tips of the fronds analyzed. This N represents uptake and storage during the previous six months and growth during March and April. The [delta][sup 15]N was uniformly high (11-13[per thousand]) in the main body of the Bay within 15 km from the sewage source. Beyond 15 km values decreased with distance to a low of 4.6[per thousand] at 35 km, where the Bay ends and the coastal waters begin. Using the 11-13 and 4.6[per thousand] as endmembers, the percentage of sewage N making up the Fucus at any point may be calculated. The [delta][sub 15]N of particulate organic matter in the offshore Baltic waters was around 0[per thousand] and Fucus had an [delta][sup 15]N about 1.5[per thousand] higher than the POM. From this and other evidence we conclude that there is a belt of coastal water with an elevated [delta][sup 15]N lying along the east coast of the Baltic. This presumably derives from sewage and perhaps from agriculture and is potentially of use as a tracer of coastal zone/pelagic zone interactions.

  2. δ15N natural abundance in permafrost soil indicates impact of fire on nitrogen cycle.

    PubMed

    Conen, Franz; Yakutin, Mikhail V; Puchnin, Alexander N; Leifeld, Jens; Alewell, Christine

    2011-03-15

    The impact of fire on the nitrogen (N) cycle of natural ecosystems is arguable. Here we report and interpret an observation from boreal ecosystems in the Lena River basin, Sakha Republic (Yakutia), Russian Federation. Different types of permafrost soil (0-30 cm depth) were sampled along transects (60-150 m length) from the forest edge towards the centre of four separate thermokarst depressions under grassland. The average values of δ(15)N were remarkably similar within three transects, but differed systematically between them. Three findings point towards fire being the cause of the observed pattern. First, the spatial extent of systematic differences in soil δ(15)N coincides with the extent of typical fire scars in the region. Second, soil enrichment in (15)N is larger in the proximity of settlements, where fire is generally more frequent than in more remote places. Third, there is a significant positive correlation between δ(15)N values and the ratio of black C to total N. These findings point towards fire having a marked impact on soil δ(15)N and, accordingly, on the N cycle of this cold and dry ecosystem. PMID:21290453

  3. Isotopic analysis of bulk, LMW, and HMW DON d15N indicates recycled nitrogen release from marine DON

    NASA Astrophysics Data System (ADS)

    Knapp, A. N.; Sigman, D. M.; Lipschultz, F.; Kustka, A.; Capone, D. G.

    2010-12-01

    Nitrogen (N) concentration and stable isotope ratio (d15N) measurements were made on bulk and size fractionated surface ocean dissolved organic nitrogen (DON) samples collected in the oligotrophic North Atlantic and Pacific Oceans. The bulk DON concentration in the upper 100 m is similar between the North Atlantic and North Pacific, between 4.5 and 5.0 uM, but the average d15N of bulk DON is significantly different, 3.9 per mil vs. air in the North Atlantic and 4.7 per mil in the North Pacific. The d15N of both bulk and HMW DON from the western tropical North Atlantic are similar to previous measurements, ~4.0 to 4.5 per mil. We report the first measurements of LMW DON d15N, which is consistently lower than HMW DON d15N. Neither the concentration nor d15N of bulk or size-fractionated DON varied with in situ N2 fixation rate, although significant variation in bulk and LMW DON d15N was observed between January and July of the same year in the western tropical North Atlantic. We propose a conceptual model to explain 1) the elevated d15N of bulk DON relative to other surface ocean N pools and fluxes, 2) the elevation of HMW DON d15N relative to LMW DON d15N, and 3) the inter-basin difference in the d15N of bulk DON. In this model, DON is produced from suspended particulate organic nitrogen (PON) without isotope fractionation because the conversion from PON to DON largely does not involve N-bearing bonds. In contrast, deamination and amide hydrolysis, with N isotope effects of 3 to 10 per mil, are major mechanisms by which DON is converted to ammonia and/or to other simple N compounds (e.g., amino acids). Thus these N-specific DON loss reactions result in an elevated d15N of residual DON relative to the parent DON and therefore also to the PON source. Moreover, the ammonium and simple organic N compounds released by microbial DON degradation are efficiently reassimilated back into the PON pool, as an integral part of the regenerated N cycle that further lowers the d15N

  4. δ15N patterns of Douglas-fir and red alder riparian forests in the Oregon Coast Range

    USGS Publications Warehouse

    Scott, E.E.; Perakis, S.S.; Hibbs, D.E.

    2008-01-01

    We used naturally occurring stable isotopes of N to compare N dynamics in near-stream and upslope environments along riparian catenas in N-fixing red alder (Alnus rubra) and Douglas-fir (Pseudotsuga menziesii) forests in the Coast Range of western Oregon. Based on the existing literature, we expected soil δ15N to be enriched closer to streams owing to inputs of isotopically heavy, marine-derived N by spawning salmon, higher rates of denitrification near the stream, or both. However, it has been unclear what effect red alder might have on soil δ15N patterns near streams. We found a consistent −1‰ δ15N signature in red alder foliage, and δ15N of total N in soils under red alder averaged 2.2‰ along sampling transects extending 20 m upslope from the stream. Surprisingly, δ15N of total N in soil under Douglas-fir was progressively depleted nearer to streams, opposite from the pattern expected from N losses by denitrification or N inputs from anadromous salmon. Instead, δ15N of total N in soil under Douglas-fir converged toward soil δ15N values typical of red alder sites. We consider that the historic presence of red alder may have contributed a legacy of lower soil δ15N nearer to streams on sites that are currently dominated by young Douglas-fir forest.

  5. Cryptic or day-to-day parts of the riverbed N cycle - new challenges for 15N

    NASA Astrophysics Data System (ADS)

    Trimmer, Mark; Ouyang, Liao; Lansdown, Katrina

    2016-04-01

    The discovery of anaerobic ammonium oxidation (anammox) not only changed our understanding of the nitrogen cycle in aquatic ecosystems but it also undermined some of the key 15N techniques used to study it. Reformulations of principle equations and the development of new 15N2 and 15N2O techniques enabled the simultaneous quantification of N2 production by anammox and denitrification in mainly soft, cohesive sediments where redox gradients are clearly defined and solute exchanged governed by diffusion. At the heart of the application of 15N, for the quantification of natural 14N cycling, is the key assumption that the respective pools of 15N and 14N are evenly mixed and that both are cycled without bias towards each other. Recent evidence, however, from a variety of aquatic ecosystems, suggests that this may not be the case. For example, organic N may be oxidised directly to N2 gas without ever mixing with the inorganic pool or inorganic intermediates (e.g. nitrite) are 'shunted' internally and also fail to mix evenly with the applied tracer pool. Our most recent work in permeable, oxic gravel riverbeds presents some particular challenges to the application of 15N. In these systems, a tight coupling between aerobic nitrification and anaerobic N2 production - in the presence of 100

  6. Intestinal renal metabolism of L-citrulline and L-arginine following enteral or parenteral infusion of L-alanyl-L-[2,15N]glutamine or L-[2,15N]glutamine in mice.

    PubMed

    Boelens, Petra G; van Leeuwen, Paul A M; Dejong, Cornelis H C; Deutz, Nicolaas E P

    2005-10-01

    Previously, we observed increased plasma arginine (ARG) concentrations after glutamine (GLN)-enriched diets, in combination with clinical benefits. GLN delivers nitrogen for ARG synthesis, and the present study was designed to quantify the interorgan relationship of exogenous L-GLN or GLN dipeptide, by enteral or parenteral route, contributing to intestinal citrulline (CIT) and renal de novo ARG synthesis in mice. To study this, we used a multicatheterized mouse model with Swiss mice (n = 43) in the postabsorptive state. Stable isotopes were infused into the jugular vein or into the duodenum {per group either free L-[2,(15)N]GLN or dipeptide L-ALA-L-[2,(15)N]GLN, all with L-[ureido-(13)C-(2)H(2)]CIT and L-[guanidino-(15)N(2)-(2)H(2)]ARG} to establish renal and intestinal ARG and CIT metabolism. Blood flow was measured using (14)C-para-aminohippuric acid. Net intestinal CIT release, renal uptake of CIT, and net renal ARG efflux was found, as assessed by arteriovenous flux measurements. Quantitatively, more de novo L-[2,(15)N]CIT was produced when free L-[2,(15)N]GLN was given than when L-ALA-L-[2,(15)N]GLN was given, whereas renal de novo L-[2,(15)N]ARG was similar in all groups. In conclusion, the intestinal-renal axis is hereby proven in mice in that L-[2,(15)N]GLN or dipeptide were both converted into de novo renal L-[2,(15)N]ARG; however, not all was derived from intestinal L-[2,(15)N]CIT production. In this model, the feeding route and form of GLN did not influence de novo renal ARG production derived from GLN.

  7. 15N chemical shift tensors and conformation of solid polypeptides containing 15N-labeled glycine residue by 15N NMR

    NASA Astrophysics Data System (ADS)

    Shoji, Akira; Ozaki, Takuo; Fujito, Teruaki; Deguchi, Kenzo; Ando, Isao; Magoshi, Jun

    1998-01-01

    The correlation between the isotropic 15N chemical shift ( δiso) and 15N chemical shift tensor components ( δ11, δ22 and δ33) and the main-chain conformation such as the polyglycine I (PGI: β-sheet), II (PGII: 3 1-helix), α-helix and β-sheet forms of solid polypeptides [Gly∗,X] n consisting of 15N-labeled glycine (Gly∗) and other amino acids (X: natural abundance of 15N) has been studied by solid-state 15N NMR method. A series of polypeptides [Gly∗,X] n (X = glycine, L-alanine, L-leucine, L-valine, L-isoleucine, β-benzyl L-aspartate, γ-benzyl L-glutamate, ɛ-carbobenzoxy L-lysine, and sarcosine) were synthesized by the α-amino acid N-carboxy anhydride (NCA) method. Conformations of these polypeptides in the solid state were characterized on the basis of conformation-dependent 13C chemical shifts in the 13C cross-polarization-magic angle spinning (CP-MAS) NMR spectra and by the characteristic bands in the IR and far-IR spectra. The δiso, δ11, δ22 and δ33 of the polypetides were determined from the 15N CP-MAS and 15N CP-static (powder pattern) spectra. It was found that the δiso, δ11, δ22 and δ33 in the PGI form (δ 83.5, 185, 40.7 and 25 ppm, resp.) are upfield from those in the PGII form (88.5, 194, 42.1 and 29 ppm, resp.), which were reproduced by the calculated 15N shielding constants using the finite perturbation theory (FPT)-INDO method. It was also found that the δ22 of the Gly∗ of [Gly∗,X] n is closely related to the main-chain conformation and the neighboring amino acid sequence, although the δiso is almost independent of the glycine content and conformation. Consequently, the δ22 value of Gly∗ containing copolypeptides is useful for the structural (main-chain conformation and neighboring amino acid sequence) analysis in the solid state by 15N NMR, if the 15N-labeled copolypeptide or natural protein can be provided. In addition, it is shown that the δiso of the glycine residue is useful for the conformational study of some

  8. Nitrate removal in two relict oxbow urban wetlands: a 15N mass-balance approach

    NASA Astrophysics Data System (ADS)

    Harrison, M. D.; Groffman, P. M.; Mayer, P. M.; Kaushal, S.

    2012-12-01

    A mass-balance approach was used to directly determine the flow of 15NO3- to plants, algae, and sediments,with unaccounted for 15N assumed to be denitrified. During the summer, plant and algal uptake accounted for 42%, of the added 15NO3 - in oxbow 1, less than 1% remained in the water column and 57% was unaccounted for. In oxbow 2 during the summer, plant and algal uptake accounted for 63% of the added 15NO3 -, with 1% remaining in the water column and 38% unaccounted for. During the early spring, plant and algal uptake were much lower in both oxbows, ranging from 0.05 to 13.3% of the 15N added, with 97 and 87% was unaccounted for in oxbow 1 and 2, respectively. The amount of unaccounted for 15N was equivalent to estimated areal denitrification rates of 12 and 6 mg N m-2 d-1 in the summer and 78 and 15 mg N m-2 d-1 in the spring, in oxbow 1 and oxbow 2, respectively. However, the uncertainty of these estimates is high as it was difficult to detect accumulation of 15N in the sediments which could have accounted for a very large percentage of the added 15N. Our results suggest that the two relict oxbow wetlands are sinks for NO3 - during both summer and spring. Plane view of Ox1 (A) and Ox2 (B) wetlands with closed contour intervals (color scale) and surrounding stream and upland elevations (labeled in black) located at Minebank Run, near Glen Arms, MD. 15N enrichment (atom %) of measured N pools prior to (Day 0) and after (Day 5) the end of the experiment in July 2009 and April 2010 for Ox1 and Ox2. Values are mean atom % (n = 2 algae, macrophytes and sediment; n = 6 for water samples).

  9. Determination of γ -ray widths in 15N using nuclear resonance fluorescence

    NASA Astrophysics Data System (ADS)

    Szücs, T.; Bemmerer, D.; Caciolli, A.; Fülöp, Zs.; Massarczyk, R.; Michelagnoli, C.; Reinhardt, T. P.; Schwengner, R.; Takács, M. P.; Ur, C. A.; Wagner, A.; Wagner, L.

    2015-07-01

    Background: The stable nucleus 15N is the mirror of 15O, the bottleneck in the hydrogen burning CNO cycle. Most of the 15N level widths below the proton emission threshold are known from just one nuclear resonance fluorescence (NRF) measurement, with limited precision in some cases. A recent experiment with the AGATA demonstrator array determined level lifetimes using the Doppler shift attenuation method in 15O. As a reference and for testing the method, level lifetimes in 15N have also been determined in the same experiment. Purpose: The latest compilation of 15N level properties dates back to 1991. The limited precision in some cases in the compilation calls for a new measurement to enable a comparison to the AGATA demonstrator data. The widths of several 15N levels have been studied with the NRF method. Method: The solid nitrogen compounds enriched in 15N have been irradiated with bremsstrahlung. The γ rays following the deexcitation of the excited nuclear levels were detected with four high-purity germanium detectors. Results: Integrated photon-scattering cross sections of 10 levels below the proton emission threshold have been measured. Partial γ -ray widths of ground-state transitions were deduced and compared to the literature. The photon-scattering cross sections of two levels above the proton emission threshold, but still below other particle emission energies have also been measured, and proton resonance strengths and proton widths were deduced. Conclusions: Gamma and proton widths consistent with the literature values were obtained, but with greatly improved precision.

  10. Potential probe for examining opiate-receptor interactions: model compound study of dynamic effects on /sup 15/N INEPT enhancements

    SciTech Connect

    Schilling, K.H.; Mikita, M.A.

    1987-10-01

    Model systems were chosen in an attempt to mimic the proton exchange environment of an agonist nitrogen in an opiate-receptor interaction. The two model systems studied were an ammonium: 18-crown-6 ether complex and a quinuclidine-trifluoroacetic acid ion pair. Each system was examined for their effects on /sup 15/N NMR INEPT enhancements. Both models were found to retard proton exchange dynamics, as observed by increased enhancements relative to free ions in neutral aqueous solutions. These results suggest that the confinement of a protonated nitrogen, such as that expected in receptor binding, may alter exchange dynamics to favor INEPT enhancements, while unbound agonists would remain unenhanced. As a result, /sup 15/N NMR INEPT enhancements from a solution of receptor subtypes with an appropriate /sup 15/N-labeled agonist may present a means of exploring the dynamics of direct opiate-receptor interactions.

  11. Community 15N isoscapes to resolve plant-plant-interactions at the spatial scale

    NASA Astrophysics Data System (ADS)

    Hellmann, Christine; Rascher, Katherine G.; Máguas, Cristina; Werner, Christiane

    2014-05-01

    Isoscapes have greatly improved our ability to understand biogeochemical processes on continental to global scales. However, the isoscapes framework may also have significant potential to resolve the spatial component of within-community interactions. For example, exotic plant invaders often exert strong impacts on ecosystem functioning, particularly regarding water-, carbon- and nutrient-cycles, but the spatial extent of such alterations is largely unknown. Here we show that massive N input by the N2-fixing exotic invasive Acacia longifolia to a Portuguese dune system can be traced using spatially resolved information on native plants' leaf δ15N. We found isotopic signatures of N to differ strongly between the native system (δ15N c. -10 o) and the atmospherically derived N in A. longifolia phyllodes (δ15N c. 0 o). Thus, sources of N for native plants could be readily distinguished. Leaf δ15N of a native, non-fixing species was increasingly enriched the closer the plant grew to the invader, indicating uptake of fixed N provided by A. longifolia. The enrichment was evident far beyond the stands of the invader, demonstrating that A. longifolia affected N budgets of native species up to a distance of 8 m exceeding the margin of the canopy. Furthermore, using the isoscapes approach, we were able to quantify the total area of N enrichment and could thus show that the area affected by invasion was at least 3.5 times larger than the area actually occupied by the invader. However, a native N2-fixing species had no such effects. Thus, downscaling isoscapes to the community level opens new frontiers in quantifying the spatial dimension of functional changes associated with plant invasions. Moreover, considering the feasibility and applicability of this approach, it may provide a promising tool to identify, quantify and monitor different types of functional plant-plant interactions within communities at a spatially explicit scale.

  12. Exploring the nitrogen ingestion of aphids--a new method using electrical penetration graph and (15)N labelling.

    PubMed

    Kuhlmann, Franziska; Opitz, Sebastian E W; Inselsbacher, Erich; Ganeteg, Ulrika; Näsholm, Torgny; Ninkovic, Velemir

    2013-01-01

    Studying plant-aphid interactions is challenging as aphid feeding is a complex process hidden in the plant tissue. Here we propose a combination of two well established methods to study nutrient acquisition by aphids focusing on the uptake of isotopically labelled nitrogen ((15)N). We combined the Electrical Penetration Graph (EPG) technique that allows detailed recording of aphid feeding behaviour and stable isotope ratio mass spectrometry (IRMS) to precisely measure the uptake of nitrogen. Bird cherry-oat aphids Rhopalosiphum padi L. (Hemiptera, Aphididae) fed for 24 h on barley plants (Hordeum vulgare L., cultivar Lina, Poaceae) that were cultivated with a (15)N enriched nutrient solution. The time aphids fed in the phloem was strongly positive correlated with their (15)N uptake. All other single behavioural phases were not correlated with (15)N enrichment in the aphids, which corroborates their classification as non-feeding EPG phases. In addition, phloem-feeding and (15)N enrichment of aphids was divided into two groups. One group spent only short time in the phloem phase and was unsuccessful in nitrogen acquisition, while the other group displayed longer phloem-feeding phases and was successful in nitrogen acquisition. This suggests that several factors such as the right feeding site, time span of feeding and individual conditions play a role for the aphids to acquire nutrients successfully. The power of this combination of methods for studying plant-aphid interactions is discussed.

  13. Variation in foliar [sup 15]N abundance and the availability of soil nitrogen on Walker Branch Watershed

    SciTech Connect

    Garten, C.T. Jr. )

    1993-10-01

    Spatial patterns in natural [sup 15]N abundance ([sigma][sup 15]N) in soil, soil solutions, and non-N[sub 2]-fixing plants were studied in the deciduous forest on Walker Branch Watershed near Oak Ridge, Tennessee. This study was undertaken to test the hypothesis that foliar [sigma][sup 15]N values are related to the availability of inorganic nitrogen in mineral soil. Soils collected in or near valley bottoms on the watershed had higher levels of net nitrogen mineralization and net nitrification potential than those sampled from ridges and slopes. More positive foliar [sigma][sup 15]N values occurred in valley bottoms, which, relative to other positions on the watershed, were characterized by greater availability of soil nitrogen and lower C-to-N ratios in the O[sub 1]-horizon, in the surface mineral soil, and in autumn leaf fall. Although leaf nitrogen concentrations changed significantly over the course of the growing season, there was little seasonal variation in foliar [sigma][sup 15]N values. A hypothesis about the relative importance of different sources of nitrogen to the forest and how nitrogen cycling varies with topography in this nitrogen-deficient ecosystem was derived, in part, from spatial patterns in natural [sup 15]N abundance. There appear to be two processes affecting the topographic patterns in foliar [sup 15]N abundance on this watershed: (1) greater uptake from isotopically heavy pools of inorganic soil nitrogen by plants in valley bottoms, and (2) uptake of isotopically light ammonium-N in atmospheric deposition by plants on ridges and slopes (where the availability of inorganic soil nitrogen to plant roots is more limited). Results from this study indicate that foliar [sigma][sup 15]N values are positively correlated with net nitrification potential in surface soil. 34 refs., 13 figs., 8 tabs.

  14. Sinks for nitrogen inputs in terrestrial ecosystems: a meta-analysis of 15N tracer field studies

    USGS Publications Warehouse

    Templer, P.H.; Mack, M.C.; Chapin, F. S.; Christenson, L.M.; Compton, J.E.; Crook, H.D.; Currie, W.S.; Curtis, C.J.; Dail, D.B.; D'Antonio, C. M.; Emmett, B.A.; Epstein, H.E.; Goodale, C.L.; Gundersen, P.; Hobbie, S.E.; Holland, K.; Hooper, D.U.; Hungate, B.A.; Lamontagne, S.; Nadelhoffer, K.J.; Osenberg, C.W.; Perakis, S.S.; Schleppi, P.; Schimel, J.; Schmidt, I.K.; Sommerkorn, M.; Spoelstra, J.; Tietema, A.; Wessel, W.W.; Zak, D.R.

    2012-01-01

    Effects of anthropogenic nitrogen (N) deposition and the ability of terrestrial ecosystems to store carbon (C) depend in part on the amount of N retained in the system and its partitioning among plant and soil pools. We conducted a meta-analysis of studies at 48 sites across four continents that used enriched 15N isotope tracers in order to synthesize information about total ecosystem N retention (i.e., total ecosystem 15N recovery in plant and soil pools) across natural systems and N partitioning among ecosystem pools. The greatest recoveries of ecosystem 15N tracer occurred in shrublands (mean, 89.5%) and wetlands (84.8%) followed by forests (74.9%) and grasslands (51.8%). In the short term (15N tracer application), total ecosystem 15N recovery was negatively correlated with fine-root and soil 15N natural abundance, and organic soil C and N concentration but was positively correlated with mean annual temperature and mineral soil C:N. In the longer term (3–18 months after 15N tracer application), total ecosystem 15N retention was negatively correlated with foliar natural-abundance 15N but was positively correlated with mineral soil C and N concentration and C: N, showing that plant and soil natural-abundance 15N and soil C:N are good indicators of total ecosystem N retention. Foliar N concentration was not significantly related to ecosystem 15N tracer recovery, suggesting that plant N status is not a good predictor of total ecosystem N retention. Because the largest ecosystem sinks for 15N tracer were below ground in forests, shrublands, and grasslands, we conclude that growth enhancement and potential for increased C storage in aboveground biomass from atmospheric N deposition is likely to be modest in these ecosystems. Total ecosystem 15N recovery decreased with N fertilization, with an apparent threshold fertilization rate of 46 kg N·ha-1·yr-1 above which most ecosystems showed net losses of applied 15N tracer in response to N fertilizer addition.

  15. Sinks for nitrogen inputs in terrestrial ecosystems: a meta-analysis of 15N tracer field studies.

    PubMed

    Templer, P H; Mack, M C; Chapin, F S; Christenson, L M; Compton, J E; Crook, H D; Currie, W S; Curtis, C J; Dail, D B; D'Antonio, C M; Emmett, B A; Epstein, H E; Goodale, C L; Gundersen, P; Hobbie, S E; Holland, K; Hooper, D U; Hungate, B A; Lamontagne, S; Nadelhoffer, K J; Osenberg, C W; Perakis, S S; Schleppi, P; Schimel, J; Schmidt, I K; Sommerkorn, M; Spoelstra, J; Tietema, A; Wessel, W W; Zak, D R

    2012-08-01

    Effects of anthropogenic nitrogen (N) deposition and the ability of terrestrial ecosystems to store carbon (C) depend in part on the amount of N retained in the system and its partitioning among plant and soil pools. We conducted a meta-analysis of studies at 48 sites across four continents that used enriched 15N isotope tracers in order to synthesize information about total ecosystem N retention (i.e., total ecosystem 15N recovery in plant and soil pools) across natural systems and N partitioning among ecosystem pools. The greatest recoveries of ecosystem 15N tracer occurred in shrublands (mean, 89.5%) and wetlands (84.8%) followed by forests (74.9%) and grasslands (51.8%). In the short term (< 1 week after 15N tracer application), total ecosystem 15N recovery was negatively correlated with fine-root and soil 15N natural abundance, and organic soil C and N concentration but was positively correlated with mean annual temperature and mineral soil C:N. In the longer term (3-18 months after 15N tracer application), total ecosystem 15N retention was negatively correlated with foliar natural-abundance 15N but was positively correlated with mineral soil C and N concentration and C:N, showing that plant and soil natural-abundance 15N and soil C:N are good indicators of total ecosystem N retention. Foliar N concentration was not significantly related to ecosystem 15N tracer recovery, suggesting that plant N status is not a good predictor of total ecosystem N retention. Because the largest ecosystem sinks for 15N tracer were below ground in forests, shrublands, and grasslands, we conclude that growth enhancement and potential for increased C storage in aboveground biomass from atmospheric N deposition is likely to be modest in these ecosystems. Total ecosystem 15N recovery decreased with N fertilization, with an apparent threshold fertilization rate of 46 kg N x ha(-1) x yr(-1) above which most ecosystems showed net losses of applied 15N tracer in response to N fertilizer

  16. Importance of Nitrate Attenuation In A Small Wetland Following Forest Harvest: 18O/16O, 15N/14N in nitrate and 15N/14N) in vegetation

    NASA Astrophysics Data System (ADS)

    Spoelstra, J.; Schiff, S. L.; Semkin, R. G.; Jeffries, D. S.; Elgood, R. J.

    2004-05-01

    Forest harvest can result in elevated nitrate concentrations in streams and groundwater affecting forest regeneration and downstream aquatic ecosystems. Turkey Lakes Watershed, located near Sault Ste Marie, Ontario (TLW), exhibits relatively high nitrate export due to naturally high rates of nitrification. During a forest harvest experiment at the TLW, stable isotope techniques were used to investigate nitrate attenuation in an intermediate position natural wetland receiving high concentrations of nitrate following forest clear-cutting. Isotopic analysis of nitrate (18O/16O, 15N/14N) and vegetation (15N/14N) demonstrated that denitrification and plant uptake of nitrate resulted in significantly lower nitrate concentrations in wetland outflow compared to incoming stream water and groundwater. The 0.2-hectare forested swamp, too small to show up on standard topographic maps, retained 65 to 100 percent of upgradient nitrate inputs, elevated due to increased nitrification in soils. The 15N/14N enrichment factor associated with nitrate attenuation in wetland surface water was lower than observed during denitrification in groundwaters, suggesting that denitrification proceeded to completion in some areas of the wetland. Even small, shallow, carbon rich pockets of organic matter in topographic depressions can significantly affect biogeochemical fluxes of C, N, S and Ca. Future forest management practices designed to recognize and preserve small wetlands could significantly reduce the potentially detrimental effects of forest harvest on aquatic systems.

  17. Cereal grain, rachis and pulse seed amino acid δ15N values as indicators of plant nitrogen metabolism.

    PubMed

    Styring, Amy K; Fraser, Rebecca A; Bogaard, Amy; Evershed, Richard P

    2014-01-01

    Natural abundance δ(15)N values of plant tissue amino acids (AAs) reflect the cycling of N into and within plants, providing an opportunity to better understand environmental and anthropogenic effects on plant metabolism. In this study, the AA δ(15)N values of barley (Hordeum vulgare) and bread wheat (Triticum aestivum) grains and rachis and broad bean (Vicia faba) and pea (Pisum sativum) seeds, grown at the experimental farm stations of Rothamsted, UK and Bad Lauchstädt, Germany, were determined by GC-C-IRMS. It was found that the δ(15)N values of cereal grain and rachis AAs could be largely attributed to metabolic pathways involved in their biosynthesis and catabolism. The relative (15)N-enrichment of phenylalanine can be attributed to its involvement in the phenylpropanoid pathway and glutamate has a δ(15)N value which is an average of the other AAs due to its central role in AA-N cycling. The relative AA δ(15)N values of broad bean and pea seeds were very different from one another, providing evidence for differences in the metabolic routing of AAs to the developing seeds in these leguminous plants. This study has shown that AA δ(15)N values relate to known AA biosynthetic pathways in plants and thus have the potential to aid understanding of how various external factors, such as source of assimilated N, influence metabolic cycling of N within plants.

  18. Compound-specific δ15N and chlorin preservation in surface sediments of the Peru Margin with implications for ancient bulk δ15N records

    NASA Astrophysics Data System (ADS)

    Junium, Christopher K.; Arthur, Michael A.; Freeman, Katherine H.

    2015-07-01

    Understanding the processes that control the preservation of paleoceanographic proxies is of clear importance. Surface sediments from the Peru Margin oxygen-minimum zone are subject to lateral and downslope transport by bottom currents that decrease organic matter (OM) quality. Indicators of bulk OM quality (pyrolysis hydrogen index, pyrolysis S1 + S2 and C/N) demonstrate significant degradation between 150 and 400 m water depth, within the oxygen-minimum zone. Concentrations of the three most abundant chlorins (chlorophyllone, pheophytin and pyropheophytin) decrease from 750 to 150 nmol g TOC-1 from 150 to 400 m water depth though the relative abundances of the chlorins in an individual sample do not change. This suggests that the three chlorins have similar reactivity over the ambient conditions. Values for δ15N of bulk sediments (δ15Nbulk) decrease by 3‰ from the inner shelf to the upper slope (1000 m) but co-occurring compound-specific δ15N values (δ15Nchlorin) do not decrease downslope. The low variability of δ15Nchlorin values supports a single source for the chlorins, and demonstrates the recalcitrance of δ15Nchlorin values despite degradation. This set of observation raises questions about which type of OM fraction best records 'primary' signatures. We assess two possible models to guide our interpretation of these disparate datasets (1) that decreasing δ15Nbulk values are the result of degradation of a 15N-enriched fraction during downslope transport, and that δ15Nchlorin values reflect primary values; (2) that δ15Nbulk values are primary and that chlorins are derived from material transported from upslope. These data reaffirm that in active sedimentary environments such as the Eastern Tropical Pacific, transport of OM can significantly alter bulk geochemical parameters of OM integrity, but the impacts on the δ15N record of bulk sediments and chlorins are less clear, and require more study to be thoroughly understood.

  19. COMPARISON OF STABLE-NITROGEN (15N/14N) ISOTOPE RATIOS IN LARGE MOUTH BASS SCALES AND MUSCLE TISSUE

    EPA Science Inventory

    Stable-nitrogen (15N/14N) isotope ratios of fish tissue are currently used to determine trophic structure, contaminant bioaccumulation, and the level of anthropogenic nitrogen enrichment in aquatic systems. The most common tissue used for these measurements is fileted dorsal musc...

  20. (15)N Heteronuclear Chemical Exchange Saturation Transfer MRI.

    PubMed

    Zeng, Haifeng; Xu, Jiadi; Yadav, Nirbhay N; McMahon, Michael T; Harden, Bradley; Frueh, Dominique; van Zijl, Peter C M

    2016-09-01

    A two-step heteronuclear enhancement approach was combined with chemical exchange saturation transfer (CEST) to magnify (15)N MRI signal of molecules through indirect detection via water protons. Previous CEST studies have been limited to radiofrequency (rf) saturation transfer or excitation transfer employing protons. Here, the signal of (15)N is detected indirectly through the water signal by first inverting selectively protons that are scalar-coupled to (15)N in the urea molecule, followed by chemical exchange of the amide proton to bulk water. In addition to providing a small sensitivity enhancement, this approach can be used to monitor the exchange rates and thus the pH sensitivity of the participating (15)N-bound protons. PMID:27548755

  1. δ15N in the turtle grass from the Mexican Caribbean

    NASA Astrophysics Data System (ADS)

    Talavera-Saenz, A.; Sanchez, A.; Ortiz-Hernandez, M.

    2013-05-01

    Nutrient inputs associated with population growth threaten the integrity of coastal ecosystems. To assess the rapid increase in tourism, we compared the δ15N from Thalassia testudinum collected at sites with different levels of tourism development and population to detect the N inputs of wastewater discharge (WD) along the coast of Quintana Roo. The contributions of nitrogen enriched in 15N are directly related to the increase of WD inputs in areas of high tourism development (Nichupte Lagoon in Cancun, >3 million tourists per year from 2007 to 2011 and 0.7 million of resident population) and decreased towards Bahia Akumal and Tulum (>3 million tourists per year from 2007 to 2011 and 0.15 million of resident population). The δ15N from T. testudinum was significantly lower at Mahahual and Puerto Morelos (about 0.4 million tourists per year in 2007 to 2011 and 0.25 million of resident population) than other the sites. In areas of the lowest development and with tourist activity restricted and small population, such as the Yum Balam Reserve and Sian Ka'an Biosphere Reserve, the δ15N values were in much higher enrichment that Mahahual and Puerto Morelos. Therefore is suggested that Mahahual and Puerto Morelos may be used for baseline isotopic monitoring, over environmental pressure on the reef lagoon ecosystem, where tourist activities and population are growing very slow rate. The anthropogenic N input has the potential to impact, both environmentally and economically, the seagrass meadows and the coral reefs along the coast of Quintana Roo and the Caribbean.

  2. In vivo uniform (15)N-isotope labelling of plants: using the greenhouse for structural proteomics.

    PubMed

    Ippel, Johannes H; Pouvreau, Laurice; Kroef, Toos; Gruppen, Harry; Versteeg, Geurt; van den Putten, Peter; Struik, Paul C; van Mierlo, Carlo P M

    2004-01-01

    Isotope labelling of proteins is important for progress in the field of structural proteomics. It enables the utilisation of the power of nuclear magnetic resonance spectroscopy (NMR) for the characterisation of the three-dimensional structures and corresponding dynamical features of proteins. The usual approach to obtain isotopically labelled protein molecules is by expressing the corresponding gene in bacterial or yeast host organisms, which grow on isotope-enriched media. This method has several drawbacks. Here, we demonstrate that it is possible to fully label a plant with (15)N-isotopes. The advantage of in vivo labelling of higher organisms is that all constituting proteins are labelled and become available as functional, post-translationally modified, correctly folded proteins. A hydroponics set-up was used to create the first example of a uniformly (15)N-labelled (> 98%) plant species, the potato plant (Solanum tuberosum L., cv. Elkana). Two plants were grown at low costs using potassium-[(15)N]-nitrate as the sole nitrogen source. At harvest time, a total of 3.6 kg of potato tubers and 1.6 kg of foliage, stolons and roots were collected, all of which were fully (15)N-labelled. Gram quantities of soluble (15)N-labelled proteins (composed mainly of the glycoprotein patatin and Kunitz-type protease inhibitors) were isolated from the tubers. NMR results on the complete proteome of potato sap and on an isolated protease inhibitor illustrate the success of the labelling procedure. The presented method of isotope labelling is easily modified to label other plants. Its envisioned impact in the field of structural proteomics of plants is discussed.

  3. Review: Mechanisms of ammonium toxicity and the quest for tolerance.

    PubMed

    Esteban, Raquel; Ariz, Idoia; Cruz, Cristina; Moran, Jose Fernando

    2016-07-01

    Ammonium sensitivity of plants is a worldwide problem, constraining crop production. Prolonged application of ammonium as the sole nitrogen source may result in physiological and morphological disorders that lead to decreased plant growth and toxicity. The main causes of ammonium toxicity/tolerance described until now include high ammonium assimilation by plants and/or low sensitivity to external pH acidification. The various ammonium transport-related components, especially the non-electrogenic influx of NH3 (related to the depletion of (15)N) and the electrogenic influx of NH4(+), may contribute to ammonium accumulation, and therefore to NH3 toxicity. However, this accumulation may be influenced by increasing K(+) concentration in the root medium. Recently, new insights have been provided by "omics" studies, leading to a suggested involvement of GDP mannose-pyrophosphorylase in the response pathways of NH4(+) stress. In this review, we highlight the cross-talk signaling between nitrate, auxins and NO, and the importance of the connection of the plants' urea cycle to metabolism of polyamines. Overall, the tolerance and amelioration of ammonium toxicity are outlined to improve the yield of ammonium-grown plants. This review identifies future directions of research, focusing on the putative importance of aquaporins in ammonium influx, and on genes involved in ammonium sensitivity and tolerance. PMID:27181951

  4. Sub-cellular localisation of a 15N-labelled peptide vector using NanoSIMS imaging

    NASA Astrophysics Data System (ADS)

    Römer, Winfried; Wu, Ting-Di; Duchambon, Patricia; Amessou, Mohamed; Carrez, Danièle; Johannes, Ludger; Guerquin-Kern, Jean-Luc

    2006-07-01

    Dynamic SIMS imaging is proposed to map sub-cellular distributions of isotopically labelled, exogenous compounds. NanoSIMS imaging allows the characterisation of the intracellular transport pathways of exogenous molecules, including peptide vectors employed in innovative therapies, using stable isotopes as molecular markers to detect the compound of interest. Shiga toxin B-subunit (STxB) was chosen as a representative peptide vector. The recombinant protein ( 15N-STxB) was synthesised in Escherichia coli using 15NH 4Cl as sole nitrogen source resulting in 15N enrichment in the molecule. Using the NanoSIMS 50 ion microprobe (Cameca), different ion species ( 12C 14N -, 12C 15N -, 31P -) originating from the same sputtered micro volume were simultaneously detected. High mass resolving power enabled the discrimination of 12C 15N - from its polyatomic isobars of mass 27. We imaged the membrane binding and internalisation of 15N-STxB in HeLa cells at spatial resolutions of less than 100 nm. Thus, the use of rare stable isotopes like 15N with dynamic SIMS imaging permits sub-cellular detection of isotopically labelled, exogenous molecules and imaging of their transport pathways at high mass and spatial resolution. Application of stable isotopes as markers can replace the large and chemically complex tags used for fluorescence microscopy, without altering the chemical and physical properties of the molecule.

  5. Phenylalanine δ15N in Paleo Archives as a New Proxy for δ15N of Exported Primary Production

    NASA Astrophysics Data System (ADS)

    McCarthy, M.; Batista, F. C.; Vokhshoori, N. L.; Brown, J. T.; Guilderson, T. P.; Ravelo, A. C.; Sherwood, O.

    2012-12-01

    Compound-specific isotope analysis of individual amino acids (CSI-AA) is emerging as a powerful new tool for studying the paleo nitrogen cycle. Because most detrital organic nitrogen is composed of amino acids, CSI-AA can reveal the mechanistic basis for organic nitrogen diagenesis, preserve a record of past food web structure, and potentially reconstruct the δ15N values of past nitrate and primary production. Within the commonly measured amino acids, the δ15N value of phenylalanine (Phe) appears uniquely promising as a new proxy that reflects the nitrogen isotopic value of the original source. Phe δ15N values remain almost unchanged with trophic transfer through food webs, and also during at least the initial stages of organic matter degradation. Here we synthesize results from both bio-archives and recent sediments, which together suggest that at least in Holocene archives the Phe δ15N value does in fact record the average inorganic nitrogen δ15N value at the base of planktonic food webs. However, several important unknowns also remain. These include the extent of variation in amino acid isotopic fractionation patterns in phylogenetically distinct algal groups. The stability of Phe δ15N values in older sediments where organic matter has undergone extensive diagenesis is also an important research area, which may ultimately establish the temporal limit for application of this approach to study past geological epochs. Together, however, results to date suggest that of Phe δ15N values in paleo archives represent a novel molecular-level proxy which is not tied to any specific organism or group, but rather can provide an integrated estimate of δ15N value of exported primary production.

  6. 15N Hyperpolarization by Reversible Exchange Using SABRE-SHEATH

    PubMed Central

    2016-01-01

    NMR signal amplification by reversible exchange (SABRE) is a NMR hyperpolarization technique that enables nuclear spin polarization enhancement of molecules via concurrent chemical exchange of a target substrate and parahydrogen (the source of spin order) on an iridium catalyst. Recently, we demonstrated that conducting SABRE in microtesla fields provided by a magnetic shield enables up to 10% 15N-polarization (Theis, T.; et al. J. Am. Chem. Soc.2015, 137, 1404). Hyperpolarization on 15N (and heteronuclei in general) may be advantageous because of the long-lived nature of the hyperpolarization on 15N relative to the short-lived hyperpolarization of protons conventionally hyperpolarized by SABRE, in addition to wider chemical shift dispersion and absence of background signal. Here we show that these unprecedented polarization levels enable 15N magnetic resonance imaging. We also present a theoretical model for the hyperpolarization transfer to heteronuclei, and detail key parameters that should be optimized for efficient 15N-hyperpolarization. The effects of parahydrogen pressure, flow rate, sample temperature, catalyst-to-substrate ratio, relaxation time (T1), and reversible oxygen quenching are studied on a test system of 15N-pyridine in methanol-d4. Moreover, we demonstrate the first proof-of-principle 13C-hyperpolarization using this method. This simple hyperpolarization scheme only requires access to parahydrogen and a magnetic shield, and it provides large enough signal gains to enable one of the first 15N images (2 × 2 mm2 resolution). Importantly, this method enables hyperpolarization of molecular sites with NMR T1 relaxation times suitable for biomedical imaging and spectroscopy. PMID:25960823

  7. Assimilation of ammonium and nitrate nitrogen by bean plants

    SciTech Connect

    Volk, R.J. ); Chaillou, S.; Morot-Gaudry, J.F. ); Mariotti, A. )

    1989-04-01

    Enhanced growth is often observed in plants growing on combined ammonium and nitrate nutrition. The physiological basis for such enhancement was examined by exposing non-nodulated bean (Phaseolus vulgaris L.) plants to {sup 15}N-labeled, 1.0 mM N solutions containing 0, 33, 67 or 100% of the N as ammonium, the balance being nitrate. Maximal total N uptake and biomass production were attained by plants receiving 33% ammonium. A higher proportion of incoming ammonium than nitrate was incorporated into root protein. This was accompanied by increased partitioning of plant biomass to roots. It was concluded that as a consequence of greater N metabolism in the root under mixed ammonium and nitrate nutrition, the root became a more active sink for photosynthate. Concurrently, the augmented supply of N to the shoot enhanced net photosynthesis as reflected in increased plant biomass.

  8. 15N2 Fixation by Bacteria Associated with Maize Roots at a Low Partial O2 Pressure

    PubMed Central

    Alexander, David B.; Zuberer, David A.

    1989-01-01

    Nitrogen fixation by bacteria associated with roots of intact maize plants was measured by exposing the roots to 15N2 at a partial O2 pressure (pO2) of 2 or 10 kPa. The plants were grown in a mixture of Weswood soil and sand and then transferred to plastic cylinders containing an N-free plant nutrient solution. The solution was sparged continuously with a mixture of air and N2 at a pO2 of 2 or 10 kPa. Acetylene reduction was measured after the roots were exposed to the low pO2 overnight. The air-N2 atmosphere in the cylinders was then replaced with an O2-He atmosphere at the same pO2, and the roots were exposed to 20 kPa of 15N2 for 20 to 22 h. Incorporation of 15N into the roots was 200 times greater at 2 kPa of O2 than at 10 kPa of O2. Adding l-malate (1 g of C liter−1) to the nutrient solution increased root-associated nitrogenase activity, producing a strong 15N label which could be traced into the shoots. Fixed 15N was detected in the shoots within 5 days after the plants were returned to unfertilized soil. In a similar experiment with undisturbed plants grown in fritted clay, movement of fixed 15N into the shoots was evident within 4 days after the roots were exposed to 15N2 at 2 kPa of O2. Inoculation with Azospirillum lipoferum yielded no significant differences in shoot dry weight, total nitrogen content, percent nitrogen, or 15N enrichment of plant tissues. Inoculated plants did exhibit greater root dry weight than uninoculated plants, however. PMID:16347968

  9. Leaf δ15N as a temporal integrator of nitrogen-cycling processes at the Mojave Desert FACE experiment

    NASA Astrophysics Data System (ADS)

    Sonderegger, D.; Koyama, A.; Jin, V.; Billings, S. A.; Ogle, K.; Evans, R. D.

    2011-12-01

    Ecosystem response to elevated carbon dioxide (CO2) in arid environments is regulated primarily by water, which may interact with nitrogen availability. Leaf nitrogen isotope composition (δ15N) can serve as an important indicator of changes in nitrogen dynamics by integrating changes in plant physiology and ecosystem biogeochemical processes. Because of this temporal integration, careful modeling of the antecedent conditions is necessary for understanding the processes driving variation in leaf δ15N. We measured leaf δ15N of Larrea tridentata (creosotebush) over the 10-year lifetime of the Nevada Desert Free-Air CO2 Enrichment (FACE) experiment. Leaf δ15N exhibited two patterns. First, elevated atmospheric CO2 significantly increased Larrea leaf δ15N by approximately 2 to 3 % compared to plants exposed to ambient CO2 concentrations Second, plants in both CO2 treatments exhibited significant seasonal cycles in leaf δ15N, with higher values during the fall and winter seasons. We modeled leaf δ15N using a hierarchical Bayesian framework that incorporated soil moisture, temperature, and the Palmer Drought Severity Index (PDSI) covariates in addition to a CO2 treatment effect and plot random effects. Antecedent moisture effects were modeled by using a combination of the previous season's aggregated conditions and a smoothly varying weighted average of the months or weeks directly preceding the observation. The time lag between the driving antecedent condition and the observed change in leaf δ15N indicates a significant and unobserved process mechanism. Preliminary results suggest a CO2 treatment interaction with the lag effect, indicating a treatment effect on the latent process.

  10. Novel labeling technique illustrates transfer of 15N2 from Sphagnum moss to vascular plants via diazotrophic nitrogen fixation

    NASA Astrophysics Data System (ADS)

    Thorp, N. R.; Vile, M. A.; Wieder, R.

    2013-12-01

    transferred to P. mariana roots (mean δ15N at 1 and 2 months of 15.26 × 3.30 and 16.19 × 1.21) more than shoots (mean δ15N at 1 and 2 months of 6.57 × 0.52 and 4.67×0.17) (initial δ15N values of roots and shoots of 2.16 × 0.37 and 5.54 × 0.35, respectively). Nitrogen also was transferred to V. oxycoccos roots (δ15N at 2 months of 21.46 × 3.61) more than shoots (δ15N 2 months of -2.17 × 0.23) (initial δ15N values of roots and shoots of -6.41 × 0.21 and -6.85 × 0.15, respectively). A two-way ANOVA and Tukey's HSD verified that both vascular plants' roots were significantly enriched with 15N (P. mariana roots; p < 0.0001, V. oxycoccus roots; p < 0.0001) after 1 month. These results indicate that bog vascular plants may derive considerable nitrogen from biological N2-fixation taking place in Sphagnum moss capitula. The experiment was subsequently repeated in-situ.

  11. Food webs in Mongolian grasslands: the analysis of 13C and 15N natural abundances.

    PubMed

    Kohzu, Ayato; Iwata, T; Kato, M; Nishikawa, J; Wada, Eitaro; Amartuvshin, N; Namkhaidorj, B; Fujita, N

    2009-09-01

    Overgrazing often lowers species richness and productivity of grassland communities. For Mongolian grassland ecosystems, a lack of detailed information about food-web structures makes it difficult to predict the effects of overgrazing on species diversity and community composition. We analysed the delta13C and delta15N signatures of herbaceous plants, arthropods (grouped by feeding habit), wild and domestic mammals, and humans in central Mongolia to understand the predominant food-web pathways in this grassland ecosystem. The delta13C and delta15N values of mammals showed little variation within species, but varied considerably with slope position for arthropods. The apparent isotopic discrimination between body tissue and hair of mammals was estimated as 2.0 per thousand for delta13C and 2.1 per thousand for delta15N, which was large enough to cause overestimation of the trophic level of mammals if not taken into account when using hair samples to measure isotopic enrichment. PMID:19507080

  12. Compound-specific 15N stable isotope probing of N assimilation by the soil microbial biomass: a new methodological paradigm in soil N cycling

    NASA Astrophysics Data System (ADS)

    Charteris, A. F.; Knowles, T. D. J.; Michaelides, K.; Evershed, R. P.

    2015-10-01

    A compound-specific nitrogen-15 stable isotope probing (15N-SIP) technique is described which allows investigation of the fate of inorganic- or organic-N amendments to soils. The technique uses gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) to determine the δ15N values of individual amino acids (AAs; determined as N-acetyl, O-isopropyl derivatives) as proxies of biomass protein production. The δ15N values are used together with AA concentrations to quantify N assimilation of 15N-labelled substrates by the soil microbial biomass. The utility of the approach is demonstrated through incubation experiments using inorganic 15N-labelled substrates ammonium (15NH4+) and nitrate (15NO3-) and an organic 15N-labelled substrate, glutamic acid (15N-Glu). Assimilation of all the applied substrates was undetectable based on bulk soil properties, i.e. % total N (% TN), bulk soil N isotope composition and AA concentrations, all of which remained relatively constant throughout the incubation experiments. In contrast, compound-specific AA δ15N values were highly sensitive to N assimilation, providing qualitative and quantitative insights into the cycling and fate of the applied 15N-labelled substrates. The utility of this 15N-AA-SIP technique is considered in relation to other currently available methods for investigating the microbially-mediated assimilation of nitrogenous substrates into the soil organic N pool. This approach will be generally applicable to the study of N cycling in any soil, or indeed, in any complex ecosystem.

  13. Paleoenvironmental implications of taxonomic variation among δ 15 N values of chloropigments

    NASA Astrophysics Data System (ADS)

    Higgins, Meytal B.; Wolfe-Simon, Felisa; Robinson, Rebecca S.; Qin, Yelun; Saito, Mak A.; Pearson, Ann

    2011-11-01

    Natural variations in the ratios of nitrogen isotopes in biomass reflect variations in nutrient sources utilized for growth. In order to use δ 15N values of chloropigments of photosynthetic organisms to determine the corresponding δ 15N values of biomass - and by extension, surface waters - the isotopic offset between chlorophyll and biomass must be constrained. Here we examine this offset in various geologically-relevant taxa, grown using nutrient sources that may approximate ocean conditions at different times in Earth's history. Phytoplankton in this study include cyanobacteria (diazotrophic and non-diazotrophic), eukaryotic algae (red and green), and anoxygenic photosynthetic bacteria (Proteobacteria), as well as environmental samples from sulfidic lake water. Cultures were grown using N 2, NO 3-, and NH 4+ as nitrogen sources, and were examined under different light regimes and growth conditions. We find surprisingly high variability in the isotopic difference (δ 15N biomass - δ 15N chloropigment) for prokaryotes, with average values for species ranging from -12.2‰ to +11.7‰. We define this difference as ɛpor, a term that encompasses diagenetic porphyrins and chlorins, as well as chlorophyll. Negative values of ɛpor reflect chloropigments that are 15N-enriched relative to biomass. Notably, this enrichment appears to occur only in cyanobacteria. The average value of ɛpor for freshwater cyanobacterial species is -9.8 ± 1.8‰, while for marine cyanobacteria it is -0.9 ± 1.3‰. These isotopic effects group environmentally but not phylogenetically, e.g., ɛpor values for freshwater Chroococcales resemble those of freshwater Nostocales but differ from those of marine Chroococcales. Our measured values of ɛpor for eukaryotic algae (range = 4.7-8.7‰) are similar to previous reports for pure cultures. For all taxa studied, values of ɛpor do not depend on the type of nitrogen substrate used for growth. The observed environmental control of

  14. Electron Shuttles Enhance Anaerobic Ammonium Oxidation Coupled to Iron(III) Reduction.

    PubMed

    Zhou, Guo-Wei; Yang, Xiao-Ru; Li, Hu; Marshall, Christopher W; Zheng, Bang-Xiao; Yan, Yu; Su, Jian-Qiang; Zhu, Yong-Guan

    2016-09-01

    Anaerobic ammonium oxidation coupled to iron(III) reduction, termed Feammox, is a newly discovered nitrogen cycling process. However, little is known about the roles of electron shuttles in the Feammox reactions. In this study, two forms of Fe(III) (oxyhydr)oxide ferrihydrite (ex situ ferrihydrite and in situ ferrihydrite) were used in dissimilatory Fe(III) reduction (DIR) enrichments from paddy soil. Evidence for Feammox in DIR enrichments was demonstrated using the (15)N-isotope tracing technique. The extent and rate of both the (30)N2-(29)N2 and Fe(II) formation were enhanced when amended with electron shuttles (either 9,10-anthraquinone-2,6-disulfonate (AQDS) or biochar) and further simulated when these two shuttling compounds were combined. Although the Feammox-associated Fe(III) reduction accounted for only a minor proportion of total Fe(II) formation compared to DIR, it was estimated that the potentially Feammox-mediated N loss (0.13-0.48 mg N L(-1) day(-1)) was increased by 17-340% in the enrichments by the addition of electron shuttles. The addition of electron shuttles led to an increase in the abundance of unclassified Pelobacteraceae, Desulfovibrio, and denitrifiers but a decrease in Geobacter. Overall, we demonstrated a stimulatory effect of electron shuttles on Feammox that led to higher N loss, suggesting that electron shuttles might play a crucial role in Feammox-mediated N loss from soils. PMID:27494694

  15. Electron Shuttles Enhance Anaerobic Ammonium Oxidation Coupled to Iron(III) Reduction.

    PubMed

    Zhou, Guo-Wei; Yang, Xiao-Ru; Li, Hu; Marshall, Christopher W; Zheng, Bang-Xiao; Yan, Yu; Su, Jian-Qiang; Zhu, Yong-Guan

    2016-09-01

    Anaerobic ammonium oxidation coupled to iron(III) reduction, termed Feammox, is a newly discovered nitrogen cycling process. However, little is known about the roles of electron shuttles in the Feammox reactions. In this study, two forms of Fe(III) (oxyhydr)oxide ferrihydrite (ex situ ferrihydrite and in situ ferrihydrite) were used in dissimilatory Fe(III) reduction (DIR) enrichments from paddy soil. Evidence for Feammox in DIR enrichments was demonstrated using the (15)N-isotope tracing technique. The extent and rate of both the (30)N2-(29)N2 and Fe(II) formation were enhanced when amended with electron shuttles (either 9,10-anthraquinone-2,6-disulfonate (AQDS) or biochar) and further simulated when these two shuttling compounds were combined. Although the Feammox-associated Fe(III) reduction accounted for only a minor proportion of total Fe(II) formation compared to DIR, it was estimated that the potentially Feammox-mediated N loss (0.13-0.48 mg N L(-1) day(-1)) was increased by 17-340% in the enrichments by the addition of electron shuttles. The addition of electron shuttles led to an increase in the abundance of unclassified Pelobacteraceae, Desulfovibrio, and denitrifiers but a decrease in Geobacter. Overall, we demonstrated a stimulatory effect of electron shuttles on Feammox that led to higher N loss, suggesting that electron shuttles might play a crucial role in Feammox-mediated N loss from soils.

  16. Selective observation of biologically important 15N-labeled metabolites in isolated rat brain and liver by 1H-detected multiple-quantum-coherence spectroscopy

    NASA Astrophysics Data System (ADS)

    Kanamori, Keiko; Ross, Brian D.; Parivar, Farhad

    Four cerebral metabolites of importance in neurotransmission, serotonin, L-tryptophan, L-glutamine, and N-acetyl- L-aspartate, and two hepatic urea-cycle intermediates, citrulline and urea, were found to be observable by 1H- 15N heteronuclear multiple-quantum-coherence (HMQC) spectroscopy in aqueous solution at physiological pH and temperature, through the protons spin-coupled to their indole, amide, or ureido nitrogen. Their 1H chemical shifts were well dispersed over a 5-10 ppm region while the 1J 15N- 1H values were 87-99 Hz. For [γ- 15N]glutamine, a 50- to 100-fold increase in sensitivity over direct 15N detection was achieved, in contrast to a 2-fold increase by the polarization-transfer method. In the isolated brain of portacaval-shunted rats, the amide protons of biologically 15N-enriched [γ- 15N]glutamine were observed in 2 min of acquisition, with suppression of proton signals from all other cerebral metabolites. In isolated liver of 15N-enriched control rats, [ 15NIurea protons were observed in 16 min. The HMQC method is likely to be effective for the in vivo study of cerebral and hepatic nitrogen metabolism.

  17. Foliar and fungal 15N:14N ratios reflect development of mycorrhizae and nitrogen supply during primary succession: testing analytical models.

    PubMed

    Hobbie, Erik A; Jumpponen, Ari; Trappe, Jim

    2005-12-01

    per thousand. Plants, fungi and soil were at least 4 per thousand higher in delta15N from the mature site than in recently exposed sites. On both the forefront and the mature site, host-specific ectomycorrhizal fungi had higher delta15N values than ectomycorrhizal fungi with a broad host range. From these isotopic patterns, we conclude: (1) large enrichments in 15N of many ectomycorrhizal fungi relative to co-occurring ectomycorrhizal plants are best explained by treating the plant-fungal-soil system as a closed system with a discrimination against 15N of 8-10 per thousand during transfer from fungi to plants, (2) based on models of 15N mass balance, ericoid and ectomycorrhizal fungi retain up to two-thirds of the N in the plant-mycorrhizal system under the N-limited conditions at forefront sites, (3) sporocarps are probably enriched in 15N by an additional 3 per thousand relative to available nitrogen, and (4) host-specific ectomycorrhizal fungi may transfer more N to plant hosts than non-host-specific ectomycorrhizal fungi. Our study confirms that nitrogen isotopes are a powerful tool for probing nitrogen dynamics between mycorrhizal fungi and associated plants.

  18. Quantification of (15)N-nitrate in urine with gas chromatography combustion isotope ratio mass spectrometry to estimate endogenous NO production.

    PubMed

    Houben, Els; Hamer, Henrike M; Luypaerts, Anja; De Preter, Vicky; Evenepoel, Pieter; Rutgeerts, Paul; Verbeke, Kristin

    2010-01-15

    The use of stable isotope labeled substrates and subsequent analysis of urinary nitrate, forms a noninvasive test for evaluation of the in vivo NO metabolism. The present paper describes a new method for simultaneous quantification of (15)N-nitrate and total nitrate with gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS). Nitrate, isolated from urine with a nitrate selective resin, was reduced to nitrite using copperized cadmium. Subsequently, Sudan I was formed by diazotation. Sudan II was added as internal standard, and both molecules were analyzed with GC-C-IRMS as tert-butyldimethylsilyl derivatives. The accuracy was determined during a recovery study of two different known nitrate concentrations and two (15)N-enrichments. A recovery of 101.6% and 103.9% for total nitrate and 107.6% and 91.2% for (15)N-nitrate was obtained, respectively. The validated method was applied on complete 72 h urine collections after intravenous administration of (15)N-nitrate and (15)N-arginine in humans. On average, 51.8% (47.0-71.0%) of administered (15)N-nitrate was excreted, while 0.68% (0.44-1.17%) of (15)N-arginine was metabolized to nitrate. In conclusion, this method can be used for accurate simultaneous determination of (15)N-nitrate and total nitrate concentrations in urine and can be applied in clinical studies for noninvasive evaluation of NO metabolism in vivo.

  19. Experimental evidence for diel δ15N-patterns in different tissues, xylem and phloem saps of castor bean (Ricinus communis L.).

    PubMed

    Peuke, A D; Gessler, A; Tcherkez, G

    2013-12-01

    Nitrogen isotope signatures in plants might give insights in the metabolism and allocation of nitrogen. To obtain a deeper understanding of the modifications of the nitrogen isotope signatures, we determined δ(15)N in transport saps and in different fractions of leaves, axes and roots during a diel course along the plant axis. The most significant diel variations were observed in xylem and phloem saps where δ(15)N was significantly higher during the day compared with during the night. However in xylem saps, this was observed only in the canopy, but not at the hypocotyl positions. In the canopy, δ(15)N was correlated fairly well between phloem and xylem saps. These variations in δ(15)N in transport saps can be attributed to nitrate reduction in leaves during the photoperiod as well as to (15)N-enriched glutamine acting as transport form of N. δ(15)N of the water soluble fraction of roots and leaves partially affected δ(15)N of phloem and xylems saps. δ(15)N patterns are likely the result of a complex set of interactions and N-fluxes between plant organs. Furthermore, the natural nitrogen isotope abundance in plant tissue is not constant during the diel course - a fact that needs to be taken into account when sampling for isotopic studies.

  20. Technical Note: Simultaneous measurement of sedimentary N2 and N2O production and new 15N isotope pairing technique

    NASA Astrophysics Data System (ADS)

    Hsu, T.-C.; Kao, S.-J.

    2013-04-01

    Dinitrogen (N2) and/or nitrous oxide (N2O) are produced through denitrification, anaerobic ammonium oxidation (anammox) or nitrification in sediments, of which entangled processes obfuscate the absolute rate estimation of gaseous nitrogen production from individual pathway. Recently, the classical isotope pairing technique (IPT), the most common 15N-nitrate enrichment method to quantify denitrification, has been modified by different researchers to (1) discriminate relative contribution of N2 production by denitrification from anammox or to (2) provide more accurate denitrification rate by considering both N2O and N2 productions. Both modified methods, however, have deficiencies such as overlooking N2O production in case 1 and neglecting anammox in case 2. In this paper, a new method was developed to refine previous methods. We installed cryogenic traps to pre-concentrate N2 and N2O separately, thus, allowing simultaneous measurement for two gases generated by one sample. The precision is better than 2% for N2 (m/z 28, m/z 29 and m/z 30), and 1.5% for N2O (m/z 44, m/z 45 and m/z 46). Based on the six m/z peaks of the two gases, we further revised IPT formulae to truthfully resolve the production rates of N2 and N2O contributed from 3 specific nitrogen removal processes, i.e. N2 and N2O from denitrification, N2 from anammox and N2O from nitrification. To validate the applicability of our new method, incubation experiments were conducted using sediment cores taken from the Danshuei estuary in Taiwan. We successfully determined the rates of aforementioned nitrogen removal processes. Moreover, N2O yield was as high as 66%, which no doubt would significantly bias previous IPT approaches when N2O was not considered. Our new method not only complements the previous IPT but also provides more comprehensive information to advance our understanding of nitrogen dynamics through the water-sediment interface.

  1. Backbone dynamics of barstar: a (15)N NMR relaxation study.

    PubMed

    Sahu, S C; Bhuyan, A K; Majumdar, A; Udgaonkar, J B

    2000-12-01

    Backbone dynamics of uniformly (15)N-labeled barstar have been studied at 32 degrees C, pH 6.7, by using (15)N relaxation data obtained from proton-detected 2D (1)H-(15)N NMR spectroscopy. (15)N spin-lattice relaxation rate constants (R(1)), spin-spin relaxation rate constants (R(2)), and steady-state heteronuclear (1)H-(15)N NOEs have been determined for 69 of the 86 (excluding two prolines and the N-terminal residue) backbone amide (15)N at a magnetic field strength of 14.1 Tesla. The primary relaxation data have been analyzed by using the model-free formalism of molecular dynamics, using both isotropic and axially symmetric diffusion of the molecule, to determine the overall rotational correlation time (tau(m)), the generalized order parameter (S(2)), the effective correlation time for internal motions (tau(e)), and NH exchange broadening contributions (R(ex)) for each residue. As per the axially symmetric diffusion, the ratio of diffusion rates about the unique and perpendicular axes (D( parallel)/D( perpendicular)) is 0.82 +/- 0.03. The two results have only marginal differences. The relaxation data have also been used to map reduced spectral densities for the NH vectors of these residues at three frequencies: 0, omega(H), and omega(N), where omega(H),(N) are proton and nitrogen Larmor frequencies. The value of tau(m) obtained from model-free analysis of the relaxation data is 5.2 ns. The reduced spectral density analysis, however, yields a value of 5.7 ns. The tau(m) determined here is different from that calculated previously from time-resolved fluorescence data (4.1 ns). The order parameter ranges from 0.68 to 0.98, with an average value of 0.85 +/- 0.02. A comparison of the order parameters with the X-ray B-factors for the backbone nitrogens of wild-type barstar does not show any considerable correlation. Model-free analysis of the relaxation data for seven residues required the inclusion of an exchange broadening term, the magnitude of which ranges from 2

  2. Synthesis of 15 alpha-hydroxyestrogen 15-N-acetylglucosaminides.

    PubMed

    Suzuki, E; Namba, S; Kurihara, H; Goto, J; Matsuki, Y; Nambara, T

    1995-03-01

    The synthesis of 15-N-acetylglucosaminides of 15 alpha-hydroxyesterone, 15 alpha-hydroxyestradiol, and 15 alpha-hydroxyestriol (estetrol) is described. The latter two were prepared by condensation of 2-acetamido-1 alpha-chloro-1,2-dideoxy-3,4,6-trio-O-acetyl-D-glucopyranose with appropriately protected 15 alpha-hydroxyestrogens by the Koenigs-Knorr reaction employing cadmium carbonate as a catalyst. Subsequent removal of protecting groups with methanolic potassium hydroxide provided the desired conjugates. 15 alpha-Hydroxyestrone 15-N-acetylglucosaminide was synthesized from the corresponding 15 alpha-hydroxyestradiol derivative by Jones oxidation followed by brief alkaline hydrolysis. These conjugates underwent enzymatic hydrolysis with beta-N-acetylglucosaminidase from Jack beans to produce 15 alpha-hydroxyestrogens. PMID:7792832

  3. A novel method for trapping and analyzing 15N in NO for tracing NO sources

    NASA Astrophysics Data System (ADS)

    Kang, Ronghua; Mulder, Jan; Dörsch, Peter

    2016-04-01

    15N isotope tracing is an effective and direct approach to investigate the biological and chemical sources of nitric oxide (NO) in soil. However, NO is highly reactive and rapidly converted to nitrogen dioxide (NO2) in the presence of ozone. Various chemical conversions of NO to the more stable solutes nitrite (NO2-) and nitrate (NO3-) have been proposed, which allow analysing the 15N abundance without major fractionation. However, NO emissions from soils are usually small, posing major challenges to conversion efficiency and background contamination. Here we present a novel method in which NO is oxidized to NO2- by chromium trioxide (CrO3) prior to conversion to NO2- and NO3- in an alkaline hydrogen peroxide (H2O2) solution. Immediately following trapping, manganese dioxide (MnO2) and 5M HCl are added to remove excess H2O2, and to adjust the pH to around 6.0-7.0, respectively. The resulting solution can be stored until analysis and is none-toxic, allowing to use a modified denitrifier method (Zhu et al., submitted), where NO2- and NO3- are reduced quantitatively to nitrous oxide (N2O). Optimum NO conversion rates of > 90% even at extremely low initial NO concentration were obtained with 4% H2O2, 0.5 M NaOH, and 0.5 L min-1 gas flow rate. In a laboratory test, using NO gas with different 15N signals produced from unlabelled and labelled NO2-, we found an overall precision of 0.4‰ for unlabelled and 49.7‰ for NO enriched with 1.0 atom% 15N, respectively. This indicates that this method can be used for both natural abundance studies of NO, as well as in labelling studies tracing NO sources. Zhu J, Yu L, Bakken LR, Mørkved PT, Mulder J, Dörsch P. Controlled induction of denitrification in Pseudomonas aureofaciens: a modified denitrifier method for 15N and 18O analysis in NO3- from natural water samples by IRMS. Submitted.

  4. 15N chemical shift referencing in solid state NMR.

    PubMed

    Bertani, Philippe; Raya, Jésus; Bechinger, Burkhard

    2014-01-01

    Solid-state NMR spectroscopy has much advanced during the last decade and provides a multitude of data that can be used for high-resolution structure determination of biomolecules, polymers, inorganic compounds or macromolecules. In some cases the chemical shift referencing has become a limiting factor to the precision of the structure calculations and we have therefore evaluated a number of methods used in proton-decoupled (15)N solid-state NMR spectroscopy. For (13)C solid-state NMR spectroscopy adamantane is generally accepted as an external standard, but to calibrate the (15)N chemical shift scale several standards are in use. As a consequence the published chemical shift values exhibit considerable differences (up to 22 ppm). In this paper we report the (15)N chemical shift of several commonly used references compounds in order to allow for comparison and recalibration of published data and future work. We show that (15)NH4Cl in its powdered form (at 39.3 ppm with respect to liquid NH3) is a suitable external reference as it produces narrow lines when compared to other reference compounds and at the same time allows for the set-up of cross-polarization NMR experiments. The compound is suitable to calibrate magic angle spinning and static NMR experiments. Finally the temperature variation of (15)NH4Cl chemical shift is reported.

  5. delta15N and delta13C diet-tissue discrimination factors for large sharks under semi-controlled conditions.

    PubMed

    Hussey, Nigel E; Brush, Jaclyn; McCarthy, Ian D; Fisk, Aaron T

    2010-04-01

    Stable isotopes (delta(15)N and delta(13)C) are being widely applied in ecological research but there has been a call for ecologists to determine species- and tissue-specific diet discrimination factors ((13)C and (15)N) for their study animals. For large sharks stable isotopes may provide an important tool to elucidate aspects of their ecological roles in marine systems, but laboratory based controlled feeding experiments are impractical. By utilizing commercial aquaria, we estimated (15)N and (13)C of muscle, liver, vertebral cartilage and a number of organs of three large sand tiger (Carcharias taurus) and one large lemon shark (Negaprion brevirostris) under a controlled feeding regime. For all sharks mean+/-SD for (15)N and (13)C in lipid extracted muscle using lipid extracted prey data were 2.29 per thousand+/-0.22 and 0.90 per thousand+/-0.33, respectively. The use of non-lipid extracted muscle and prey resulted in very similar (15)N and (13)C values but mixing of lipid and non-lipid extracted data produced variable estimates. Values of (15)N and (13)C in lipid extracted liver and prey were 1.50 per thousand+/-0.54 and 0.22 per thousand+/-1.18, respectively. Non-lipid extracted diet discrimination factors in liver were highly influenced by lipid content and studies that examine stable isotopes in shark liver, and likely any high lipid tissue, should strive to remove lipid effects through standardising C:N ratios, prior to isotope analysis. Mean vertebral cartilage (15)N and (13)C values were 1.45 per thousand+/-0.61 and 3.75 per thousand+/-0.44, respectively. Organ (15)N and (13)C values were more variable among individual sharks but heart tissue was consistently enriched by approximately 1-2.5 per thousand. Minimal variability in muscle and liver delta(15)N and delta(13)C sampled at different intervals along the length of individual sharks and between liver lobes suggests that stable isotope values are consistent within tissues of individual animals. To our

  6. Spatial Patterns of Plant δ13C and δ15N Along a Topoedaphic Gradient in a Subtropical Savanna Landscape

    NASA Astrophysics Data System (ADS)

    Bai, E.; Boutton, T. W.; Liu, F.; Wu, B.; Archer, S. R.

    2005-12-01

    relationship between δ15N and leaf [N] and soil water may reflect the fact that plants with high δ15N values occurred on lower-lying portions of the landscape with relatively high N-transformation rates where soil mineral N was both relatively available and 15N-enriched. The lack of variation in δ15N values of Prosopis suggests that it may be somewhat uncoupled from these processes because it is an N-fixer. Results of this study indicate that N-availability plays an important role in landscale scale variation in plant δ13C and δ15N.

  7. (15)N methodologies for quantifying the response of N2-fixing associations to elevated [CO2]: A review.

    PubMed

    Chalk, Phillip M; Lam, Shu K; Chen, Deli

    2016-11-15

    Methodologies based on (15)N enrichment (E) and (15)N natural abundance (NA) have been used to obtain quantitative estimates of the response of biological N2 fixation (BNF) of legumes (woody, grain and forage) and actinorhizal plants grown in artificial media or in soil exposed to elevated atmospheric concentrations of carbon dioxide e[CO2] for extended periods of time, in growth rooms, greenhouses, open top chambers or free-air CO2 enrichment (FACE) facilities. (15)N2 has also been used to quantify the response of endophytic and free-living diazotrophs to e[CO2]. The primary criterion of response was the proportional dependence of the N2-fixing system on the atmosphere as a source of N. i.e. the symbiotic dependence (Patm). The unique feature of (15)N-based methods is their ability to provide time-integrated and yield-independent estimates of Patm. In studies conducted in artificial media or in soil using the E methodology there was either no response or a positive response of Patm to e[CO2]. The interpretation of results obtained in artificial media or with (15)N2 is straight forward, not being subject to the assumptions on which the E and NA soil-cultured methods are based. A variety of methods have been used to estimate isotopic fractionation attendant on the NA technique, the so-called 'B value', which attaches a degree of uncertainty to the results obtained. Using the NA technique, a suite of responses of Patm to e[CO2] has been published, from positive to neutral to sometimes negative effects. Several factors which interact with the response of N2-fixing species to e[CO2] were identified.

  8. (15)N methodologies for quantifying the response of N2-fixing associations to elevated [CO2]: A review.

    PubMed

    Chalk, Phillip M; Lam, Shu K; Chen, Deli

    2016-11-15

    Methodologies based on (15)N enrichment (E) and (15)N natural abundance (NA) have been used to obtain quantitative estimates of the response of biological N2 fixation (BNF) of legumes (woody, grain and forage) and actinorhizal plants grown in artificial media or in soil exposed to elevated atmospheric concentrations of carbon dioxide e[CO2] for extended periods of time, in growth rooms, greenhouses, open top chambers or free-air CO2 enrichment (FACE) facilities. (15)N2 has also been used to quantify the response of endophytic and free-living diazotrophs to e[CO2]. The primary criterion of response was the proportional dependence of the N2-fixing system on the atmosphere as a source of N. i.e. the symbiotic dependence (Patm). The unique feature of (15)N-based methods is their ability to provide time-integrated and yield-independent estimates of Patm. In studies conducted in artificial media or in soil using the E methodology there was either no response or a positive response of Patm to e[CO2]. The interpretation of results obtained in artificial media or with (15)N2 is straight forward, not being subject to the assumptions on which the E and NA soil-cultured methods are based. A variety of methods have been used to estimate isotopic fractionation attendant on the NA technique, the so-called 'B value', which attaches a degree of uncertainty to the results obtained. Using the NA technique, a suite of responses of Patm to e[CO2] has been published, from positive to neutral to sometimes negative effects. Several factors which interact with the response of N2-fixing species to e[CO2] were identified. PMID:27424117

  9. Technical Note: Simultaneous measurement of sedimentary N2 and N2O production and a modified 15N isotope pairing technique

    NASA Astrophysics Data System (ADS)

    Hsu, T.-C.; Kao, S.-J.

    2013-12-01

    Dinitrogen (N2) and/or nitrous oxide (N2O) are produced through denitrification, anaerobic ammonium oxidation (anammox) or nitrification in sediments, of which entangled processes complicate the absolute rate estimations of gaseous nitrogen production from individual pathways. The classical isotope pairing technique (IPT), the most common 15N nitrate enrichment method to quantify denitrification, has recently been modified by different researchers to (1) discriminate between the N2 produced by denitrification and anammox or to (2) provide a more accurate denitrification rate under considering production of both N2O and N2. In case 1, the revised IPT focused on N2 production being suitable for the environments of a low N2O-to-N2 production ratio, while in case 2, anammox was neglected. This paper develops a modified method to refine previous versions of IPT. Cryogenic traps were installed to separately preconcentrate N2 and N2O, thus allowing for subsequent measurement of the two gases generated in one sample vial. The precision is better than 2% for N2 (m/z 28, m/z 29 and m/z 30), and 1.5% for N2O (m/z 44, m/z 45 and m/z 46). Based on the six m/z peaks of the two gases, the 15N nitrate traceable processes including N2 and N2O from denitrification and N2 from anammox were estimated. Meanwhile, N2O produced by nitrification was estimated via the production rate of unlabeled 44N2O. To validate the applicability of our modified method, incubation experiments were conducted using sediment cores taken from the Danshuei Estuary in Taiwan. Rates of the aforementioned nitrogen removal processes were successfully determined. Moreover, N2O yield was as high as 66%, which would significantly bias previous IPT approaches if N2O was not considered. Our modified method not only complements previous versions of IPT but also provides more comprehensive information to advance our understanding of nitrogen dynamics of the water-sediment interface.

  10. Multi-Isotope Secondary Ion Mass Spectrometry Combining Heavy Water 2H with 15N Labeling As Complementary Tracers for Metabolic Heterogeneity at the Single-Cell Level

    NASA Astrophysics Data System (ADS)

    Kopf, S.; McGlynn, S.; Cowley, E.; Green, A.; Newman, D. K.; Orphan, V. J.

    2014-12-01

    Metabolic rates of microbial communities constitute a key physiological parameter for understanding the in situ growth constraints for life in any environment. Isotope labeling techniques provide a powerful approach for measuring such biological activity, due to the use of isotopically enriched substrate tracers whose incorporation into biological materials can be detected with high sensitivity by isotope-ratio mass spectrometry. Nano-meter scale secondary ion mass spectrometry (NanoSIMS) combined with stable isotope labeling provides a unique tool for studying the spatiometabolic activity of microbial populations at the single cell level in order to assess both community structure and population diversity. However, assessing the distribution and range of microbial activity in complex environmental systems with slow-growing organisms, diverse carbon and nitrogen sources, or heterotrophic subpopulations poses a tremendous technical challenge because the introduction of isotopically labeled substrates frequently changes the nutrient availability and can inflate or bias measures of activity. Here, we present the use of hydrogen isotope labeling with deuterated water as an important new addition to the isotopic toolkit and apply it for the determination of single cell microbial activities by NanoSIMS imaging. This tool provides a labeling technique that minimally alters any aquatic chemical environment, can be administered with strong labels even in minimal addition (natural background is very low), is an equally universal substrate for all forms of life even in complex, carbon and nitrogen saturated systems, and can be combined with other isotopic tracers. The combination of heavy water labeling with the most commonly used NanoSIMS tracer, 15N, is technically challenging but opens up a powerful new set of multi-tracer experiments for the study of microbial activity in complex communities. We present the first truly simultaneous single cell triple isotope system

  11. 15N Tracing Studies on In Vitro Reactions of Ferredoxin-Dependent Nitrite Reductase and Glutamate Synthase Using Reconstituted Electron Donation Systems.

    PubMed

    Yoneyama, Tadakatsu; Fujimori, Tamaki; Yanagisawa, Shuichi; Hase, Toshiharu; Suzuki, Akira

    2015-06-01

    It is known that plants contain ferredoxin (Fd)-dependent nitrite reductase (NiR) and glutamate synthase (GOGAT). The Fd-NiR reaction produces ammonia from nitrite, and the activity is usually measured by nitrite disappearance. The Fd-GOGAT reaction forms two glutamates of different origin, from glutamine and 2-oxoglutarate, and the activity is measured by the oxidation of reductant (NADPH) or by formation of total glutamate. Here, a quantitative probe of the products and efficiency of the process was conducted using (15)N tracing techniques on these reactions in vitro. We quantified the reduction of (15)N-labeled [Formula: see text] to [Formula: see text] and the formation of [(15)N]glutamate and [(14)N]glutamate from [5-(15)N-amide]glutamine plus 2-oxoglutarate by NiR and GOGAT, respectively, with the reductant-Fd-NADP(+) oxidoreductase (FNR)-Fd system as the sequential electron donors. The supply of dithionite or NADPH to recombinant cyanobacterial NiR led to electron donation system-dependent formation of [(15)N]ammonium from [(15)N]nitrite. Addition of 20 mM NaCl and 20 mM Na-ascorbate accelerated nitrite reduction under high concentrations of NADPH. A sufficient supply of NADPH to recombinant Zea mays Fd-GOGAT generated complete GOGAT activity (transferring the [5-(15)N]amide of glutamine to 2-oxoglutarate to form [(15)N]glutamate), whereas a shortage of NADPH resulted in glutaminase activity only, which removed the amide from glutamine and released ammonia and [(14)N]glutamate. We conclude that although the recombinant Fd-GOGAT enzyme has two forms of glutamate synthesis, the first by glutaminase (ammonia release by glutamine amidotransferase) and the second by glutamate synthase (coupling of the ammonia and exogenously applied 2-oxoglutarate), the first works without NADPH, while the second is strictly dependent on NADPH availability.

  12. Development and application of 15N-tracer substances for measuring the whole-body protein turnover rates in the human, especially in neonates: a review.

    PubMed

    Wutzke, Klaus D

    2012-06-01

    Our research group of the Children's Hospital of the University of Rostock (Rostock group) has long-time experience in (15)N-labelling and in using yeast protein and its hydrolysates for tracer kinetic studies to evaluate parameters of the whole-body protein metabolism in premature infants. The particular advantage of applying an economically convenient, highly (15)N-enriched, and completely labelled yeast protein for evaluating protein turnover rates is the fact that the (15)N dose is spread among all proteinogenic amino acids. The absorption has been improved by hydrolysing [(15)N]yeast protein with thermitase into a mixture of amino acids, dipeptides and tripeptides so that faecal analysis becomes unnecessary when determining turnover rates. The review shows that, in contrast to the application of single (15)N-labelled amino acids with resulting overestimation of protein turnover rates, the (15)N-labelled yeast protein thermitase hydrolysate represents the amino acid metabolism more closely without causing amino acid imbalances. The (15)N-labelled yeast protein thermitase hydrolysate leads to the estimation of reliable protein turnover rates, particularly in premature infants.

  13. Incorporation of (15)N-TNT transformation products into humifying plant organic matter as revealed by one- and two-dimensional solid state NMR spectroscopy.

    PubMed

    Knicker, Heike

    2003-06-01

    Solid-state double cross polarization magic angle spinning (DCPMAS) 15N 13C nuclear magnetic resonance (NMR) spectroscopy was applied to study the incorporation of TNT transformation products into humifying plant organic matter. For this approach, 13C-enriched plant material (Lolium perenne) was mixed with quartz sand and aerobically incubated for 11 months after addition of 15N(3)-2,4,6-trinitrotoluene (TNT). After successive extraction of the incubate with water, methanol and ethyl acetate, approximately 60% of the 15N added as 15N(3)-TNT (15N(add)) remained in the solid organic residue (SOR-fraction). The acid insoluble fraction (AI) obtained after NaOH and HCl extractions contained approximately 20% of 15N(add). For both fractions, 15N NMR spectroscopy revealed an almost complete reduction of the TNT after 11 months of aerobic incubation. Most of the reduced nitrogen groups underwent further condensation. The corresponding DCPMAS NMR spectra allowed the identification of amides that are further substituted by alkyl groups that resist even acid hydrolysis. This assigns them to relatively stable compounds rather than to newly synthesized microbial peptides. The results of this study suggest further that the covalent binding of TNT transformation products to plant derived organic matter is mediated by alkylation and acetylation reactions, rather than by 1,4 addition of TNT-derived nitrogenous groups to quinones of the humic material.

  14. Nitrogen Dynamics in Stream Wood Samples Incubated with [14C]Lignocellulose and Potassium [15N]Nitrate †

    PubMed Central

    Aumen, Nicholas G.; Bottomley, Peter J.; Gregory, Stan V.

    1985-01-01

    Surface wood samples obtained from a Douglas fir log (Pseudotsuga menziesii) in a Pacific Northwest stream were incubated in vitro with [14C]lignocellulose in a defined mineral salts medium supplemented with 10 mg of N liter−1 of 15N-labeled NO3− (50 atom% 15N). Evolution of 14CO2, distribution and isotopic dilution of 15N, filtrate N concentrations, and the rates of denitrification, N2 fixation, and respiration were measured at 6, 12, and 18 days of incubation. The organic N content of the lignocellulose-wood sample mixture had increased from 132 μg of N to a maximum of 231 μg of N per treatment after 6 days of incubation. Rates of [14C]lignocellulose decomposition were greatest during the first 6 days and then began to decline over the remaining 12 days. Total CO2 evolution was also highest at day 6 and declined steadily over the remaining duration of the incubation. Filtrate NH4+-N increased from background levels to a final value of 57 μg of N per treatment. Filtrate NO3− N completely disappeared by day 6, and organic N showed a slight decline between days 12 and 18. The majority of the 15N that could be recovered appeared in the particulate organic fraction by day 6 (41 μg of N), and the filtrate NH4+ N fraction contained 11 μg of 15N by day 18. The 15N enrichment values of the filtrate NH4+ and the inorganic N associated with the particulate fraction had increased to approximately 20 atom% 15N by 18 days of incubation, whereas the particulate organic fraction reached its highest enrichment by day 6. Measurements of N2 fixation and denitrification indicated an insignificant gain or loss of N from the experimental system by these processes. The data show that woody debris in stream ecosystems might function as a rapid and efficient sink for exogenous N, resulting in stimulation of wood decomposition and subsequent activation of other N cycling processes. PMID:16346785

  15. Non-homogeneity of isotopic labelling in 15N gas flux studies: theory, some observations and possible lessons

    NASA Astrophysics Data System (ADS)

    Well, Reinhard; Buchen, Caroline; Deppe, Marianna; Eschenbach, Wolfram; Gattinger, Andreas; Giesemann, Anette; Krause, Hans-Martin; Lewicka-Szczebak, Dominika

    2015-04-01

    Quantifying dinitrogen (N2) and nitrous oxide (N2O) fluxes from different soil N pools and processes can be accomplished using the 15N tracer technique but this is subject to four different sources of bias (i. - iv.). This approach includes 15N labelling of selected N pools in soil and subsequent isotope analysis of all relevant N pools as well as of gas samples from enclosures, i.e. mixtures of soil-derived and atmospheric N2 and N2O. Depending on the processes of interest, there may be 15N labelling of one or several N pools, were several labelling treatment are needed in the latter case (e.g. Müller et al., 2004). Measuring pool-derived N2 or N2O has been shown to include two calculation problems, (i.) arising from multiple pools (e.g. Arah, 1992) and (ii.) dealing with the non-random distribution of N2 and N2O mole masses (Hauck et al., 1958). Non-randomness can be solved if m/z 28, 29 and 30 are correctly analysed and the 15N enrichment of one (to distinguish two pools, i.e. soil and atmosphere) or two pools (in case of three pools) is known (Spott & Stange, 2008). Moreover (iii.), NO3- pools generating N2 and N2O via denitrification can be identical or different, e.g. if N2O evolved from higher enriched NO3- in deeper soil was more reduced to N2 compared to N2O evolved from N2O from shallow soil with lower enrichment, or vice versa. Apportioning N2O fluxes to NH4+ (nitrification and/or nitrifier denitrification) and NO3- (denitrification) is often conducted by NO3-labeling, measuring δ15N of emitted N2O and applying mixing equations were the measured 15N enrichment of NH4+and NO3-pool is used. However, this assumes that the average 15N enrichment of NH4+and NO3-in the soil is identical to the enrichment in the active soil domain producing N2 and/or N2O. Violation of this precondition must lead to bias in source apportionment (iv.), but to our knowledge this has not been investigated until now. Here we present conceptual models and model calculations

  16. Synthesis and biosynthesis of {sup 13}C-, {sup 15}N-labeled deoxynucleosides useful for biomolecular structural determinations

    SciTech Connect

    Ashburn, D.A.; Garcia, K.; Hanners, J.L.; Silks, L.A. III; Unkefer, C.J.

    1994-12-01

    Currently, there is a great emphasis on elucidating the structure, function, and dynamics of DNA. Much of the research involved in this study uses nuclear magnetic resonance (NMR) spectroscopy. Effective use of NMR spectroscopy for DNA molecules with mw > 10,000 requires stable isotope enrichment. We present strategies for site-specific isotopic labeling of the purine bases adenosine and guanosine and the biosynthesis of (U-{sup 13}C, {sup 15}N) DNA from methylotropic bacteria. With commercially available 6-chloropurine, an effective two-step route leads to 2{prime}-deoxy-(amino-{sup 15}N)adenosine (dA). The resulting d(amino-{sup 15}N)A is used in a series of reactions to synthesize 2{prime}-deoxy-(2-{sup 13}C,1,amino-{sup 15}N{sub 2})guanosine or any combination thereof. An improved biosynthesis of labeled DNA has been accomplished using Methylobacterium extorquens AS1. Each liter of growth medium contains 4 g of methanol to yield 1 g of lyophilized cells. As much as 200 mg of RNA per liter of culture has been obtained. We are currently developing large-scale isolation protocols. General synthetic pathways to oligomeric DNA will be presented.

  17. 15N/14N Variation in CalCOFI Zooplankton: A 51 year history of Marine Nitrogen Dynamics and Climate Variability off Central California

    NASA Astrophysics Data System (ADS)

    Rau, G. H.; Ohman, M. D.; Pierrot-Bults, A.

    2003-12-01

    Long-term variability in marine zooplankton 15N/14N was investigated in two species of calanoid copepods (Calanus pacificus and Eucalanus californicus) and two chaetognaths (Sagitta bierii and Sagitta euneritica) sampled in the spring of selected years from 1951 to 2001 near Monterey Bay, California. No statistically significant trend in 15N/14N was detected for any of the four species, with isotopic ratios in 2001 resembling those in copepods and chaetognaths sampled from the same area five decades earlier. With respect to proposed oceanographic regime shifts in this region, heterogeneity in 15N/14N was detected only for S. bierii when comparing the periods 1951-1975, 1978-1998, and 1999-2001. In this species the 15N/14N in the most recent, brief period (1999-2001) averaged slightly lower than in the previous period. Three of the four species (C. pacificus, S. bierii, and S. euneritica) showed significant increases in 15N/14N during major El Ninos. El Nino-related enrichment in 15N could arise as a consequence of increased nitrate demand/supply at the base of the food web or advection of 15N-enriched nitrate from more southerly waters. While a range of physical and climate indices were evaluated, anomalies of 15N/14N from the long-term mean were found to be significantly related only to; i) the Southern Oscillation Index in the case of both chaetognath species, ii) a regional surface water temperature record (S. bierii only), iii) an index of wind-driven coastal upwelling for the surface-dwelling C. pacificus, and iv) variability in the Pacific Decadal Oscillation for the somewhat deeper-dwelling E. californicus.

  18. Nitrogen-15 spin-rotation relaxation in ammonium perchlorate

    NASA Astrophysics Data System (ADS)

    Harnden, Anne M. C.; Hunter, Brian K.; Brown, R. Julian C.

    The spin-lattice relaxation time has been measured for 15N in ammonium perchlorate in the temperature range 240 to 292 K. The temperature dependence of T, suggests that spin-rotation is the dominant relaxation mechanism, and this is confirmed by calculation and by nuclear Overhauser effect measurements. The spin-rotation coupling constant for 15NH 4+ is estimated to be 11.1 ± 0.2 kHz.

  19. Uptake of stormwater nitrogen in bioretention systems demonstrated from 15N tracer techniques

    NASA Astrophysics Data System (ADS)

    Houdeshel, D.; Hultine, K. R.; Pomeroy, C. A.

    2012-12-01

    Bioretention stormwater management systems are engineered ecosystems that capture urban stormwater in order to reduce the harmful effects of stormwater pollution on receiving waters. Bioretention systems have been shown to be effective at reducing the volume of runoff, and thereby reduce the nutrient loading to receiving waters from urban areas. However, little work has been done to evaluate the treatment processes that are responsible for reductions in effluent nitrogen (N). We hypothesize that the pulses of inorganic nitrogen associated with urban runoff events are captured in the plat tissues within these systems and not adsorbed to the soil media, thus creating a long-term, sustainable treatment approach to reducing the total nutrient loading to receiving waters. Nitrogen treatment performance was tested on two bioretention systems in Salt Lake City, UT: 1) an upland native community that does not require irrigation in semi-arid climates, and 2) a wetland community that requires 250 l of daily irrigation to offset the relatively high evaporative demand in the region. Each cell is sized to treat a 2.5 cm storm from a 140 m2 impervious surface: the area of the bioretention system is 10 m2. To test the N removal performance of each system, runoff events were simulated to represent an average precipitation regime using a synthetic stormwater blend starting in January, 2012. Effluent was collected from an underdrain and analyzed for total nitrogen (TN); mass removal was calculated for each month by subtracting the TN mass added to the garden minus the TN mass that flowed out of the garden. To test the hypothesis that plants assimilate stormwater N, 4 g of 100 atom% 15N NH4NO3 tracer was used as the N source in the synthetic stormwater during the first 2,000 l synthetic storm event in May. This isotopic label was calculated to enrich the total N pool of each garden to 100‰ 15N/14Nair. New growth was harvested from each plant in both cells and analyzed for 15N

  20. Paleoenvironmental implications of taxonomic variation among δ15N values of chloropigments

    USGS Publications Warehouse

    Higgins, Meytal B.; Wolfe-Simon, Felisa; Robinson, Rebecca S.; Qin, Yelun; Saito, Mark A.; Pearson, Ann

    2011-01-01

    Natural variations in the ratios of nitrogen isotopes in biomass reflect variations in nutrient sources utilized for growth. In order to use δ15N values of chloropigments of photosynthetic organisms to determine the corresponding δ15N values of biomass – and by extension, surface waters – the isotopic offset between chlorophyll and biomass must be constrained. Here we examine this offset in various geologically-relevant taxa, grown using nutrient sources that may approximate ocean conditions at different times in Earth’s history. Phytoplankton in this study include cyanobacteria (diazotrophic and non-diazotrophic), eukaryotic algae (red and green), and anoxygenic photosynthetic bacteria (Proteobacteria), as well as environmental samples from sulfidic lake water. Cultures were grown using N2, NO3−, and NH4+ as nitrogen sources, and were examined under different light regimes and growth conditions. We find surprisingly high variability in the isotopic difference (δ15Nbiomass − δ15Nchloropigment) for prokaryotes, with average values for species ranging from −12.2‰ to +11.7‰. We define this difference as εpor, a term that encompasses diagenetic porphyrins and chlorins, as well as chlorophyll. Negative values of εpor reflect chloropigments that are 15N-enriched relative to biomass. Notably, this enrichment appears to occur only in cyanobacteria. The average value of εpor for freshwater cyanobacterial species is −9.8 ± 1.8‰, while for marine cyanobacteria it is −0.9 ± 1.3‰. These isotopic effects group environmentally but not phylogenetically, e.g., εpor values for freshwater Chroococcales resemble those of freshwater Nostocales but differ from those of marine Chroococcales. Our measured values of εpor for eukaryotic algae (range = 4.7–8.7‰) are similar to previous reports for pure cultures. For all taxa studied, values of εpor do not depend on the type of nitrogen substrate used for growth. The observed

  1. Ammonium nitrate explosive systems

    DOEpatents

    Stinecipher, Mary M.; Coburn, Michael D.

    1981-01-01

    Novel explosives which comprise mixtures of ammonium nitrate and an ammonium salt of a nitroazole in desired ratios are disclosed. A preferred nitroazole is 3,5-dinitro-1,2,4-triazole. The explosive and physical properties of these explosives may readily be varied by the addition of other explosives and oxidizers. Certain of these mixtures have been found to act as ideal explosives.

  2. Use of a {sup 15}N isotope dilution method to assess contaminant effects on soil nitrification

    SciTech Connect

    Nason, G.E.; Dinwoodie, G.D.

    1995-12-31

    Ecologically relevant bioassays are needed to assess effects of contaminants on soil processes such as decomposition and nutrient cycling. This study was conducted to assess the potential of a soil-based nitrification bioassay. Soil samples adjusted to 0.03 MPa moisture content were amended with 0.1, 1.0, 10 and 100 mg kg{sup {minus}1} PCP or PCB, and 0.05, 0.5, 5 and 50 mg kg{sup {minus}1} Hg and preincubated for 7 days. A 2-d incubation was then started by addition of 10 mg kg{sup {minus}1} {sup 15}NO{sub 3}-N. Diethyl ether used as a carrier for PCP addition had little effect on inorganic nitrogen concentrations during the incubation. Net nitrogen mineralization and nitrification were unaffected by PCB. Higher amendment levels of both PCP and Hg resulted in increases in ammonium concentrations and decreases in net nitrification. {sup 15}N-nitrate pool dilution was sensitive to contamination and showed some gross nitrification was occurring even when net nitrification had ceased. Recoveries of Hg and PCB at the end of the study were greater than 90%. Recovery of PCP was 5%. Incubations carried out under sterile and non-sterile conditions indicated that both sorption and biological degradation were factors in the low PCP recovery.

  3. Toward hyperpolarized molecular imaging of HIV: synthesis and longitudinal relaxation properties of 15N-Azidothymidine

    PubMed Central

    Shchepin, Roman V.; Chekmenev, Eduard Y.

    2015-01-01

    Previously unreported 15N labeled Azidothymidine (AZT) was prepared as an equimolar mixture of two isotopomers: 1-15N-AZT and 3-15N-AZT. Polarization decay of 15N NMR signal was studied in high (9.4 T) and low (~50 mT) magnetic fields. 15N T1 values were 45 ± 5 s (1-15N-AZT) and 37 ± 2 s (3-15N-AZT) at 9.4 T, and 140 ± 16 s (3-15N-AZT) at 50 mT. 15N-AZT can be potentially 15N hyperpolarized by several methods. These sufficiently long 15N-AZT T1 values potentially enable hyperpolarized in vivo imaging of 15N-AZT, because of the known favorable efficient (i.e., of the time scale shorter than the longest reported here 15N T1) kinetics of uptake of injected AZT. Therefore, 3-15N-AZT can be potentially used for HIV molecular imaging using hyperpolarized magnetic resonance imaging. PMID:25156931

  4. Comparison of three 15N methods to correct for microbial contamination when assessing in situ protein degradability of fresh forages.

    PubMed

    Kamoun, M; Ammar, H; Théwis, A; Beckers, Y; France, J; López, S

    2014-11-01

    The use of stable (15)N as a marker to determine microbial contamination in nylon bag incubation residues to estimate protein degradability was investigated. Three methods using (15)N were compared: (15)N-labeled forage (dilution method, LF), (15)N enrichment of rumen solids-associated bacteria (SAB), and (15)N enrichment of rumen liquid-associated bacteria (LAB). Herbage from forages differing in protein and fiber contents (early-cut Italian ryegrass, late-cut Italian ryegrass, and red clover) were freeze-dried and ground and then incubated in situ in the rumen of 3 steers for 3, 6, 12, 24, and 48 h using the nylon bag technique. The (15)N-labeled forages were obtained by fertilizing the plots where herbage was grown with (15)NH4 (15)NO3. Unlabeled forages (obtained from plots fertilized with NH4NO3) were incubated at the same time that ((15)NH4)2SO4 was continuously infused into the rumen of the steers, and then pellets of labeled SAB and LAB were isolated by differential centrifugation of samples of ruminal contents. The proportion of bacterial N in the incubation residues increased from 0.09 and 0.45 g bacterial N/g total N at 3 h of incubation to 0.37 and 0.85 g bacterial N/g total N at 48 h of incubation for early-cut and late-cut ryegrass, respectively. There were differences (P < 0.001) between uncorrected N degradability values and those corrected for microbial contamination with all of the methods. Apparent N degradability of the low-N, high-fiber forage (late-cut ryegrass) was 0.51, whereas the corrected values were 0.85, 0.84, and 0.77 for the LF, SAB, and LAB methods, respectively. With early-cut ryegrass and red clover, the differences between uncorrected and corrected values ranged between 6% and 13%, with small differences among the labeling methods. Generally, methods using labeled forage or labeled SAB and LAB provided similar corrected degradability values. The accuracy in estimating the extent of degradation of protein in the rumen from in situ

  5. Population of Nitrifying Bacteria and Nitrification in Ammonium Saturated Clinoptilolite

    NASA Technical Reports Server (NTRS)

    McGilloway, R. L.; Weaver, R. W.; Ming, Douglas W.; Gruener, J.

    1999-01-01

    As humans begin to spend longer periods of time in space, plants will be incorporated into life support systems. Ammonium saturated clinoptilolite is one plant growth substrate but a balance between ammonium and nitrate is needed. A laboratory study was conducted to determine effects of nitrifying bacteria on ammonium concentrations and kinetics of nitrification. Columns containing clinoptilolite substrate amended with nitrifying bacteria obtained from soil enrichment were analyzed weekly for a 90 day period. The enrichment culture initially contained 1 x 10(exp 5) ammonium oxidizing bacteria and 1 x 10(exp 2) nitrite oxidizing bacteria per gram of substrate. Populations of ammonium oxidizing bacteria increased to 1 x 10(exp 6) and nitrite oxidizing bacteria increased to 1 x 10(exp 3) per gram of substrate. The nitrification rate was approximately 0.25mg NO3(-)-N/kg.hr. Experiments were also conducted to enumerate nitrifying bacteria in a clinoptilolite substrate used to grow wheat (Triticum aestivum L.). Seventy days following the initial inoculation with an unknown number of commercial nitrifying bacteria, 1 x 10(exp 5) ammonium oxidizing bacteria per gram of substrate were present. The number of nitrite oxidizing bacteria was between 1 x 10(exp 3) to 10(exp 4) per gram of substrate as measured by the most probable number method. Nitrification rates were approximately 0.20mg NO3(-)-N/kg.hr. Clinoptilolite readily exchanged sufficient concentrations of ammonium to support nitrifying bacteria and they survived well in this medium.

  6. Aerosol isotopic ammonium signatures over the remote Atlantic Ocean

    NASA Astrophysics Data System (ADS)

    Lin, C. T.; Jickells, T. D.; Baker, A. R.; Marca, A.; Johnson, M. T.

    2016-05-01

    We report aerosol ammonium 15N signatures for samples collected from research cruises on the South Atlantic and Caribbean using a new high sensitivity method. We confirm a pattern of isotopic signals from generally light (δ15N -5 to -10‰), for aerosols with very low (<2  nmol m-3) ammonium concentrations from the remote high latitude ocean, to generally heavier values (δ15N +5 to +10‰), for aerosols collected in temperate and tropical latitudes and with higher ammonium concentrations (>2  nmol m-3). We discuss whether this reflects a mixing of aerosols from two end-members (polluted continental and remote marine emissions), or isotopic fractionation during aerosol transport.

  7. Primary productivity, bacterial productivity and nitrogen uptake in response to iron enrichment during the SEEDS II

    NASA Astrophysics Data System (ADS)

    Kudo, Isao; Noiri, Yoshifumi; Cochlan, William P.; Suzuki, Koji; Aramaki, Takafumi; Ono, Tsuneo; Nojiri, Yukihiro

    2009-12-01

    Primary productivity (PP), bacterial productivity (BP) and the uptake rates of nitrate and ammonium were measured using isotopic methods ( 13C, 3H, 15N) during a mesoscale iron (Fe)-enrichment experiment conducted in the western subarctic Pacific Ocean in 2004 (SEEDS II). PP increased following Fe enrichment, reached maximal rates 12 days after the enrichment, and then declined to the initial level on day 17. During the 23-day observation period, we observed the development and decline of the Fe-induced bloom. The surface mixed layer (SML) integrated PP increased by 3-fold, but was smaller than the 5-fold increase observed in the previous Fe-enrichment experiment conducted at almost the same location and season during 2001 (SEEDS). Nitrate uptake rates were enhanced by Fe enrichment but decreased after day 5, and became lower than ammonium uptake rates after day 17. The total nitrogenous nutrient uptake rate declined after the peak of the bloom, and accumulation of ammonium was obvious in the euphotic layer. Nitrate utilization accounted for all the requirements of N for the massive bloom development during SEEDS, whereas during SEEDS II, nitrate accounted for >90% of total N utilization on day 5, declining to 40% by the end of the observation period. The SML-integrated BP increased after day 2 and peaked twice on days 8 and 21. Ammonium accumulation and the delayed heterotrophic activity suggested active regeneration occurred after the peak of the bloom. The SML-integrated PP between days 0 and 23 was 19.0 g C m -2. The SML-integrated BP during the same period was 2.6 g C m -2, which was 14% of the SML-integrated PP. Carbon budget calculation for the whole experimental period indicated that 33% of the whole (particulate plus dissolved) PP (21.5 g C m -2) was exported below the SML and 18% was transferred to the meso-zooplankton (growth). The bacterial carbon consumption (43% of the whole PP) was supported by DOC or POC release from phytoplankton, zooplankton

  8. The Titan 14N/ 15N and 12C/ 13C isotopic ratios in HCN from Cassini/CIRS

    NASA Astrophysics Data System (ADS)

    Vinatier, Sandrine; Bézard, Bruno; Nixon, Conor A.

    2007-11-01

    We report the detection of H 13CN and HC 15N in mid-infrared spectra recorded by the Composite Infrared Spectrometer (CIRS) aboard Cassini, along with the determination of the 12C/ 13C and 14N/ 15N isotopic ratios. We analyzed two sets of limb spectra recorded near 13-15° S (Tb flyby) and 83° N (T4 flyby) at 0.5 cm -1 resolution. The spectral range 1210-1310 cm -1 was used to retrieve the temperature profile in the range 145-490 km at 13° S and 165-300 km at 83° N. These two temperature profiles were then incorporated in the atmospheric model to retrieve the abundance profile of H 12C 14N, H 13CN and HC 15N from their bands at 713, 706 and 711 cm -1, respectively. The HCN abundance profile was retrieved in the range 90-460 km at 15° S and 165-305 km at 83° N. There is no evidence for vertical variations of the isotopic ratios. Constraining the isotopic abundance profiles to be proportional to the HCN one, we find C12/C13=89-18+22 at 15° S, and 68-12+16 at 83° N, two values that are statistically consistent. A combination of these results yields a 12C/ 13C value equal to 75±12. This global result, as well as the 15° S one, envelop the value in Titan's methane ( 82.3±1) [Niemann, H.B., and 17 colleagues, 2005. Nature 438, 779-784] measured at 10° S and is slightly lower than the terrestrial inorganic standard value (89). The 14N/ 15N isotopic ratio is found equal to 56-13+16 at 15° S and 56-9+10 at 83° N. Combining the two values yields 14N/ 15N = 56 ± 8, which corresponds to an enrichment in 15N of about 4.9 compared with the terrestrial ratio. These results agree with the values obtained from previous ground-based millimeter observations [Hidayat, T., Marten, A., Bézard, B., Gautier, D., Owen, T., Matthews, H.E., Paubert, G., 1997. Icarus 126, 170-182; Marten, A., Hidayat, T., Biraud, Y., Moreno, R., 2002. Icarus 158, 532-544]. The 15N/ 14N ratio found in HCN is ˜3 times higher than in N 2 [Niemann, H.B., and 17 colleagues, 2005. Nature 438, 779

  9. FIRST MEASUREMENTS OF {sup 15}N FRACTIONATION IN N{sub 2}H{sup +} TOWARD HIGH-MASS STAR-FORMING CORES

    SciTech Connect

    Fontani, F.; Caselli, P.; Bizzocchi, L.; Palau, A.; Ceccarelli, C.

    2015-08-01

    We report on the first measurements of the isotopic ratio {sup 14}N/{sup 15}N in N{sub 2}H{sup +} toward a statistically significant sample of high-mass star-forming cores. The sources belong to the three main evolutionary categories of the high-mass star formation process: high-mass starless cores, high-mass protostellar objects, and ultracompact H ii regions. Simultaneous measurements of the {sup 14}N/{sup 15}N ratio in CN have been made. The {sup 14}N/{sup 15}N ratios derived from N{sub 2}H{sup +} show a large spread (from ∼180 up to ∼1300), while those derived from CN are in between the value measured in the terrestrial atmosphere (∼270) and that of the proto-solar nebula (∼440) for the large majority of the sources within the errors. However, this different spread might be due to the fact that the sources detected in the N{sub 2}H{sup +} isotopologues are more than those detected in the CN ones. The {sup 14}N/{sup 15}N ratio does not change significantly with the source evolutionary stage, which indicates that time seems to be irrelevant for the fractionation of nitrogen. We also find a possible anticorrelation between the {sup 14}N/{sup 15}N (as derived from N{sub 2}H{sup +}) and the H/D isotopic ratios. This suggests that {sup 15}N enrichment could not be linked to the parameters that cause D enrichment, in agreement with the prediction by recent chemical models. These models, however, are not able to reproduce the observed large spread in {sup 14}N/{sup 15}N, pointing out that some important routes of nitrogen fractionation could be still missing in the models.

  10. δ15N of nitrate derived from explosive sources in a karst aquifer beneath the Ammunition Burning Ground, Crane Naval Surface Warfare Center, Indiana, USA

    NASA Astrophysics Data System (ADS)

    DiGnazio, Frank J.; Krothe, Noel C.; Baedke, Steve J.; Spalding, Roy F.

    1998-05-01

    Military institutions involved in the production and demolition of explosives, propellants, and pyrotechnics have the potential to degrade groundwater aquifers through the addition of numerous contaminants including nitrate. A nitrate plume has been identified in a karst aquifer beneath the Ammunition Burning Ground (ABG) at the Crane Naval Surface Warfare Center, Indiana, USA. Wells located in the vicinity of surface impoundments and burn pans used for treatment of explosive materials show the highest concentrations of nitrate ranging from 11.2 to 19.6 mg 1 -1 as NO 3-. Little is known about the isotopic composition of nitrates originating from these processes. Eight wells within the ABG were sampled and analyzed for nitrogen isotopic composition of nitrate. An enrichment in the δ15N ( δ15N = +8.9, +12.0, +13.1, and +13.5‰) occurred at four wells located near the primary areas of disposal activities within the ABG. Four wells located near the outer limits of the ABG had δ15N values significantly lower than those observed in the central area of the ABG ( δ15N = +4.0, +4.1, +4.6, and +2.0‰). Soil samples and burn-pan ash samples were collected and analyzed for the nitrogen isotopic composition of nitrate. Three soil nitrate samples had low δ15N values of -1.7, -1.8, and +2.2‰. The burn-pan ash sample produced nitrate with a δ15N value of +2.9‰. The observed enrichment in δ15N from samples taken from wells located near the ABG has been postulated to be a result of photodegradation or biochemical modification of RDX and TNT contaminated sludges and volatilization of NH 3 in storage lagoons within the ABG.

  11. Preferential flow, nitrogen transformations and 15N balance under urine-affected areas of irrigated and non-irrigated clover-based pastures

    NASA Astrophysics Data System (ADS)

    Pakro, Naser; Dillon, Peter

    1995-12-01

    Urine-affected areas can lead to considerable losses of N by leaching, ammonia volatilisation and denitrification from dairy pastures in the southeast of South Australia. Potable groundwater supplies are considered to have become contaminated by nitrate as a result of leaching from these leguminous pastures. Dairy cow urine, labelled with 15N urea, was applied to micro-plots and mini-lysimeters installed in two adjacent irrigated (white clover-rye grass) and non-irrigated (subterranean clover-annual grasses) paddocks of a dairy farm on four occasions representing different seasonal conditions. These experiments allowed measurement of nitrogen transformations, recovery of 15N in the pasture and soil, and leaching below various depths. Gaseous losses were calculated from the nitrogen balance. The results of the four experiments showed that within a day of urine application up to 40% of the applied urinary-N was leached below a depth of 150 mm as a result of macropore flow in the irrigated paddock, and up to 24% in the non-irrigated one. After application to the irrigated paddock 17% of the urinary-N moved immediately below 300 mm but only 2% below the 450-mm depth. The urinary-N remaining in the soil was converted from urea to ammonium within a day regardless of season. Within the first 7 days of application six times more nitrate was produced in summer than in winter. This has obvious implications for leaching potential. Leaching of 15N from the top 150 mm of soil, following urine applications in all seasons, was between 41% and 62% of the applied 15N in the irrigated paddock and 25-51% in the non-irrigated paddock. However, leaching losses measured at depths of 300 or 450 mm were smaller by a factor of 2-4. The leaching loss of 15N applied in spring in both paddocks was 41% below 150 mm and 12% below 450 mm. Recovery of 15N from the soil-plant system in the 450-nm deep lysimeters was ˜60% of that applied. Estimated ammonia was ˜9% of applied 15N with no paddock

  12. The effect of noncollinearity of 15N-1H dipolar and 15N CSA tensors and rotational anisotropy on 15N relaxation, CSA/dipolar cross correlation, and TROSY.

    PubMed

    Fushman, D; Cowburn, D

    1999-02-01

    Current approaches to 15N relaxation in proteins assume that the 15N-1H dipolar and 15N CSA tensors are collinear. We show theoretically that, when there is significant anisotropy of molecular rotation, different orientations of the two tensors, experimentally observed in proteins, nucleic acids, and small peptides, will result in differences in site-specific correlation functions and spectral densities. The standard treatments of the rates of longitudinal and transverse relaxation of amide 15N nuclei, of the 15N CSA/15N-1H dipolar cross correlation, and of the TROSY experiment are extended to account for the effect of noncollinearity of the 15N-1H dipolar and 15N CSA (chemical shift anisotropy) tensors. This effect, proportional to the degree of anisotropy of the overall motion, (D parallel/D perpendicular - 1), is sensitive to the relative orientation of the two tensors and to the orientation of the peptide plane with respect to the diffusion coordinate frame. The effect is negligible at small degrees of anisotropy, but is predicted to become significant for D parallel/D perpendicular > or = 1.5, and at high magnetic fields. The effect of noncollinearity of 15N CSA and 15N-1H dipolar interaction is sensitive to both gross (hydrodynamic) properties and atomic-level details of protein structure. Incorporation of this effect into relaxation data analysis is likely to improve both precision and accuracy of the derived characteristics of protein dynamics, especially at high magnetic fields and for molecules with a high degree of anisotropy of the overall motion. The effect will also make TROSY efficiency dependent on local orientation in moderately anisotropic systems.

  13. Infrared band intensities in ammonium hydroxide and ammonium salts

    NASA Technical Reports Server (NTRS)

    Sethna, P. P.; Downing, H. D.; Pinkley, L. W.; Williams, D.

    1978-01-01

    We have applied Kramers-Kronig analysis to reflection spectra to determine the optical constants of ammonium hydroxide and of aqueous solutions of ammonium chloride and bromide. From considerations of the absorption indices k(nu) we conclude that ammonium hydroxide consists of a solution of NH3 in water, in which NH3 molecules are hydrogen bonded to neighboring water molecules. The spectrum of ammonium hydroxide differs from the spectra of ammonium salts, in which bands characteristic of NH4(+) ions are prominent. The existence of ammonium hydroxide as an aerosol in planetary atmospheres is briefly discussed

  14. Linking nitrogen dynamics to climate variability off central California: a 51 year record based on 15N/ 14N in CalCOFI zooplankton

    NASA Astrophysics Data System (ADS)

    Rau, Greg H.; Ohman, Mark D.; Pierrot-Bults, Annelies

    2003-08-01

    Long-term variability in zooplankton 15N/ 14N was investigated in two species of calanoid copepods ( Calanus pacificus and Eucalanus californicus) and two chaetognaths ( Sagitta bierii and Sagitta euneritica) sampled in the spring of selected years from 1951 to 2001 off the central California coast. No statistically significant trend in 15N/ 14N was detected for any of the four species, with isotopic ratios in 2001 resembling those in copepods and chaetognaths sampled five decades earlier. Zooplankton body lengths also showed no long-term trends. With respect to proposed regime shifts in this region, heterogeneity in 15N/ 14N was detected only for S. bierii when comparing the periods 1951-1975, 1978-1998, and 1999-2001. In this species the 15N/ 14N in the most recent, brief period (1999-2001) averaged slightly lower than in the previous period. Three of the four species ( C. pacificus, S. bierii, and S. euneritica) showed significant increases in 15N/ 14N during major El Niños. El Niño-related enrichment in 15N could arise as a consequence of increased nitrate demand:supply at the base of the food web or advection of 15N-enriched nitrate from more southerly waters. While a range of physical and climate indices were evaluated, anomalies of 15N/ 14N from the long-term mean were found to be significantly related only to: (i) the Southern Oscillation Index in the case of both chaetognath species, (ii) a regional surface water temperature record ( S. bierii only), (iii) an index of wind-driven coastal upwelling for the surface-dwelling C. pacificus, and (iv) variability in the Pacific Decadal Oscillation for the somewhat deeper-dwelling E. californicus. The relationships among each species' 15N/ 14N averaged over the total sampling period was: E. californicus≈C. pacificus≪S. euneritica < S. bierii, consistent with trophic 15N biomagnification and the predatory nature of Sagitta.

  15. The fate of 15N-nitrate in mesocosms from five European peatlands differing in long-term nitrogen deposition rate

    NASA Astrophysics Data System (ADS)

    Zając, K.; Blodau, C.

    2016-02-01

    Elevated nitrogen (N) deposition changes the retention, transformation, and fluxes of N in ombrotrophic peatlands. To evaluate such effects we applied a 15N tracer (NH4 15NO3) at a rate of 2.3 g N m-2 yr-1 to mesocosms of five European peatlands with differing long-term N deposition rates for a period of 76 days of dry and 90 days of wet conditions. We determined background N content and moss length growth, and recovered the 15N tracer from the mosses, graminoids, shrubs, the peat, and dissolved N. Background N contents in Sphagnum mosses increased from 5.5 (Degerö Stormyr, deposition < 0.2 g N m-2 yr-1) up to 12.2 mg g-1 (Frölichshaier Sattelmoor, 4.7-6.0 g N m-2 yr-1). In peat from Degerö, nitrate and ammonium concentrations were below 3 mg L-1, whereas up to 30 (nitrate) and 11 mg L-1 (ammonium) was found in peat from Frölichshaier Sattelmoor. Sphagnum mosses (down to 5 cm below surface) generally intercepted large amounts of 15N (0.2-0.35 mg g-1) and retained the tracer most effectively relative to their biomass. Similar quantities of the 15N were recovered from the peat, followed by shrubs, graminoids, and the dissolved pool. At the most polluted sites we recovered more 15N from shrubs (up to 12.4 %) and from nitrate and ammonium (up to 0.7 %). However, no impact of N deposition on 15N retention by Sphagnum could be identified and their length growth was highest under high N background deposition. Our experiment suggests that the decline in N retention at levels above ca. 1.5 g m-2 yr-1, as expressed by elevated near-surface peat N content and increased dissolved N concentrations, is likely more modest than previously thought. This conclusion is related to the finding that Sphagnum species can apparently thrive at elevated long-term N deposition rates in European peatlands.

  16. Measurement of multiple psi torsion angles in uniformly 13C,15N-labeled alpha-spectrin SH3 domain using 3D 15N-13C-13C-15N MAS dipolar-chemical shift correlation spectroscopy.

    PubMed

    Ladizhansky, Vladimir; Jaroniec, Christopher P; Diehl, Annette; Oschkinat, Hartmut; Griffin, Robert G

    2003-06-01

    We demonstrate the simultaneous measurement of several backbone torsion angles psi in the uniformly (13)C,(15)N-labeled alpha-Spectrin SH3 domain using two different 3D 15N-13C-13C-15N dipolar-chemical shift magic-angle spinning (MAS) NMR experiments. The first NCCN experiment utilizes double quantum (DQ) spectroscopy combined with the INADEQUATE type 13C-13C chemical shift correlation. The decay of the DQ coherences formed between 13C'(i) and 13C(alphai) spin pairs is determined by the "correlated" dipolar field due to 15N(i)-13C(alphai) and 13C'(i)-15N(i+1) dipolar couplings and is particularly sensitive to variations of the torsion angle in the regime |psi| > 140 degrees. However, the ability of this experiment to constrain multiple psi-torsion angles is limited by the resolution of the 13C(alpha)-(13)CO correlation spectrum. This problem is partially addressed in the second approach described here, which is an NCOCA NCCN experiment. In this case the resolution is enhanced by the superior spectral dispersion of the 15N resonances present in the 15N(i+1)-13C(alphai) part of the NCOCA chemical shift correlation spectrum. For the case of the 62-residue alpha-spectrin SH3 domain, we determined 13 psi angle constraints with the INADEQUATE NCCN experiment and 22 psi constraints were measured in the NCOCA NCCN experiment.

  17. 70. INTERIOR VIEW OF AMMONIUM NITRATE HOUSE, LOOKING AT AMMONIUM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    70. INTERIOR VIEW OF AMMONIUM NITRATE HOUSE, LOOKING AT AMMONIUM NITRATE IN STORAGE. APRIL 18, 1919. - United States Nitrate Plant No. 2, Reservation Road, Muscle Shoals, Muscle Shoals, Colbert County, AL

  18. Nitrogen Transformations in Wetland Soil Cores Measured by (sup15)N Isotope Pairing and Dilution at Four Infiltration Rates

    PubMed Central

    Stepanauskas, R.; Davidsson, E. T.; Leonardson, L.

    1996-01-01

    The effect of water infiltration rate (IR) on nitrogen cycling in a saturated wetland soil was investigated by applying a (sup15)N isotope dilution and pairing method. Water containing [(sup15)N]nitrate was infiltrated through 10-cm-long cores of sieved and homogenized soil at rates of 72, 168, 267, and 638 mm day(sup-1). Then the frequencies of (sup30)N(inf2), (sup29)N(inf2), (sup15)NO(inf3)(sup-), and (sup15)NH(inf4)(sup+) in the outflow water were measured. This method allowed simultaneous determination of nitrification, coupled and uncoupled denitrification, and nitrate assimilation rates. From 3% (at the highest IR) to 95% (at the lowest IR) of nitrate was removed from the water, mainly by denitrification. The nitrate removal was compensated for by the net release of ammonium and dissolved organic nitrogen. Lower oxygen concentrations in the soil at lower IRs led to a sharper decrease in the nitrification rate than in the ammonification rate, and, consequently, more ammonium leaked from the soil. The decreasing organic-carbon-to-nitrogen ratio (from 12.8 to 5.1) and the increasing light A(inf250)/A(inf365) ratio (from 4.5 to 5.2) indicated an increasing bioavailability of the outflowing dissolved organic matter with increasing IR. The efflux of nitrous oxide was also very sensitive to IR and increased severalfold when a zone of low oxygen concentration was close to the outlet of the soil cores. N(inf2)O then constituted 8% of the total gaseous N lost from the soil. PMID:16535352

  19. Glufosinate-ammonium

    Integrated Risk Information System (IRIS)

    Glufosinate - ammonium ; CASRN 77182 - 82 - 2 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarci

  20. Exploring the ammonium and nitrate transport of marine phytoplankton with nutrient analogues

    SciTech Connect

    Balch, W.M.

    1985-01-01

    Radiolabelled methylamine, an ammonium analogue and chlorate, a nitrate analogue, were transported constitutely by laboratory and field populations of phytoplakton. There was no effect of light on the transport of methylamine or chlorate which is contrary to similar measurements made with /sup 15/N-NH/sub 4//sup +/ and /sup 15/N-NO/sub 3//sup -/. The discrepancy appears to result from the fact that the analogues are only transported, while /sup 15/N-NH/sub 4/ and /sup 15/N-NO/sub 3//sup -/ are both transported and assimilated. Transport of ammonium and nitrate appeared to be active; it was against typical values of algal electrochemical gradients. Influx and efflux rates of methylamine and chlorate were measured in pulse-chase experiments; efflux rates increased as intracellular pools filled and net uptake slowed after approximately one to six hours. The pulse-chase experiments indicated that methylamine and chlorate (hence ammonium and nitrate) were stored in two intracellular compartments of diatoms, probably the vacuole and cytoplasm. Laboratory and field experiments demonstrated that chlorate transport by phytoplankton was inhibited when ambient ammonium or nitrite concentrations were high.

  1. 15N2 formation and fast oxygen isotope exchange during pulsed 15N18O exposure of MnOx/CeO2

    SciTech Connect

    Kwak, Ja Hun; Szanyi, Janos

    2014-12-23

    Pulsing 15N18O onto an annealed 1% Mn16Ox/Ce16O2 catalyst resulted in very fast oxygen isotope exchange and 15N2 formation at 295 K. In the 1st 15N18O pulse, due to the presence of large number of surface oxygen defects, extensive 15N218O and 15N2 formations were observed. In subsequent pulses oxygen isotope exchange dominated as a result of highly labile oxygen in the oxide. We gratefully acknowledge the US Department of Energy (DOE), Office of Energy Efficiency and Renewable Energy/Vehicle Technologies Program for the support of this work. The research described in this paper was performed at the Environmental Molecular Sciences Laboratory (EMSL), a national scientific user facility sponsored by the DOE’s Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory (PNNL). PNNL is operated for the US DOE by Battelle.

  2. Acid-base interactions and secondary structures of poly-L-lysine probed by 15N and 13C solid state NMR and Ab initio model calculations.

    PubMed

    Dos, Alexandra; Schimming, Volkmar; Tosoni, Sergio; Limbach, Hans-Heinrich

    2008-12-11

    The interactions of the 15N-labeled amino groups of dry solid poly-L-lysine (PLL) with various halogen and oxygen acids HX and the relation to the secondary structure have been studied using solid-state 15N and 13C CPMAS NMR spectroscopy (CP = cross polarization and MAS = magic angle spinning). For comparison, 15N NMR spectra of an aqueous solution of PLL were measured as a function of pH. In order to understand the effects of protonation and hydration on the 15N chemical shifts of the amino groups, DFT and chemical shielding calculations were performed on isolated methylamine-acid complexes and on periodic halide clusters of the type (CH3NH3(+)X(-))n. The combined experimental and computational results reveal low-field shifts of the amino nitrogens upon interaction with the oxygen acids HX = HF, H2SO4, CH3COOH, (CH3)2POOH, H3PO4, HNO3, and internal carbamic acid formed by reaction of the amino groups with gaseous CO2. Evidence is obtained that only hydrogen-bonded species of the type (Lys-NH2***H-X)n are formed in the absence of water. 15N chemical shifts are maximum when H is located in the hydrogen bond center and then decrease again upon full protonation, as found for aqueous solution at low pH. By contrast, halogen acids interact in a different way. They form internal salts of the type (Lys-NH3(+)X(-))n via the interaction of many acid-base pairs. This salt formation is possible only in the beta-sheet conformation. By contrast, the formation of hydrogen-bonded complexes can occur both in beta-sheet domains as well as in alpha-helical domains. The 15N chemical shifts of the protonated ammonium groups increase when the size of the interacting halogen anions is increased from chloride to iodide and when the number of the interacting anions is increased. Thus, the observed high-field 15N shift of ammonium groups upon hydration is the consequence of replacing interacting halogen atoms by oxygen atoms.

  3. What can Δ 15N and Δ 18O isotopes tell us about sources, transport, and fate of nitrate in the Mississippi River Basin?

    NASA Astrophysics Data System (ADS)

    Battaglin, W. A.

    2003-12-01

    Water and nutrients, primarily nitrate (NO3) in Mississippi River discharge, affect the size and severity of the Gulf of Mexico hypoxic (depleted dissolved oxygen) zone. Approximately 120 water samples were collected from 16 sites on small streams and 6 sites on large rivers within the Mississippi River Basin in 1997-98 to see if NO3 sources and transformations can be identified using the stable isotopic ratios Δ 15N and Δ 18O. Results from Lagrangian sampling at the large river sites indicate that nitrate mass decreases slightly, while Δ 15N and Δ 18O isotope ratios are unchanged in the 1500 river kilometers between the Upper Mississippi-Ohio River confluence and the Gulf of Mexico. Results also show that Δ 15N and Δ 18O values from small streams draining lands dominated by row crops or livestock tended to be distinct from those dominated by urban or undeveloped land. Mean Δ 15N values at the 16 sites on small streams were most strongly correlated (Pearson's r) with manure production rate (0.64), percent residential land use (-0.45), and urea use rate (0.43). The best multiple linear regression (MLR) model for mean Δ 15N values (r2=0.69) used manure production rate and ammonium nitrate use rate as explanatory variables. Mean Δ 18O values were most strongly correlated with percent wetlands (0.72), mean NO3 concentration (-0.71), and percent residential land use (0.58). The best MLR model for mean Δ 18O values (r2=0.85) used percent residential land use, percent wetlands, and ammonium nitrate use rate as explanatory variables. Mean NO3 concentrations were most strongly correlated with percent row-crops land use (0.84), nitrogen-fertilizer use rate (0.74), and hog-manure production rate (0.66). The best MLR model for mean NO3 concentration (r2=0.85) used percent row-crops land use and percent grain-crops land use as explanatory variables. MLR equations developed from the 16 smaller streams were used to predict mean Δ 15N and Δ 18O values and NO3

  4. Millipedes and earthworms increase the decomposition rate of 15N-labelled winter rape litter in an arable field.

    PubMed

    Martens, H; Alphei, J; Schaefer, M; Scheu, S

    2001-01-01

    Effects of millipedes and earthworms on the decomposition of 15N-labelled litter of winter oilseed rape were investigated in a microcosm field experiment over a period of 264 days on an oat field near Göttingen managed by integrated farming. A total of 32 microcosms were filled with defaunated soil. 15N-labelled rape litter was placed either on top of the soil or buried into the soil simulating mulching and ploughing, respectively. To the microcosms nine adult individuals of Blaniulus guttulatus (Diplopoda) and two of Aporrectodea caliginosa (Lumbricidae) were added separately or in combination. In general, the presence of the animals accelerated the decomposition rate of the litter material. The effects were most pronounced in the presence of Aporrectodea caliginosa. The total amount of nitrate, ammonium and the amount of 35N leached from the microcosms was increased in the presence of earthworms or of both earthworms and millipedes. Both species proved to be important members of the detritus food web of the agricultural system studied.

  5. Synthesis of 13C and 15N labeled 2,4-dinitroanisole.

    PubMed

    Jagadish, Bhumasamudram; Field, Jim A; Chorover, Jon; Sierra-Alvarez, Reyes; Abrell, Leif; Mash, Eugene A

    2014-05-30

    Syntheses of [(13)C6]-2,4-dinitroanisole (ring-(13)C6) from [(13)C6]-anisole (ring-(13)C6) and [(15)N2]-2,4-dinitroanisole from anisole using in situ generated acetyl nitrate and [(15)N]-acetyl nitrate, respectively, are described. Treatment of [(13)C6]-anisole (ring-(13)C6) with acetyl nitrate generated in 100% HNO3 gave [(13)C6]-2,4-dinitroanisole (ring-(13)C6) in 83% yield. Treatment of anisole with [(15)N]-acetyl nitrate generated in 10 N [(15)N]-HNO3 gave [(15)N2 ]-2,4-dinitroanisole in 44% yield after two cycles of nitration. Byproducts in the latter reaction included [(15)N]-2-nitroanisole and [(15)N]-4-nitroanisole.

  6. The “Speedy” Synthesis of Atom-Specific 15N Imino/Amido-Labeled RNA

    PubMed Central

    Kreutz, Christoph; Micura, Ronald

    2016-01-01

    Although numerous reports on the synthesis of atom-specific 15N-labeled nucleosides exist, fast and facile access to the corresponding phosphoramidites for RNA solid-phase synthesis is still lacking. This situation represents a severe bottleneck for NMR spectroscopic investigations on functional RNAs. Here, we present optimized procedures to speed up the synthesis of 15N(1) adenosine and 15N(1) guanosine amidites, which are the much needed counterparts of the more straightforward-to-achieve 15N(3) uridine and 15N(3) cytidine amidites in order to tap full potential of 1H/15N/15N-COSY experiments for directly monitoring individual Watson–Crick base pairs in RNA. Demonstrated for two preQ1 riboswitch systems, we exemplify a versatile concept for individual base-pair labeling in the analysis of conformationally flexible RNAs when competing structures and conformational dynamics are encountered. PMID:26237536

  7. Quantification of the contribution of biological nitrogen fixation to tropical green manure crops and the residual benefit to a subsequent maize crop using 15N-isotope techniques.

    PubMed

    Ramos, M G; Villatoro, M A; Urquiaga, S; Alves, B J; Boddey, R M

    2001-10-01

    In this study the contribution of biological N2 fixation (BNF) to leguminous green manures was quantified in the field at different sites with different 15N methodologies. In the first experiment, conducted on a Terra Roxa soil in Cuba, the BNF contribution to three legumes (Crotalaria juncea, Mucuna aterrima and Canavalia ensiformis) was quantified by applying 15N-labelled ammonium sulphate to the soil. The second experiment was planted in a very low fertility sandy soil near Rio de Janeiro, and the 15N natural abundance technique was applied to quantify BNF in C. juncea, M. niveum and soybean. In both studies the advantages of using several non-N2-fixing reference plants was apparent and despite the much greater accumulation of the C. juncea in the experiment performed on the fertile soil of Cuba, the above ground contributions of BNF at both sites were similar (40-80 kg N x ha(-1)) and greater than for the other legumes. In a further experiment the possible contribution of root-derived N to the soil/plant system of two of the legumes was quantified using a 15N-leaf-labelling technique performed in pots. The results of this study suggested that total below-ground N could constitute as much as 39 to 49% of the total N accumulated by the legume crops. PMID:11566383

  8. Fate of nitrogen deposition and decomposed nitrogen from litter in a 15N-tracer mesocosm experiment

    NASA Astrophysics Data System (ADS)

    Nair, R.; Perks, M.; Mencuccini, M.

    2013-12-01

    Atmospheric deposition of anthropogenic-derived nitrogen may be a major driver of the 0.6-0.7 Pg y-1 increase in the carbon sink in historically N-limited northern and boreal forests, but the magnitude of its effect is still uncertain. A strong effect depends on the allocation of N to trees, because of their high C:N ratio in woody tissues, and isotope tracer experiments have shown that the majority of 15N tracers applied directly to the soil are lost via leeching or retained in soil pools rather than being acquired by tree root systems. However, ambient anthropogenic inputs of N to these systems are transported in the atmosphere and intercepted by foliage before they reach the soil system, while labelled fertilization experiments also can only explicitly trace the fate of the 15N-tracer from deposition, as opposed to changes in the fate of N from litter, where decomposition rates may be enhanced at low ambient levels of deposition, affecting the availability of N from this pool for tree nutrition. We present initial results from a potted Sitka Spruce mesocosm 15N-tracer experiment where ambient nitrogen deposition was supplemented with a minor (0.4 kg ha-1 y-1) input of additional N, applied to either the soil or the foliage. Either this deposition, or litter in the pots, was enriched in 15N, allowing the fate of the isotope from two different methods of deposition to be compared with that of nitrogen released from the litter under the deposition treatment.

  9. Assignment of the sup 1 H and sup 15 N NMR spectra of Rhodobacter capsulatus ferrocytochrome c sub 2

    SciTech Connect

    Gooley, P.R.; Caffrey, M.S.; Cusanovich, M.A.; MacKenzie, N.E. )

    1990-03-06

    The peptide resonances of the {sup 1}H and {sup 15}N nuclear magnetic resonance spectra of ferrocytochrome c{sub 2} from Rhodobacter capsulatus are sequentially assigned by a combination of 2D {sup 1}H-{sup 1}H and {sup 1}H-{sup 15}N spectroscopy, the latter performed on {sup 15}N-enriched protein. Short-range nuclear Overhauser effect (NOE) data show {alpha}-helices from residues 3-17, 55-65, 69-88, and 103-115. Within the latter two {alpha}-helices, there are three single 3{sub 10} turns, 70-72, 76-78, and 107-109. In addition {alpha}H-NH{sub i+1} and {alpha}H-NH{sub i+2} NOEs indicate that the N-terminal helix (3-17) is distorted. Compared to horse or tuna cytochrome c and cytochrome c{sub 2} of Rhodospirillium rubrum, there is a 6-residue insertion at residues 23-29 in R. capsulatus cytochrome c{sub 2}. The NOE data show that this insertion forms a loop, probably an {Omega} loop. {sup 1}H-{sup 15}N heteronuclear multiple quantum correlation experiments are used to follow NH exchange over a period of 40 h. As the 2D spectra are acquired in short time periods (30 min), rates for intermediate exchanging protons can be measured. Comparison of the NH exchange data for the N-terminal helix of cytochrome c{sub 2} of R. capsulatus with the highly homologous horse heart cytochrome c shows that this helix is less stable in cytochrome c{sub 2}.

  10. Prevalence of anaerobic ammonium-oxidizing bacteria in contaminated groundwater.

    PubMed

    Moore, Tara A; Xing, Yangping; Lazenby, Brent; Lynch, Michael D J; Schiff, Sherry; Robertson, William D; Timlin, Robert; Lanza, Sadia; Ryan, M Cathryn; Aravena, Ramon; Fortin, Danielle; Clark, Ian D; Neufeld, Josh D

    2011-09-01

    Anaerobic ammonium-oxidizing (anammox) bacteria perform an important step in the global nitrogen cycle: anaerobic oxidation of ammonium and reduction of nitrite to form dinitrogen gas (N(2)). Anammox organisms appear to be widely distributed in natural and artificial environments. However, their roles in groundwater ammonium attenuation remain unclear and only limited biomarker-based data confirmed their presence prior to this study. We used complementary molecular and isotope-based methods to assess anammox diversity and activity occurring at three ammonium-contaminated groundwater sites: quantitative PCR, denaturing gradient gel electrophoresis, sequencing of 16S rRNA genes, and (15)N-tracer incubations. Here we show that anammox performing organisms were abundant bacterial community members. Although all sites were dominated by Candidatus Brocadia-like sequences, the community at one site was particularly diverse, possessing four of five known genera of anammox bacteria. Isotope data showed that anammox produced up to 18 and 36% of N(2) at these sites. By combining molecular and isotopic results we have demonstrated the diversity, abundance, and activity of these autotrophic bacteria. Our results provide strong evidence for their important biogeochemical role in attenuating groundwater ammonium contamination.

  11. Heavy water and 15N labeling with NanoSIMS analysis reveals growth-rate dependent metabolic heterogeneity in chemostats

    PubMed Central

    McGlynn, Shawn E.; Green-Saxena, Abigail

    2015-01-01

    To measure single cell microbial activity and substrate utilization patterns in environmental systems, we employ a new technique using stable isotope labeling of microbial populations with heavy water (a passive tracer) and 15N ammonium in combination with multi-isotope imaging mass spectrometry. We demonstrate simultaneous NanoSIMS analysis of hydrogen, carbon and nitrogen at high spatial and mass resolution, and report calibration data linking single cell isotopic compositions to the corresponding bulk isotopic equivalents for Pseudomonas aeruginosa and Staphylococcus aureus. Our results show that heavy water is capable of quantifying in situ single cell microbial activities ranging from generational time scales of minutes to years, with only light isotopic incorporation (∼0.1 atom % 2H). Applying this approach to study the rates of fatty acid biosynthesis by single cells of S. aureus growing at different rates in chemostat culture (∼6 hours, 1 day and 2 week generation times), we observe the greatest anabolic activity diversity in the slowest growing populations. By using heavy water to constrain cellular growth activity, we can further infer the relative contributions of ammonium vs. amino acid assimilation to the cellular nitrogen pool. The approach described here can be applied to disentangle individual cell activities even in nutritionally complex environments. PMID:25655651

  12. Comparing the Influence of Wildfire and Prescribed Burns on Watershed Nitrogen Biogeochemistry Using 15N Natural Abundance in Terrestrial and Aquatic Ecosystem Components

    PubMed Central

    Stephan, Kirsten; Kavanagh, Kathleen L.; Koyama, Akihiro

    2015-01-01

    We evaluated differences in the effects of three low-severity spring prescribed burns and four wildfires on nitrogen (N) biogeochemistry in Rocky Mountain headwater watersheds. We compared paired (burned/unburned) watersheds of four wildfires and three spring prescribed burns for three growing seasons post-fire. To better understand fire effects on the entire watershed ecosystem, we measured N concentrations and δ15N in both the terrestrial and aquatic ecosystems components, i.e., soil, understory plants in upland and riparian areas, streamwater, and in-stream moss. In addition, we measured nitrate reductase activity in foliage of Spiraea betulifolia, a dominant understory species. We found increases of δ15N and N concentrations in both terrestrial and aquatic ecosystem N pools after wildfire, but responses were limited to terrestrial N pools after prescribed burns indicating that N transfer from terrestrial to aquatic ecosystem components did not occur in low-severity prescribed burns. Foliar δ15N differed between wildfire and prescribed burn sites; the δ15N of foliage of upland plants was enriched by 2.9 ‰ (difference between burned and unburned watersheds) in the first two years after wildfire, but only 1.3 ‰ after prescribed burns. In-stream moss δ15N in wildfire-burned watersheds was enriched by 1.3 ‰, but there was no response by moss in prescription-burned watersheds, mirroring patterns of streamwater nitrate concentrations. S. betulifolia showed significantly higher nitrate reductase activity two years after wildfires relative to corresponding unburned watersheds, but no such difference was found after prescribed burns. These responses are consistent with less altered N biogeochemistry after prescribed burns relative to wildfire. We concluded that δ15N values in terrestrial and aquatic plants and streamwater nitrate concentrations after fire can be useful indicators of the magnitude and duration of fire effects and the fate of post

  13. Comparing the influence of wildfire and prescribed burns on watershed nitrogen biogeochemistry using 15N natural abundance in terrestrial and aquatic ecosystem components.

    PubMed

    Stephan, Kirsten; Kavanagh, Kathleen L; Koyama, Akihiro

    2015-01-01

    We evaluated differences in the effects of three low-severity spring prescribed burns and four wildfires on nitrogen (N) biogeochemistry in Rocky Mountain headwater watersheds. We compared paired (burned/unburned) watersheds of four wildfires and three spring prescribed burns for three growing seasons post-fire. To better understand fire effects on the entire watershed ecosystem, we measured N concentrations and δ15N in both the terrestrial and aquatic ecosystems components, i.e., soil, understory plants in upland and riparian areas, streamwater, and in-stream moss. In addition, we measured nitrate reductase activity in foliage of Spiraea betulifolia, a dominant understory species. We found increases of δ15N and N concentrations in both terrestrial and aquatic ecosystem N pools after wildfire, but responses were limited to terrestrial N pools after prescribed burns indicating that N transfer from terrestrial to aquatic ecosystem components did not occur in low-severity prescribed burns. Foliar δ15N differed between wildfire and prescribed burn sites; the δ15N of foliage of upland plants was enriched by 2.9 ‰ (difference between burned and unburned watersheds) in the first two years after wildfire, but only 1.3 ‰ after prescribed burns. In-stream moss δ15N in wildfire-burned watersheds was enriched by 1.3 ‰, but there was no response by moss in prescription-burned watersheds, mirroring patterns of streamwater nitrate concentrations. S. betulifolia showed significantly higher nitrate reductase activity two years after wildfires relative to corresponding unburned watersheds, but no such difference was found after prescribed burns. These responses are consistent with less altered N biogeochemistry after prescribed burns relative to wildfire. We concluded that δ15N values in terrestrial and aquatic plants and streamwater nitrate concentrations after fire can be useful indicators of the magnitude and duration of fire effects and the fate of post

  14. Comparing the influence of wildfire and prescribed burns on watershed nitrogen biogeochemistry using 15N natural abundance in terrestrial and aquatic ecosystem components.

    PubMed

    Stephan, Kirsten; Kavanagh, Kathleen L; Koyama, Akihiro

    2015-01-01

    We evaluated differences in the effects of three low-severity spring prescribed burns and four wildfires on nitrogen (N) biogeochemistry in Rocky Mountain headwater watersheds. We compared paired (burned/unburned) watersheds of four wildfires and three spring prescribed burns for three growing seasons post-fire. To better understand fire effects on the entire watershed ecosystem, we measured N concentrations and δ15N in both the terrestrial and aquatic ecosystems components, i.e., soil, understory plants in upland and riparian areas, streamwater, and in-stream moss. In addition, we measured nitrate reductase activity in foliage of Spiraea betulifolia, a dominant understory species. We found increases of δ15N and N concentrations in both terrestrial and aquatic ecosystem N pools after wildfire, but responses were limited to terrestrial N pools after prescribed burns indicating that N transfer from terrestrial to aquatic ecosystem components did not occur in low-severity prescribed burns. Foliar δ15N differed between wildfire and prescribed burn sites; the δ15N of foliage of upland plants was enriched by 2.9 ‰ (difference between burned and unburned watersheds) in the first two years after wildfire, but only 1.3 ‰ after prescribed burns. In-stream moss δ15N in wildfire-burned watersheds was enriched by 1.3 ‰, but there was no response by moss in prescription-burned watersheds, mirroring patterns of streamwater nitrate concentrations. S. betulifolia showed significantly higher nitrate reductase activity two years after wildfires relative to corresponding unburned watersheds, but no such difference was found after prescribed burns. These responses are consistent with less altered N biogeochemistry after prescribed burns relative to wildfire. We concluded that δ15N values in terrestrial and aquatic plants and streamwater nitrate concentrations after fire can be useful indicators of the magnitude and duration of fire effects and the fate of post

  15. Comparing isotope signatures of prey fish: does gut removal affect δ13C or δ15N?

    USGS Publications Warehouse

    Chipps, Steven R.; Fincel, Mark J.; VanDeHey, Justin A.; Wuestewald, Andrew

    2011-01-01

    Stable isotope analysis is a quick and inexpensive method to monitor the effects of food web changes on aquatic communities. Traditionally, whole specimens have been used when determining isotope composition of prey fish or age-0 recreational fishes. However, gut contents of prey fish could potentially alter isotope composition of the specimen, especially when recent foraging has taken place or when the gut contains non-assimilated material that would normally pass through fishes undigested. To assess the impacts of gut content on prey fish isotope signatures, we examined the differences in isotopic variation of five prey fish species using whole fish, whole fish with the gut contents removed, and dorsal muscle only. We found significant differences in both δ15N and δ13C between the three tissue treatments. In most cases, muscle tissue was enriched compared to whole specimens or gut-removed specimens. Moreover, differences in mean δ15N within a species were up to 2‰ among treatments. This would result in a change of over half a trophic position (TP) based on a 3.4‰ increase per trophic level. However, there were no apparent relationships between tissue isotope values in fish with increased gut fullness (more prey tissue present). We suggest that muscle tissue should be used as the standard tissue for determining isotope composition of prey fish or age-0 recreational fishes, especially when determining enrichment for mixing models, calculating TP, or constructing aquatic food webs.

  16. Numerical evaluation of subsoil diffusion of (15) N labelled denitrification products during employment of the (15) N gas flux method in the field

    NASA Astrophysics Data System (ADS)

    Well, Reinhard; Buchen, Caroline; Lewicka-Szczebak, Dominika; Ruoss, Nicolas

    2016-04-01

    Common methods for measuring soil denitrification in situ include monitoring the accumulation of 15N labelled N2 and N2O evolved from 15N labelled soil nitrate pool in soil surface chambers. Gas diffusion is considered to be the main accumulation process. Because accumulation of the gases decreases concentration gradients between soil and chamber over time, gas production rates are underestimated if calculated from chamber concentrations. Moreover, concentration gradients to the non-labelled subsoil exist, inevitably causing downward diffusion of 15N labelled denitrification products. A numerical model for simulating gas diffusion in soil was used in order to determine the significance of this source of error. Results show that subsoil diffusion of 15N labelled N2 and N2O - and thus potential underestimation of denitrification derived from chamber fluxes - increases with cover closure time as well as with increasing diffusivity. Simulations based on the range of typical gas diffusivities of unsaturated soils show that the fraction of subsoil diffusion after chamber closure for 1 hour is always significant with values up to >30 % of total production of 15N labelled N2 and N2O. Field experiments for measuring denitrification with the 15N gas flux method were conducted. The ability of the model to predict the time pattern of gas accumulation was evaluated by comparing measured 15N2 concentrations and simulated values.

  17. A 115-year δ15N record of cumulative nitrogen pollution in California serpentine grasslands

    NASA Astrophysics Data System (ADS)

    Vallano, D.; Zavaleta, E. S.

    2010-12-01

    Until the 1980s, California’s biodiverse serpentine grasslands were threatened primarily by development and protected by reserve creation. However, nitrogen (N) fertilization due to increasing fossil fuel emissions in the expanding Bay Area is thought to be contributing to rapid, recent invasion of these ecosystems by exotic annual grasses that are displacing rare and endemic serpentine species. Documenting the cumulative effects of N deposition in this ecosystem can direct policy and management actions to mitigate the role of N deposition in its transformation. Natural abundance stable isotopes of N in vegetation have been increasingly used as bio-indicators of N deposition patterns and subsequent changes to plant N cycling and assimilation. However, the long-term record of atmospheric reactive N enrichment and the resulting changes in ecosystem N dynamics have yet to be adequately reconstructed in many ecosystems. Museum archives of vascular plant tissue are valuable sources of materials to reconstruct temporal and spatial isotopic patterns of N inputs to ecosystems. Here, we present N stable isotope data from archived and current specimens of an endemic California serpentine grassland species, leather oak (Quercus durata), since 1895 across the greater San Francisco Bay region. We measured spatial and temporal trends in stable isotope composition (δ15N and δ13C) and concentration (%N and %C) of historical and current samples of leather oak leaves from sites within the Bay Area, impacted by increasing development, and sites northeast of the Bay Area, with significantly lower rates of urbanization and industrialization. Specifically, we sampled dry museum and fresh leaf specimens from serpentine sites within Lake (n=27) and Santa Clara (n=30) counties dating from 1895 to 2010. Leaf δ15N values were stable from 1895 to the 1950s and then decreased strongly throughout the last 50 years as fossil fuel emissions rapidly increased in the Bay Area, indicating that

  18. Millimeter-Wave Observations of Circumstellar 14N/15N and 12C/13C Ratios: New Insights into J-Type Stars

    NASA Astrophysics Data System (ADS)

    Adande, Gilles; Ziurys, Lucy M.; Woolf, Neville

    2016-06-01

    Measurements of 14N/15N and 12C/13C isotopic ratios have been conducted towards circumstellar envelopes of a sample of evolved stars using the J = 3→2 rotational transitions of the isotopologues of HCN, observed with the Submillimeter Telescope (SMT) of the Arizona Radio Observatory (ARO). Towards the J-type stars Y CVn and RY Dra, where 12C/13C ~ 3, the 14N/15N ratios were found to be 120-180 and 225, respectively. The 14N/15N ratio is thus anomalously low relative to interstellar values and a factor ~100 lower than equilibrium values predicted from the CNO cycle. Combining these results with previous chemical and isotopic prior observations of these stars, we conclude that two anomalous behaviors are likely to have occurred in Y CVn and RY Dra. First, the stellar envelope failed to participate in the normal mixing seen in low mass red giants, in which C and then O are substantially converted to N. Secondly, both the carbon enrichment and anomalous isotopic composition of both 13C and15N could have been caused by a plume of hot gas, hydrogen poor but enriched in 12C, from a helium flash mixing into the envelope.

  19. Root Environment Acidity as a Regulatory Factor in Ammonium Assimilation by the Bean Plant 1

    PubMed Central

    Barker, A. V.; Volk, R. J.; Jackson, W. A.

    1966-01-01

    Previous experiments have revealed a much greater efficiency of ammonium utilization by bean plants (Phaseolus vulgaris L.) when the acidity of the ambient medium was maintained at near-neutral conditions with carbonates or hydroxides. The present investigation, in which 15N-labeled ammonium was used, permitted an assessment of the origin of nitrogen in tissue nitrogen pools with and without acidity control (CaCO3 treated and untreated, respectively) in the root environment. Control of acidity resulted in greater ammonium uptake and greater incorporation into the amino fraction, amide, and ethanol-insoluble nitrogen by the root tissue. These differences were clearly evident by the fifth day after ammonium nitrogen had been applied. Shoots of the untreated plants rapidly accumulated free ammonium and amino nitrogen. A substantial portion of both fractions came from pre-existing nitrogen in the plants, indicating significant protein degradation. No evidence was found for such degradation in the roots of the untreated plants or in either roots or shoots of CaCO3 treated plants. The data indicate that control of ambient acidity in the root environment during ammonium absorption enhanced the conversion of entering ammonium to organic nitrogen compounds in the root tissue thereby restricting movement of free ammonium to shoots. Consequently, the detrimental effects of high ammonium concentrations in the leaves were largely prevented. PMID:16656383

  20. Ammonia 15N/14N Isotope Ratio in the Jovian Atmosphere

    NASA Technical Reports Server (NTRS)

    Mahaffy, P.R.; Niemann, H. B.; Atreya, S. K.; Wong, M. H.; Owen, T. C; Einaudi, Franco (Technical Monitor)

    2000-01-01

    Data from the Galileo Probe Mass Spectrometer has been used to derive the N-15/N-14 isotope ratio in ammonia at Jupiter. Although the mass spectral interference from the water contribution to 18 amu makes an accurate derivation of the (N-15)H3/(N-14)H3 ratio difficult from measurements of the singly ionized signals at 18 and 17 amu, this interference is not present in the doubly charged 8.5 and 9.0 amu signals from (N-14)H3++ and (N-15)H3++ respectively. Although the count rate from the 9 amu signal is low during the direct sampling of the atmosphere, the ammonia signal was considerably enhanced during the first enrichment cell (EC1) experiment that measured gas sampled between 0.8 and 2.8 bar. Count rates at 9 amu in the EC1 experiment reach 60/second and measure ammonia sampled from 0.88 to 2.8 bar. In the EC1 measurements the 8.5 amu signal is not measured directly, but can be calculated from the ammonia contribution to 17 amu and the ratio of NH3 ions of a double to single charged observed during a high resolution mass scan taken near the end of the descent. The high resolution scan gives this ratio from ammonia sampled much deeper in the atmosphere. These results are described and compared with Infrared Space Observatory-Short Wavelength Spectrometer (ISO-SWS) observations that give this ratio at 400 mbar.

  1. δ13C and δ15N changes after dietary shift in veliger larvae of the slipper limpet Crepidula fornicata: an experimental evidence

    NASA Astrophysics Data System (ADS)

    Comtet, T.; Riera, P.

    2006-12-01

    δ13C and δ15N measurements are still poorly conducted in benthic invertebrate larvae. To assess the δ13C and δ15N changes occurring after a dietary shift, experiments were conducted on veliger larvae of Crepidula fornicata fed with two cultured microalgae ( Isochrysis galbana and Pavlova lutheri) of known isotopic composition, 13C-enriched and 15N-depleted compared to the initial values of the larvae. Rapid changes in larval δ13C and δ15N were observed after the dietary shift, with an increase in δ13C and a decrease in δ15N. After 19 days of feeding, isotopic equilibrium was still not reached, a period which is close to the duration of the pelagic life of the larvae. This implies that the isotopic composition measured in field-collected larvae might only partly reflect actual larval feeding but also the parental isotopic signature, especially during the early developmental stages. Isotopic measurements in marine invertebrate larvae should thus be interpreted cautiously. In planktonic food web investigations, the study of field-collected larvae of different size/developmental stage may reduce potential misinterpretations.

  2. Nitrogen retention across a gradient of 15N additions to an unpolluted temperate forest soil in Chile

    USGS Publications Warehouse

    Perakis, Steven S.; Compton, J.E.; Hedin, L.O.

    2005-01-01

    Accelerated nitrogen (N) inputs can drive nonlinear changes in N cycling, retention, and loss in forest ecosystems. Nitrogen processing in soils is critical to understanding these changes, since soils typically are the largest N sink in forests. To elucidate soil mechanisms that underlie shifts in N cycling across a wide gradient of N supply, we added 15NH415NO3 at nine treatment levels ranging in geometric sequence from 0.2 kg to 640 kg NA? ha-1A? yr-1 to an unpolluted old-growth temperate forest in southern Chile. We recovered roughly half of tracers in 0-25 cm of soil, primarily in the surface 10 cm. Low to moderate rates of N supply failed to stimulate N leaching, which suggests that most unrecovered 15N was transferred from soils to unmeasured sinks above ground. However, soil solution losses of nitrate increased sharply at inputs > 160 kg NA? ha-1A? yr-1, corresponding to a threshold of elevated soil N availability and declining 15N retention in soil. Soil organic matter (15N in soils at the highest N inputs and may explain a substantial fraction of the 'missing N' often reported in studies of fates of N inputs to forests. Contrary to expectations, N additions did not stimulate gross N cycling, potential nitrification, or ammonium oxidizer populations. Our results indicate that the nonlinearity in N retention and loss resulted directly from excessive N supply relative to sinks, independent of plant-soil-microbial feedbacks. However, N additions did induce a sharp decrease in microbial biomass C:N that is predicted by N saturation theory, and which could increase long-term N storage in soil organic matter by lowering the critical C:N ratio for net N mineralization. All measured sinks accumulated 15N tracers across the full gradient of N supply, suggesting that short-term nonlinearity in N retention resulted from saturation of uptake kinetics, not uptake capacity, in plant, soil, and microbial pools.

  3. Variability of the 15N Chemical Shielding Tensors in the B3 Domain of Protein G from 15N Relaxation Measurements at Several Fields

    PubMed Central

    Hall, Jennifer B.; Fushman, David

    2008-01-01

    We applied a combination of 15N relaxation and CSA/dipolar cross-correlation measurements at five magnetic fields (9.4, 11.7, 14.1, 16.4, and 18.8 Tesla) to determine the 15N chemical shielding tensors for backbone amides in protein G in solution. The data were analyzed using various model-independent approaches and those based on Lipari-Szabo approximation, all of them yielding similar results. The results indicate a range of site-specific values of the anisotropy (CSA) and orientation of the 15N chemical shielding tensor, similar to those in ubiquitin. Assuming a Gaussian distribution of the 15N CSA values, the mean anisotropy is -173.9 to -177.2 ppm (for 1.02-Å NH-bond length) and the site-so-site CSA variability is ±17.6 to ±21.4 ppm, depending on the method used. This CSA variability is significantly larger than derived previously for ribonuclease H or recently, using “meta-analysis” for ubiquitin. Standard interpretation of 15N relaxation studies of backbone dynamics in proteins involves an a priori assumption of a uniform 15N CSA. We show that this assumption leads to a significant discrepancy between the order parameters obtained at different fields. Using the site-specific CSAs obtained from our study removes this discrepancy and allows simultaneous fit of relaxation data at all five fields to Lipari-Szabo spectral densities. These findings emphasize the necessity of taking into account the variability of 15N CSA for accurate analysis of protein dynamics from 15N relaxation measurements. PMID:16771499

  4. Soil processes drive seasonal variation in retention of 15N tracers in a deciduous forest catchment.

    PubMed

    Goodale, Christine L; Fredriksen, Guinevere; Weiss, Marissa S; McCalley, K; Sparks, Jed P; Thomas, Steven A

    2015-10-01

    Seasonal patterns of stream nitrate concentration have long been interpreted as demonstrating the central role of plant uptake in regulating stream nitrogen loss from forested catchments. Soil processes are rarely considered as important drivers of these patterns. We examined seasonal variation in N retention in a deciduous forest using three whole-ecosystem 15N tracer additions: in late April (post-snowmelt, pre-leaf-out), late July (mid-growing- season), and late October (end of leaf-fall). We expected that plant 15N uptake would peak in late spring and midsummer, that immobilization in surface litter and soil would peak the following autumn leaf-fall, and that leaching losses would vary inversely with 15N retention. Similar to most other 15N tracer studies, we found that litter and soils dominated ecosystem retention of added 15N. However, 15N recovery in detrital pools varied tremendously by season, with > 90% retention in spring and autumn and sharply reduced 15N retention in late summer. During spring, over half of the 15N retained in soil occurred within one day in the heavy (mineral-associated) soil fraction. During summer, a large decrease in 15N retention one week after addition coincided with increased losses of 15NO3- to soil leachate and seasonal increases in soil and stream NO3- concentrations, although leaching accounted for only a small fraction of the lost 15N (< 0.2%). Uptake of 15N into roots did not vary by season and accounted for < 4% of each tracer addition. Denitrification or other processes that lead to N gas loss may have consumed the rest. These measurements of 15N movement provide strong evidence for the dominant role of soil processes in regulating seasonal N retention and losses in this catchment and perhaps others with similar soils.

  5. Isotopic evidence for a marine ammonium source in rainwater at Bermuda

    NASA Astrophysics Data System (ADS)

    Altieri, K. E.; Hastings, M. G.; Peters, A. J.; Oleynik, S.; Sigman, D. M.

    2014-10-01

    Emissions of anthropogenic nitrogen (N) to the atmosphere have increased tenfold since preindustrial times, resulting in increased N deposition to terrestrial and coastal ecosystems. The sources of N deposition to the ocean, however, are poorly understood. Two years of event-based rainwater samples were collected on the island of Bermuda in the western North Atlantic, which experiences both continent- and ocean-influenced air masses. The rainwater ammonium concentration ranged from 0.36 to 24.6 μM, and the ammonium δ15N from -12.5 to 0.7‰; and neither has a strong relationship with air mass history (6.0 ± 4.2 μM, -4.1 ± 2.6‰ in marine air masses and 5.9 ± 3.2 μM, -5.8 ± 2.5‰ in continental air masses; numerical average ± standard deviation). A simple box model suggests that the ocean can account for the concentration and isotopic composition of ammonium in marine rainwater, consistent with the lack of correlation between ammonium δ15N and air mass history. If so, ammonium deposition reflects the cycling of N between the ocean and the atmosphere, rather than representing a net input to the ocean. The δ15N data appear to require that most of the ammonium/a flux to the ocean is by dissolution in surface waters rather than atmospheric deposition. This suggests that the atmosphere and surface ocean are near equilibrium with respect to air/sea gas exchange, implying that anthropogenic ammonia will equilibrate near the coast and not reach the open marine atmosphere. Whereas ~90% of the ammonium deposition to the global ocean has previously been attributed to anthropogenic sources, the evidence at Bermuda suggests that the anthropogenic contribution could be much smaller.

  6. Through-space (19) F-(15) N couplings for the assignment of stereochemistry in flubenzimine.

    PubMed

    Ghiviriga, Ion; Rubinski, Miles A; Dolbier, William R

    2016-07-01

    Through-space (19) F-(15) N couplings revealed the configuration of flubenzimine, with the CF3 group on N4 pointing towards the lone pair of N5. The (19) F-(15) N coupling constants were measured at natural abundance using a spin-state selective indirect-detection pulse sequence. As (15) N-labelled proteins are routinely synthesized for NMR studies, through-space (19) F-(15) N couplings have the potential to probe the stereochemistry of these proteins by (19) F labelling of some amino acids or can reveal the site of docking of fluorine-containing drugs. Copyright © 2016 John Wiley & Sons, Ltd.

  7. Fermentation and Cost-Effective 13C/15N Labeling of the Nonribosomal Peptide Gramicidin S for Nuclear Magnetic Resonance Structure Analysis

    PubMed Central

    Berditsch, Marina; Afonin, Sergii; Steineker, Anna; Orel, Nataliia; Jakovkin, Igor; Weber, Christian

    2015-01-01

    Gramicidin S (GS) is a nonribosomally synthesized decapeptide from Aneurinibacillus migulanus. Its pronounced antibiotic activity is attributed to amphiphilic structure and enables GS interaction with bacterial membranes. Despite its medical use for over 70 years, the peptide-lipid interactions of GS and its molecular mechanism of action are still not fully understood. Therefore, a comprehensive structural analysis of isotope-labeled GS needs to be performed in its biologically relevant membrane-bound state, using advanced solid-state nuclear magnetic resonance (NMR) spectroscopy. Here, we describe an efficient method for producing the uniformly 13C/15N-labeled peptide in a minimal medium supplemented by selected amino acids. As GS is an intracellular product of A. migulanus, we characterized the producer strain DSM 5759 (rough-convex phenotype) and examined its biosynthetic activity in terms of absolute and biomass-dependent peptide accumulation. We found that the addition of either arginine or ornithine increases the yield only at very high supplementing concentrations (1% and 0.4%, respectively) of these expensive 13C/15N-labeled amino acids. The most cost-effective production of 13C/15N-GS, giving up to 90 mg per gram of dry cell weight, was achieved in a minimal medium containing 1% 13C-glycerol and 0.5% 15N-ammonium sulfate, supplemented with only 0.025% of 13C/15N-phenylalanine. The 100% efficiency of labeling is corroborated by mass spectrometry and preliminary solid-state NMR structure analysis of the labeled peptide in the membrane-bound state. PMID:25795666

  8. Fermentation and Cost-Effective 13C/15N Labeling of the Nonribosomal Peptide Gramicidin S for Nuclear Magnetic Resonance Structure Analysis.

    PubMed

    Berditsch, Marina; Afonin, Sergii; Steineker, Anna; Orel, Nataliia; Jakovkin, Igor; Weber, Christian; Ulrich, Anne S

    2015-06-01

    Gramicidin S (GS) is a nonribosomally synthesized decapeptide from Aneurinibacillus migulanus. Its pronounced antibiotic activity is attributed to amphiphilic structure and enables GS interaction with bacterial membranes. Despite its medical use for over 70 years, the peptide-lipid interactions of GS and its molecular mechanism of action are still not fully understood. Therefore, a comprehensive structural analysis of isotope-labeled GS needs to be performed in its biologically relevant membrane-bound state, using advanced solid-state nuclear magnetic resonance (NMR) spectroscopy. Here, we describe an efficient method for producing the uniformly (13)C/(15)N-labeled peptide in a minimal medium supplemented by selected amino acids. As GS is an intracellular product of A. migulanus, we characterized the producer strain DSM 5759 (rough-convex phenotype) and examined its biosynthetic activity in terms of absolute and biomass-dependent peptide accumulation. We found that the addition of either arginine or ornithine increases the yield only at very high supplementing concentrations (1% and 0.4%, respectively) of these expensive (13)C/(15)N-labeled amino acids. The most cost-effective production of (13)C/(15)N-GS, giving up to 90 mg per gram of dry cell weight, was achieved in a minimal medium containing 1% (13)C-glycerol and 0.5% (15)N-ammonium sulfate, supplemented with only 0.025% of (13)C/(15)N-phenylalanine. The 100% efficiency of labeling is corroborated by mass spectrometry and preliminary solid-state NMR structure analysis of the labeled peptide in the membrane-bound state.

  9. Ammonium-oxidizing bacteria facilitate aerobic degradation of sulfanilic acid in activated sludge.

    PubMed

    Chen, Gang; Ginige, Maneesha P; Kaksonen, Anna H; Cheng, Ka Yu

    2014-01-01

    Sulfanilic acid (SA) is a toxic sulfonated aromatic amine commonly found in anaerobically treated azo dye contaminated effluents. Aerobic acclimatization of SA-degrading mixed microbial culture could lead to co-enrichment of ammonium-oxidizing bacteria (AOB) because of the concomitant release of ammonium from SA oxidation. To what extent the co-enriched AOB would affect SA oxidation at various ammonium concentrations was unclear. Here, a series of batch kinetic experiments were conducted to evaluate the effect of AOB on aerobic SA degradation in an acclimatized activated sludge culture capable of oxidizing SA and ammonium simultaneously. To account for the effect of AOB on SA degradation, allylthiourea was used to inhibit AOB activity in the culture. The results indicated that specific SA degradation rate of the mixed culture was negatively correlated with the initial ammonium concentration (0-93 mM, R²= 0.99). The presence of AOB accelerated SA degradation by reducing the inhibitory effect of ammonium (≥ 10 mM). The Haldane substrate inhibition model was used to correlate substrate concentration (SA and ammonium) and oxygen uptake rate. This study revealed, for the first time, that AOB could facilitate SA degradation at high concentration of ammonium (≥ 10 mM) in an enriched activated sludge culture.

  10. Hydrothermal synthesis of ammonium illite

    USGS Publications Warehouse

    Sucha, V.; Elsass, F.; Eberl, D.D.; Kuchta, L'.; Madejova, J.; Gates, W.P.; Komadel, P.

    1998-01-01

    Synthetic gel and glass of illitic composition, natural kaolinite, and mixed-layer illite-smectite were used as starting materials for hydrothermal synthesis of ammonium illite. Ammonium illite was prepared from synthetic gel by hydrothermal treatment at 300??C. The onset of crystallization began within 3 h, and well-crystallized ammonium illite appeared at 24 h. Increasing reaction time (up to four weeks) led to many illite layers per crystal. In the presence of equivalent proportions of potassium and ammonium, the gel was transformed to illite with equimolar contents of K and NH4. In contrast, synthesis using glass under the same conditions resulted in a mixture of mixed-layer ammonium illite-smectite with large expandability and discrete illite. Hydrothermal treatments of the fine fractions of natural kaolinite and illite-smectite produced ammonium illite from kaolinite but the illite-smectite remained unchanged.

  11. Factors Controlling the Stable Nitrogen Isotopic Composition (δ15N) of Lipids in Marine Animals

    PubMed Central

    Svensson, Elisabeth; Schouten, Stefan; Hopmans, Ellen C.; Middelburg, Jack J.; Sinninghe Damsté, Jaap S.

    2016-01-01

    Lipid extraction of biomass prior to stable isotope analysis is known to cause variable changes in the stable nitrogen isotopic composition (δ15N) of residual biomass. However, the underlying factors causing these changes are not yet clear. Here we address this issue by comparing the δ15N of bulk and residual biomass of several marine animal tissues (fish, crab, cockle, oyster, and polychaete), as well as the δ15N of the extracted lipids. As observed previously, lipid extraction led to a variable offset in δ15N of biomass (differences ranging from -2.3 to +1.8 ‰). Importantly, the total lipid extract (TLE) was highly depleted in 15N compared to bulk biomass, and also highly variable (differences ranging from -14 to +0.7 ‰). The TLE consisted mainly of phosphatidylcholines, a group of lipids with one nitrogen atom in the headgroup. To elucidate the cause for the 15N-depletion in the TLE, the δ15N of amino acids was determined, including serine because it is one of the main sources of nitrogen to N-containing lipids. Serine δ15N values differed by -7 to +2 ‰ from bulk biomass δ15N, and correlated well with the 15N depletion in TLEs. On average, serine was less depleted (-3‰) than the TLE (-7 ‰), possibly due to fractionation during biosynthesis of N-containing headgroups, or that other nitrogen-containing compounds, such as urea and choline, or recycled nitrogen contribute to the nitrogen isotopic composition of the TLE. The depletion in 15N of the TLE relative to biomass increased with the trophic level of the organisms. PMID:26731720

  12. Factors Controlling the Stable Nitrogen Isotopic Composition (δ15N) of Lipids in Marine Animals.

    PubMed

    Svensson, Elisabeth; Schouten, Stefan; Hopmans, Ellen C; Middelburg, Jack J; Sinninghe Damsté, Jaap S

    2016-01-01

    Lipid extraction of biomass prior to stable isotope analysis is known to cause variable changes in the stable nitrogen isotopic composition (δ15N) of residual biomass. However, the underlying factors causing these changes are not yet clear. Here we address this issue by comparing the δ15N of bulk and residual biomass of several marine animal tissues (fish, crab, cockle, oyster, and polychaete), as well as the δ15N of the extracted lipids. As observed previously, lipid extraction led to a variable offset in δ15N of biomass (differences ranging from -2.3 to +1.8 ‰). Importantly, the total lipid extract (TLE) was highly depleted in 15N compared to bulk biomass, and also highly variable (differences ranging from -14 to +0.7 ‰). The TLE consisted mainly of phosphatidylcholines, a group of lipids with one nitrogen atom in the headgroup. To elucidate the cause for the 15N-depletion in the TLE, the δ15N of amino acids was determined, including serine because it is one of the main sources of nitrogen to N-containing lipids. Serine δ15N values differed by -7 to +2 ‰ from bulk biomass δ15N, and correlated well with the 15N depletion in TLEs. On average, serine was less depleted (-3‰) than the TLE (-7 ‰), possibly due to fractionation during biosynthesis of N-containing headgroups, or that other nitrogen-containing compounds, such as urea and choline, or recycled nitrogen contribute to the nitrogen isotopic composition of the TLE. The depletion in 15N of the TLE relative to biomass increased with the trophic level of the organisms.

  13. Factors Controlling the Stable Nitrogen Isotopic Composition (δ15N) of Lipids in Marine Animals.

    PubMed

    Svensson, Elisabeth; Schouten, Stefan; Hopmans, Ellen C; Middelburg, Jack J; Sinninghe Damsté, Jaap S

    2016-01-01

    Lipid extraction of biomass prior to stable isotope analysis is known to cause variable changes in the stable nitrogen isotopic composition (δ15N) of residual biomass. However, the underlying factors causing these changes are not yet clear. Here we address this issue by comparing the δ15N of bulk and residual biomass of several marine animal tissues (fish, crab, cockle, oyster, and polychaete), as well as the δ15N of the extracted lipids. As observed previously, lipid extraction led to a variable offset in δ15N of biomass (differences ranging from -2.3 to +1.8 ‰). Importantly, the total lipid extract (TLE) was highly depleted in 15N compared to bulk biomass, and also highly variable (differences ranging from -14 to +0.7 ‰). The TLE consisted mainly of phosphatidylcholines, a group of lipids with one nitrogen atom in the headgroup. To elucidate the cause for the 15N-depletion in the TLE, the δ15N of amino acids was determined, including serine because it is one of the main sources of nitrogen to N-containing lipids. Serine δ15N values differed by -7 to +2 ‰ from bulk biomass δ15N, and correlated well with the 15N depletion in TLEs. On average, serine was less depleted (-3‰) than the TLE (-7 ‰), possibly due to fractionation during biosynthesis of N-containing headgroups, or that other nitrogen-containing compounds, such as urea and choline, or recycled nitrogen contribute to the nitrogen isotopic composition of the TLE. The depletion in 15N of the TLE relative to biomass increased with the trophic level of the organisms. PMID:26731720

  14. Nitrate turnover in a peat soil under drained and rewetted conditions: results from a [(15)N]nitrate-bromide double-tracer study.

    PubMed

    Russow, Rolf; Tauchnitz, Nadine; Spott, Oliver; Mothes, Sibylle; Bernsdorf, Sabine; Meissner, Ralph

    2013-01-01

    Under natural conditions, peatlands are generally nitrate-limited. However, recent concerns about an additional N input into peatlands by atmospheric N deposition have highlighted the risk of an increased denitrification activity and hence the likelihood of a rise of emissions of the greenhouse gas nitrous oxide. Therefore, the aim of the present study was to investigate the turnover of added nitrate in a drained and a rewetted peatland using a [(15)N]nitrate-bromide double-tracer method. The double-tracer method allows a separation between physical effects (dilution, dispersion and dislocation) and microbial and chemical nitrate transformation by comparing with the conservative Br(-) tracer. In the drained peat site, low NO3(-) consumption rates have been observed. In contrast, NO3(-) consumption at the rewetted peat site rises rapidly to about 100% within 4 days after tracer application. Concomitantly, the (15)N abundances of nitrite and ammonium in soil water increased and lead to the conclusion that, besides commonly known NO3(-) reduction to nitrite (i.e. denitrification), a dissimilatory nitrate reduction to ammonium has simultaneously taken place. The present study reveals that increasing NO3(-) inputs into rewetted peatlands via atmospheric deposition results in a rapid NO3(-) consumption, which could lead to an increase in N2O emissions into the atmosphere.

  15. Uptake of nitrate, ammonium and glycine by plants of Tasmanian wet eucalypt forests.

    PubMed

    Warren, C R; Adams, P R

    2007-03-01

    A central assumption of ecosystem N cycling has been that organic N must be converted to inorganic N to be available for plant uptake, but this has been questioned by recent studies. We examined uptake of nitrate, ammonium and the amino acid glycine in three species from Eucalyptus obliqua L'Her. wet forest in Tasmania, south-eastern Australia, to test the hypothesis that all three species can take up glycine, and to compare rates of glycine uptake with rates of uptake of nitrate and ammonium uptake. The alternative hypothesis that species vary in their preference for nitrate, ammonium and glycine ("niche differentiation") was also examined. Measurements were made on the canopy dominant Eucalyptus obliqua, and two rain forest tree species found in the understory or as sub-dominants of the canopy, Nothofagus cunninghamii (Hook.) Oerst. and Phyllocladus aspleniifolius (Labill.) Hook.f. Nitrogen uptake was examined in situ with attached roots placed in uptake solutions containing equimolar concentrations (100 micromol l(-1)) of (15)N-nitrate, (15)N-ammonium and 2-(13)C(2) (15)N-glycine. Species did not differ in their preference for different forms of N (species x N form interaction, P > 0.05), and thus there was no evidence of niche differentiation. In all species, rates of uptake were highest for ammonium (11 +/- 5 micromol g(DM) (-1) h(-1); mean +/- SD, n = 108), uptake of glycine occurred at less than half this rate (4.4 +/- 2.6 micromol g(DM) (-1) h(-1)), whereas uptake of nitrate occurred at one-tenth of this rate (0.9 +/- 1.2 micromol g(DM) (-1) h(-1)). The strong positive relationship between (15)N and (13)C uptake indicated that at least 72% of glycine-N was taken up intact. These findings indicate the potential for considerable uptake of organic N in the field.

  16. Disturbance and topography shape nitrogen availability and δ15N over long-term forest succession

    EPA Science Inventory

    Forest disturbance and long-term succession can promote open N cycling that increases N loss and soil δ15N values. We examined soil and foliar patterns in N and δ15N, and soil N mineralization, across a topographically complex montane forest landscape influenced by human logging ...

  17. δ 15 N constraints on long-term nitrogen balances in temperate forests

    EPA Science Inventory

    Natural abundance δ15N of ecosystems integrates nitrogen (N) inputs and losses, and thus reflects factors that control the long-term development of ecosystem N balances. We here report N and carbon (C) content of forest vegetation and soils, and associated δ15N, across nine Doug...

  18. δ(15) N from soil to wine in bulk samples and proline.

    PubMed

    Paolini, Mauro; Ziller, Luca; Bertoldi, Daniela; Bontempo, Luana; Larcher, Roberto; Nicolini, Giorgio; Camin, Federica

    2016-09-01

    The feasibility of using δ(15) N as an additional isotopic marker able to link wine to its area of origin was investigated. The whole production chain (soil-leaves-grape-wine) was considered. Moreover, the research included evaluation of the effect of the fermentation process, the use of different types of yeast and white and red vinification, the addition of nitrogen adjuvants and ultrasound lysis simulating wine ageing. The δ(15) N of grapes and wine was measured in bulk samples and compounds, specifically in proline, for the first time. Despite isotopic fractionation from soil to wine, the δ(15) N values of leaves, grapes, wine and particularly must and wine proline conserved the variability of δ(15) N in the growing soil. Fermentation and ultrasound treatment did not affect the δ(15) N values of grape must, which was therefore conserved in wine. The addition of inorganic or organic adjuvants was able to influence the δ(15) N of bulk wine, depending on the amount and the difference between the δ(15) N of must and that of the adjuvant. The δ(15) N of wine proline was not influenced by adjuvant addition and is therefore the best marker for tracing the geographical origin of wine. Copyright © 2016 John Wiley & Sons, Ltd.

  19. δ(15) N from soil to wine in bulk samples and proline.

    PubMed

    Paolini, Mauro; Ziller, Luca; Bertoldi, Daniela; Bontempo, Luana; Larcher, Roberto; Nicolini, Giorgio; Camin, Federica

    2016-09-01

    The feasibility of using δ(15) N as an additional isotopic marker able to link wine to its area of origin was investigated. The whole production chain (soil-leaves-grape-wine) was considered. Moreover, the research included evaluation of the effect of the fermentation process, the use of different types of yeast and white and red vinification, the addition of nitrogen adjuvants and ultrasound lysis simulating wine ageing. The δ(15) N of grapes and wine was measured in bulk samples and compounds, specifically in proline, for the first time. Despite isotopic fractionation from soil to wine, the δ(15) N values of leaves, grapes, wine and particularly must and wine proline conserved the variability of δ(15) N in the growing soil. Fermentation and ultrasound treatment did not affect the δ(15) N values of grape must, which was therefore conserved in wine. The addition of inorganic or organic adjuvants was able to influence the δ(15) N of bulk wine, depending on the amount and the difference between the δ(15) N of must and that of the adjuvant. The δ(15) N of wine proline was not influenced by adjuvant addition and is therefore the best marker for tracing the geographical origin of wine. Copyright © 2016 John Wiley & Sons, Ltd. PMID:27479606

  20. Application of Nitrogen and Carbon Stable Isotopes (δ15N and δ13C) to Quantify Food Chain Length and Trophic Structure

    PubMed Central

    Perkins, Matthew J.; McDonald, Robbie A.; van Veen, F. J. Frank; Kelly, Simon D.; Rees, Gareth; Bearhop, Stuart

    2014-01-01

    Increasingly, stable isotope ratios of nitrogen (δ15N) and carbon (δ13C) are used to quantify trophic structure, though relatively few studies have tested accuracy of isotopic structural measures. For laboratory-raised and wild-collected plant-invertebrate food chains spanning four trophic levels we estimated nitrogen range (NR) using δ15N, and carbon range (CR) using δ13C, which are used to quantify food chain length and breadth of trophic resources respectively. Across a range of known food chain lengths we examined how NR and CR changed within and between food chains. Our isotopic estimates of structure are robust because they were calculated using resampling procedures that propagate variance in sample means through to quantified uncertainty in final estimates. To identify origins of uncertainty in estimates of NR and CR, we additionally examined variation in discrimination (which is change in δ15N or δ13C from source to consumer) between trophic levels and among food chains. δ15N discrimination showed significant enrichment, while variation in enrichment was species and system specific, ranged broadly (1.4‰ to 3.3‰), and importantly, propagated variation to subsequent estimates of NR. However, NR proved robust to such variation and distinguished food chain length well, though some overlap between longer food chains infers a need for awareness of such limitations. δ13C discrimination was inconsistent; generally no change or small significant enrichment was observed. Consequently, estimates of CR changed little with increasing food chain length, showing the potential utility of δ13C as a tracer of energy pathways. This study serves as a robust test of isotopic quantification of food chain structure, and given global estimates of aquatic food chains approximate four trophic levels while many food chains include invertebrates, our use of four trophic level plant-invertebrate food chains makes our findings relevant for a majority of ecological systems

  1. Application of nitrogen and carbon stable isotopes (δ(15)N and δ(13)C) to quantify food chain length and trophic structure.

    PubMed

    Perkins, Matthew J; McDonald, Robbie A; van Veen, F J Frank; Kelly, Simon D; Rees, Gareth; Bearhop, Stuart

    2014-01-01

    Increasingly, stable isotope ratios of nitrogen (δ(15)N) and carbon (δ(13)C) are used to quantify trophic structure, though relatively few studies have tested accuracy of isotopic structural measures. For laboratory-raised and wild-collected plant-invertebrate food chains spanning four trophic levels we estimated nitrogen range (NR) using δ(15)N, and carbon range (CR) using δ(13)C, which are used to quantify food chain length and breadth of trophic resources respectively. Across a range of known food chain lengths we examined how NR and CR changed within and between food chains. Our isotopic estimates of structure are robust because they were calculated using resampling procedures that propagate variance in sample means through to quantified uncertainty in final estimates. To identify origins of uncertainty in estimates of NR and CR, we additionally examined variation in discrimination (which is change in δ(15)N or δ(13)C from source to consumer) between trophic levels and among food chains. δ(15)N discrimination showed significant enrichment, while variation in enrichment was species and system specific, ranged broadly (1.4‰ to 3.3‰), and importantly, propagated variation to subsequent estimates of NR. However, NR proved robust to such variation and distinguished food chain length well, though some overlap between longer food chains infers a need for awareness of such limitations. δ(13)C discrimination was inconsistent; generally no change or small significant enrichment was observed. Consequently, estimates of CR changed little with increasing food chain length, showing the potential utility of δ(13)C as a tracer of energy pathways. This study serves as a robust test of isotopic quantification of food chain structure, and given global estimates of aquatic food chains approximate four trophic levels while many food chains include invertebrates, our use of four trophic level plant-invertebrate food chains makes our findings relevant for a majority of

  2. 15N and13C NMR investigation of hydroxylamine-derivatized humic substances

    USGS Publications Warehouse

    Thorn, K.A.; Arterburn, J.B.; Mikita, M.A.

    1992-01-01

    Five fulvic and humic acid samples of diverse origins were derivatized with 15N-labeled hydroxylamine and analyzed by liquid-phase 15N NMR spectrometry. The 15N NMR spectra indicated that hydroxylamine reacted similarly with all samples and could discriminate among carbonyl functional groups. Oximes were the major derivatives; resonances attributable to hydroxamic acids, the reaction products of hydroxylamine with esters, and resonances attributable to the tautomeric equilibrium position between the nitrosophenol and monoxime derivatives of quinones, the first direct spectroscopic evidence for quinones, also were evident. The 15N NMR spectra also suggested the presence of nitriles, oxazoles, oxazolines, isocyanides, amides, and lactams, which may all be explained in terms of Beckmann reactions of the initial oxime derivatives. INEPT and ACOUSTIC 15N NMR spectra provided complementary information on the derivatized samples. 13C NMR spectra of derivatized samples indicated that the ketone/quinone functionality is incompletely derivatized with hydroxylamine. ?? 1991 American Chemical Society.

  3. 13C and 15N fractionation of CH4/N2 mixtures during photochemical aerosol formation: Relevance to Titan

    NASA Astrophysics Data System (ADS)

    Sebree, Joshua A.; Stern, Jennifer C.; Mandt, Kathleen E.; Domagal-Goldman, Shawn D.; Trainer, Melissa G.

    2016-05-01

    The ratios of the stable isotopes that comprise each chemical species in Titan's atmosphere provide critical information towards understanding the processes taking place within its modern and ancient atmosphere. Several stable isotope pairs, including 12C/13C and 14N/15N, have been measured in situ or probed spectroscopically by Cassini-borne instruments, space telescopes, or through ground-based observations. Current attempts to model the observed isotope ratios incorporate fractionation resulting from atmospheric diffusion, hydrodynamic escape, and primary photochemical processes. However, the effect of a potentially critical pathway for isotopic fractionation - organic aerosol formation and subsequent deposition onto the surface of Titan - has not been considered due to insufficient data regarding fractionation during aerosol formation. To better understand the nature of this process, we have conducted a laboratory study to measure the isotopic fractionation associated with the formation of Titan aerosol analogs, commonly referred to as 'tholins', via far-UV irradiation of several methane (CH4) and dinitrogen (N2) mixtures. Analysis of the δ13C and δ15N isotopic signatures of the photochemical aerosol products using an isotope ratio mass spectrometer (IRMS) show that fractionation direction and magnitude are dependent on the initial bulk composition of the gas mixture. In general, the aerosols showed enrichment in 13C and 14N, and the observed fractionation trends can provide insight into the chemical mechanisms controlling photochemical aerosol formation.

  4. Pathways for nitrate release from an alpine watershed: determination using d15N and d18O

    USGS Publications Warehouse

    Campbell, Donald H.; Kendall, Carol; Chang, Cecily C.Y.; Silva, Steven R.; Tonnessen, Kathy A.

    2002-01-01

    [1] Snowpack, snowmelt, precipitation, surface water, and groundwater samples from the Loch Vale watershed in Colorado were analyzed for ??15N and ??18O of nitrate to determine the processes controlling the release of atmospherically deposited nitrogen from alpine and subalpine ecosystems. Although overlap was found between the ??15N(NO3) values for all water types (-4 to +6???), the ??18O(NO3) values for surface water and groundwater (+10 to +30???) were usually distinct from snowpack, snowmelt, and rainfall values (+40 to +70???). During snowmelt, ??18O(NO3) indicated that about half of the nitrate in stream water was the product of microbial nitrification; at other times that amount was greater than half. Springs emerging from talus deposits had high nitrate concentrations and a seasonal pattern in ??18O(NO3) that was similar to the pattern in the streams, indicating that shallow groundwater in talus deposits is a likely source of stream water nitrate. Only a few samples of surface water and groundwater collected during early snowmelt and large summer rain events had isotopic compositions that indicated most of the nitrate came directly from atmospheric deposition with no biological assimilation and release. This study demonstrates the value of the nitrate double-isotope technique for determining nitrogen-cycling processes and sources of nitrate in small, undisturbed watersheds that are enriched with inorganic nitrogen.

  5. Disturbance and topography shape nitrogen availability and δ15 N over long-term forest succession

    USGS Publications Warehouse

    Perakis, Steven; Tepley, Alan J.; Compton, Jana

    2015-01-01

    Forest disturbance and long-term succession towards old-growth are thought to increase nitrogen (N) availability and N loss, which should increase soil δ15N values. We examined soil and foliar patterns in N and δ15N, and soil N mineralization, across 800 years of forest succession in a topographically complex montane landscape influenced by human logging and wildfire. In contrast to expectations, we found that disturbance caused declines in surface mineral soil δ15N values, both in logged forests measured 40–50 years after disturbance, and in unlogged forests disturbed by severe wildfire within the last 200 years. Both symbiotic N fixation and N transfers from disturbed vegetation and detritus could lower soil δ15N values after disturbance. A more important role for symbiotic N fixation is suggested by lower soil δ15N values in slow-successional sites with slow canopy closure, which favors early-successional N fixers. Soil δ15N values increased only marginally throughout 800 years of succession, reflecting soil N uptake by vegetation and strong overall N retention. Although post-disturbance N inputs lowered surface soil δ15N values, steady-state mass balance calculations suggest that wildfire combustion of vegetation and detritus can dominate long-term N loss and increase whole-ecosystem δ15N. On steeper topography, declining soil δ15N values highlight erosion and accelerated soil turnover as an additional abiotic control on N balances. We conclude for N-limited montane forests that soil δ15N and N availability are less influenced by nitrate leaching and denitrification loss than by interactions between disturbance, N fixation, and erosion.

  6. Sedimentary records of δ(13)C, δ(15)N and organic matter accumulation in lakes receiving nutrient-rich mine waters.

    PubMed

    Widerlund, Anders; Chlot, Sara; Öhlander, Björn

    2014-07-01

    Organic C and total N concentrations, C/N ratios, δ(15)N and δ(13)C values in (210)Pb-dated sediment cores were used to reconstruct historical changes in organic matter (OM) accumulation in three Swedish lakes receiving nutrient-rich mine waters. Ammonium-nitrate-based explosives and sodium cyanide (NaCN) used in gold extraction were the major N sources, while lesser amounts of P originated from apatite and flotation chemicals. The software IsoSource was used to model the relative contribution of soil, terrestrial and littoral vegetation, and phytoplankton detritus in the lake sediments. In one lake the IsoSource modelling failed, suggesting the presence of additional, unknown OM sources. In two of the lakes sedimentary detritus of littoral vegetation and phytoplankton had increased by 15-20% and 20-35%, respectively, since ~1950, when N- and P-rich mine waters began to reach the lakes. Today, phytoplankton is the dominating OM component in these lake sediments, which appears to be a eutrophication effect related to mining operations. Changes in the N isotopic composition of biota, lake water, and sediments related to the use of ammonium-nitrate-based explosives and NaCN were evident in the two studied systems. However, N isotope signals in the receiving waters (δ(15)N~+9‰ to +19‰) were clearly shifted from the primary signal in explosives (δ(15)N-NO3=+3.4±0.3‰; δ(15)N-NH4=-8.0±0.3‰) and NaCN (δ(15)N=+1.1±0.5‰), and direct tracing of the primary N isotope signals in mining chemicals was not possible in the receiving waters. Systems where mine waters with a well known discharge history are a major point source of N with well-defined isotopic composition should, however, be suitable for further studies of processes controlling N isotope signatures and their transformation in aquatic systems receiving mine waters. PMID:24727038

  7. Sedimentary records of δ(13)C, δ(15)N and organic matter accumulation in lakes receiving nutrient-rich mine waters.

    PubMed

    Widerlund, Anders; Chlot, Sara; Öhlander, Björn

    2014-07-01

    Organic C and total N concentrations, C/N ratios, δ(15)N and δ(13)C values in (210)Pb-dated sediment cores were used to reconstruct historical changes in organic matter (OM) accumulation in three Swedish lakes receiving nutrient-rich mine waters. Ammonium-nitrate-based explosives and sodium cyanide (NaCN) used in gold extraction were the major N sources, while lesser amounts of P originated from apatite and flotation chemicals. The software IsoSource was used to model the relative contribution of soil, terrestrial and littoral vegetation, and phytoplankton detritus in the lake sediments. In one lake the IsoSource modelling failed, suggesting the presence of additional, unknown OM sources. In two of the lakes sedimentary detritus of littoral vegetation and phytoplankton had increased by 15-20% and 20-35%, respectively, since ~1950, when N- and P-rich mine waters began to reach the lakes. Today, phytoplankton is the dominating OM component in these lake sediments, which appears to be a eutrophication effect related to mining operations. Changes in the N isotopic composition of biota, lake water, and sediments related to the use of ammonium-nitrate-based explosives and NaCN were evident in the two studied systems. However, N isotope signals in the receiving waters (δ(15)N~+9‰ to +19‰) were clearly shifted from the primary signal in explosives (δ(15)N-NO3=+3.4±0.3‰; δ(15)N-NH4=-8.0±0.3‰) and NaCN (δ(15)N=+1.1±0.5‰), and direct tracing of the primary N isotope signals in mining chemicals was not possible in the receiving waters. Systems where mine waters with a well known discharge history are a major point source of N with well-defined isotopic composition should, however, be suitable for further studies of processes controlling N isotope signatures and their transformation in aquatic systems receiving mine waters.

  8. Sequence-specific sup 1 H and sup 15 N resonance assignments for human dihydrofolate reductase in solution

    SciTech Connect

    Stockman, B.J.; Nirmala, N.R.; Wagner, G. ); Delcamp, T.J.; DeYarman, M.T.; Freisheim, J.H. )

    1992-01-14

    Dihydrofolate reductase is an intracellular target enzyme for folate antagonists, including the anticancer drug methotrexate. In order to design novel drugs with altered binding properties, a detailed description of protein-drug interactions in solution is desirable to understand the specificity of drug binding. As a first step in this process, heteronuclear three-dimensional NMR spectroscopy has been used to make sequential resonance assignments for more than 90% of the residues in human dihydrofolate reductase complexed with methotrexate. Uniform enrichment of the 21.5-kDa protein with {sup 15}N was required to obtain the resonance assignments via heteronuclear 3D NMR spectroscopy since homonuclear 2D spectra did not provide sufficient {sup 1}H resonance dispersion. Medium- and long-range NOE's have been used to characterize the secondary structure of the binary ligand-enzyme complex in solution.

  9. Nitrogen addition regulates soil nematode community composition through ammonium suppression.

    PubMed

    Wei, Cunzheng; Zheng, Huifen; Li, Qi; Lü, Xiaotao; Yu, Qiang; Zhang, Haiyang; Chen, Quansheng; He, Nianpeng; Kardol, Paul; Liang, Wenju; Han, Xingguo

    2012-01-01

    Nitrogen (N) enrichment resulting from anthropogenic activities has greatly changed the composition and functioning of soil communities. Nematodes are one of the most abundant and diverse groups of soil organisms, and they occupy key trophic positions in the soil detritus food web. Nematodes have therefore been proposed as useful indicators for shifts in soil ecosystem functioning under N enrichment. Here, we monitored temporal dynamics of the soil nematode community using a multi-level N addition experiment in an Inner Mongolia grassland. Measurements were made three years after the start of the experiment. We used structural equation modeling (SEM) to explore the mechanisms regulating nematode responses to N enrichment. Across the N enrichment gradient, significant reductions in total nematode abundance, diversity (H' and taxonomic richness), maturity index (MI), and the abundance of root herbivores, fungivores and omnivores-predators were found in August. Root herbivores recovered in September, contributing to the temporal variation of total nematode abundance across the N gradient. Bacterivores showed a hump-shaped relationship with N addition rate, both in August and September. Ammonium concentration was negatively correlated with the abundance of total and herbivorous nematodes in August, but not in September. Ammonium suppression explained 61% of the variation in nematode richness and 43% of the variation in nematode trophic group composition. Ammonium toxicity may occur when herbivorous nematodes feed on root fluid, providing a possible explanation for the negative relationship between herbivorous nematodes and ammonium concentration in August. We found a significantly positive relationship between fungivores and fungal phospholipid fatty acids (PLFA), suggesting bottom-up control of fungivores. No such relationship was found between bacterivorous nematodes and bacterial PLFA. Our findings contribute to the understanding of effects of N enrichment in

  10. Standing Out From the Crowd: New Fish Recruits Have Unique δ15N Signatures in a Heavily Polluted Intermittently Open Estuary.

    NASA Astrophysics Data System (ADS)

    Hadwen, W. L.; Arthington, A. H.

    2005-05-01

    Intermittently open estuaries (IOEs) are the dominant coastal ecosystems of the Australian coastline. Conditions in IOEs range from freshwater when closed, to marine when open to the ocean, during which time marine fish often migrate into these systems. Many IOEs are facing increasing pressure from coastal development and ongoing nutrient inputs from industrial and urban sources have been shown to increase the δ15N isotope signatures of biota in receiving waters. In this study, we examined the food web structure of Tallows Creek, a relatively small IOE in northern NSW, Australia, which receives ongoing sewage treatment plant inputs. We first sampled Tallows Creek when it was closed to the ocean and found that all components of the food web had enriched δ15N signatures relative to biota sampled from a relatively undisturbed neighbouring system. However, samples collected immediately following an entrance opening event revealed some individuals of highly mobile taxa with δ15N signatures more typical of unpolluted estuaries. We suggest that these isotopically light individuals were recent migrants into Tallows Creek and that δ15N signatures can act as a guide to the importance of heavily polluted systems as nursery grounds for mobile taxa, owing to the distinction between resident and migrant signatures.

  11. sup 13 C and sup 15 N nuclear magnetic resonance evidence of the ionization state of substrates bound to bovine dihydrofolate reductase

    SciTech Connect

    Selinsky, B.S.; Perlman, M.E.; London, R.E. ); Unkefer, C.J. ); Mitchell, J. ); Blakley, R.L. Univ. of Tennessee, Memphis )

    1990-02-06

    The state of protonation of substrates bound to mammalian dihydrofolate reductase (DHFR) has significance for the mechanism of catalysis. To investigate this, dihydrofolate and dihydropteroylpentaglutamate have been synthesized with {sup 15}N enrichment at N-5. {sup 15}N NMR studies have been performed on the binary complexes formed by bovine DHFR with these compounds and with (5-{sup 15}N)dihydrobiopterin. The results indicate that there is no protonation at N-5 in the binary complexes, and this was confirmed by {sup 13}C NMR studies with folate and dihydrofolate synthesized with {sup 13}C enrichment at C-6. The chemical shift displacements produced by complex formation are in the same direction as those which result from deprotonation of the N-3/C-4-O amide group and are consistent with at least partial loss of the proton from N-3. This would be possible if, as crystallographic data indicate, there is interaction of N-3 and the 2-amino group of the bound ligands with the carboxylate of the active site glutamate residue (Glu{sup 30}).

  12. Steroselective synthesis and application of L-( sup 15 N) amino acids

    SciTech Connect

    Unkefer, C.J. ); Lodwig, S.N. . Div. of Science)

    1991-01-01

    We have developed two general approaches to the stereoselective synthesis of {sup 15}N- and {sup 13}C-labeled amino acids. First, labeled serine, biosynthesized using the methylotrophic bacterium M. extorquens AM1, serves as a chiral precursor for the synthesis of other amino acids. For example, pyridoxal phosphate enzymes can be used for the conversion of L-({alpha}-{sup 15}N)serine to L-({alpha}-{sup 15}N)tyrosine, L-({alpha}-{sup 15}N)tryptophan, and L-({alpha}-{sup 15}N)cysteine. In the second approach, developed by Oppolzer and Tamura, an electrophilic amination'' reagent, 1-chloro-1-nitrosocyclohexane, was used to convert chiral enolates into L-{alpha}-amino acids. We prepared 1-chloro-1-({sup 15}N) nitrosocyclohexane and used it to aminate chiral enolates to produce L-({alpha}-{sup 15}N)amino acids. The stereoselectivity of this scheme using the Oppolzer sultam chiral auxiliary is remarkable, producing enantiomer ratios of 200 to 1. 22 refs., 4 figs.

  13. Nitrogen stable isotope composition (δ15N) of vehicle-emitted NOx.

    PubMed

    Walters, Wendell W; Goodwin, Stanford R; Michalski, Greg

    2015-02-17

    The nitrogen stable isotope ratio of NOx (δ(15)N-NOx) has been proposed as a regional indicator for NOx source partitioning; however, knowledge of δ(15)N values from various NOx emission sources is limited. This study presents a detailed analysis of δ(15)N-NOx emitted from vehicle exhaust, the largest source of anthropogenic NOx. To accomplish this, NOx was collected from 26 different vehicles, including gasoline and diesel-powered engines, using a modification of a NOx collection method used by the United States Environmental Protection Agency, and δ(15)N-NOx was analyzed. The vehicles sampled in this study emitted δ(15)N-NOx values ranging from -19.1 to 9.8‰ that negatively correlated with the emitted NOx concentrations (8.5 to 286 ppm) and vehicle run time because of kinetic isotope fractionation effects associated with the catalytic reduction of NOx. A model for determining the mass-weighted δ(15)N-NOx from vehicle exhaust was constructed on the basis of average commute times, and the model estimates an average value of -2.5 ± 1.5‰, with slight regional variations. As technology improvements in catalytic converters reduce cold-start emissions in the future, it is likely to increase current δ(15)N-NOx values emitted from vehicles. PMID:25621737

  14. Nitrogen stable isotope composition (δ15N) of vehicle-emitted NOx.

    PubMed

    Walters, Wendell W; Goodwin, Stanford R; Michalski, Greg

    2015-02-17

    The nitrogen stable isotope ratio of NOx (δ(15)N-NOx) has been proposed as a regional indicator for NOx source partitioning; however, knowledge of δ(15)N values from various NOx emission sources is limited. This study presents a detailed analysis of δ(15)N-NOx emitted from vehicle exhaust, the largest source of anthropogenic NOx. To accomplish this, NOx was collected from 26 different vehicles, including gasoline and diesel-powered engines, using a modification of a NOx collection method used by the United States Environmental Protection Agency, and δ(15)N-NOx was analyzed. The vehicles sampled in this study emitted δ(15)N-NOx values ranging from -19.1 to 9.8‰ that negatively correlated with the emitted NOx concentrations (8.5 to 286 ppm) and vehicle run time because of kinetic isotope fractionation effects associated with the catalytic reduction of NOx. A model for determining the mass-weighted δ(15)N-NOx from vehicle exhaust was constructed on the basis of average commute times, and the model estimates an average value of -2.5 ± 1.5‰, with slight regional variations. As technology improvements in catalytic converters reduce cold-start emissions in the future, it is likely to increase current δ(15)N-NOx values emitted from vehicles.

  15. Protein expression and isotopic enrichment based on induction of the Entner-Doudoroff pathway in Escherichia coli

    SciTech Connect

    Refaeli, Bosmat; Goldbourt, Amir

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer The Entner-Doudoroff pathway is induced during protein expression in E. coli. Black-Right-Pointing-Pointer 1-{sup 13}C-gluconate and {sup 15}NH{sub 4}Cl provide a carbonyl-amide protein backbone labeling scheme. Black-Right-Pointing-Pointer The enrichment pattern is determined by nuclear magnetic resonance. -- Abstract: The Entner-Doudoroff pathway is known to exist in many organisms including bacteria, archea and eukarya. Although the common route for carbon catabolism in Escherichia coli is the Embden-Meyerhof-Parnas pathway, it was shown that gluconate catabolism in E. coli occurs via the Entner-Doudoroff pathway. We demonstrate here that by supplying BL21(DE3) competent E.coli cells with gluconate in a minimal growth medium, protein expression can be induced. Nuclear magnetic resonance data of over-expressed ubiquitin show that by using [1-{sup 13}C]-gluconate as the only carbon source, and {sup 15}N-enriched ammonium chloride, sparse isotopic enrichment in the form of a spin-pair carbonyl-amide backbone enrichment is obtained. The specific amino acid labeling pattern is analyzed and is shown to be compatible with Entner-Doudoroff metabolism. Isotopic enrichment serves as a key factor in the biophysical characterization of proteins by various methods including nuclear magnetic resonance, mass spectrometry, infrared spectroscopy and more. Therefore, the method presented here can be applied to study proteins by obtaining sparse enrichment schemes that are not based on the regular glycolytic pathway, or to study the Entner-Doudoroff metabolism during protein expression.

  16. Influence of roots and mycorrhiza on the internal nitrogen cycle in an organic forest soil ­revealed by a 15N tracing experiment

    NASA Astrophysics Data System (ADS)

    Holz, M.; Rutting, T.; Klemedtsson, L.; Kuzyakov, Y.

    2014-12-01

    The cycle of nitrogen in soil is complex, consisting of many simultaneous occurring transformation processes. So far, microorganisms have been thought to govern N cycling in soil. Nevertheless, plant roots and their associated mycorrhizal symbionts may exert control on N turnover for example by input of labile C to soil. However, studies investigating the effect of roots on gross N turnover rates are scarce. We conducted a 15N tracer study under field conditions to reveal the effect of plants on soil N cycle. The experiment includes three treatments: (a) control, (b) excluding roots and (c) excluding roots + mycorrhiza. On the study site, exclusion of roots + mycorrhiza has previously been shown to increase N2O emissions which indicate that plants affect internal N cycling. 15NH4NO3 and NH415NO3 were given to the soil and traced for a period of 10 days. Gross N turnover rates were determined applying a numerical 15N tracing model. Results on N turnover rates showed that roots and their fungal symbionts increased N cycling probably by input of labile C to soil which may results in an activation of the microbial biomass. While gross N mineralization increased by 270 and 313 % compared to the treatment excluding roots + mycorrhiza, NH4+ immobilization increased by 402 and 489 %. Differences in ammonium and nitrate immobilization further indicated that ammonium was the preferred N source for roots and microorganisms. While ammonium availability decreased with trenching (0.59 compared to -0.47 and -0.96 μg N g-1 d-1), the opposite was true for nitrate (0.50 compared to 2.08 and 2.18 μg N g-1 d-1), explaining the increased N2O emissions which were likely caused by denitrification. Further, plants increased dissimilarity nitrate reduction to ammonium (DNRA) and affected autotrophic nitrification probably by the release of nitrification inhibitors and by influencing ammonium availability. We conclude that plants and their mycorrhizal symbionts actively control N cycling

  17. Thaumarchaeal ammonium oxidation and evidence for a nitrogen cycle in a subsurface radioactive thermal spring in the Austrian Central Alps.

    PubMed

    Gerbl, Friedrich W; Weidler, Gerhard W; Wanek, Wolfgang; Erhardt, Angelika; Stan-Lotter, Helga

    2014-01-01

    Previous studies had suggested the presence of ammonium oxidizing Thaumarchaeota as well as nitrite oxidizing Bacteria in the subsurface spring called Franz Josef Quelle (FJQ), a slightly radioactive thermal mineral spring with a temperature of 43.6-47°C near the alpine village of Bad Gastein, Austria. The microbiological consortium of the FJQ was investigated for its utilization of nitrogen compounds and the putative presence of a subsurface nitrogen cycle. Microcosm experiments made with samples from the spring water, containing planktonic microorganisms, or from biofilms, were used in this study. Three slightly different media, enriched with vitamins and trace elements, and two incubation temperatures (30 and 40°C, respectively) were employed. Under aerobic conditions, high rates of conversion of ammonium to nitrite, as well as nitrite to nitrate were measured. Under oxygen-limited conditions nitrate was converted to gaseous compounds. Stable isotope probing with (15)NH4Cl or ((15)NH4)2SO4as sole energy sources revealed incorporation of (15)N into community DNA. Genomic DNA as well as RNA were extracted from all microcosms. The following genes or fragments of genes were successfully amplified, cloned and sequenced by standard PCR from DNA extracts: Ammonia monooxygenase subunit A (amoA), nitrite oxidoreductase subunits A and B (nxrA and nxrB), nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductases (cnorB and qnorB), nitrous oxide reductase (nosZ). Reverse transcription of extracted total RNA and real-time PCR suggested the expression of each of those genes. Nitrogen fixation (as probed with nifH and nifD) was not detected. However, a geological origin of NH(+) 4 in the water of the FJQ cannot be excluded, considering the silicate, granite and gneiss containing environment. The data suggested the operation of a nitrogen cycle in the subsurface environment of the FJQ. PMID:24904540

  18. Thaumarchaeal ammonium oxidation and evidence for a nitrogen cycle in a subsurface radioactive thermal spring in the Austrian Central Alps

    PubMed Central

    Gerbl, Friedrich W.; Weidler, Gerhard W.; Wanek, Wolfgang; Erhardt, Angelika; Stan-Lotter, Helga

    2014-01-01

    Previous studies had suggested the presence of ammonium oxidizing Thaumarchaeota as well as nitrite oxidizing Bacteria in the subsurface spring called Franz Josef Quelle (FJQ), a slightly radioactive thermal mineral spring with a temperature of 43.6–47°C near the alpine village of Bad Gastein, Austria. The microbiological consortium of the FJQ was investigated for its utilization of nitrogen compounds and the putative presence of a subsurface nitrogen cycle. Microcosm experiments made with samples from the spring water, containing planktonic microorganisms, or from biofilms, were used in this study. Three slightly different media, enriched with vitamins and trace elements, and two incubation temperatures (30 and 40°C, respectively) were employed. Under aerobic conditions, high rates of conversion of ammonium to nitrite, as well as nitrite to nitrate were measured. Under oxygen-limited conditions nitrate was converted to gaseous compounds. Stable isotope probing with 15NH4Cl or (15NH4)2SO4as sole energy sources revealed incorporation of 15N into community DNA. Genomic DNA as well as RNA were extracted from all microcosms. The following genes or fragments of genes were successfully amplified, cloned and sequenced by standard PCR from DNA extracts: Ammonia monooxygenase subunit A (amoA), nitrite oxidoreductase subunits A and B (nxrA and nxrB), nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductases (cnorB and qnorB), nitrous oxide reductase (nosZ). Reverse transcription of extracted total RNA and real-time PCR suggested the expression of each of those genes. Nitrogen fixation (as probed with nifH and nifD) was not detected. However, a geological origin of NH+4 in the water of the FJQ cannot be excluded, considering the silicate, granite and gneiss containing environment. The data suggested the operation of a nitrogen cycle in the subsurface environment of the FJQ. PMID:24904540

  19. Thaumarchaeal ammonium oxidation and evidence for a nitrogen cycle in a subsurface radioactive thermal spring in the Austrian Central Alps.

    PubMed

    Gerbl, Friedrich W; Weidler, Gerhard W; Wanek, Wolfgang; Erhardt, Angelika; Stan-Lotter, Helga

    2014-01-01

    Previous studies had suggested the presence of ammonium oxidizing Thaumarchaeota as well as nitrite oxidizing Bacteria in the subsurface spring called Franz Josef Quelle (FJQ), a slightly radioactive thermal mineral spring with a temperature of 43.6-47°C near the alpine village of Bad Gastein, Austria. The microbiological consortium of the FJQ was investigated for its utilization of nitrogen compounds and the putative presence of a subsurface nitrogen cycle. Microcosm experiments made with samples from the spring water, containing planktonic microorganisms, or from biofilms, were used in this study. Three slightly different media, enriched with vitamins and trace elements, and two incubation temperatures (30 and 40°C, respectively) were employed. Under aerobic conditions, high rates of conversion of ammonium to nitrite, as well as nitrite to nitrate were measured. Under oxygen-limited conditions nitrate was converted to gaseous compounds. Stable isotope probing with (15)NH4Cl or ((15)NH4)2SO4as sole energy sources revealed incorporation of (15)N into community DNA. Genomic DNA as well as RNA were extracted from all microcosms. The following genes or fragments of genes were successfully amplified, cloned and sequenced by standard PCR from DNA extracts: Ammonia monooxygenase subunit A (amoA), nitrite oxidoreductase subunits A and B (nxrA and nxrB), nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductases (cnorB and qnorB), nitrous oxide reductase (nosZ). Reverse transcription of extracted total RNA and real-time PCR suggested the expression of each of those genes. Nitrogen fixation (as probed with nifH and nifD) was not detected. However, a geological origin of NH(+) 4 in the water of the FJQ cannot be excluded, considering the silicate, granite and gneiss containing environment. The data suggested the operation of a nitrogen cycle in the subsurface environment of the FJQ.

  20. Radiative p 15N Capture in the Region of Astrophysical Energies

    NASA Astrophysics Data System (ADS)

    Dubovichenko, S. B.; Burtebaev, N.; Dzhazairov-Kakhramanov, A. V.; Alimov, D. K.

    2016-06-01

    Within the framework of the modified potential cluster model with classification of orbital states according to the Young schemes, the possibility of describing experimental data for the astrophysical S-factor of p 15N radiative capture at energies from 50 to 1500 keV is considered. It is shown that on the basis of M1 and E1 transitions from various p 15N scattering states to the ground state of the 16O nucleus in the p 15N channel it is entirely possible to successfully explain the overall behavior of the S-factor in the considered energy region in the presence of two resonances.

  1. Continuous field measurement of N2O isotopologues using FTIR spectroscopy following 15N addition

    NASA Astrophysics Data System (ADS)

    Phillips, R. L.; Griffith, D. W.; Dijkstra, F. A.; Lugg, G.; Lawrie, R.; Macdonald, B.

    2012-12-01

    Anthropogenic additions of fertilizer nitrogen (N) have significantly increased the mole fraction of nitrous oxide (N2O) in the troposphere. Tracking the fate of fertilizer N and its transformation to N2O is important to advance knowledge of greenhouse gas emissions from soils. Transport and transformations are frequently studied using 15N labeling experiments, but instruments capable of continuous measurements of 15N-N2O at the surface of soil have only recently come to the fore. Our primary aim was to quantify emissions of N2O and the fraction of 15N emitted as N2O from an agricultural soil following 15N addition using a mobile Fourier Transform Infrared (FTIR) spectrometer. We set up a short-term field experiment on a coastal floodplain site near Nowra, New South Wales. We deployed an automated chamber system connected to a multi-pass cell (optical pathlength 24 m) and low resolution FTIR spectrometer to measure fluxes of all N2O isotopologues collected from five 0.25 m2 chambers every three hours. We measured N2O fluxes pre and post-application of 15N-labeled substrate as potassium nitrate (KNO3) or urea [CO(NH2)2] to the soil surface. Root mean square uncertainties for all isotopologue measurements were less than 0.3 nmol mol-1 for 1 minute average concentration measurements, and minimum detectable fluxes for each isotopologue were <0.1 ng N m-2 s-1. Emissions of all N2O isotopologues were evident immediately following 15N addition. Emissions of 14N15NO, 15N14NO and 15N15NO isotopologues subsided within 10 d, but 14N14NO fluxes were evident over the entire experiment. The figure provides an overview of the emissions. Cumulative 15N-N2O fluxes (sum of the three 15N isotopologues) per chamber for the 14 days following 15N addition ranged from 1.5 to 10.3 mg 15N-N2O m-2. The chambers were destructively sampled after 2 weeks and 15N analyzed in soil and plant material using isotope ratio mass spectrometry. Approximately 1% (range 0.7 - 1.9%) of the total amount of

  2. Carbon-rich presolar grains from massive stars. Subsolar 12 C/ 13 C and 14 N/ 15 N ratios and the mystery of 15 N

    DOE PAGES

    Pignatari, M.; Zinner, E.; Hoppe, P.; Jordan, C. J.; Gibson, B. K.; Trappitsch, R.; Herwig, F.; Fryer, C.; Hirschi, R.; Timmes, F. X.

    2015-07-30

    We compared carbon-rich grains with isotopic anomalies to the Sun are found in primitive meteorites. They were made by stars, and carry the original stellar nucleosynthesis signature. Silicon carbide grains of Type X and C and low-density (LD) graphites condensed in the ejecta of core-collapse supernovae. Furthermore, we present a new set of models for the explosive He shell and compare them with the grains showing 12C/13C and 14N/15N ratios lower than solar. In the stellar progenitor H was ingested into the He shell and not fully destroyed before the explosion. All of the explosion energies and H concentrations aremore » considered. If the supernova shock hits the He-shell region with some H still present, the models can reproduce the C and N isotopic signatures in C-rich grains. Hot-CNO cycle isotopic signatures are obtained, including a large production of 13C and 15N. The short-lived radionuclides 22Na and 26Al are increased by orders of magnitude. The production of radiogenic 22Ne from the decay of 22Na in the He shell might solve the puzzle of the Ne-E(L) component in LD graphite grains. This scenario is attractive for the SiC grains of type AB with 14N/15N ratios lower than solar, and provides an alternative solution for SiC grains originally classified as nova grains. Finally, this process may contribute to the production of 14N and 15N in the Galaxy, helping to produce the 14N/15N ratio in the solar system.« less

  3. Carbon-rich Presolar Grains from Massive Stars: Subsolar 12C/13C and 14N/15N Ratios and the Mystery of 15N

    NASA Astrophysics Data System (ADS)

    Pignatari, M.; Zinner, E.; Hoppe, P.; Jordan, C. J.; Gibson, B. K.; Trappitsch, R.; Herwig, F.; Fryer, C.; Hirschi, R.; Timmes, F. X.

    2015-08-01

    Carbon-rich grains with isotopic anomalies compared to the Sun are found in primitive meteorites. They were made by stars, and carry the original stellar nucleosynthesis signature. Silicon carbide grains of Type X and C and low-density (LD) graphites condensed in the ejecta of core-collapse supernovae. We present a new set of models for the explosive He shell and compare them with the grains showing 12C/13C and 14N/15N ratios lower than solar. In the stellar progenitor H was ingested into the He shell and not fully destroyed before the explosion. Different explosion energies and H concentrations are considered. If the supernova shock hits the He-shell region with some H still present, the models can reproduce the C and N isotopic signatures in C-rich grains. Hot-CNO cycle isotopic signatures are obtained, including a large production of 13C and 15N. The short-lived radionuclides 22Na and 26Al are increased by orders of magnitude. The production of radiogenic 22Ne from the decay of 22Na in the He shell might solve the puzzle of the Ne-E(L) component in LD graphite grains. This scenario is attractive for the SiC grains of type AB with 14N/15N ratios lower than solar, and provides an alternative solution for SiC grains originally classified as nova grains. Finally, this process may contribute to the production of 14N and 15N in the Galaxy, helping to produce the 14N/15N ratio in the solar system.

  4. Covalent binding of reduced metabolites of [{sup 15}N{sub 3}]TNT to soil organic matter during a bioremediation process analyzed by {sup 15}N NMR spectroscopy

    SciTech Connect

    Achtnich, C.; Fernandes, E.; Bollag, J.M.; Knackmuss, H.J.; Lenke, H.

    1999-12-15

    Evidence is presented for the covalent binding of biologically reduced metabolites of 2,4,6-{sup 15}N{sub 3}-trinitrotoluene (TNT) to different soil fractions, using liquid {sup 15}N NMR spectroscopy. A silylation procedure was used to release soil organic matter from humin and whole soil for spectroscopic measurements. TNT-contaminated soil was spiked with 2,4,6-{sup 15}N{sub 3}-trinitrotoluene and {sup 14}C-ring labeled TNT, before treatment in a soil slurry reactor. During the anaerobic/aerobic incubation the amount of radioactivity detected in the fulvic and humic acid fractions did not change significantly whereas the radioactivity bound to humin increased to 71%. The {sup 15}N NMR spectra of the fulvic acid samples were dominated by a large peak that corresponded to aliphatic amines or ammonia. In the early stages of incubation, {sup 15}N NMR analysis of the humic acids indicated bound azoxy compounds. The signals arising from nitro and azoxy groups disappeared with further anaerobic treatment. At the end of incubation, the NMR shifts showed that nitrogen was covalently bound to humic acid as substituted amines and amides. The NMR spectra of the silylated humin suggest formation of azoxy compounds and imine linkages. Bound metabolites possessing nitro groups were also detected. Primary amines formed during the anaerobic incubation disappeared during the aerobic treatment. Simultaneously, the amount of amides and tertiary amines increased. Nitro and azoxy groups of bound molecules were still present in humin at the end of the incubation period. Formation of azoxy compounds from partially reduced TNT followed by binding and further reduction appears to be an important mechanism for the immobilization of metabolites of TNT to soil.

  5. δ(13)C and δ(15)N in deep-living fishes and shrimps after the Deepwater Horizon oil spill, Gulf of Mexico.

    PubMed

    Quintana-Rizzo, Ester; Torres, Joseph J; Ross, Steve W; Romero, Isabel; Watson, Kathleen; Goddard, Ethan; Hollander, David

    2015-05-15

    The blowout of the Deepwater Horizon (DWH) drill-rig produced a surface oil layer, dispersed micro-droplets throughout the water column, and sub-surface plumes. We measured stable carbon and nitrogen isotopes in mesopelagic fishes and shrimps in the vicinity of DWH collected prior to, six weeks after, and one year after the oil spill (2007, 2010 and 2011). In 2010, the year of the oil spill, a small but significant depletion of δ(13)C was found in two mesopelagic fishes (Gonostoma elongatum and Chauliodus sloani) and one shrimp (Systellaspis debilis); a significant δ(15)N enrichment was identified in the same shrimp and in three fish species (G. elongatum, Ceratoscopelus warmingii, and Lepidophanes guentheri). The δ(15)N change did not suggest a change of trophic level, but did indicate a change in diet. The data suggest that carbon from the Deepwater Horizon oil spill was incorporated into the mesopelagic food web of the Gulf of Mexico.

  6. Tracing industrial ammonium in atmospheric deposition in the Athabasca Oil Sands Region, Alberta, Canada

    NASA Astrophysics Data System (ADS)

    Mayer, B.; Proemse, B. C.; Fenn, M. E.

    2013-12-01

    The expanding industrial development in the Athabasca oil sands region (AOSR) in northeastern Alberta, Canada, has raised concerns about increasing nitrogen (N) emissions from oil sands operations and their potential effects on the surrounding terrestrial and aquatic ecosystems. Stable isotope techniques may help to trace industrial emissions provided that they are isotopically distinct from background isotope ratios of atmospheric N compounds. Ammonium deposition rates (NH4-N) typically exceed nitrate deposition rates (NO3-N) in the AOSR (Proemse et al., 2013), suggesting that emissions of reduced nitrogen compounds play a significant role for the atmospheric nitrogen budget in the AOSR. We collected atmospheric ammonium in open field bulk deposition and throughfall using ion exchange resins over ~6 months time periods from summer 2007 to summer 2011 located at distances between 3 to 113 km to one of the major oil sands developments in the AOSR. Ammonium deposition rates and δ15N-NH4 values were determined using ion chromatography and the ammonium diffusion method (Sebilo et al., 2004) on resin extracts. Atmospheric ammonium deposition rates in open field bulk collectors and throughfall collectors ranged from 1.0 to 4.7 kg ha-1 yr-1 NH4-N, and from 1.0 to 18.3 kg ha-1 yr-1 NH4-N, respectively. δ15N-NH4 values varied from -6.3 to +14.8‰ with the highest δ15N values typically associated with elevated NH4-N deposition rates. δ15N-NH4 values of up to +20.1‰ were observed for industrially emitted NH4 in particulate matter (PM2.5) emissions (Proemse et al., 2012) suggesting that industrial NH3 and NH4 emissions are associated with elevated δ15N values providing a potential tracer. Applying a two-end-member mixing analysis using a background δ15N-NH4 value of -3.6‰ for summer and -3.2‰ for winter periods revealed that particularly sites within ~30 km radius from the main oil sands developments are significantly affected by industrial contributions to

  7. Eastern oyster (Crassostrea virginica) δ15N as a bioindicator of nitrogen sources: Observations and modeling

    PubMed Central

    Fertig, B.; Carruthers, T.J.B.; Dennison, W.C.; Fertig, E.J.; Altabet, M.A.

    2013-01-01

    Stable nitrogen isotopes (δ15N) in bioindicators are increasingly employed to identify nitrogen sources in many ecosystems and biological characteristics of the eastern oyster (Crassostrea virginica) make it an appropriate species for this purpose. To assess nitrogen isotopic fractionation associated with assimilation and baseline variations in oyster mantle, gill, and muscle tissue δ15N, manipulative fieldwork in Chesapeake Bay and corresponding modeling exercises were conducted. This study (1) determined that five individuals represented an optimal sample size; (2) verified that δ15N in oysters from two locations converged after shared deployment to a new location reflecting a change in nitrogen sources; (3) identified required exposure time and temporal integration (four months for muscle, two to three months for gill and mantle); and (4) demonstrated seasonal δ15N increases in seston (summer) and oysters (winter). As bioindicators, oysters can be deployed for spatial interpolation of nitrogen sources, even in areas lacking extant populations. PMID:20381097

  8. Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction

    DOEpatents

    Chen, Xian; Gupta, Goutam; Bradbury, E. Morton

    2001-01-01

    Preparation of .sup.13 C/.sup.15 N-labeled DNA oligomers using the polymerase chain reaction (PCR). A PCR based method for uniform (.sup.13 C/.sup.15 N)-labeling of DNA duplexes is described. Multiple copies of a blunt-ended duplex are cloned into a plasmid, each copy containing the sequence of interest and restriction Hinc II sequences at both the 5' and 3' ends. PCR using bi-directional primers and uniformly .sup.13 C/.sup.15 N-labeled dNTP precursors generates labeled DNA duplexes containing multiple copies of the sequence of interest. Twenty-four cycles of PCR, followed by restriction and purification, gave the uniformly .sup.13 C/.sup.15 N-labeled duplex sequence with a 30% yield. Such labeled duplexes find significant applications in multinuclear magnetic resonance spectroscopy.

  9. An economical method for (15)N/(13)C isotopic labeling of proteins expressed in Pichia pastoris.

    PubMed

    Rodriguez, E; Krishna, N R

    2001-07-01

    We report a new and cost-effective approach to prepare (15)N/(13)C labeled proteins for NMR using the Pichia pastoris expression system. Four protocols (P1 to P4) were defined and compared using recombinant Ovine interferon-tau (rOvIFN-tau). Our results demonstrate that in order to get full incorporation of (15)N and (13)C, the isotopes are not totally required during the initial growth phase of P. pastoris culture. The addition of small amounts of (15)N and (13)C compounds 6 h prior to the methanol induction phase is sufficient to obtain 99% incorporation of heavy isotopes into the protein. Our optimized protocol P4 is two-thirds less costly than the classical method using (15)N and (13)C isotopes during the entire growth phase.

  10. Distribution of 15N-chlorocholine chloride in eggs of laying hens.

    PubMed

    Songsang, A; Chakeredza, S; Thinggaard, G; Vearasilp, T; ter Meulen, U

    2002-06-01

    The distribution of Chlorocholine chloride (CCC) in the eggs of laying hens was studied using 15N-CCC. Twelve layers (37 weeks old) were divided into four groups and used in this study consisting of three feeding phases. In phase one (7 days), all the hens received a CCC-free diet [165 g CP/kg dry matter (DM); 11.58 MJ ME/kg DM]. In phase two (11 days), four levels of 15N-CCC: 0, 5, 50 and 250 ppm were added to the respective diets, while in phase three (7 days), CCC-free feed was again offered. Egg samples were taken and the 15N content of egg yolk and albumin were determined. At the end of phase two, there was a significant (p < 0.05) increase in 15N content in egg yolk from hens fed the 50 and 250 ppm CCC diets and in albumin from hens fed the 250 ppm CCC diet. The estimated 15N-CCC residue was 1.71, 6.64, 28.80 ppm in egg yolk and 1.58, 1.08 and 4.50 ppm in albumin from hens fed 5, 50 and 250 ppm CCC, respectively. The CCC residue, from quantitative analysis ranged from 0.21 to 0.93 and 0.93 to 2.43 ppm in yolk of hens fed 50 and 250 ppm CCC, respectively, whereas a range of 0.40-1.46 ppm, was found in the albumin of hens fed 250 ppm. The difference in measured CCC in yolk and albumin and that estimated from 15N-CCC could have been due to breakdown products of 15N-CCC. Seven days after withdrawal of 15N-CCC, the estimated 15N-CCC residue in egg yolk decreased to 0.43, 2.45 and 15.59 ppm, on 5, 50 and 250 ppm CCC dietary treatments, respectively, and to 2.46 ppm in albumin from hens fed 250 ppm CCC. The higher increase in 15N content could have been due to a higher incorporation of 15N-CCC into yolk than albumin during the process of rapid yolk deposition. This experiment showed that consumed CCC is distributed both into yolk and albumin in a dose dependent manner and that CCC is metabolized in laying hens. However, the level of CCC in the diet which could lead to accumulation of detectable CCC levels in eggs as observed in this study, is much higher than the

  11. Partition of the relative contribution of nitrification and denitrification from Amazon forest soils using a model based on bulk 15N of N2O natural abundance determinations.

    NASA Astrophysics Data System (ADS)

    Perez, T. J.; Trumbore, S. E.; Tyler, S. C.; Park, S.; Boering, K.; Decamargo, P.

    2004-12-01

    Most of the available methods for the determination of the relative contribution of nitrification and denitrification to the soil emitted N2O are invasive. Therefore, they could produce biased results due to the change in soil structure, alteration to the microbial community and substrates. However, the soil community bacterial activity has intrinsic properties such as isotopic fractionation factors that are relative constant through different sets of soil conditions. We took advantage of these bacterial properties and devised a mass balance method for partitioning the relative contribution of each process by using: (1) The 15N enrichment factors for N2O production via nitrification and denitrification for soils (determined previously by acetylene addition soil incubation methods) and (2) the δ 15N-N2O soil emission values from the selected studied soils. We selected soils from a forest soil texture gradient from the Tapajos National Forest (TNF), in the Amazon Basin, Pará State, Brazil and Nova Vida Farm (NV), Rondonia State, Brazil where we had determined the 15N enrichment factors for each microbial process and collected N2O soil emissions for bulk stable isotope analysis during the rainy season of 2002. The soils selected were Oxisol (clay) and Ultison (sandy) at TNF and Latosol (sandy loam) at NV. We found that for all studied soils, the relative contribution of nitrification was smaller than 40 %. This corroborates the assumption that the N2O emitted from Amazon forest is mostly denitrification-derived. The advantage of this method is that is non invasive. However, the uncertainties associated with the method increase when δ 15N-N2O values of emitted N2O are smaller than -25 per mil.

  12. Water proton spin saturation affects measured protein backbone 15 N spin relaxation rates

    NASA Astrophysics Data System (ADS)

    Chen, Kang; Tjandra, Nico

    2011-12-01

    Protein backbone 15N NMR spin relaxation rates are useful in characterizing the protein dynamics and structures. To observe the protein nuclear-spin resonances a pulse sequence has to include a water suppression scheme. There are two commonly employed methods, saturating or dephasing the water spins with pulse field gradients and keeping them unperturbed with flip-back pulses. Here different water suppression methods were incorporated into pulse sequences to measure 15N longitudinal T1 and transversal rotating-frame T1ρ spin relaxation. Unexpectedly the 15N T1 relaxation time constants varied significantly with the choice of water suppression method. For a 25-kDa Escherichiacoli. glutamine binding protein (GlnBP) the T1 values acquired with the pulse sequence containing a water dephasing gradient are on average 20% longer than the ones obtained using a pulse sequence containing the water flip-back pulse. In contrast the two T1ρ data sets are correlated without an apparent offset. The average T1 difference was reduced to 12% when the experimental recycle delay was doubled, while the average T1 values from the flip-back measurements were nearly unchanged. Analysis of spectral signal to noise ratios ( s/ n) showed the apparent slower 15N relaxation obtained with the water dephasing experiment originated from the differences in 1H N recovery for each relaxation time point. This in turn offset signal reduction from 15N relaxation decay. The artifact becomes noticeable when the measured 15N relaxation time constant is comparable to recycle delay, e.g., the 15N T1 of medium to large proteins. The 15N relaxation rates measured with either water suppression schemes yield reasonable fits to the structure. However, data from the saturated scheme results in significantly lower Model-Free order parameters (< S2> = 0.81) than the non-saturated ones (< S2> = 0.88), indicating such order parameters may be previously underestimated.

  13. Ecosystem N distribution and δ15N during a century of forest regrowth after agricultural abandonment

    USGS Publications Warehouse

    Compton, J.E.; Hooker, T.D.; Perakis, S.S.

    2007-01-01

    Stable isotope ratios of terrestrial ecosystem nitrogen (N) pools reflect internal processes and input–output balances. Disturbance generally increases N cycling and loss, yet few studies have examined ecosystem δ15N over a disturbance-recovery sequence. We used a chronosequence approach to examine N distribution and δ15N during forest regrowth after agricultural abandonment. Site ages ranged from 10 to 115 years, with similar soils, climate, land-use history, and overstory vegetation (white pine Pinus strobus). Foliar N and δ15N decreased as stands aged, consistent with a progressive tightening of the N cycle during forest regrowth on agricultural lands. Over time, foliar δ15N became more negative, indicating increased fractionation along the mineralization–mycorrhizal–plant uptake pathway. Total ecosystem N was constant across the chronosequence, but substantial internal N redistribution occurred from the mineral soil to plants and litter over 115 years (>25% of ecosystem N or 1,610 kg ha−1). Temporal trends in soil δ15N generally reflected a redistribution of depleted N from the mineral soil to the developing O horizon. Although plants and soil δ15N are coupled over millennial time scales of ecosystem development, our observed divergence between plants and soil suggests that they can be uncoupled during the disturbance-regrowth sequence. The approximate 2‰ decrease in ecosystem δ15N over the century scale suggests significant incorporation of atmospheric N, which was not detected by traditional ecosystem N accounting. Consideration of temporal trends and disturbance legacies can improve our understanding of the influence of broader factors such as climate or N deposition on ecosystem N balances and δ15N.

  14. High performance ammonium nitrate propellant

    NASA Technical Reports Server (NTRS)

    Anderson, F. A. (Inventor)

    1979-01-01

    A high performance propellant having greatly reduced hydrogen chloride emission is presented. It is comprised of: (1) a minor amount of hydrocarbon binder (10-15%), (2) at least 85% solids including ammonium nitrate as the primary oxidizer (about 40% to 70%), (3) a significant amount (5-25%) powdered metal fuel, such as aluminum, (4) a small amount (5-25%) of ammonium perchlorate as a supplementary oxidizer, and (5) optionally a small amount (0-20%) of a nitramine.

  15. Acetylene inhibition of N2O reduction in laboratory soil and groundwater denitrification assays: evaluation by 15N tracer and 15N site preference of N2O

    NASA Astrophysics Data System (ADS)

    Weymann, Daniel; Well, Reinhard; Lewicka-Szczebak, Dominika; Lena, Rohe

    2013-04-01

    The measurement of denitrification in soils and aquifers is still challenging and often enough associated with considerable experimental effort and high costs. Against this background, the acetylene inhibition technique (AIT) applied in laboratory soil and groundwater denitrification assays is by far the most effective approach. However, this method has been largely criticized, as it is susceptible to underestimate denitrification rates and adds an additional carbon source to the substrates to be investigated. Here we provide evidence that the AIT is not necessarily an inappropriate approach to measure denitrification, that its reliability depends on the drivers governing the process, and that the 15N site preference of N2O (SP) may serve as a tool to assess this reliability. Two laboratory batch experiments were conducted, where sandy aquifer material and a peat soil were incubated as slurries. We established (i) a standard anaerobic treatment by adding KNO3 (10 mg N L-1), (ii) an oxygen treatment by adding KNO3 and O2 (5 mg L-1), and (iii) a glucose treatment by adding KNO3 supplemented with glucose (200 mg C L-1). Both experiments were run under 10 % (v/v) acetylene atmosphere and as 15N tracer treatments using labeled K15NO3 (60 atom % 15N). In the case of the standard anaerobic treatments, we found a very good agreement of denitrification potential obtained by the AIT and 15N tracer methods. SP of N2O of the AIT samples from this treatment ranged between -4.8 and 2.6 ‰ which is indicative for N2O production during bacterial denitrification but not for N2O reduction to N2. In contrast, we observed substantial underestimation of denitrification by AIT for the glucose treatments compared to the 15N method, i.e. denitrification was underestimated by 36 % (sandy aquifer material) and 47 % (peat soil). SP of N2O of the AIT samples from this treatment ranged between 4.5 and 9.6 ‰, which suggests occurrence of bacterial N2O reduction. In the case of the oxygen

  16. Soil N and 15N variation with time in a California annual grassland ecosystem

    USGS Publications Warehouse

    Brenner, D.L.; Amundson, Ronald; Baisden, W. Troy; Kendall, C.; Harden, J.

    2001-01-01

    The %N and ??15N values of soils and plants were measured along a chronosequence spanning 3 to 3000 Ky in a California annual grassland. Total soil N decreased with increasing soil age (1.1 to 0.4 kg N m-2) while the mean ?? 15N values of the soil N increased by several ??? from the youngest to oldest sites (+3.5 to +6.2 ???). The ?? 15N values of plants varied along the gradient, reflecting changing soil N pools and differences in the form of N uptake. The decline in total N storage with time is hypothesized to be due to a shift from N to P limitation with increasing soil age. The general increase in ?? 15N values with time is interpreted using a N mass balance model, and appears to reflect a shift toward an increasing proportional losses of inorganic mineral forms of N (vs. organic forms) with increasing soil age. We develop a quantitative index of this trend (mineral vs. organic forms of N loss) using mass balance considerations and parameters. The %N and ?? 15N values along the California age gradient were compared to the published data for a comparably aged chronosequence in Hawaii. Most striking in this comparison is the observation that the California soil and plant ?? 15N values are several ??? greater than those on comparably aged Hawaiian sites. Multiple explanations are plausible, but assuming the sites have a similar range in ?? 15N values of atmospheric inputs, the isotopic differences suggest that N may be, at least seasonally, in greater excess in the strongly seasonal, semi-arid, California grassland. Copyright ?? 2001 Elsevier Science Ltd.

  17. The Five-Year Fate of a 15N Tracer in a Mixed Deciduous Forest: Retention, Redistribution, and Differences by Mycorrhizal Association

    NASA Astrophysics Data System (ADS)

    Goodale, C. L.

    2015-12-01

    The impact of nitrogen deposition on forest ecosystems depends in large part on its fate: uptake by trees can stimulate growth, while gaseous or leaching losses contribute to air and water pollution and represent the loss of a limiting nutrient. Past tracer studies have shown that soils dominate the short-term fate of added 15N, but its longer-term term fate remains uncertain. This study examined how much 15N tracer moved plant or soil pools or was lost over 5-6 years. In 2007, a 15N tracer (0.21 kg/ha as 99% enriched 15N-KNO3) was added to 0.25 ha mixed hardwood forest in central NY. All of the tracer was recovered in the days after its addition, but recovery fell to 78% by the end of this year (25% surface litter, 48% 0-10 cm soil, 5% roots). One year later, recovery in these pools fell (to 51%), with losses from surface litter (-11%) and the 0-10 cm soils (-15%), including losses from the "heavy" soil fraction. Additional tracer moved to other plant pools (+5%) and to deeper soil (+13%; up to 30 cm), for a total recovery of 69% of the added tracer. Between years 1 and 5-6, only total tracer recovery decreased by only 1.4%. Recovery decreased in foliage (-0.2%), all roots (-3.5%), and surface litter (-9.8%), while increasing in woody biomass (+0.9%), 0-10 cm soil (+8.9%), and deep soil (+2.3%; up to 50 cm). Tracer recovery in live and dead plant N pools (11%) did not change, as 3% moved from roots into aboveground plant tissues and 3% moved from live plant pools into leaf litter; these results imply no net transfer of 15N from soil to plants during this period. Over all 5-6 years, only 1.6% of the tracer moved into bark or wood, a small but important sink because of its high C:N ratio; however, roughly one-third of this total was in wood formed prior to the start of the tracer addition. Tree species differed in their recovery of 15N: the six species with ectomycorrhizal associations showed more enrichment than the four species with arbuscular mycorrhizae. It is

  18. Oak loss increases foliar nitrogen, δ(15)N and growth rates of Betula lenta in a northern temperate deciduous forest.

    PubMed

    Falxa-Raymond, Nancy; Patterson, Angelica E; Schuster, William S F; Griffin, Kevin L

    2012-09-01

    Oak forests dominate much of the eastern USA, but their future is uncertain due to a number of threats and widespread failure of oak regeneration. A sudden loss of oaks (Quercus spp.) could be accompanied by major changes in forest nitrogen (N) cycles with important implications for plant nutrient uptake and tree species composition. In this study, we measured the changes in N use and growth rates of black birch trees (Betula lenta L.) following oak girdling at the Black Rock Forest in southeastern New York, USA. Data were collected from nine experimental plots composed of three treatments: 100% oaks girdled (OG), 50% oaks girdled (O50) and control (C). Foliar N concentration and foliar (15)N abundance increased significantly in the oak-girdled plots relative to the control, indicating that the loss of oaks significantly altered N cycling dynamics. As mineralization and nitrification rates increase following oak loss, black birch trees increase N absorption as indicated by higher foliar N content and increased growth rates. Foliar N concentration increased by 15.5% in the O50 and 30.6% in the OG plots relative to the control, while O50 and OG plots were enriched in (15)N by 1.08‰ and 3.33‰, respectively (P < 0.0001). A 641% increase in black birch growth rates in OG plots suggests that this species is able to respond to additional N availability and/or increased light availability. The loss of oaks and subsequent increase in black birch productivity may have a lasting impact on ecosystem form and function.

  19. Oak loss increases foliar nitrogen, δ(15)N and growth rates of Betula lenta in a northern temperate deciduous forest.

    PubMed

    Falxa-Raymond, Nancy; Patterson, Angelica E; Schuster, William S F; Griffin, Kevin L

    2012-09-01

    Oak forests dominate much of the eastern USA, but their future is uncertain due to a number of threats and widespread failure of oak regeneration. A sudden loss of oaks (Quercus spp.) could be accompanied by major changes in forest nitrogen (N) cycles with important implications for plant nutrient uptake and tree species composition. In this study, we measured the changes in N use and growth rates of black birch trees (Betula lenta L.) following oak girdling at the Black Rock Forest in southeastern New York, USA. Data were collected from nine experimental plots composed of three treatments: 100% oaks girdled (OG), 50% oaks girdled (O50) and control (C). Foliar N concentration and foliar (15)N abundance increased significantly in the oak-girdled plots relative to the control, indicating that the loss of oaks significantly altered N cycling dynamics. As mineralization and nitrification rates increase following oak loss, black birch trees increase N absorption as indicated by higher foliar N content and increased growth rates. Foliar N concentration increased by 15.5% in the O50 and 30.6% in the OG plots relative to the control, while O50 and OG plots were enriched in (15)N by 1.08‰ and 3.33‰, respectively (P < 0.0001). A 641% increase in black birch growth rates in OG plots suggests that this species is able to respond to additional N availability and/or increased light availability. The loss of oaks and subsequent increase in black birch productivity may have a lasting impact on ecosystem form and function. PMID:22851552

  20. Screening enoxaparin tetrasaccharide SEC fractions for 3-O-sulfo-N-sulfoglucosamine residues using [(1)H,(15)N] HSQC NMR.

    PubMed

    Beecher, Consuelo N; Manighalam, Matthew S; Nwachuku, Adanma F; Larive, Cynthia K

    2016-02-01

    Heparin and heparan sulfate (HS) are important in mediating a variety of biological processes through binding to myriad different proteins. Specific structural elements along the polysaccharide chains are essential for high affinity protein binding, such as the 3-O-sulfated N-sulfoglucosamine (GlcNS3S) residue, a relatively rare modification essential for heparin's anticoagulant activity. The isolation of 3-O-sulfated oligosaccharides from complex mixtures is challenging because of their low abundance. Although methods such as affinity chromatography are useful in isolating oligosaccharides that bind specific proteins with high affinity, other important 3-O-sulfated oligosaccharides may easily be overlooked. Screening preparative-scale size-exclusion chromatography (SEC) fractions of heparin or HS digests using [(1)H,(15)N] HSQC NMR allows the identification of fractions containing 3-O-sulfated oligosaccharides through the unique (1)H and (15)N chemical shifts of the GlcNS3S residue. Those SEC fractions containing 3-O-sulfated oligosaccharides can then be isolated using strong anion-exchange (SAX)-HPLC. Compared with the results obtained by pooling the fractions comprising a given SEC peak, SAX-HPLC analysis of individual SEC fractions produces a less complicated chromatogram in which the 3-O-sulfated oligosaccharides are enriched relative to more abundant components. The utility of this approach is demonstrated for tetrasaccharide SEC fractions of the low molecular weight heparin drug enoxaparin facilitating the isolation and characterization of an unsaturated 3-O-sulfated tetrasaccharide containing a portion of the antithrombin-III binding sequence.

  1. Solubility of the Sodium and Ammonium Salts of Oxalic Acid in Water with Ammonium Sulfate.

    PubMed

    Buttke, Lukas G; Schueller, Justin R; Pearson, Christian S; Beyer, Keith D

    2016-08-18

    The solubility of the sodium and ammonium salts of oxalic acid in water with ammonium sulfate present has been studied using differential scanning calorimetry, X-ray crystallography, and infrared spectroscopy. The crystals that form from aqueous mixtures of ammonium sulfate/sodium hydrogen oxalate were determined to be sodium hydrogen oxalate monohydrate under low ammonium sulfate conditions and ammonium hydrogen oxalate hemihydrate under high ammonium sulfate conditions. Crystals from aqueous mixtures of ammonium sulfate/sodium oxalate were determined to be ammonium oxalate monohydrate under moderate to high ammonium sulfate concentrations and sodium oxalate under low ammonium sulfate concentrations. It was also found that ammonium sulfate enhances the solubility of the sodium oxalate salts (salting in effect) and decreases the solubility of the ammonium oxalate salts (salting out effect). In addition, a partial phase diagram for the ammonium hydrogen oxalate/water system was determined.

  2. Solubility of the Sodium and Ammonium Salts of Oxalic Acid in Water with Ammonium Sulfate.

    PubMed

    Buttke, Lukas G; Schueller, Justin R; Pearson, Christian S; Beyer, Keith D

    2016-08-18

    The solubility of the sodium and ammonium salts of oxalic acid in water with ammonium sulfate present has been studied using differential scanning calorimetry, X-ray crystallography, and infrared spectroscopy. The crystals that form from aqueous mixtures of ammonium sulfate/sodium hydrogen oxalate were determined to be sodium hydrogen oxalate monohydrate under low ammonium sulfate conditions and ammonium hydrogen oxalate hemihydrate under high ammonium sulfate conditions. Crystals from aqueous mixtures of ammonium sulfate/sodium oxalate were determined to be ammonium oxalate monohydrate under moderate to high ammonium sulfate concentrations and sodium oxalate under low ammonium sulfate concentrations. It was also found that ammonium sulfate enhances the solubility of the sodium oxalate salts (salting in effect) and decreases the solubility of the ammonium oxalate salts (salting out effect). In addition, a partial phase diagram for the ammonium hydrogen oxalate/water system was determined. PMID:27482644

  3. Hydrogen Bonds in Crystalline Imidazoles Studied by 15N NMR and ab initio MO Calculations

    NASA Astrophysics Data System (ADS)

    Ueda, Takahiro; Nagatomo, Shigenori; Masui, Hirotsugu; Nakamura, Nobuo; Hayashi, Shigenobu

    1999-07-01

    Intermolecular hydrogen bonds of the type N-H...N in crystals of imidazole and its 4-substituted and 4,5-disubstituted derivatives were studied by 15N CP/MAS NMR and an ab initio molecular orbital (MO) calculation. In the 15N CP/MAS NMR spectrum of each of the imidazole derivatives, two peaks due to the two different functional groups, >NH and =N-, were observed. The value of the 15N isotropic chemical shift for each nitrogen atom depends on both the length of the intermolecular hydrogen bond and the kind of the substituent or substituents. It was found that the difference between the experimen-tal chemical shifts of >NH and =N-varies predominantly with the hydrogen bond length but does not show any systematic dependence on the kind of substituent. The ab initio MO calculations suggest that the hydrogen bond formation influences the 15N isotropic chemical shift predominantly, and that the difference between the 15N isotropic chemical shift of >NH and =N-varies linearly with the hydrogen bond length.

  4. Rivermouth Alteration of Agricultural Impacts on Consumer Tissue δ15N

    PubMed Central

    Larson, James H.; Richardson, William B.; Vallazza, Jon M.; Nelson, John C.

    2013-01-01

    Terrestrial agricultural activities strongly influence riverine nitrogen (N) dynamics, which is reflected in the δ15N of riverine consumer tissues. However, processes within aquatic ecosystems also influence consumer tissue δ15N. As aquatic processes become more important terrestrial inputs may become a weaker predictor of consumer tissue δ15N. In a previous study, this terrestrial-consumer tissue δ15N connection was very strong at river sites, but was disrupted by processes occurring in rivermouths (the ‘rivermouth effect’). This suggested that watershed indicators of N loading might be accurate in riverine settings, but could be inaccurate when considering N loading to the nearshore of large lakes and oceans. In this study, the rivermouth effect was examined on twenty-five sites spread across the Laurentian Great Lakes. Relationships between agriculture and consumer tissue δ15N occurred in both upstream rivers and at the outlets where rivermouths connect to the nearshore zone, but agriculture explained less variation and had a weaker effect at the outlet. These results suggest that rivermouths may sometimes be significant sources or sinks of N, which would cause N loading estimates to the nearshore zone that are typically made at discharge gages further upstream to be inaccurate. Identifying definitively the controls over the rivermouth effect on N loading (and other nutrients) will require integration of biogeochemical and hydrologic models. PMID:23935980

  5. Rivermouth alteration of agricultural impacts on consumer tissue δ15N

    USGS Publications Warehouse

    Larson, James H.; Richardson, William B.; Vallazza, Jonathan M.; Nelson, J. C.

    2013-01-01

    Terrestrial agricultural activities strongly influence riverine nitrogen (N) dynamics, which is reflected in the δ15N of riverine consumer tissues. However, processes within aquatic ecosystems also influence consumer tissue δ15N. As aquatic processes become more important terrestrial inputs may become a weaker predictor of consumer tissue δ15N. In a previous study, this terrestrial-consumer tissue δ15N connection was very strong at river sites, but was disrupted by processes occurring in rivermouths (the ‘rivermouth effect’). This suggested that watershed indicators of N loading might be accurate in riverine settings, but could be inaccurate when considering N loading to the nearshore of large lakes and oceans. In this study, the rivermouth effect was examined on twenty-five sites spread across the Laurentian Great Lakes. Relationships between agriculture and consumer tissue δ15N occurred in both upstream rivers and at the outlets where rivermouths connect to the nearshore zone, but agriculture explained less variation and had a weaker effect at the outlet. These results suggest that rivermouths may sometimes be significant sources or sinks of N, which would cause N loading estimates to the nearshore zone that are typically made at discharge gages further upstream to be inaccurate. Identifying definitively the controls over the rivermouth effect on N loading (and other nutrients) will require integration of biogeochemical and hydrologic models.

  6. Organically treated biochar increases plant production and reduces N2O emissions: mechanistic insights by 15N tracing

    NASA Astrophysics Data System (ADS)

    Kammann, Claudia; Messerschmidt, Nicole; Clough, Tim; Schmidt, Hans-Peter; Marhan, Sven; Koyro, Hans-Werner; Steffens, Diedrich; Müller, Christoph

    2014-05-01

    Pyrogenic carbon (biochar) offers considerable potential for carbon capture and soil storage (CCSS) compared to other, less recalcitrant soil-C additives. Recent meta-analysis demonstrated that it can significantly reduce agricultural N2O emissions. Freshly produced biochars, however, do not always have yield-improving effects, i.e. there is no immediate economic incentive for using it. Hence, combining biochar with organic nutrient-rich amendments may be a promising agricultural strategy to accelerate CCSS, but it is unclear if biochar still reduces N2O emissions, in particular when it may act as nutrient carrier. We explored the potential of biochar to improve the GHG-cost/yield ratio and thereby its socio-economic value as soil amendment in two subsequent studies under controlled conditions: (1) A proof-of-concept study where the effects of untreated biochar were compared to those of co-composted biochar combined with stepwise improved nutritional regimes (+/- compost; +/- mineral-N application), and (2) a 15N-labeling-tracing study to unravel N exchange on biochar particles and N2O production and reduction mechanisms. Both studies were carried out in nutrient-poor sandy soils, the most likely initial target soils for biochar-CCSS strategies. While the untreated biochar reduced plant growth under N-limiting conditions, or at best did not reduce it, the co-composted biochar always significantly stimulated plant growth. The relative stimulation was largest with the lowest nutrient additions (305% versus 61% of control with untreated biochar). Electro-ultra-filtration analyses revealed that the co-composted but not the untreated biochar carried considerable amounts of easily extractable as well as more strongly sorbed plant nutrients, in particular nitrate and phosphorus. The subsequent 15N labelling-tracing study revealed that the co-composted biochar still (i) acted as a mineral-N exchange site for nitrate and ammonium despite its N-preloading, (ii) reduced N2O

  7. Anaerobic ammonium oxidation for treatment of ammonium-rich wastewaters*

    PubMed Central

    Zhang, Lei; Zheng, Ping; Tang, Chong-jian; Jin, Ren-cun

    2008-01-01

    The concept of anaerobic ammonium oxidation (ANAMMOX) is presently of great interest. The functional bacteria belonging to the Planctomycete phylum and their metabolism are investigated by microbiologists. Meanwhile, the ANAMMOX is equally valuable in treatment of ammonium-rich wastewaters. Related processes including partial nitritation-ANAMMOX and completely autotrophic nitrogen removal over nitrite (CANON) have been developed, and lab-scale experiments proved that both processes were quite feasible in engineering with appropriate control. Successful full-scale practice in the Netherlands will accelerate application of the process in future. This review introduces the microbiology and more focuses on application of the ANAMMOX process. PMID:18500782

  8. Source apportionment of ammonium and nitrate ion using nitrogen stable isotope

    NASA Astrophysics Data System (ADS)

    Kawashima, H.

    2013-12-01

    Suspended particulate matter (SPM), defined to particle size as 100 % cut-off aerodynamic diameter at 10 μm, has adverse effects on human health. In these years, stable isotope ratio of small sample volume can be analyzed high precision by isotope ratio mass spectrometry coupling with elemental analyzer. Recently some fields are using stable isotope ratio. For environmental field, it is expected such as powerful tool for source identification and understanding mechanism. But the existed researches intended for stable nitrogen isotope (δ15N) of particulate matter have been limited. We try to analysis δ15N-ammonium15N-NH4+) and nitrate (δ15N-NO3-) of SPM, to estimate source of NH4+ and NO3- of SPM. Average δ15N-NH4+ and δ15N-NO3- of SPM in Akita prefecture, Japan were 15.9 ‰ (1.3‰ to 38.5 ‰) and - 0.7 ‰ (-4.6 ‰ to 4.8 ‰), respectively. Although δ15N-NH4+ do not show seasonal trend, δ15N-NO3- increased in winter markedly and decreasing in summer. In generally, the dominant origin of NO3- of SPM is produced from NOx emitted by combustion of some fuel and NO by agriculture source. Heaton (1990) summarized that δ15N-NOx is very different by temperature of combustion. They insisted that δ15N-NOx values are between -13 ‰ to -2 ‰ over 2000 °C (e.g. vehicle engine) and 6 ‰ to 13 ‰ under 1300 °C (e.g. coal combustion). Therefore, the reason of the winter high trend in this study might be combustion process such as coal combustion source. Moreover, the baseline might be made by vehicle sources. In addition, the reason of decreasing in summer seemed to be affected very low δ15N-NO of fertilizer and urea indicated by Li and Wang (2008). Bacteria were activated in summer, and NO from fertilizer and urea was emitted. This summary seemed to be very reasonable.

  9. Hyperpolarized 15N-pyridine Derivatives as pH-Sensitive MRI Agents

    PubMed Central

    Jiang, Weina; Lumata, Lloyd; Chen, Wei; Zhang, Shanrong; Kovacs, Zoltan; Sherry, A. Dean; Khemtong, Chalermchai

    2015-01-01

    Highly sensitive MR imaging agents that can accurately and rapidly monitor changes in pH would have diagnostic and prognostic value for many diseases. Here, we report an investigation of hyperpolarized 15N-pyridine derivatives as ultrasensitive pH-sensitive imaging probes. These molecules are easily polarized to high levels using standard dynamic nuclear polarization (DNP) techniques and their 15N chemical shifts were found to be highly sensitive to pH. These probes displayed sharp 15N resonances and large differences in chemical shifts (Δδ >90 ppm) between their free base and protonated forms. These favorable features make these agents highly suitable candidates for the detection of small changes in tissue pH near physiological values. PMID:25774436

  10. Synthesis and NMR of {sup 15}N-labeled DNA fragments

    SciTech Connect

    Jones, R.A.

    1994-12-01

    DNA fragments labeled with {sup 15}N at the ring nitrogens and at the exocyclic amino groups can be used to obtain novel insight into interactions such as base pairing, hydration, drug binding, and protein binding. A number of synthetic routes to {sup 15}N-labeled pyrimidine nucleosides, purines, and purine nucleosides have been reported. Moreover, many of these labeled bases or monomers have been incorporated into nucleic acids, either by chemical synthesis or by biosynthetic procedures. The focus of this chapter will be on the preparation of {sup 15}N-labeled purine 2{prime}-deoxynucleosides, their incorporation into DNA fragments by chemical synthesis, and the results of NMR studies using these labeled DNA fragments.

  11. Intraspecific variation in hair delta(13)C and delta(15)N values of ring-tailed lemurs (Lemur catta) with known individual histories, behavior, and feeding ecology.

    PubMed

    Loudon, James E; Sponheimer, Matt; Sauther, Michelle L; Cuozzo, Frank P

    2007-07-01

    Stable carbon and nitrogen isotope compositions were analyzed from hair samples of 30 sympatric ring-tailed lemurs (Lemur catta) inhabiting the Beza Mahafaly Special Reserve, Madagascar. All lemurs were known individuals involved in a longitudinal study, which allowed us to explore the degree to which group membership, sex, health status, and migration influenced their stable isotope compositions. The differences in delta(13)C and delta(15)N values between groups were small (<1.5 per thousand) but highly significant. In fact, each group was tightly clustered, and discriminant function analysis of the stable isotope data assigned individuals to the group in which they were originally collared with over 90% accuracy. In general, the differences between groups reflected the degree to which they utilized forested versus open habitats. As open habitats at Beza Mahafaly often correspond to areas of anthropogenic disturbance, these data suggest that isotopic data can be useful for addressing questions of lemur conservation. There were few sex differences, but significant differences did occur between individuals of normal and suboptimal health, with those in poor health (especially those in the worst condition) being enriched in (15)N and to a lesser degree (13)C compared with healthy individuals. Moreover, lemurs that had emigrated between 2003 and 2004 had different delta(13)C and delta(15)N compositions than their original groups. PMID:17455284

  12. Inoculation of Bacillus sphaericus UPMB-10 to Young Oil Palm and Measurement of Its Uptake of Fixed Nitrogen Using the 15N Isotope Dilution Technique

    PubMed Central

    Zakry, Fitri Abdul Aziz; Shamsuddin, Zulkifli H.; Rahim, Khairuddin Abdul; Zakaria, Zin Zawawi; Rahim, Anuar Abdul

    2012-01-01

    There are increasing applications of diazotrophic rhizobacteria in the sustainable agriculture system. A field experiment on young immature oil palm was conducted to quantify the uptake of N derived from N2 fixation by the diazotroph Bacillus sphaericus strain UPMB-10, using the 15N isotope dilution method. Eight months after 15N application, young immature oil palms that received 67% of standard N fertilizer application together with B. sphaericus inoculation had significantly lower 15N enrichment than uninoculated palms that received similar N fertilizers. The dilution of labeled N served as a marker for the occurrence of biological N2 fixation. The proportion of N uptake that was derived from the atmosphere was estimated as 63% on the whole plant basis. The inoculation process increased the N and dry matter yields of the palm leaflets and rachis significantly. Field planting of young, immature oil palm in soil inoculated with B. sphaericus UPMB-10 might mitigate inorganic fertilizer-N application through supplementation by biological nitrogen fixation. This could be a new and important source of nitrogen biofertilizer in the early phase of oil palm cultivation in the field. PMID:22446306

  13. Stable Isotope Evidence for Abiotic Ammonium Production in the Hydrothermal Vent Fluids from the Mid-Cayman Rise

    NASA Astrophysics Data System (ADS)

    Charoenpong, C.; Wankel, S. D.; Seewald, J.

    2015-12-01

    The Mid-Cayman Rise hosts the world's deepest (up to 4,987 meters) hydrothermal vent field, Piccard. Under the tremendous pressure, the vent fluid from Piccard can be as hot as 398°C. Here, the concentration of ammonium (35 μmol/kg) is much higher than that of the bottom water nitrate (22 μmol/kg). The undetectable nitrate in the vent fluid suggests that nitrate is completely reduced to ammonium in the reaction zone and there has to be an additional source for ammonium production. Because Piccard is unsedimented (i.e., lacking significant sedimentary organic matter), the other possible source of ammonium is the reduction of nitrogen gas (N2). We demonstrated that the isotopic composition of ammonium15N-NH4+) for the Piccard vent fluid end-member (3.4 ± 0.1 ‰) supports the mixing between two ammonium sources: the reduction of nitrate and the reduction of N2. However, at Von Damm, shallower depth (up to 2,300 meters) causes the vent fluid to be relatively cooler (138°C) compared to Piccard and it is very likely that the reduction of nitrate is the only source of ammonium in the vent fluid. Studying the vent fluids from these sites where the temperatures are well above the upper limit for life can be useful in assessing the conditions and abiotic processes that might have given rise to the ammonium production in the early prebiotic ocean.

  14. δ 13C and δ 15N biogeographic trends in rocky intertidal communities along the coast of South Africa: Evidence of strong environmental signatures

    NASA Astrophysics Data System (ADS)

    Hill, Jaclyn M.; McQuaid, Christopher D.

    2008-11-01

    Ecosystem dynamics driven by top-down controls have been well documented in rocky intertidal communities, while the effects of bottom-up influences are comparatively poorly understood. We hypothesized that large-scale signatures of the physical environment may be identifiable along the South African coastline as it is subject to two very different current systems (Benguela and Agulhas Currents) that profoundly influence primary production and thus both food type and availability. Through stable isotope analysis, we examined biogeographic patterns in multiple trophic levels at four sites along a 1400-km stretch of South African coastline and investigated the dietary role of macroalgal-derived organic carbon in rocky intertidal communities. The general positioning of trophic groups was comparable across all sites, with animals from the same trophic levels grouping together and with a δ 15N fractionation of 1-2‰ between levels. The species found at all sites demonstrated east-west δ 15N enrichment, presumably reflecting a biogeographic shift in nitrogen sources linked to upwelling on the west coast. Filter-feeders gave particularly clear results. Using discriminant analysis, mussels could be categorized into four geographic groups based on carbon and nitrogen signatures: east coast, southeast coast, south-west coast and west coast. Barnacles and polychaetes showed similar geographic groupings to mussels, but with shifts in actual values (1‰ depletion in δ 13C and 3‰ enrichment in δ 15N relative to mussels). This suggests that fractionation varies between species within a trophic level. IsoSource models showed that Ulva sp. made large contributions to the diets of two microalgal grazers ( Siphonaria capensis and Scutellastra granularis) and this dietary dependence increased when moving from west to east coast, along the shoreline. Additionally, IsoSource models determined that relative to phytoplankton, macroalgae accounted for upwards of 60% of suspended

  15. TREATMENT OF AMMONIUM NITRATE SOLUTIONS

    DOEpatents

    Boyer, T.W.; MacHutchin, J.G.; Yaffe, L.

    1958-06-10

    The treatment of waste solutions obtained in the processing of neutron- irradiated uranium containing fission products and ammonium nitrate is described. The object of this process is to provide a method whereby the ammonium nitrate is destroyed and removed from the solution so as to permit subsequent concentration of the solution.. In accordance with the process the residual nitrate solutions are treated with an excess of alkyl acid anhydride, such as acetic anhydride. Preferably, the residual nitrate solution is added to an excess of the acetic anhydride at such a rate that external heat is not required. The result of this operation is that the ammonium nitrate and acetic anhydride react to form N/sub 2/ O and acetic acid.

  16. 15N Fractionation in Star-Forming Regions and Solar System Objects

    NASA Technical Reports Server (NTRS)

    Wirstrom, Eva; Milam, Stefanie; Adande, GIlles; Charnley, Steven; Cordiner, Martin

    2015-01-01

    A central issue for understanding the formation and evolution of matter in the early Solar System is the relationship between the chemical composition of star-forming interstellar clouds and that of primitive Solar System materials. The pristinemolecular content of comets, interplanetary dust particles and carbonaceous chondrites show significant bulk nitrogen isotopic fractionation relative to the solar value, 14N15N 440. In addition, high spatial resolution measurements in primitive materials locally show even more extreme enhancements of 14N15N 100.

  17. Stable isotope enrichment in stratospheric nitrous oxide

    SciTech Connect

    Rahn, T.; Wahlen, M.

    1997-12-05

    Nitrous oxide is a greenhouse gas that also plays a role in the cycling of stratospheric ozone. Air samples from the lower stratosphere exhibit {sup 15}N/{sup 14}N and {sup 18}O/{sup 16}O enrichment in nitrous oxide, which can be accounted for with a simple model describing an irreversible destruction process. The observed enrichments are quite large and incompatible with those determined for the main stratospheric nitrous oxide loss processes of photolysis and reaction with excited atomic oxygen. Thus, although no stratospheric source needs to be invoked, the data indicate that present understanding of stratospheric nitrous oxide chemistry is incomplete. 21 refs., 1 fig., 1 tab.

  18. Stellar Origins of C-13 and N-15-Enriched Presolar SiC Grains

    NASA Technical Reports Server (NTRS)

    Liu, Nan; Nittler, Larry R.; Alexander, Conel M. O’D.; Wang, Jianhua; Pignatari, Marco; Jose, Jordi; Nguyen, Ann

    2016-01-01

    Extreme excesses of 13 C ( C (12 C/ 13 C<10) and 15 N ( N (14 N/ 15 N< 20) in rare presolar SiC 20) in rare presolar SiClar SiC grains have been considered diagnostic of an origin in classical novae [1], though an origin in core-collapse supernovae (CCSNe) has also been proposed [2]. We report multi-element isotopic data for 19 13 C- and 15 N-enriched presolar SiC grains(12 C/13 C<16 and 14 N/ 15 N<150) from an acid resistant residue of the Murchison meteorite. These grains are enriched in 13 C and15 N, but with quite diverse Si isotopic signatures. Four grains with isotopic signatures. Four grains with isotopic signatures. Four grains with isotopic signatures. Four grains with isotopic signatures.

  19. Using δ15 N- and δ18 O-NO to Evaluate Mechanisms of Nitric Oxide Production Following the Wetting of Dry Soil

    NASA Astrophysics Data System (ADS)

    Homyak, P. M.; Schimel, J.; Sickman, J. O.

    2014-12-01

    In xeric environments, where soils can remain dry for more than 6 months, abrupt transitions from dry-to-wet conditions produce NO pulses within seconds after soils wet up. During these periods of intense gaseous N production, biological processes (nitrification and denitrification) are known to control NO fluxes, but it is not clear how soil microbes can recover from drought-induced stress within seconds after soils wet up. Are NO pulses immediately following rewetting more so controlled by abiotic NO-producing reactions? Because biotic and abiotic mechanisms can occur simultaneously, distinguishing between these processes can be problematic. To understand the contribution of biotic and abiotic processes to NO pulses, and to better inform biogeochemical models, we measured the δ15N- and δ18O-NO following a field soil rewetting experiment in a California annual grassland. In October, during the end of the dry season, we artificially watered soils and captured NO emissions for up to 15 minutes, 1 hour, 1 day, and 3 days after wet-up. Pulses of NO following the wetting of dry soil were explained by a two-component mixing model, where two distinct sources or processes produced NO. Within 15 minutes after soil wet-up, the isotopic composition of soil NO (δ15N =-8.95 ‰, δ18O=14.28 ‰) was similar to that of atmospheric samples (δ15N =-4.45 ‰, δ18O=15.20 ‰), but became increasingly depleted after 1 hour (δ15N =-21.08 ‰, δ18O=0.53 ‰), and more so after 1 day (δ15N =-37.44 ‰, δ18O=-9.45 ‰). After 3 days, the isotopic composition of NO (δ15N =-28.31 ‰, δ18O=-2.07 ‰) began to return to pre-wet-up conditions closely following the two-component mixing line. We conclude that NO-producing reactions immediately after the wetting of dry soil (up to 15 min) are different than those occurring after 1 hour post-wetting. We hypothesize that abiotic processes control the initial response to wetting, but that biological processes, which discriminate

  20. First results on the incorporation and excretion of 15N from orally administered urea in lactating pony mares.

    PubMed

    Schubert, R; Zander, R; Gruhn, K; Hennig, A

    1991-05-01

    Two lactating pony mares were given oral offers of 20 g 15N urea [95 atom-% 15N-excess (15N')] on 6 subsequent days. About 80% of the consumed 15N' were excreted via urine and faeces, but only about 2% via milk. The 15N' secreted via milk-lysine only amounted to 0.04% of the 15N' intake. The recovery was about 90% in each case. Tissues with active metabolism had an unexpectedly high labelling (greater than 0.3 atom-% 15N'). The low extent of the conversion of oral urea N into milk-lysine speaks against an essential participation of the enteral synthesis in meeting the amino acid requirement of lactating mares. It was already concluded from this results that the determination of the amino acid requirement will be necessary for this group of performance. PMID:1888274

  1. The degree of urbanization across the globe is not reflected in the δ(15)N of seagrass leaves.

    PubMed

    Christiaen, Bart; Bernard, Rebecca J; Mortazavi, Behzad; Cebrian, Just; Ortmann, Alice C

    2014-06-30

    Many studies show that seagrass δ(15)N ratios increase with the amount of urbanization in coastal watersheds. However, there is little information on the relationship between urbanization and seagrass δ(15)N ratios on a global scale. We performed a meta-analysis on seagrass samples from 79 independent locations to test if seagrass δ(15)N ratios correlate with patterns of population density and fertilizer use within a radius of 10-200 km around the sample locations. Our results show that seagrass δ(15)N ratios are more influenced by intergeneric and latitudinal differences than the degree of urbanization or the amount of fertilizer used in nearby watersheds. The positive correlation between seagrass δ(15)N ratios and latitude hints at an underlying pattern in discrimination or a latitudinal gradient in the (15)N isotopic signature of nitrogen assimilated by the plants. The actual mechanisms responsible for the correlation between δ(15)N and latitude remain unknown.

  2. Using 15N, 17O, and 18O to determine nitrate sources in the Yellow River, China.

    PubMed

    Liu, Ting; Wang, Fan; Michalski, Greg; Xia, Xinghui; Liu, Shaoda

    2013-01-01

    Many previous studies have used δ(15)N and δ(18)O of nitrate (δ(15)NNO3 and δ(18)ONO3) to determine the nitrate sources in rivers but were subject to substantial uncertainties and limitations, especially associated with evaluating the atmospheric contribution. The Δ(17)O of nitrate (Δ(17)ONO3) has been suggested as an unambiguous tracer of atmospheric NO3(-) and may serve as an additional nitrate source constraint. In the present study, triple nitrate isotopes (δ(15)NNO3, Δ(17)ONO3, and δ(18)ONO3) were used for the first time to assess the sources and sinks of nitrate in the Yellow River (YR) basin, which is the second longest river in China. Results showed that the Δ(17)ONO3 of the water from the YR ranged from 0‰ to 1.6‰ during two normal-water seasons. This suggested that unprocessed atmospheric nitrate accounted for 0-7% of the total nitrate in the YR. The corrected δ(15)NNO3 and δ(18)ONO3 values with atmospheric imprints being removed indicated that the main terrestrial sources of nitrate were sewage/manure effluents in the upstream of the YR and manure/sewage effluents and ammonium/urea-containing fertilizer in the middle and lower reaches which made comparable contributions to the nitrate. In addition, there was a significant positive relationship between δ(15)NNO3 and δ(18)ONO3 values of river water (p < 0.01) which may signal the presence of denitrification. This study indicates that the triple nitrate isotope method is useful for assessing the nitrate sources in rivers, especially for the measurements of atmospheric nitrate contribution.

  3. Importance of bacterivory and preferential selection toward diatoms in larvae of Crepidula fornicata (L.) assessed by a dual stable isotope (13C, 15N) labeling approach

    NASA Astrophysics Data System (ADS)

    Leroy, Fanny; Riera, Pascal; Jeanthon, Christian; Edmond, Frédérique; Leroux, Cédric; Comtet, Thierry

    2012-05-01

    In Europe, the gastropod Crepidula fornicata is an invasive species characterized by a long reproductive period (from February to November). Thus, its larvae are exposed to variations in available food sources (in terms of quantity and quality). We aimed to investigate if bacteria could contribute to larval food both in presence or absence of phytoplankton, and to compare these results to seasonal variations of bacteria and phytoplankton abundances at a coastal site in the English Channel. First, ingestion of fluorescent beads of 0.5 to 2 μm diameter, showed that larvae were able to ingest particles of typical bacterial size. Then we used a dual stable isotope labeling approach which consisted in labeling a bacterial pelagic community with 15N and a diatom (Chaetoceros gracilis) culture with 13C, and supplying larvae with 15N-labeled bacteria, 13C-labeled diatoms, and both labeled sources. This technique has, to our knowledge, never been applied to invertebrate larvae. After 24 h of experiment, larvae were significantly enriched in all treatments: + 21.5‰ (∆δ13C) when supplied with diatoms, + 1364‰ (∆δ15N) when supplied with bacteria, and + 24‰ (∆δ13C) and + 135‰ (∆δ15N) when supplied with the two mixed sources. These results indicated that bacteria can contribute to the larval nutrition in C. fornicata, even in the presence of phytoplankton. Our results however suggested that larvae of C. fornicata preferentially used diatoms and showed that the supply of free bacteria did not alter the uptake of diatoms. Considering the seasonal variations of bacteria and phytoplankton abundances at the study site, these results suggested that bacteria may constitute a complementary resource for the larvae of C. fornicata when phytoplankton is abundant and may become a substitute resource when phytoplankton is less available. This approach offers promising perspectives to trace food sources and assess nitrogen and carbon fluxes between planktotrophic larvae

  4. /sup 15/N-Ammonia assimilation, 2-oxoglutarate transport, and glutamate export in spinach chloroplasts in the presence of dicarboxylates in the light

    SciTech Connect

    Woo, K.C.; Boyle, F.A.; Flugge, I.U.; Heldt, H.W.

    1987-11-01

    The direct incorporation of /sup 15/NH/sub 4/Cl into amino acids in illuminated spinach (Spinacia oleracea L.) chloroplasts in the presence of 2-oxoglutarate plus malate was determined. The amido-N of glutamine was the most highly labeled N-atom during /sup 15/NH/sub 4/ assimilation in the presence of malate. In 4 minutes the /sup 15/N-label of the amido-N of glutamine was 37% enriched. In contrast, values obtained for both the N-atom of glutamate and the amino-N of glutamine were only about 20% while that of the N-atom of aspartate was only 3%. The addition of malate during the assimilation of /sup 15/NH/sub 4/Cl and Na/sup 15/NO/sub 2/ greatly increased the /sup 15/N-label into glutamine but did not qualitatively change the order of the incorporation of /sup 15/N-label into all the amino acids examined. This evidence indicates the direct involvement of the glutamine synthetase/glutamate synthase pathway for ammonia and nitrite assimilation in isolated chloroplasts. The addition of malate or succinate during ammonia assimilation also led to more than 3-fold increase in (/sup 14/C)2-oxoglutarate transport into the chloroplast as well as an increase in the export of (/sup 14/C)glutamate out of the chloroplast. Little (/sup 14/C)glutamine was detected in the medium of the chloroplast preparations. The stimulation of /sup 15/N-incorporation and (/sup 14/C)glutamate export by malate could be directly attributed to the increase in 2-oxoglutarate transport activity (via the 2-oxoglutarate translocator) observed in the presence of exogenous malate.

  5. Changes in leaf δ13C and δ15N for three Mediterranean tree species in relation to soil water availability

    NASA Astrophysics Data System (ADS)

    Ogaya, Romà; Peñuelas, Josep

    2008-11-01

    A rain exclusion experiment simulating drought conditions expected in Mediterranean areas for the following decades (15% decrease in soil moisture) was conducted in a Mediterranean holm oak forest to study the response of leaf δ13C, δ15N, and N concentrations to the predicted climatic changes for the coming decades. Plant material was sampled in 2000, 2003, 2004, and 2005 in eight plots: four of them were control plots and the other four plots received the rain exclusion treatment. Although there was a negative relationship between δ13C and soil moisture, for each species and year, the rain exclusion treatment did not have any significant effect on δ13C, and therefore on the intrinsic water use efficiency (iWUE) of the three dominant species: Phillyrea latifolia, Arbutus unedo, and Quercus ilex. On the other hand, rain exclusion clearly increased the δ15N values in the three species studied, probably indicating higher N losses at the soil level leading to a 15N enrichment of the available N. It suggested that rain exclusion exerted a greater effect on the nitrogen biogeochemical cycle than on the carbon assimilation process. δ15N values were inversely correlated with summer soil moisture in Q. ilex and A. unedo, but no relationship was observed in P. latifolia. This latter species showed the lowest iWUE values, but it was the only species with no decrease in annual basal increment in response to the rain exclusion treatment, and it also had the highest resistance to the hot and dry conditions projected for the Mediterranean basin in the coming decades. The different strategies to resist rain exclusion conditions of these species could induce changes in their competitive ability and future distribution. The losses of N from the ecosystem may further limit plant growth and ecosystem functioning.

  6. Spectroscopic observations of 14N/15N ratios in both NH2 and CN in comet C/2013 US10 (Catalina)

    NASA Astrophysics Data System (ADS)

    Shinnaka, Yoshiharu; Kawakita, Hideyo

    2016-10-01

    Comet is one of the primordial small bodies in the solar system and probably it has kept the information about the evolution of materials from the pre-solar molecular cloud to the solar nebula.Isotopic ratio in volatiles is one of the primordial properties of comets. A heavier isotopes trend to be captured into a molecule by chemical reactions under very low-temperature conditions (called as fractionation). For instance, D/H ratio of water (HDO/H2O) in comet is enriched in D atom than the elemental abundance ratios of D/H in entire solar system [1]. Based on the observed D/H ratios in cometary water, a presumed temperature is ~20–50 K as the formation temperature of water (most abundant volatiles in cometary nucleus), by assuming water formed in gas-phase chemistry [2].Besides, the nitrogen isotopic ratios (14N/15N) have been determined from CN and HCN (which is believed a dominant "parent" species of CN in the coma) in >20 comets [3,4]. They demonstrated cometary HCN and CN show high 15N-fractionation with respect to the proto-solar value by a factor of ~3 and with a small diversity. Moreover, 14N/15N ratios in NH3 in comets has been determined from intensity ratios of NH2 isotopologues [5,6,7], and both 15N-fractionation as much as HCN in comets and a small diversity are seen in those 14N/15N ratios in NH3. However, there is a few reports about 14N/15N ratios in both HCN and NH3 in the same comets, and discussions about the relationship between these 14N/15N ratios have not been yet.We present 14N/15N ratios in both NH2 and CN in comet C/2013 US10 (Catalina). High-resolution optical spectra of the comet were taken with the HDS spectrograph mounted on the Subaru Telescope (Hawaii) on UT 2016 January 2–3. We will discuss about the origins of these volatiles based on the 14N/15N ratios.This work was supported by Graint-in-Aid for JSPS Fellows, 15J10864 (YS).References:[1] Lis et al., 2013, ApJ 774, L3[2] Millar et al., 1989, ApJ 340, 906[3] Bockelée-Morvan et

  7. Food web structure of the epibenthic and infaunal invertebrates on the Catalan slope (NW Mediterranean): Evidence from δ 13C and δ 15N analysis

    NASA Astrophysics Data System (ADS)

    Fanelli, E.; Papiol, V.; Cartes, J. E.; Rumolo, P.; Brunet, C.; Sprovieri, M.

    2011-01-01

    The food-web structure of the epibenthic and infaunal invertebrates on the continental slope of the Catalan Sea (Balearic basin, NW Mediterranean) was investigated using carbon and nitrogen stable isotopes on a total of 34 species, and HPLC pigment analyses for three key species. Samples were collected close to Barcelona (NE Iberian Peninsula), between 650 and 800 m depth and between February 2007 and February 2008. Mean δ 13C values ranged from -21.0‰ (small Calocaris macandreae and Amphipholis squamata) to -14.5‰ ( Sipunculus norvegicus). Values of δ 15N ranged from 4.0‰ ( A. squamata) to 12.1‰ ( Molpadia musculus). The stable isotope ratios of benthic fauna displayed a continuum of values (e.g. δ 15N range of 8‰), confirming a wide spectrum of feeding strategies (from active suspension feeders to predators) and complex food webs. According to the available information on diets of benthic fauna, the lowest values were found for surface deposit feeders (small C. macandrae and the two ophiuroids A. squamata and Amphiura chiajei) and active suspension feeders ( Abra longicallus and Scalpellum scalpellum) feeding on different sizes of particulate organic matter (POM), among which small particles may exhibit lower δ 15N. High annual mean δ 15N values were found among sub-surface deposit feeders, exploiting refractory or frequently recycled organic matter that is enriched in δ 15N. Carnivorous polychaetes ( Nephtys spp., Oenonidae and Polynoidae) and large decapods ( Geryon longipes and Paromola cuvieri) also displayed high δ 15N values. δ 13C ranges were particularly wide among surface deposit feeders (ranging from -21.0‰ to -16.4‰), suggesting exploitation of POM of both terrigenous and oceanic origins. Correlation between δ 13C and δ 15N was generally weak, indicating multiple carbon sources, likely due to the consumption of different kinds of sinking particles (e.g. marine snow, phytodetritus, etc.), sedimented and frequently recycled POM

  8. Nitrate Removal in Two Relict Oxbow Urban Wetlands: A 15N Mass-balance Approach

    EPA Science Inventory

    A 15N-tracer method was used to quantify nitrogen (N) removal processes in two relict oxbow wetlands located adjacent to the Minebank Run restored stream reach in Baltimore County (Maryland, USA) during summer 2009 and early spring 2010. A mass-balance approach was used to determ...

  9. Using a Macroalgal δ15N Bioassay to Detect Cruise Ship Waste Water Effluent Inputs

    EPA Science Inventory

    Nitrogen stable isotopes are a powerful tool for tracking sources of N to marine ecosystems. I used green macroalgae as a bioassay organism to evaluate if the δ15N signature of cruise ship waste water effluent (CSWWE) could be detected in Skagway Harbor, AK. Opportunistic green...

  10. δ15N as a proxy for historic anthropogenic nitrogen loading in Charleston Harbor, SC, USA

    NASA Astrophysics Data System (ADS)

    Payne, T. N.; Andrus, C. F. T.

    2015-12-01

    Bivalve shell geochemistry can serve as a useful indicator of changes in coastal environments. There is increasing interest in developing paleoenvironmental proxies from mollusk shell organic components. Numerous studies have focused on how the δ15N obtained from bivalve tissues can be used to trace present-day wastewater input into estuaries. However, comparatively little attention has been paid to tracing the impact of anthropogenic nitrogen loading into estuaries over time. By measuring historic levels of δ15N in the organic fraction of oyster shells (Crassostrea virginica) from archaeological sites around Charleston Harbor and comparing those levels to the δ15N content of modern shells, it is possible to assess how nitrogen has fluctuated historically in the area. Whole-shell samples from the Late Archaic Period (~3000-4000 BP, Late Woodland Period (~1400-800 BP), 18th and 19th centuries, and modern controls were measured for %N and d15N. Evidence of increased anthropogenic input of N is expected to begin in the early historic period based on similar analysis in Chesapeake Bay. More ancient samples may give insight into baseline conditions prior to recent population growth and industrialization. This information could help understand how large-scale anthropogenic nitrogen loading has affected coastal ecosystems over time and guide future remediation. Furthermore, this project will help refine and improve this novel proxy of past environmental conditions.

  11. Monitoring the refinement of crystal structures with (15)N solid-state NMR shift tensor data.

    PubMed

    Kalakewich, Keyton; Iuliucci, Robbie; Mueller, Karl T; Eloranta, Harriet; Harper, James K

    2015-11-21

    The (15)N chemical shift tensor is shown to be extremely sensitive to lattice structure and a powerful metric for monitoring density functional theory refinements of crystal structures. These refinements include lattice effects and are applied here to five crystal structures. All structures improve based on a better agreement between experimental and calculated (15)N tensors, with an average improvement of 47.0 ppm. Structural improvement is further indicated by a decrease in forces on the atoms by 2-3 orders of magnitude and a greater similarity in atom positions to neutron diffraction structures. These refinements change bond lengths by more than the diffraction errors including adjustments to X-Y and X-H bonds (X, Y = C, N, and O) of 0.028 ± 0.002 Å and 0.144 ± 0.036 Å, respectively. The acquisition of (15)N tensors at natural abundance is challenging and this limitation is overcome by improved (1)H decoupling in the FIREMAT method. This decoupling dramatically narrows linewidths, improves signal-to-noise by up to 317%, and significantly improves the accuracy of measured tensors. A total of 39 tensors are measured with shifts distributed over a range of more than 400 ppm. Overall, experimental (15)N tensors are at least 5 times more sensitive to crystal structure than (13)C tensors due to nitrogen's greater polarizability and larger range of chemical shifts. PMID:26590548

  12. The use of delta(15)N in assessing sewage stress on coral reefs.

    PubMed

    Risk, Michael J; Lapointe, Brian E; Sherwood, Owen A; Bedford, Bradley J

    2009-06-01

    While coral reefs decline, scientists argue, and effective strategies to manage land-based pollution lag behind the extent of the problem. There is need for objective, cost-effective, assessment methods. The measurement of stable nitrogen isotope ratios, delta(15)N, in tissues of reef organisms shows promise as an indicator of sewage stress. The choice of target organism will depend upon study purpose, availability, and other considerations such as conservation. Algae are usually plentiful and have been shown faithfully to track sewage input. The organic matrix of bivalve shells can provide time series spanning, perhaps, decades. Gorgonians have been shown to track sewage, and can provide records potentially centuries-long. In areas where baseline data are lacking, which is almost everywhere, delta(15)N in gorgonians can provide information on status and trends. In coral tissue, delta(15)N combined with insoluble residue determination can provide information on both sewage and sediment stress in areas lacking baseline data. In the developed world, delta(15)N provides objective assessment in a field complicated by conflicting opinions. Sample handling and processing are simple and analysis costs are low. This is a method deserving widespread application.

  13. Tracing Nitrogen through Landscapes to Coastal Wetlands using d15N of Larval Fish

    EPA Science Inventory

    Our objective was to evaluate the use of the nitrogen stable isotope value (d15N) of larval fish as an indicator of incipient anthropogenic nitrogen loading to coastal wetlands in the Great Lakes. We sampled coastal wetlands in five Lake Superior south shore tributaries that had ...

  14. Monitoring the refinement of crystal structures with (15)N solid-state NMR shift tensor data.

    PubMed

    Kalakewich, Keyton; Iuliucci, Robbie; Mueller, Karl T; Eloranta, Harriet; Harper, James K

    2015-11-21

    The (15)N chemical shift tensor is shown to be extremely sensitive to lattice structure and a powerful metric for monitoring density functional theory refinements of crystal structures. These refinements include lattice effects and are applied here to five crystal structures. All structures improve based on a better agreement between experimental and calculated (15)N tensors, with an average improvement of 47.0 ppm. Structural improvement is further indicated by a decrease in forces on the atoms by 2-3 orders of magnitude and a greater similarity in atom positions to neutron diffraction structures. These refinements change bond lengths by more than the diffraction errors including adjustments to X-Y and X-H bonds (X, Y = C, N, and O) of 0.028 ± 0.002 Å and 0.144 ± 0.036 Å, respectively. The acquisition of (15)N tensors at natural abundance is challenging and this limitation is overcome by improved (1)H decoupling in the FIREMAT method. This decoupling dramatically narrows linewidths, improves signal-to-noise by up to 317%, and significantly improves the accuracy of measured tensors. A total of 39 tensors are measured with shifts distributed over a range of more than 400 ppm. Overall, experimental (15)N tensors are at least 5 times more sensitive to crystal structure than (13)C tensors due to nitrogen's greater polarizability and larger range of chemical shifts.

  15. Benzylic Ammonium Ylide Mediated Epoxidations

    PubMed Central

    Roiser, Lukas; Robiette, Raphaël; Waser, Mario

    2016-01-01

    A high yielding synthesis of stilbene oxides using ammonium ylides has been developed. It turned out that the amine leaving group plays a crucial role as trimethylamine gives higher yields than DABCO or quinuclidine. The amine group also influences the diastereoselectivity, and detailed DFT calculations to understand the key parameters of these reactions have been carried out. PMID:27766017

  16. Closed-loop 15N measurement of N2O and its isotopomers for real-time greenhouse gas tracing

    NASA Astrophysics Data System (ADS)

    Slaets, Johanna; Mayr, Leopold; Heiling, Maria; Zaman, Mohammad; Resch, Christian; Weltin, Georg; Gruber, Roman; Dercon, Gerd

    2016-04-01

    Quantifying sources of nitrous oxide is essential to improve understanding of the global N cycle and to develop climate-smart agriculture, as N2O has a global warming potential 300 times higher than CO2. The isotopic signature and the intramolecular distribution (site preference) of 15N are powerful tools to trace N2O, but the application of these methods is limited as conventional methods cannot provide continuous and in situ data. Here we present a method for closed-loop, real time monitoring of the N2O flux, the isotopic signature and the intramolecular distribution of 15N by using off-axis integrated cavity output spectroscopy (ICOS, Los Gatos Research). The developed method was applied to a fertilizer inhibitor experiment, in which N2O emissions were measured on undisturbed soil cores for three weeks. The treatments consisted of enriched urea-N (100 kg urea-N/ha), the same fertilizer combined with the nitrification inhibitor nitrapyrin (375 g/100 kg urea), and control cores. Monitoring the isotopic signature makes it possible to distinguish emissions from soil and fertilizer. Characterization of site preference could additionally provide a tool to identify different microbial processes leading to N2O emissions. Furthermore, the closed-loop approach enables direct measurement on site and does not require removal of CO2 and H2O. Results showed that 75% of total N2O emissions (total=11 346 μg N2O-N/m2) in the fertilized cores originated from fertilizer, while only 55% of total emissions (total=2 450 μg N2ON/m2) stemmed from fertilizer for the cores treated with nitrapyrin. In the controls, N2O derived from soil was only 40% of the size of the corresponding pool from the fertilized cores, pointing towards a priming effect on the microbial community from the fertilizer and demonstrating the bias that could be introduced by relying on non-treated cores to estimate soil emission rates, rather than using the isotopic signature. The site preference increased linearly

  17. Evaluating δ(15)N-body size relationships across taxonomic levels using hierarchical models.

    PubMed

    Reum, Jonathan C P; Marshall, Kristin N

    2013-12-01

    Ecologists routinely set out to estimate the trophic position of individuals, populations, and species composing food webs, and nitrogen stable isotopes (δ(15)N) are a widely used proxy for trophic position. Although δ(15)N values are often sampled at the level of individuals, estimates and confidence intervals are frequently sought for aggregations of individuals. If individual δ(15)N values are correlated as an artifact of sampling design (e.g., clustering of samples in space or time) or due to intrinsic groupings (e.g., life history stages, social groups, taxonomy), such estimates may be biased and exhibit overly optimistic confidence intervals. However, these issues can be accommodated using hierarchical modeling methods. Here, we demonstrate how hierarchical models offer an additional quantitative tool for investigating δ(15)N variability and we explicitly evaluate how δ(15)N varies with body size at successively higher levels of taxonomic aggregation in a diverse fish assemblage. The models take advantage of all available data, better account for uncertainty in parameters estimates, may improve inferences on coefficients corresponding to groups with small to moderate sample sizes, and partition variation across model levels, which provides convenient summaries of the 'importance' of each level in terms of unexplained heterogeneity in the data. These methods can easily be applied to diet-based studies of trophic position. Although hierarchical models are well-understood and established tools, their benefits have yet to be fully reaped by stable isotope and food web ecologists. We suggest that hierarchical models can provide a robust framework for conceptualizing and statistically modeling trophic position at multiple levels of aggregation. PMID:23812110

  18. Stable isotopes (δ13C and δ15N) of organic matrix from coral skeleton

    PubMed Central

    Muscatine, Leonard; Goiran, Claire; Land, Lynton; Jaubert, Jean; Cuif, Jean-Pierre; Allemand, Denis

    2005-01-01

    The evolutionary success of reef-building corals in nutrient-poor tropical waters is attributed to endosymbiotic dinoflagellates. The algae release photosynthetic products to the coral animal cells, augment nutrient flux, and enhance the rate of coral calcification. Natural abundance of stable isotopes (δ13C and δ18O) provides answers to modern and paleobiological questions about the effect of photosymbiosis on sources of carbon and oxygen in coral skeletal calcium carbonate. Here we compare 17 species of symbiotic and nonsymbiotic corals to determine whether evidence for photosymbiosis appears in stable isotopes (δ13C and δ15N) of an organic skeletal compartment, the coral skeletal organic matrix (OM). Mean OM δ13C in symbiotic and nonsymbiotic corals was similar (-26.08‰ vs. -24.31‰), but mean OM δ15N was significantly depleted in 15N in the former (4.09‰) relative to the latter (12.28‰), indicating an effect of the algae on OM synthesis and revealing OM δ15N as a proxy for photosymbiosis. To answer an important paleobiological question about the origin of photosymbiosis in reef-building corals, we applied this proxy test to a fossil coral (Pachythecalis major) from the Triassic (240 million years ago) in which OM is preserved. Mean OM δ15N was 4.66‰, suggesting that P. major was photosymbiotic. The results show that symbiotic algae augment coral calcification by contributing to the synthesis of skeletal OM and that they may have done so as early as the Triassic. PMID:15671164

  19. Variable δ15N Diet-Tissue Discrimination Factors among Sharks: Implications for Trophic Position, Diet and Food Web Models

    PubMed Central

    Olin, Jill A.; Hussey, Nigel E.; Grgicak-Mannion, Alice; Fritts, Mark W.; Wintner, Sabine P.; Fisk, Aaron T.

    2013-01-01

    The application of stable isotopes to characterize the complexities of a species foraging behavior and trophic relationships is dependent on assumptions of δ15N diet-tissue discrimination factors (∆15N). As ∆15N values have been experimentally shown to vary amongst consumers, tissues and diet composition, resolving appropriate species-specific ∆15N values can be complex. Given the logistical and ethical challenges of controlled feeding experiments for determining ∆15N values for large and/or endangered species, our objective was to conduct an assessment of a range of reported ∆15N values that can hypothetically serve as surrogates for describing the predator-prey relationships of four shark species that feed on prey from different trophic levels (i.e., different mean δ15N dietary values). Overall, the most suitable species-specific ∆15N values decreased with increasing dietary-δ15N values based on stable isotope Bayesian ellipse overlap estimates of shark and the principal prey functional groups contributing to the diet determined from stomach content analyses. Thus, a single ∆15N value was not supported for this speciose group of marine predatory fishes. For example, the ∆15N value of 3.7‰ provided the highest percent overlap between prey and predator isotope ellipses for the bonnethead shark (mean diet δ15N = 9‰) whereas a ∆15N value < 2.3‰ provided the highest percent overlap between prey and predator isotope ellipses for the white shark (mean diet δ15N = 15‰). These data corroborate the previously reported inverse ∆15N-dietary δ15N relationship when both isotope ellipses of principal prey functional groups and the broader identified diet of each species were considered supporting the adoption of different ∆15N values that reflect the predators’ δ15N-dietary value. These findings are critical for refining the application of stable isotope modeling approaches as inferences regarding a species’ ecological role in their

  20. Determination of Multimodal Isotopic Distributions: The Case of a (15)N Labeled Protein Produced into Hairy Roots.

    PubMed

    Trouillard, Romain; Hubert-Roux, Marie; Tognetti, Vincent; Guilhaudis, Laure; Plasson, Carole; Menu-Bouaouiche, Laurence; Coquet, Laurent; Guerineau, François; Hardouin, Julie; Ele Ekouna, Jean-Pierre; Cosette, Pascal; Lerouge, Patrice; Boitel-Conti, Michèle; Afonso, Carlos; Ségalas-Milazzo, Isabelle

    2015-06-16

    Isotopic labeling is widely used in various fields like proteomics, metabolomics, fluxomics, as well as in NMR structural studies, but it requires an efficient determination of the isotopic enrichment. Mass spectrometry is the method of choice for such analysis. However, when complex expression systems like hairy roots are used for production, multiple populations of labeled proteins may be obtained. If the isotopic incorporation determination is actually well-known for unimodal distributions, the multimodal distributions have scarcely been investigated. Actually, only a few approaches allow the determination of the different labeled population proportions from multimodal distributions. Furthermore, they cannot be used when the number of the populations and their respective isotope ratios are unknown. The present study implements a new strategy to measure the (15)N labeled populations inside a multimodal distribution knowing only the peptide sequence and peak intensities from mass spectrometry analyses. Noteworthy, it could be applied to other elements, like carbon and hydrogen, and extended to a larger range of biomolecules.

  1. Influence of open ocean nitrogen supply on the skeletal δ15N of modern shallow-water scleractinian corals

    NASA Astrophysics Data System (ADS)

    Wang, Xingchen T.; Sigman, Daniel M.; Cohen, Anne L.; Sinclair, Daniel J.; Sherrell, Robert M.; Cobb, Kim M.; Erler, Dirk V.; Stolarski, Jarosław; Kitahara, Marcelo V.; Ren, Haojia

    2016-05-01

    The isotopic composition of skeleton-bound organic nitrogen in shallow-water scleractinian corals (hereafter, CS-δ15N) is an emerging tool for studying the marine nitrogen cycle in the past. The CS-δ15N has been shown to reflect the δ15N of nitrogen (N) sources to corals, with most applications to date focusing on the anthropogenic/terrestrial N inputs to reef environments. However, many coral reefs receive their primary N sources from the open ocean, and the CS-δ15N of these corals may provide information on past changes in the open ocean regional and global N cycle. Using a recently developed persulfate/denitrifier-based method, we measured CS-δ15N in modern shallow-water scleractinian corals from 8 sites proximal to the open ocean. At sites with low open ocean surface nitrate concentrations typical of the subtropics and tropics, measured CS-δ15N variation on seasonal and annual timescales is most often less than 2‰. In contrast, a broad range in CS-δ15N (of ∼10‰) is measured across these sites, with a strong correlation between CS-δ15N and the δ15N of the deep nitrate supply to the surface waters near the reefs. While CS-δ15N can be affected by other N sources as well and can vary in response to local reef conditions as well as coral/symbiont physiological changes, this survey indicates that, when considering corals proximal to the open ocean, the δ15N of the subsurface nitrate supply to surface waters drives most of the CS-δ15N variation across the global ocean. Thus, CS-δ15N is a promising proxy for reconstructing the open ocean N cycle in the past.

  2. 21 CFR 582.1135 - Ammonium bicarbonate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Additives § 582.1135 Ammonium bicarbonate. (a) Product. Ammonium bicarbonate. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or...

  3. 21 CFR 582.1137 - Ammonium carbonate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Additives § 582.1137 Ammonium carbonate. (a) Product. Ammonium carbonate. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or...

  4. 21 CFR 582.1139 - Ammonium hydroxide.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Additives § 582.1139 Ammonium hydroxide. (a) Product. Ammonium hydroxide. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or...

  5. 21 CFR 582.1135 - Ammonium bicarbonate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Additives § 582.1135 Ammonium bicarbonate. (a) Product. Ammonium bicarbonate. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or...

  6. 21 CFR 582.1139 - Ammonium hydroxide.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Additives § 582.1139 Ammonium hydroxide. (a) Product. Ammonium hydroxide. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or...

  7. 21 CFR 582.1143 - Ammonium sulfate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Additives § 582.1143 Ammonium sulfate. (a) Product. Ammonium sulfate. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or feeding practice....

  8. 21 CFR 582.1137 - Ammonium carbonate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Additives § 582.1137 Ammonium carbonate. (a) Product. Ammonium carbonate. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or...

  9. 21 CFR 582.1143 - Ammonium sulfate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Additives § 582.1143 Ammonium sulfate. (a) Product. Ammonium sulfate. (b) Conditions of use. This substance is generally recognized as safe when used in accordance with good manufacturing or feeding practice....

  10. 21 CFR 556.375 - Maduramicin ammonium.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... residues of maduramicin ammonium in chickens as follows: (a) A tolerance for maduramicin ammonium (marker... concentration of marker residues in the target tissue used to monitor for total drug residues in the...

  11. Isotopic fractionation of nitrogen and carbon in Paleoarchean cherts from Pilbara craton, Western Australia: Origin of 15N-depleted nitrogen

    NASA Astrophysics Data System (ADS)

    Pinti, Daniele L.; Hashizume, Ko; Sugihara, Akiyo; Massault, Marc; Philippot, Pascal

    2009-07-01

    Nitrogen and carbon isotopic compositions, together with mineralogy and trace element geochemistry, were studied in a few kerogen-rich Paleoarchean cherts, a barite and a dolomitic stromatolite belonging to the eastern (Dixon Island Formation) and western (Dresser and Strelley Pool Chert Formations; North Pole Dome and Marble Bar) terranes of Pilbara Craton, Western Australia. The aim of the study was to search for 15N-depleted isotopic signatures, often found in kerogens of this period, and explain the origin of these anomalies. Trace elements suggest silica precipitation by hydrothermal fluids as the main process of chert formation with a contamination from volcanoclastic detritus. This is supported by the occurrence of hydrothermal-derived minerals in the studied samples indicating precipitation temperatures up to 350 °C. Only a dolomitic stromatolite from Strelley Pool shows a superchondritic Y/Ho ratio of 72 and a positive Eu/Eu * anomaly of 1.8, characteristic of chemical precipitates from the Archean seawater. The bulk δ 13C vs. δ 15N values measured in the cherts show a roughly positive co-variation, except for one sample from the North Pole (PI-85-00). The progressive enrichment in 15N and 13C from a pristine source having δ 13C ⩽ -36‰ and δ 15N ⩽ -4‰ is correlated with a progressive depletion in N content and to variations in Ba/La and Co/As ratios. These trends have been interpreted as a progressive hydrothermal alteration of the cherts by metamorphic fluids. Isotopic exchange at 350 °C between NH 4+(rock) and N 2(fluid) may explain the isotopic and elemental composition of N in the studied cherts. However, we need to assume isotopic exchange at 350 °C between carbonate C and graphite to explain the large 13C enrichment recorded. Only sample PI-85-00 shows a large N loss (90%) with a positive δ 15N value (+11‰), while C (up to 120 ppm and δ 13C -38‰) seems to be unaffected. This pattern has been interpreted as the result of

  12. NITROGEN UPTAKE AND ASSIMILATION IN ENTEROMORPHA INTESTINALIS (L.) LINK (CHLOROPHYTA): USING 15N TO DETERMINE PREFERENCE DURING SIMULTANEOUS PULSES OF NITRATE AND AMMONIUM. (R827637)

    EPA Science Inventory

    We investigated the ability of Enteromorpha intestinalis (L.) Link to take up pulses of different species of nitrogen simultaneously, as this would be an important mechanism to enhance bloom ability in estuaries. Uptake rates and preference for NH4+

  13. Ammonium first: natural mosses prefer atmospheric ammonium but vary utilization of dissolved organic nitrogen depending on habitat and nitrogen deposition.

    PubMed

    Liu, Xue-Yan; Koba, Keisuke; Makabe, Akiko; Li, Xiao-Dong; Yoh, Muneoki; Liu, Cong-Qiang

    2013-07-01

    Mosses, among all types of terrestrial vegetation, are excellent scavengers of anthropogenic nitrogen (N), but their utilization of dissolved organic N (DON) and their reliance on atmospheric N remain uncharacterized in natural environments, which obscures their roles in N cycles. Natural (15) N abundance of N sources (nitrate (NO(3)(-)), ammonium (NH(4)(+)) and DON in deposition and soil) for epilithic and terricolous mosses was analyzed at sites with different N depositions at Guiyang, China. Moss NO(3)(-) assimilation was inhibited substantially by the high supply of NH(4)(+) and DON. Therefore, contributions of NH(4)(+) and DON to moss N were partitioned using isotopic mass-balance methods. The N contributions averaged 56% and 46% from atmospheric NH(4)(+), and 44% and 17% from atmospheric DON in epilithic and terricolous mosses, respectively. In terricolous mosses, soil NH(4)(+) and soil DON accounted for 16% and 21% of bulk N, which are higher than current estimations obtained using (15) N-labeling methods. Moreover, anthropogenic NH(4)(+) deposition suppressed utilization of DON and soil N because of the preference of moss for NH(4)(+) under elevated NH(4)(+) deposition. These results underscore the dominance of, and preference for, atmospheric NH(4)(+) in moss N utilization, and highlight the importance of considering DON and soil N sources when estimating moss N sequestration and the impacts of N deposition on mosses.

  14. Isotopic variability of cave bears (δ15N, δ13C) across Europe during MIS 3

    NASA Astrophysics Data System (ADS)

    Krajcarz, Magdalena; Pacher, Martina; Krajcarz, Maciej T.; Laughlan, Lana; Rabeder, Gernot; Sabol, Martin; Wojtal, Piotr; Bocherens, Hervé

    2016-01-01

    Collagen, the organic fraction of bone, records the isotopic parameters of consumed food for carbon (δ13C) and nitrogen (δ15N). This relationship of isotopic signature between diet and tissue is an important tool for the study of dietary preferences of modern and fossil animal species. Since the first information on the isotopic signature of cave bear was reported, numerous data from Europe have become available. The goal of this work is to track the geographical variation of cave bear collagen isotopic values in Europe during Marine Isotopic Stage 3 (about 60,000-25,000 yr BP). In this study the results of new δ13C and δ15N isotopic analyses of cave bear collagen from four Central-Eastern European sites are presented, as well as a review of all published isotopic data for cave bears of the same period. The main conclusion is a lack of geographical East-West pattern in the variations of δ13C and δ15N values of cave bear collagen. Moreover, no relationship was found between cave bear taxonomy and isotopic composition. The cave bears from Central-Eastern Europe exhibit δ13C and δ15N values near the average of the range of Central, Western and Southern European cave bears. Despite the fact that most cave bear sites follow an altitudinal gradient, separate groups of sites exhibit shift in absolute values of δ13C, what disturbs an altitude-related isotopic pattern. The most distinct groups are: high Alpine sites situated over 1500 m a.s.l. - in terms of δ13C; and two Romanian sites Peştera cu Oase and Urşilor - in case of δ15N. Although the cave bear isotopic signature is driven by altitude, the altitudinal adjustment of isotopic data is not enough to explain the isotopic dissimilarity of these cave bears. The unusually high δ15N signature of mentioned Romanian sites is an isolated case in Europe. Cave bears from relatively closely situated Central-Eastern European sites and other Romanian sites are more similar to Western European than to Romanian

  15. 21 CFR 184.1138 - Ammonium chloride.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... from the filtrate on cooling. Alternatively, hydrogen chloride formed by the burning of hydrogen in... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Ammonium chloride. 184.1138 Section 184.1138 Food... Specific Substances Affirmed as GRAS § 184.1138 Ammonium chloride. (a) Ammonium chloride (NH4Cl, CAS...

  16. 21 CFR 184.1138 - Ammonium chloride.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... from the filtrate on cooling. Alternatively, hydrogen chloride formed by the burning of hydrogen in... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ammonium chloride. 184.1138 Section 184.1138 Food... Specific Substances Affirmed as GRAS § 184.1138 Ammonium chloride. (a) Ammonium chloride (NH4Cl, CAS...

  17. 21 CFR 184.1138 - Ammonium chloride.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... from the filtrate on cooling. Alternatively, hydrogen chloride formed by the burning of hydrogen in... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Ammonium chloride. 184.1138 Section 184.1138 Food... Specific Substances Affirmed as GRAS § 184.1138 Ammonium chloride. (a) Ammonium chloride (NH4Cl, CAS...

  18. 21 CFR 184.1138 - Ammonium chloride.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... from the filtrate on cooling. Alternatively, hydrogen chloride formed by the burning of hydrogen in... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Ammonium chloride. 184.1138 Section 184.1138 Food... Specific Substances Affirmed as GRAS § 184.1138 Ammonium chloride. (a) Ammonium chloride (NH4Cl, CAS...

  19. 21 CFR 184.1138 - Ammonium chloride.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    .... Alternatively, hydrogen chloride formed by the burning of hydrogen in chlorine is dissolved in water and then... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Ammonium chloride. 184.1138 Section 184.1138 Food... GRAS § 184.1138 Ammonium chloride. (a) Ammonium chloride (NH4Cl, CAS Reg. No. 12125-02-9) is...

  20. 21 CFR 582.1141 - Ammonium phosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ammonium phosphate. 582.1141 Section 582.1141 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1141 Ammonium phosphate. (a) Product. Ammonium phosphate (mono- and dibasic). (b)...

  1. 21 CFR 556.375 - Maduramicin ammonium.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... residues of maduramicin ammonium in chickens as follows: (a) A tolerance for maduramicin ammonium (marker residue) in chickens is 0.38 parts per million in fat (target tissue). A tolerance refers to the... animals. (b) The safe concentrations for total maduramicin ammonium residues in uncooked edible...

  2. 21 CFR 556.375 - Maduramicin ammonium.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... residues of maduramicin ammonium in chickens as follows: (a) A tolerance for maduramicin ammonium (marker residue) in chickens is 0.38 parts per million in fat (target tissue). A tolerance refers to the... animals. (b) The safe concentrations for total maduramicin ammonium residues in uncooked edible...

  3. 21 CFR 582.1135 - Ammonium bicarbonate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ammonium bicarbonate. 582.1135 Section 582.1135 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1135 Ammonium bicarbonate. (a) Product. Ammonium bicarbonate. (b) Conditions of use....

  4. 21 CFR 582.1143 - Ammonium sulfate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ammonium sulfate. 582.1143 Section 582.1143 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1143 Ammonium sulfate. (a) Product. Ammonium sulfate. (b) Conditions of use. This...

  5. 21 CFR 582.1137 - Ammonium carbonate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ammonium carbonate. 582.1137 Section 582.1137 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1137 Ammonium carbonate. (a) Product. Ammonium carbonate. (b) Conditions of use....

  6. 21 CFR 582.1139 - Ammonium hydroxide.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ammonium hydroxide. 582.1139 Section 582.1139 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1139 Ammonium hydroxide. (a) Product. Ammonium hydroxide. (b) Conditions of use....

  7. 21 CFR 582.7133 - Ammonium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ammonium alginate. 582.7133 Section 582.7133 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Ammonium alginate. (a) Product. Ammonium alginate. (b) Conditions of use. This substance is...

  8. 21 CFR 582.7133 - Ammonium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ammonium alginate. 582.7133 Section 582.7133 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Ammonium alginate. (a) Product. Ammonium alginate. (b) Conditions of use. This substance is...

  9. 21 CFR 582.1141 - Ammonium phosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ammonium phosphate. 582.1141 Section 582.1141 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1141 Ammonium phosphate. (a) Product. Ammonium phosphate (mono- and dibasic). (b)...

  10. Extraction of urea and ammonium ion

    NASA Technical Reports Server (NTRS)

    Anselmi, R. T.; Husted, R. R.; Schulz, J. R.

    1977-01-01

    Water purification system keeps urea and ammonium ion concentration below toxic limits in recirculated water of closed loop aquatic habitat. Urea is first converted to ammonium ions and carbon dioxide by enzygmatic action. Ammonium ions are removed by ion exchange. Bioburden is controlled by filtration through 0.45 micron millipore filters.

  11. Nitrogen cycling in an extreme hyperarid environment inferred from δ(15)N analyses of plants, soils and herbivore diet.

    PubMed

    Díaz, Francisca P; Frugone, Matías; Gutiérrez, Rodrigo A; Latorre, Claudio

    2016-01-01

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ(15)N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ(15)N and δ(13)C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ(15)N values span the entire gradient, soil δ(15)N values show a positive correlation with aridity as expected. In contrast, foliar δ(15)N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ(15)N values. PMID:26956399

  12. Nitrogen cycling in an extreme hyperarid environment inferred from δ15N analyses of plants, soils and herbivore diet

    NASA Astrophysics Data System (ADS)

    Díaz, Francisca P.; Frugone, Matías; Gutiérrez, Rodrigo A.; Latorre, Claudio

    2016-03-01

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ15N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ15N and δ13C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ15N values span the entire gradient, soil δ15N values show a positive correlation with aridity as expected. In contrast, foliar δ15N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ15N values.

  13. Nitrogen cycling in an extreme hyperarid environment inferred from δ15N analyses of plants, soils and herbivore diet

    PubMed Central

    Díaz, Francisca P.; Frugone, Matías; Gutiérrez, Rodrigo A.; Latorre, Claudio

    2016-01-01

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ15N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ15N and δ13C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ15N values span the entire gradient, soil δ15N values show a positive correlation with aridity as expected. In contrast, foliar δ15N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ15N values. PMID:26956399

  14. In situ measurement of ammonium utilization by phytoplankton in the Sacramento River to determine the impacts of high nutrient loading on the base of the food web

    NASA Astrophysics Data System (ADS)

    Schmidt, C.; Kendall, C.; Silva, S. R.; Young, M. B.

    2012-12-01

    High concentrations of ammonium in the Sacramento River and San Francisco Bay-Delta Estuary (SFE) have been hypothesized to inhibit the growth of larger phytoplankton such as diatoms, which are an important food source to zooplankton at the base of the pelagic food web. The goal of this research is to use stable isotope approaches to distinguish ammonium and nitrate utilization at the base of the food web in a range of habitats extending from freshwater portions of Sacramento River through San Pablo Bay. Preliminary results from transects sampled over a range of hydrologic conditions between 2007 and 2010 indicate shifts in the δ15N composition of the bulk particulate organic matter (POM) where ammonium concentrations increase downstream of the Sacramento Regional Wastewater Treatment Plant and again where ammonium concentrations decrease near the confluence of the Sacramento and San Joaquin Rivers. However, interpretation of δ15N of bulk POM as a proxy for δ15N of phytoplankton is complicated by the fact that the C:N ratios of bulk POM often increase to values greater than 8 downstream of the confluence of the Sacramento and San Joaquin Rivers, suggesting that in this region of the estuary bulk POM includes a significant proportion of terrestrial detrital material. Given the interest in understanding nitrogen source utilization in this portion of the estuary, work is currently underway to isolate algae from bulk POM using flow cytometry prior to isotopic analysis. δ15N vales from isolated algal samples will allow downstream changes in nutrient source utilization to be distinguished from mixing of different POM sources. Because considerable bulk POM isotopic data have been collected over the last ten years, greater understanding of the relationship between δ15N-POM and δ15N of isolated algal and terrestrial fractions will allow for improved use of the bulk POM isotope record to understand the potential link between historical changes in phytoplankton nutrient

  15. Interaction between Ammonium Toxicity and Green Tide Development Over Seagrass Meadows: A Laboratory Study.

    PubMed

    Moreno-Marín, Francisco; Vergara, Juan J; Pérez-Llorens, J Lucas; Pedersen, Morten F; Brun, Fernando G

    2016-01-01

    Eutrophication affects seagrasses negatively by increasing light attenuation through stimulation of biomass of fast-growing, bloom-forming algae and because high concentrations of ammonium in the water can be toxic to higher plants. We hypothesized nevertheless, that moderate amounts of nitrophilic macroalgae that coexists with seagrasses under eutrophic conditions, can alleviate the harmful effects of eutrophication on seagrasses by reducing ammonium concentrations in the seawater to non-toxic levels because such algae have a very large capacity to take up inorganic nutrients. We studied therefore how combinations of different ammonium concentrations (0, 25 and 50 μM) and different standing stocks of macroalgae (i.e. 0, 1 and 6 layers of Ulva sp.) affected survival, growth and net production of the seagrass Zostera noltei. In the absence of Ulva sp., increasing ammonium concentrations had a negative influence on the performance of Z. noltei. The presence of Ulva sp. without ammonium supply had a similar, but slightly smaller, negative effect on seagrass fitness due to light attenuation. When ammonium enrichment was combined with presence of Ulva sp., Ulva sp. ameliorated some of negative effects caused by high ammonium availability although Ulva sp. lowered the availability of light. Benthic microalgae, which increased in biomass during the experiment, seemed to play a similar role as Ulva sp.--they contributed to remove ammonium from the water, and thus, aided to keep the ammonium concentrations experienced by Z. noltei at relatively non-toxic levels. Our findings show that moderate amounts of drift macroalgae, eventually combined with increasing stocks of benthic microalgae, may aid seagrasses to alleviate toxic effects of ammonium under eutrophic conditions, which highlights the importance of high functional diversity for ecosystem resistance to anthropogenic disturbance.

  16. Interaction between Ammonium Toxicity and Green Tide Development Over Seagrass Meadows: A Laboratory Study

    PubMed Central

    Moreno-Marín, Francisco; Vergara, Juan J.; Pérez-Llorens, J. Lucas; Pedersen, Morten F.; Brun, Fernando G.

    2016-01-01

    Eutrophication affects seagrasses negatively by increasing light attenuation through stimulation of biomass of fast-growing, bloom-forming algae and because high concentrations of ammonium in the water can be toxic to higher plants. We hypothesized nevertheless, that moderate amounts of nitrophilic macroalgae that coexists with seagrasses under eutrophic conditions, can alleviate the harmful effects of eutrophication on seagrasses by reducing ammonium concentrations in the seawater to non-toxic levels because such algae have a very large capacity to take up inorganic nutrients. We studied therefore how combinations of different ammonium concentrations (0, 25 and 50 μM) and different standing stocks of macroalgae (i.e. 0, 1 and 6 layers of Ulva sp.) affected survival, growth and net production of the seagrass Zostera noltei. In the absence of Ulva sp., increasing ammonium concentrations had a negative influence on the performance of Z. noltei. The presence of Ulva sp. without ammonium supply had a similar, but slightly smaller, negative effect on seagrass fitness due to light attenuation. When ammonium enrichment was combined with presence of Ulva sp., Ulva sp. ameliorated some of negative effects caused by high ammonium availability although Ulva sp. lowered the availability of light. Benthic microalgae, which increased in biomass during the experiment, seemed to play a similar role as Ulva sp.–they contributed to remove ammonium from the water, and thus, aided to keep the ammonium concentrations experienced by Z. noltei at relatively non-toxic levels. Our findings show that moderate amounts of drift macroalgae, eventually combined with increasing stocks of benthic microalgae, may aid seagrasses to alleviate toxic effects of ammonium under eutrophic conditions, which highlights the importance of high functional diversity for ecosystem resistance to anthropogenic disturbance. PMID:27035662

  17. Interaction between Ammonium Toxicity and Green Tide Development Over Seagrass Meadows: A Laboratory Study.

    PubMed

    Moreno-Marín, Francisco; Vergara, Juan J; Pérez-Llorens, J Lucas; Pedersen, Morten F; Brun, Fernando G

    2016-01-01

    Eutrophication affects seagrasses negatively by increasing light attenuation through stimulation of biomass of fast-growing, bloom-forming algae and because high concentrations of ammonium in the water can be toxic to higher plants. We hypothesized nevertheless, that moderate amounts of nitrophilic macroalgae that coexists with seagrasses under eutrophic conditions, can alleviate the harmful effects of eutrophication on seagrasses by reducing ammonium concentrations in the seawater to non-toxic levels because such algae have a very large capacity to take up inorganic nutrients. We studied therefore how combinations of different ammonium concentrations (0, 25 and 50 μM) and different standing stocks of macroalgae (i.e. 0, 1 and 6 layers of Ulva sp.) affected survival, growth and net production of the seagrass Zostera noltei. In the absence of Ulva sp., increasing ammonium concentrations had a negative influence on the performance of Z. noltei. The presence of Ulva sp. without ammonium supply had a similar, but slightly smaller, negative effect on seagrass fitness due to light attenuation. When ammonium enrichment was combined with presence of Ulva sp., Ulva sp. ameliorated some of negative effects caused by high ammonium availability although Ulva sp. lowered the availability of light. Benthic microalgae, which increased in biomass during the experiment, seemed to play a similar role as Ulva sp.--they contributed to remove ammonium from the water, and thus, aided to keep the ammonium concentrations experienced by Z. noltei at relatively non-toxic levels. Our findings show that moderate amounts of drift macroalgae, eventually combined with increasing stocks of benthic microalgae, may aid seagrasses to alleviate toxic effects of ammonium under eutrophic conditions, which highlights the importance of high functional diversity for ecosystem resistance to anthropogenic disturbance. PMID:27035662

  18. Stable Isotope (δ13C, δ15N, δ34S) Analysis and Satellite Telemetry Depict the Complexity of Gray Wolf (Canis lupus) Diets in Southwest Alaska

    NASA Astrophysics Data System (ADS)

    Stanek, A.; Watts, D. E.; Cohn, B. R.; Spencer, P.; Mangipane, B.; Welker, J. M.

    2010-12-01

    Throughout Alaska, gray wolves (Canis lupus) are a top predator of large ungulates. While they primarily rely on ungulates such as moose (Alces alces) and caribou (Rangifer tarandus) as food, they are opportunistic and use alternative resources. The variation and supplemental protein sources in wolf diet has not been studied extensively on live animals currently using the landscape. With large seasonal influxes of Pacific salmon (Oncorhynchus sp.) into Alaska, terrestrial carnivore use of marine species is of particular interest. Using stable isotope (δ13C, δ15N, δ34S) analysis of wolf guard hair and blood, this study aims to determine the proportion of marine derived nutrients (MDN) in the diet of wolf packs within and surrounding Lake Clark National Park and Preserve and Alaska Peninsula and Becharof National Wildlife Refuges in Southwest Alaska. Satellite telemetry from the animals sampled facilitates quantification of landscape use patterns in correspondence with isotopic traits. Wolf pack territories within and surrounding the Lake Clark region appear to vary in spatial extent and in availability of MDN, such as salmon. Initial analysis shows that two packs with smaller home ranges, centrally located around areas with greater salmon availability, have enriched δ15N values compared to packs that have larger home ranges not centralized around salmon spawning waters. This pattern of isotopic enrichment is found in red blood cells, blood serum and hair, representing diets over different time scales. The enrichment in both blood and hair indicates a sustained use of MDN over the previous six to nine months. In the Lake Clark region, simple mixing model estimates suggest that up to 30% of wolf pack diets may be from marine sources. In contrast, packs with larger home ranges and less access to salmon have stable isotope values representative of a terrestrial diet.

  19. Phase diagram of ammonium nitrate

    NASA Astrophysics Data System (ADS)

    Dunuwille, M.; Yoo, C. S.

    2014-05-01

    Ammonium Nitrate (AN) has often subjected to uses in improvised explosive devices, due to its wide availability as a fertilizer and its capability of becoming explosive with slight additions of organic and inorganic compounds. Yet, the origin of enhanced energetic properties of impure AN (or AN mixtures) is neither chemically unique nor well understood -resulting in rather catastrophic disasters in the past1 and thereby a significant burden on safety in using ammonium nitrates even today. To remedy this situation, we have carried out an extensive study to investigate the phase stability of AN at high pressure and temperature, using diamond anvil cells and micro-Raman spectroscopy. The present results confirm the recently proposed phase IV-to-IV' transition above 17 GPa2 and provide new constraints for the melting and phase diagram of AN to 40 GPa and 400 °C.

  20. Catalytic Roles of βLys87 in Tryptophan Synthase: 15N Solid State NMR Studies

    PubMed Central

    Caulkins, Bethany G.; Yang, Chen; Hilario, Eduardo; Fan, Li; Dunn, Michael F.; Mueller, Leonard J.

    2015-01-01

    The proposed mechanism for tryptophan synthase shows βLys87 playing multiple catalytic roles: it bonds to the PLP cofactor, activates C4′ for nucleophilic attack via a protonated Schiff base nitrogen, and abstracts and returns protons to PLP-bound substrates (i.e. acid-base catalysis). ε-15N-lysine TS was prepared to access the protonation state of βLys87 using 15N solid-state nuclear magnetic resonance (SSNMR) spectroscopy for three quasi-stable intermediates along the reaction pathway. These experiments establish that the protonation state of the ε-amino group switches between protonated and neutral states as the β-site undergoes conversion from one intermediate to the next during catalysis, corresponding to mechanistic steps where this lysine residue has been anticipated to play alternating acid and base catalytic roles that help steer reaction specificity in tryptophan synthase catalysis. PMID:25688830

  1. Natural abundance 14N and 15N solid-state NMR of pharmaceuticals and their polymorphs

    DOE PAGES

    Veinberg, Stanislav L.; Johnston, Karen E.; Jaroszewicz, Michael J.; Kispal, Brianna M.; Mireault, Christopher R.; Kobayashi, Takeshi; Pruski, Marek; Schurko, Robert W.

    2016-06-08

    14N ultra-wideline (UW), 1H{15N} indirectly-detected HETCOR (idHETCOR) and 15N dynamic nuclear polarization (DNP) solid-state NMR (SSNMR) experiments, in combination with plane-wave density functional theory (DFT) calculations of 14N EFG tensors, were utilized to characterize a series of nitrogen-containing active pharmaceutical ingredients (APIs), including HCl salts of scopolamine, alprenolol, isoprenaline, acebutolol, dibucaine, nicardipine, and ranitidine. Here, a case study applying these methods for the differentiation of polymorphs of bupivacaine HCl is also presented. All experiments were conducted upon samples with naturally-abundant nitrogen isotopes. For most of the APIs, it was possible to acquire frequency-stepped UW 14N SSNMR spectra of stationarymore » samples, which display powder patterns corresponding to pseudo-tetrahedral (i.e., RR'R"NH+ and RR'NH2+) or other (i.e., RNH2 and RNO2) nitrogen environments.« less

  2. 1H, 13C and 15N NMR assignments of phenazopyridine derivatives.

    PubMed

    Burgueño-Tapia, Eleuterio; Mora-Pérez, Yolanda; Morales-Ríos, Martha S; Joseph-Nathan, Pedro

    2005-03-01

    Phenazopyridine hydrochloride (1), a drug in clinical use for many decades, and some derivatives were studied by one- and two-dimensional (1)H, (13)C and (15)N NMR methodology. The assignments, combined with DFT calculations, reveal that the preferred protonation site of the drug is the pyridine ring nitrogen atom. The chemoselective acetylation of phenazopyridine (2) and its influence on the polarization of the azo nitrogen atoms were evidenced by the (15)N NMR spectra. Molecular calculations of the phenazopyridines 2-4 show that the pyridine and phenyl groups are oriented in an antiperiplanar conformation with intramolecular hydrogen bonding between the N-b atom and the C-2 amino group preserving the E-azo stereochemistry.

  3. Using delta15N values to characterise the nitrogen nutrient pathways from intensive animal units.

    PubMed

    Skinner, R A; Ineson, P; Jones, H; Sleep, D; Rank, R

    2006-01-01

    Previous studies on foliar delta15N values, in certain bryophytes, have indicated signature similarities to source pollutants. The object of this study was to investigate the effect further, by examining the mechanisms whereby isotopic fractionation occurs in systems such as atmospheric ammonia (NH3), throughfall, vegetation and soil. Measurements taken in and around point emission sources will then be used to characterise the various fractionation effects associated with these N transformations, as well as to demonstrate some of the issues associated with using delta15N values as pollution indicators. The atmospheric dispersion model UK-ADMS has also been used to model atmospheric delta15NH3 emissions, with signatures exhibiting marked negative shifts immediately downwind of an agricultural NH3 source. Similar dispersion patterns were mapped for NH3 concentration data illustrating the link between these two forms of measurement.

  4. Determination of the δ15N of total nitrogen in solids; RSIL lab code 2893

    USGS Publications Warehouse

    Revesz, Kinga; Qi, Haiping; Coplen, Tyler B.

    2006-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2893 is to determine the δ(15N/14N), abbreviated as δ15N , of total nitrogen in solid samples. A Carlo Erba NC 2500 elemental analyzer (EA) is used to convert total nitrogen in a solid sample into N2 gas. The EA is connected to a continuous flow isotope-ratio mass spectrometer (CF-IRMS), which determines relative difference in the isotope-amount ratios of stable nitrogen isotopes (15N/14N)of the product N2 gas. The combustion is quantitative; no isotopic fractionation is involved. Samples are placed in a tin capsule and loaded into the Costech Zero Blank Autosampler of the EA. Under computer control, samples are dropped into a heated reaction tube that contains an oxidant, where the combustion takes place in a helium atmosphere containing an excess of oxygen gas. Combustion products are transported by a helium carrier through a reduction tube to remove excess oxygen and convert all nitrous oxides into N2 and through a drying tube to remove water. The gas-phase products, mainly CO2 and N2, are separated by a gas chromatograph. The gas is then introduced into the isotope-ratio mass spectrometer (IRMS) through a Finnigan MAT (now Thermo Scientific) ConFlo II interface, which also is used to inject N2 reference gas and helium for sample dilution. The IRMS is a Thermo Scientific Delta V Plus CF-IRMS. It has a universal triple collector, two wide cups with a narrow cup in the middle, capable of measuring mass/charge (m/z) 28, 29, 30, simultaneously. The ion beams from N2 are as follows: m/z 28 = N2 = 14N14N; m/z 29 = N2 = 14N15N primarily; m/z 30 = NO = 14N16O primarily, which is a sign of contamination or incomplete reduction.

  5. High irradiance improves ammonium tolerance in wheat plants by increasing N assimilation.

    PubMed

    Setién, Igor; Fuertes-Mendizabal, Teresa; González, Azucena; Aparicio-Tejo, Pedro Ma; González-Murua, Carmen; González-Moro, María Begoña; Estavillo, José María

    2013-05-15

    Ammonium is a paradoxical nutrient ion. Despite being a common intermediate in plant metabolism whose oxidation state eliminates the need for its reduction in the plant cell, as occurs with nitrate, it can also result in toxicity symptoms. Several authors have reported that carbon enrichment in the root zone enhances the synthesis of carbon skeletons and, accordingly, increases the capacity for ammonium assimilation. In this work, we examined the hypothesis that increasing the photosynthetic photon flux density is a way to increase plant ammonium tolerance. Wheat plants were grown in a hydroponic system with two different N sources (10mM nitrate or 10mM ammonium) and with two different light intensity conditions (300 μmol photon m(-2)s(-1) and 700 μmol photon m(-2)s(-1)). The results show that, with respect to biomass yield, photosynthetic rate, shoot:root ratio and the root N isotopic signature, wheat behaves as a sensitive species to ammonium nutrition at the low light intensity, while at the high intensity, its tolerance is improved. This improvement is a consequence of a higher ammonium assimilation rate, as reflected by the higher amounts of amino acids and protein accumulated mainly in the roots, which was supported by higher tricarboxylic acid cycle activity. Glutamate dehydrogenase was a key root enzyme involved in the tolerance to ammonium, while glutamine synthetase activity was low and might not be enough for its assimilation.

  6. Impact of seaweed beachings on dynamics of δ(15)N isotopic signatures in marine macroalgae.

    PubMed

    Lemesle, Stéphanie; Mussio, Isabelle; Rusig, Anne-Marie; Menet-Nédélec, Florence; Claquin, Pascal

    2015-08-15

    A fine-scale survey of δ(15)N, δ(13)C, tissue-N in seaweeds was conducted using samples from 17 sampling points at two sites (Grandcamp-Maisy (GM), Courseulles/Mer (COU)) along the French coast of the English Channel in 2012 and 2013. Partial triadic analysis was performed on the parameter data sets and revealed the functioning of three areas: one estuary (EstA) and two rocky areas (GM(∗), COU(∗)). In contrast to oceanic and anthropogenic reference points similar temporal dynamics characterized δ(15)N signatures and N contents at GM(∗) and COU(∗). Nutrient dynamics were similar: the N-concentrations in seawater originated from the River Seine and local coastal rivers while P-concentrations mainly from these local rivers. δ(15)N at GM(∗) were linked to turbidity suggesting inputs of autochthonous organic matter from large-scale summer seaweed beachings made up of a mixture of Rhodophyta, Phaeophyta and Chlorophyta species. This study highlights the coupling between seaweed beachings and nitrogen sources of intertidal macroalgae.

  7. 13N,15N isotope and kinetic evidence against hyponitrite as an intermediate in dentrification.

    PubMed

    Hollocher, T C; Garber, E; Cooper, A J; Reiman, R E

    1980-06-10

    13N- and 15N-labeling experiments were carried out with Paracoccus denitrificans, grown anaerobically on nitrate, to determine whether hyponitrite might be an obligatory intermediate in denitrification and a precursor of nitrous oxide. From experiments designed to trap [13N]- or [15N,15N]hyponitrite by dilution into authentic hyponitrite it was calculated that the intracellular concentration of a presumptive hyponitrite pool must be less than 0.4 mM. In order for a pool of this size to turn over rapidly enough to handle the flux of nitrogen during dentrifucation, the spontaneous rate of hyponitrite dehydration must be enhanced by a factor of several thousand through enzyme catalysis. Cell extracts failed to catalyze this reaction under a variety of conditions. It is concluded that hyponitrite cannot be an intermediate in dentrification. In addition, the assimilation of inorganic nitrogen was studied in P. denitrificans using 13N as tracer. At low concentrations (less than 10(-8) M) of labeled nitrate and nitrite 5 to 10% of the label was assimilated into non-volatile metabolites and 90 to 95% was reduced to N2. Similarly, with 15 mM [13N]nitrate, 5% of the label went into metabolites and 95% to N2. High pressure liquid chromatography analysis of the labeled metabolites indicated that the major pathway for assimilation of inorganic nitrogen in P. denitrificans under these conditions is through ammonia incorporation via the aspartase reaction. PMID:7372623

  8. Solid-state /sup 15/N NMR of oriented lipid bilayer bound gramicidin A'

    SciTech Connect

    Nicholson, L.K.; Moll, F.; Mixon, T.E.; LoGrasso, P.V.; Lay, J.C.; Cross, T.A.

    1987-10-20

    Highly oriented samples of lipid and gramicidin A' (8:1 molar ratio) have been prepared with the samples extensively hydrated (approximately 70% water v/w). These preparations have been shown to be completely in a bilayer phase with a transition temperature of 28/sup 0/C, and evidence is presented indicating that the gramicidin is in the channel conformation. An estimate of the disorder in the alignment of the bilayers parallel with the glass plates used to align the bilayers can be made from the asymmetry of the nuclear magnetic resonances (NMR). Such an analysis indicates a maximal range of disorder of +-3/sup 0/. Uniformly /sup 15/N-labeled gramicidin has been biosynthesized by Bacillus brevis grown in a media containing /sup 15/N-labeled Escherichia coli cells as the only nitrogen source. When prepared with labeled gramicidin, the oriented samples result in high-resolution /sup 15/N NMR spectra showing 12 resonances for the 20 nitrogen sites of the polypeptide. The frequency of the three major multiple resonance peaks has been interpreted to yield the approximate orientation of the N-H bonds in the peptide linkages with respect to the magnetic field. The bond orientations are only partially consistent with the extant structural models of gramicidin.

  9. Affordable uniform isotope labeling with (2)H, (13)C and (15)N in insect cells.

    PubMed

    Sitarska, Agnieszka; Skora, Lukasz; Klopp, Julia; Roest, Susan; Fernández, César; Shrestha, Binesh; Gossert, Alvar D

    2015-06-01

    For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80% can be achieved for (15)N and (13)C with yields comparable to expression in full media. For (2)H,(15)N and (2)H,(13)C,(15)N labeling, incorporation is only slightly lower with 75 and 73%, respectively, and yields are typically twofold reduced. The media were optimized for isotope incorporation, reproducibility, simplicity and cost. High isotope incorporation levels for all labeling patterns are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The final formulation consists of just five commercially available components, at costs 12-fold lower than labeling media from vendors. The approach was applied to several cytosolic and secreted target proteins. PMID:25929326

  10. Natural abundance (14)N and (15)N solid-state NMR of pharmaceuticals and their polymorphs.

    PubMed

    Veinberg, Stanislav L; Johnston, Karen E; Jaroszewicz, Michael J; Kispal, Brianna M; Mireault, Christopher R; Kobayashi, Takeshi; Pruski, Marek; Schurko, Robert W

    2016-06-29

    (14)N ultra-wideline (UW), (1)H{(15)N} indirectly-detected HETCOR (idHETCOR) and (15)N dynamic nuclear polarization (DNP) solid-state NMR (SSNMR) experiments, in combination with plane-wave density functional theory (DFT) calculations of (14)N EFG tensors, were utilized to characterize a series of nitrogen-containing active pharmaceutical ingredients (APIs), including HCl salts of scopolamine, alprenolol, isoprenaline, acebutolol, dibucaine, nicardipine, and ranitidine. A case study applying these methods for the differentiation of polymorphs of bupivacaine HCl is also presented. All experiments were conducted upon samples with naturally-abundant nitrogen isotopes. For most of the APIs, it was possible to acquire frequency-stepped UW (14)N SSNMR spectra of stationary samples, which display powder patterns corresponding to pseudo-tetrahedral (i.e., RR'R''NH(+) and RR'NH2(+)) or other (i.e., RNH2 and RNO2) nitrogen environments. Directly-excited (14)N NMR spectra were acquired using the WURST-CPMG pulse sequence, which incorporates WURST (wideband, uniform rate, and smooth truncation) pulses and a CPMG (Carr-Purcell Meiboom-Gill) refocusing protocol. In certain cases, spectra were acquired using (1)H → (14)N broadband cross-polarization, via the BRAIN-CP (broadband adiabatic inversion - cross polarization) pulse sequence. These spectra provide (14)N electric field gradient (EFG) tensor parameters and orientations that are particularly sensitive to variations in local structure and intermolecular hydrogen-bonding interactions. The (1)H{(15)N} idHETCOR spectra, acquired under conditions of fast magic-angle spinning (MAS), used CP transfers to provide (1)H-(15)N chemical shift correlations for all nitrogen environments, except for two sites in acebutolol and nicardipine. One of these two sites (RR'NH2(+) in acebutolol) was successfully detected using the DNP-enhanced (15)N{(1)H} CP/MAS measurement, and one (RNO2 in nicardipine) remained elusive due to the absence of

  11. A holistic approach to understanding the N isotopic composition (d15N) of deep-sea sediments: diatom-bound, foraminifera-bound, whole sediment and modern nitrate d15N from the equatorial Pacific

    NASA Astrophysics Data System (ADS)

    Rafter, P. A.; Charles, C. D.; Sigman, D. M.; Haug, G. H.

    2010-12-01

    The nitrogen (N) isotopic composition (d15N) of sediment is well established as a proxy of nitrate consumption and the d15N of nitrate in the surface ocean, but it is commonly assumed that post-depositional fractionation during organic matter degradation provides an additional unconstrained variable in sediments from the deep-sea. This mistrust of deep-sea sediments essentially renders most of the global ocean off-limits to the application of this powerful proxy of nutrient dynamics. Here we address this issue with new measurements of diatom-bound, foraminifera-bound, and whole sediment d15N from deep-sea sediments of the equatorial Pacific—a region where we have also investigated the processes influencing modern nitrate characteristics. With the results of these new records and previously published measurements relevant to the composition of sedimentary nitrogen, we are confident that whole sediment d15N from the deep-sea is not altered after deposition on the sea floor and that it accurately records surface ocean conditions. With these new constraints on whole sediment d15N, we discuss the exciting implications for alternative sediment d15N measurements (such as diatom- and foraminifera-bound d15N) and present evidence for a long-term increase in equatorial Pacific d15N of nitrate.

  12. The signatures of stable isotopes δ 15N and δ 13C in anadromous and non-anadromous Coilia nasus living in the Yangtze River, and the adjacent sea waters

    NASA Astrophysics Data System (ADS)

    Wang, Lei; Tang, Wenqiao; Dong, Wenxia

    2015-12-01

    Stable isotopes are increasingly used to investigate seasonal migrations of aquatic organisms. This study employed stable isotopes ( δ 13C and δ 15N) for Coilia nasus from the lower Yangtze River and the adjacent East China Sea to distinguish different ecotypic groups, ascertain trophic nutrition positions, and reflect environmental influences on C. nasus. δ 13C signatures of C. nasus sampled from Zhoushan (ZS), Chongming (CM), and Jingjiang (JJ) waters were significantly higher than those from the Poyang Lake (PYL) ( P < 0.05). By contrast, δ 15N signatures of C. nasus in ZS, CM, and JJ groups were significantly lower than those in PYL group ( P < 0.05). Basing on δ 13C and δ 15N signatures, we could distinguish anadromous (ZS, CM, and JJ) and non-anadromous (PYL) groups. The trophic level (TL) of anadromous C. nasus ranged from 2.90 to 3.04, whereas that of non-anadromous C. nasus was 4.38. C. nasus occupied the middle and top nutrition positions in the marine and Poyang Lake food webs, respectively. C. nasus in Poyang Lake were significantly more enriched in δ 15N but depleted in δ 13C, suggesting that anthropogenic nutrient inputs and terrigenous organic carbon are important to the Poyang Lake food web. This study is the first to apply δ 15N and δ 13C to population assignment studies of C. nasus in the Yangtze River and its affiliated waters. Analysis of stable isotopes ( δ 15N and δ 13C) is shown to be a useful tool for discriminating anadromous and non-anadromous C. nasus.

  13. Establishing spatial trends in water chemistry and stable isotopes (δ15N and δ13C) in the Elwha River prior to dam removal and salmon recolonization

    USGS Publications Warehouse

    Duda, J.J.; Coe, H.J.; Morley, S.A.; Kloehn, K.K.

    2011-01-01

    Two high-head dams on the Elwha River in Washington State (USA) have changed the migratory patterns of resident and anadromous fish, limiting Pacific salmon to the lower 7.9 km of a river that historically supported large Pacific salmon runs. To document the effects of the dams prior to their removal, we measured carbon and nitrogen stable isotope ratios of primary producers, benthic macroinvertebrates, and fish, and water chemistry above, between and below the dams. We found that δ15N was significantly higher in fish, stoneflies, black flies, periphyton and macroalgae where salmon still have access. Fish and chloroperlid stoneflies were enriched in δ13C, but the values were more variable than in δ15N. For some taxa, there were also differences between the two river sections that lack salmon, suggesting that factors other than marine-derived nutrients are structuring longitudinal isotopic profiles. Consistent with trophic theory, macroalgae had the lowest δ15N, followed by periphyton, macroinvertebrates and fish, with a range of 6.9, 6.2 and 7.7‰ below, between, and above the dams, respectively. Water chemistry analyses confirmed earlier reports that the river is oligotrophic. Phosphorous levels in the Elwha were lower than those found in other regional rivers, with significant differences among regulated, unregulated and reference sections. The removal of these dams, among the largest of such projects ever attempted, is expected to facilitate the return of salmon and their marine-derived nutrients (MDN) throughout the watershed, possibly altering the food web structure, nutrient levels and stable isotope values that we documented.

  14. Dependence of in vivo glutamine synthetase activity on ammonia concentration in rat brain studied by 1H - 15N heteronuclear multiple-quantum coherence-transfer NMR.

    PubMed Central

    Kanamori, K; Ross, B D; Kuo, E L

    1995-01-01

    The dependence of the in vivo rate of glutamine synthesis on the substrate ammonia concentration was studied in rat brain by 1H-15N heteronuclear multiple-quantum coherence-transfer NMR in combination with biochemical techniques. In vivo rates were measured at various steady-state blood and brain ammonia concentrations within the ranges 0.4-0.55 mumol/g and 0.86-0.98 mumol/g respectively, after low-rate intravenous 15NH4+ infusion (isotope chase). The rate of glutamine synthesis at steady state was determined from the change in brain [5-15N]glutamine levels during isotope chase, observed selectively through the amide proton by NMR, and 15N enrichments of brain glutamine and of blood and brain ammonia measured byN gas chromatography-MS. The in vivo rate (v) was 3.3-4.5 mumol/h per g of brain at blood ammonia concentrations (s) of 0.40-0.55 mumol/g. A linear increase of 1/v with 1/s permitted estimation of the in vivo glutamine synthetase (GS) activity at a physiological blood ammonia concentration to be 0.4-2.1 mumol/h per g. The observed ammonia-dependence strongly suggests that, under physiological conditions, in vivo GS activity is kinetically limited by sub-optimal in situ concentrations of ammonia as well as glutamate and ATP. Comparison of the observed in vivo GS activity with the reported in vivo rates of glutaminase and of gamma-aminobutyrate (GABA) synthesis suggests that, under mildly hyperammonaemic conditions, glutamine is synthesized at a sufficiently high rate to serve as a precursor of GABA, but glutaminase-catalysed hydrolysis of glutamine is too slow to be the sole provider of glutamate used for GABA synthesis. PMID:7487913

  15. A 15N-poor isotopic composition for the solar system as shown by Genesis solar wind samples.

    PubMed

    Marty, B; Chaussidon, M; Wiens, R C; Jurewicz, A J G; Burnett, D S

    2011-06-24

    The Genesis mission sampled solar wind ions to document the elemental and isotopic compositions of the Sun and, by inference, of the protosolar nebula. Nitrogen was a key target element because the extent and origin of its isotopic variations in solar system materials remain unknown. Isotopic analysis of a Genesis Solar Wind Concentrator target material shows that implanted solar wind nitrogen has a (15)N/(14)N ratio of 2.18 ± 0.02 × 10(-3) (that is, ≈40% poorer in (15)N relative to terrestrial atmosphere). The (15)N/(14)N ratio of the protosolar nebula was 2.27 ± 0.03 × 10(-3), which is the lowest (15)N/(14)N ratio known for solar system objects. This result demonstrates the extreme nitrogen isotopic heterogeneity of the nascent solar system and accounts for the (15)N-depleted components observed in solar system reservoirs.

  16. A 15N-poor isotopic composition for the solar system as shown by Genesis solar wind samples.

    PubMed

    Marty, B; Chaussidon, M; Wiens, R C; Jurewicz, A J G; Burnett, D S

    2011-06-24

    The Genesis mission sampled solar wind ions to document the elemental and isotopic compositions of the Sun and, by inference, of the protosolar nebula. Nitrogen was a key target element because the extent and origin of its isotopic variations in solar system materials remain unknown. Isotopic analysis of a Genesis Solar Wind Concentrator target material shows that implanted solar wind nitrogen has a (15)N/(14)N ratio of 2.18 ± 0.02 × 10(-3) (that is, ≈40% poorer in (15)N relative to terrestrial atmosphere). The (15)N/(14)N ratio of the protosolar nebula was 2.27 ± 0.03 × 10(-3), which is the lowest (15)N/(14)N ratio known for solar system objects. This result demonstrates the extreme nitrogen isotopic heterogeneity of the nascent solar system and accounts for the (15)N-depleted components observed in solar system reservoirs. PMID:21700869

  17. Natural attenuation potential of tricholoroethene in wetland plant roots: role of native ammonium-oxidizing microorganisms.

    PubMed

    Qin, Ke; Struckhoff, Garrett C; Agrawal, Abinash; Shelley, Michael L; Dong, Hailiang

    2015-01-01

    Bench-scale microcosms with wetland plant roots were investigated to characterize the microbial contributions to contaminant degradation of trichloroethene (TCE) with ammonium. The batch system microcosms consisted of a known mass of wetland plant roots in aerobic growth media where the roots provided both an inoculum of root-associated ammonium-oxidizing microorganisms and a microbial habitat. Aqueous growth media, ammonium, and TCE were replaced weekly in batch microcosms while retaining roots and root-associated biomass. Molecular biology results indicated that ammonium-oxidizing bacteria (AOB) were enriched from wetland plant roots while analysis of contaminant and oxygen concentrations showed that those microorganisms can degrade TCE by aerobic cometabolism. Cometabolism of TCE, at 29 and 46 μg L(-1), was sustainable over the course of 9 weeks, with 20-30 mg L(-1) ammonium-N. However, at 69 μg L(-1) of TCE, ammonium oxidation and TCE cometabolism were completely deactivated in two weeks. This indicated that between 46 and 69 μg L(-1) TCE with 30 mg L(-1) ammonium-N there is a threshold [TCE] below which sustainable cometabolism can be maintained with ammonium as the primary substrate. However, cometabolism-induced microbial deactivation of ammonium oxidation and TCE degradation at 69 μg L(-1) TCE did not result in a lower abundance of the amoA gene in the microcosms, suggesting that the capacity to recover from TCE inhibition was still intact, given time and removal of stress. Our study indicates that microorganisms associated with wetland plant roots can assist in the natural attenuation of TCE in contaminated aquatic environments, such as urban or treatment wetlands, and wetlands impacted by industrial solvents.

  18. Worms from venus and mars: proteomics profiling of sexual differences in Caenorhabditis elegans using in vivo 15N isotope labeling.

    PubMed

    Tops, Bastiaan B J; Gauci, Sharon; Heck, Albert J R; Krijgsveld, Jeroen

    2010-01-01

    Hermaphrodites of the nematode Caenorhabditis elegans produce both sperm and oocytes in the same germline. To investigate the process underlying spermatogenesis and oogenesis separately, we used a quantitative proteomics approach applied to two mutant worm lines (fem-3(q20) and fem-1(hc17)) developing only male and female germlines, respectively. We used stable isotopic labeling of whole animals by feeding them either (14)N or (15)N labeled Escherichia coli. This way, we could confidently identify and quantify 1040 proteins in two independent experiments. Of these, approximately 400 proteins showed significant differential expression between female-like and male-like animals. As expected, proteins linked to oogenesis were found to be highly upregulated in the feminized worms, whereas proteins involved in spermatogenesis were found to be highly upregulated in the masculinized worms. This was complemented by many proteins strongly enriched in either mutant. Although the function of the majority of these proteins is unknown, their expression profile indicates that they have an as yet unrecognized role in the development and/or function of the female- and male germline in C. elegans. We show that members of several protein complexes as well as functionally similar proteins show comparable abundance ratios, indicating coregulation of protein expression. Additional analysis comparing our protein data to a previously published microarray data set shows that mRNA and protein expression are poorly correlating. We provide one of the first examples of a large-scale quantitative proteomics experiment in C. elegans and show the potential and feasibility of an approach enabling system-wide accurate quantitative proteomics experiments in this model organism. PMID:19916504

  19. Mammalian production of an isotopically enriched outer domain of the HIV-1 gp120 glycoprotein for NMR spectroscopy.

    PubMed

    Sastry, Mallika; Xu, Ling; Georgiev, Ivelin S; Bewley, Carole A; Nabel, Gary J; Kwong, Peter D

    2011-07-01

    NMR spectroscopic characterization of the structure or the dynamics of proteins generally requires the production of samples isotopically enriched in (15)N, (13)C, or (2)H. The bacterial expression systems currently in use to obtain isotopic enrichment, however, cannot produce a number of eukaryotic proteins, especially those that require post-translational modifications such as N-linked glycosylation for proper folding or activity. Here, we report the use of an adenovirus vector-based mammalian expression system to produce isotopically enriched (15)N or (15)N/(13)C samples of an outer domain variant of the HIV-1 gp120 envelope glycoprotein with 15 sites of N-linked glycosylation. Yields for the (15)N- and (15)N/(13)C-labeled gp120s after affinity chromatography were 45 and 44 mg/l, respectively, with an average of over 80% isotope incorporation. Recognition of the labeled gp120 by cognate antibodies that recognize complex epitopes showed affinities comparable to the unlabeled protein. NMR spectra, including (1)H-(15)N and (1)H-(13)C HSQCs, (15)N-edited NOESY-HSQC, and 3D HNCO, were of high quality, with signal-to-noise consistent with an efficient level of isotope incorporation, and with chemical shift dispersion indicative of a well-folded protein. The exceptional protein yields, good isotope incorporation, and ability to obtain well-folded post-translationally modified proteins make this mammalian system attractive for the production of isotopically enriched eukaryotic proteins for NMR spectroscopy.

  20. Localization of 15N uptake in a Tibetan alpine Kobresia pasture

    NASA Astrophysics Data System (ADS)

    Schleuß, Per-Marten; Kuzyakov, Yakov

    2014-05-01

    The Kobresia Pygmea ecotone covers approximately 450.000 km2 and is of large global and regional importance due several socio-ecological aspects. For instance Kobresia pastures store high amounts of carbon, nitrogen and other nutrients, represent large grazing areas for herbivores, provide a fast regrowth after grazing events and protect against mechanical degradation and soil erosion. However, Kobresia pastures are assumed to be a grazing induced and are accompanied with distinct root mats varying in thickness between 5-30 cm. Yet, less is known about the morphology and the functions of this root mats, especially in the background of a progressing degradation due to changes of climate and management. Thus we aimed to identify the importance of single soil layers for plant nutrition. Accordingly, nitrogen uptake from different soil depths and its remain in above-ground biomass (AGB), belowground biomass (BGB) and soil were determined by using a 15N pulse labeling approach during the vegetation period in summer 2012. 15N urea was injected into six different soil depths (0.5 cm, 2.5 cm, 7.5 cm, 12.5 cm, 17.5 cm, 22.5 cm / for each 4 replicates) and plots were sampled 45 days after the labeling. For soil and BGB samples were taken in strict sample intervals of 0-1 cm, 1-5 cm, 5-10 cm, 10-15 cm, 15-20 cm, 20-25 cm. Results indicate that total recovery (including AGB, BGB and soil) was highest, if tracer was injected into the top 5 cm and subsequently decreased with decreasing injection depth. This is especially the case for the 15N recovery of BGB, which is clearly attributed to the root density and strongly decreased with soil depth. In contrast, the root activity derived from the 15N content of roots increased with soil depth, which is primary associated to a proportionate increase of living roots related to dead roots. However, most 15N was captured in plant biomass (67.5-85.3 % of total recovery), indicating high 15N uptake efficiency possibly due to N limitation

  1. Particulate δ15N in laminated marine sediments as a proxy for mixing between the California Undercurrent and the California Current: A proof of concept

    NASA Astrophysics Data System (ADS)

    Tems, Caitlin E.; Berelson, William M.; Prokopenko, Maria G.

    2015-01-01

    measurements of particulate δ15N in coastal marine laminated sediments provide a high-resolution proxy for fluctuations in the intensity of denitrification in the water column. In the eastern tropical North Pacific oxygen minimum zone, this denitrification signal is transported northward by the California Undercurrent, thus serving as a tracer of ocean circulation. This is verified through comparisons between salinity in the thermocline off Southern California (Santa Monica Basin) and the difference between δ15Nsed within age equivalent sediments from a southern (Pescadero Slope) and northern (Santa Monica Basin) site. Trends in this parameter, Δδ15Nsed, relate to Pacific Decadal Oscillation (PDO) phase changes between 1900 and 1990. We hypothesize that the decline in Δδ15Nsed during warm PDO phases is due to a strengthening of the California Undercurrent transporting 15N-enriched nitrate from the eastern tropical North Pacific northward. The deviation from this trend after 1990 suggests recent changes in circulation and/or California Current water nutrient biogeochemistry.

  2. δ(13)C and δ(15)N in deep-living fishes and shrimps after the Deepwater Horizon oil spill, Gulf of Mexico.

    PubMed

    Quintana-Rizzo, Ester; Torres, Joseph J; Ross, Steve W; Romero, Isabel; Watson, Kathleen; Goddard, Ethan; Hollander, David

    2015-05-15

    The blowout of the Deepwater Horizon (DWH) drill-rig produced a surface oil layer, dispersed micro-droplets throughout the water column, and sub-surface plumes. We measured stable carbon and nitrogen isotopes in mesopelagic fishes and shrimps in the vicinity of DWH collected prior to, six weeks after, and one year after the oil spill (2007, 2010 and 2011). In 2010, the year of the oil spill, a small but significant depletion of δ(13)C was found in two mesopelagic fishes (Gonostoma elongatum and Chauliodus sloani) and one shrimp (Systellaspis debilis); a significant δ(15)N enrichment was identified in the same shrimp and in three fish species (G. elongatum, Ceratoscopelus warmingii, and Lepidophanes guentheri). The δ(15)N change did not suggest a change of trophic level, but did indicate a change in diet. The data suggest that carbon from the Deepwater Horizon oil spill was incorporated into the mesopelagic food web of the Gulf of Mexico. PMID:25778549

  3. In vivo, large-scale preparation of uniformly (15)N- and site-specifically (13)C-labeled homogeneous, recombinant RNA for NMR studies.

    PubMed

    Le, My T; Brown, Rachel E; Simon, Anne E; Dayie, T Kwaku

    2015-01-01

    Knowledge of how ribonucleic acid (RNA) structures fold to form intricate, three-dimensional structures has provided fundamental insights into understanding the biological functions of RNA. Nuclear magnetic resonance (NMR) spectroscopy is a particularly useful high-resolution technique to investigate the dynamic structure of RNA. Effective study of RNA by NMR requires enrichment with isotopes of (13)C or (15)N or both. Here, we present a method to produce milligram quantities of uniformly (15)N- and site-specifically (13)C-labeled RNAs using wild-type K12 and mutant tktA Escherichia coli in combination with a tRNA-scaffold approach. The method includes a double selection protocol to obtain an E. coli clone with consistently high expression of the recombinant tRNA-scaffold. We also present protocols for the purification of the tRNA-scaffold from a total cellular RNA extract and the excision of the RNA of interest from the tRNA-scaffold using DNAzymes. Finally, we showcase NMR applications to demonstrate the benefit of using in vivo site-specifically (13)C-labeled RNA. PMID:26577743

  4. [Isotopic signature (15N/14N and 13C/12C) confirms similarity of trophic niches of millipedes (Myriapoda, Diplopoda) in a temperate deciduous forest].

    PubMed

    Semeniuk, I I; Tiunov, A V

    2011-01-01

    The species composition, abundance, and isotopic signature of millipedes (Myriapoda, Diplopoda) were investigated in seven biotopes of Kaluzhskie Zaseki State Nature Reserve. Nine Diplopoda species were found in total, and the local species diversity (within a sampling plot) reached seven species. The Diplopoda tissues were similar to the plant litter in the isotopic composition of nitrogen (delta15N was by 0.4% per hundred higher, on average), but were more strongly enriched in heavy carbon (delta13C was by 4% per hundred higher, on average). Removal of mineral carbon from the cuticle reduced delta13C of Diplopoda by about 1.4% per hundred on average. Differences in the delta15N and delta13C values between the species did not exceed 2.5 per hundred. Differences in the isotopic compositions of the considered species are insignificant, and thus, it is impossible to distinguish particular trophic guilds in the Diplopoda community. Analysis of the published data confirmed that isotopic differentiation of millipedes was much less pronounced than in other investigated groups of soil animals. Hence, millipedes of the deciduous forest form a uniform trophic group.

  5. Retrieving nitrogen isotopic signatures from fresh leaf reflectance spectra: disentangling δ(15)N from biochemical and structural leaf properties.

    PubMed

    Hellmann, Christine; Große-Stoltenberg, André; Lauströ, Verena; Oldeland, Jens; Werner, Christiane

    2015-01-01

    Linking remote sensing methodology to stable isotope ecology provides a promising approach to study ecological processes from small to large spatial scales. Here, we show that δ(15)N can be detected in fresh leaf reflectance spectra of field samples along a spatial gradient of increasing nitrogen input from an N2-fixing invasive species. However, in field data it is unclear whether δ(15)N directly influences leaf reflectance spectra or if the relationship is based on covariation between δ(15)N and foliar nitrogen content or other leaf properties. Using a (15)N-labeling approach, we experimentally varied δ(15)N independently of any other leaf properties in three plant species across different leaf developmental and physiological states. δ(15)N could successfully be modeled by means of partial least squares (PLSs) regressions, using leaf reflectance spectra as predictor variables. PLS models explained 53-73% of the variation in δ(15)N within species. Several wavelength regions important for predicting δ(15)N were consistent across species and could furthermore be related to known absorption features of N-containing molecular bonds. By eliminating covariation with other leaf properties as an explanation for the relationship between reflectance and δ(15)N, our results demonstrate that (15)N itself has an inherent effect on leaf reflectance spectra. Thus, our study substantiates the use of spectroscopic measurements to retrieve isotopic signatures for ecological studies and encourages future development. Furthermore, our results highlight the great potential of optical measurements for up-scaling isotope ecology to larger spatial scales.

  6. Retrieving nitrogen isotopic signatures from fresh leaf reflectance spectra: disentangling δ15N from biochemical and structural leaf properties

    PubMed Central

    Hellmann, Christine; Große-Stoltenberg, André; Lauströ, Verena; Oldeland, Jens; Werner, Christiane

    2015-01-01

    Linking remote sensing methodology to stable isotope ecology provides a promising approach to study ecological processes from small to large spatial scales. Here, we show that δ15N can be detected in fresh leaf reflectance spectra of field samples along a spatial gradient of increasing nitrogen input from an N2-fixing invasive species. However, in field data it is unclear whether δ15N directly influences leaf reflectance spectra or if the relationship is based on covariation between δ15N and foliar nitrogen content or other leaf properties. Using a 15N-labeling approach, we experimentally varied δ15N independently of any other leaf properties in three plant species across different leaf developmental and physiological states. δ15N could successfully be modeled by means of partial least squares (PLSs) regressions, using leaf reflectance spectra as predictor variables. PLS models explained 53–73% of the variation in δ15N within species. Several wavelength regions important for predicting δ15N were consistent across species and could furthermore be related to known absorption features of N-containing molecular bonds. By eliminating covariation with other leaf properties as an explanation for the relationship between reflectance and δ15N, our results demonstrate that 15N itself has an inherent effect on leaf reflectance spectra. Thus, our study substantiates the use of spectroscopic measurements to retrieve isotopic signatures for ecological studies and encourages future development. Furthermore, our results highlight the great potential of optical measurements for up-scaling isotope ecology to larger spatial scales. PMID:25983740

  7. Leaf Urea Metabolism in Potato. Urease Activity Profile and Patterns of Recovery and Distribution of 15N after Foliar Urea Application in Wild-Type and Urease-Antisense Transgenics1

    PubMed Central

    Witte, Claus-Peter; Tiller, Sarah A.; Taylor, Mark A.; Davies, Howard V.

    2002-01-01

    The influence of urease activity on N distribution and losses after foliar urea application was investigated using wild-type and transgenic potato (Solanum tuberosum cv Désirée) plants in which urease activity was down-regulated. A good correlation between urease activity and 15N urea metabolism (NH3 accumulation) was found. The general accumulation of ammonium in leaves treated with urea indicated that urease activity is not rate limiting, at least initially, for the assimilation of urea N by the plant. It is surprising that there was no effect of urease activity on either N losses or 15N distribution in the plants after foliar urea application. Experiments with wild-type plants in the field using foliar-applied 15N urea demonstrated an initial rapid export of N from urea-treated leaves to the tubers within 48 h, followed by a more gradual redistribution during the subsequent days. Only 10% to 18% of urea N applied was lost (presumably because of NH3 volatilization) in contrast to far greater losses reported in several other studies. The pattern of urease activity in the canopy was investigated during plant development. The activity per unit protein increased up to 10-fold with leaf and plant age, suggesting a correlation with increased N recycling in senescing tissues. Whereas several reports have claimed that plant urease is inducible by urea, no evidence for urease induction could be found in potato. PMID:11891267

  8. Nitrate Reduction in a Groundwater Microcosm Determined by 15N Gas Chromatography-Mass Spectrometry

    PubMed Central

    Bengtsson, Göran; Annadotter, Heléne

    1989-01-01

    Aerobic and anaerobic groundwater continuous-flow microcosms were designed to study nitrate reduction by the indigenous bacteria in intact saturated soil cores from a sandy aquifer with a concentration of 3.8 mg of NO3−-N liter−1. Traces of 15NO3− were added to filter-sterilized groundwater by using a Darcy flux of 4 cm day−1. Both assimilatory and dissimilatory reduction rates were estimated from analyses of 15N2, 15N2O, 15NH4+, and 15N-labeled protein amino acids by capillary gas chromatography-mass spectrometry. N2 and N2O were separated on a megabore fused-silica column and quantified by electron impact-selected ion monitoring. NO3− and NH4+ were analyzed as pentafluorobenzoyl amides by multiple-ion monitoring and protein amino acids as their N-heptafluorobutyryl isobutyl ester derivatives by negative ion-chemical ionization. The numbers of bacteria and their [methyl-3H]thymidine incorporation rates were simultaneously measured. Nitrate was completely reduced in the microcosms at a rate of about 250 ng g−1 day−1. Of this nitrate, 80 to 90% was converted by aerobic denitrification to N2, whereas only 35% was denitrified in the anaerobic microcosm, where more than 50% of NO3− was reduced to NH4+. Assimilatory reduction was recorded only in the aerobic microcosm, where N appeared in alanine in the cells. The nitrate reduction rates estimated for the aquifer material were low in comparison with rates in eutrophic lakes and coastal sediments but sufficiently high to remove nitrate from an uncontaminated aquifer of the kind examined in less than 1 month. PMID:16348048

  9. Utilizing the charge field effect on amide (15)N chemical shifts for protein structure validation.

    PubMed

    Bader, Reto

    2009-01-01

    Of all the nuclei in proteins, the nuclear magnetic resonance (NMR) chemical shifts of nitrogen are the theoretically least well understood. In this study, quantum chemical methods are used in combination with polarizable-continuum models in order to show that consideration of the effective electric field, including charge screening due to solvation, improves considerably the consistencies of statistical relationships between experimental and computed amide (15)N shifts between various sets of charged and uncharged oligopeptides and small organic molecules. A single conversion scheme between shielding parameters from first principles using density functional theory (DFT) and experimental shifts is derived that holds for all classes of compounds examined here. This relationship is then used to test the accuracy of such (15)N chemical shift predictions in the cyclic decapeptide antibiotic gramicidin S (GS). GS has previously been studied in great detail, both by NMR and X-ray crystallography. It adopts a well-defined backbone conformation, and hence, only a few discrete side chain conformational states need to be considered. Moreover, a charge-relay effect of the two cationic ornithine side chains to the protein backbone has been described earlier by NMR spectroscopy. Here, DFT-derived backbone amide nitrogen chemical shifts were calculated for multiple conformations of GS. Overall, the structural dynamics of GS is revisited in view of chemical shift behavior along with energetic considerations. Together, the study demonstrates proof of concept that (15)N chemical shift information is particularly useful in the analysis and validation of protein conformational states in a charged environment.

  10. A spectral correlation function for efficient sequential NMR assignments of uniformly (15)N-labeled proteins.

    PubMed

    Bartels, C; Wüthrich, K

    1994-11-01

    A new computer-based approach is described for efficient sequence-specific assignment of uniformly (15)N-labeled proteins. For this purpose three-dimensional (15)N-correlated [(1)H, (1)H]-NOESY spectra are divided up into two-dimensional (1)H-(1)H strips which extend over the entire spectral width along one dimension and have a width of ca. 100 Hz, centered about the amide proton chemical shifts along the other dimension. A spectral correlation function enables sorting of these strips according to proximity of the corresponding residues in the amino acid sequence. Thereby, starting from a given strip in the spectrum, the probability of its corresponding to the C-terminal neighboring residue is calculated for all other strips from the similarity of their peak patterns with a pattern predicted for the sequentially adjoining residue, as manifested in the scalar product of the vectors representing the predicted and measured peak patterns. Tests with five different proteins containing both α-helices and β-sheets, and ranging in size from 58 to 165 amino acid residues show that the discrimination achieved between the sequentially neighboring residue and all other residues compares well with that obtained with an unguided interactive search of pairs of sequentially neighboring strips, with important savings in the time needed for complete analysis of 3D (15)N-correlated [(1)H, (1)H]-NOESY spectra. The integration of this routine into the program package XEASY ensures that remaining ambiguities can be resolved by visual inspection of the strips, combined with reference to the amino acid sequence and information on spin-system types obtained from additional NMR spectra.

  11. Determination of the δ15N of nitrate in solids; RSIL lab code 2894

    USGS Publications Warehouse

    Coplen, Tyler B.; Qi, Haiping; Revesz, Kinga; Casciotti, Karen; Hannon, Janet E.

    2007-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2894 is to determine the δ15N of nitrate (NO3-) in solids. The nitrate fraction of the nitrogen species is dissolved by water (called leaching) and can be analyzed by the bacterial method covered in RSIL lab code 2899. After leaching, the δ15N of the dissolved NO3- is analyzed by conversion of the NO3- to nitrous oxide (N2O), which serves as the analyte for mass spectrometry. A culture of denitrifying bacteria is used in the enzymatic conversion of NO3- to N2O, which follows the pathway shown in equation 1: NO3- → NO2- → NO → 1/2 N2O (1) Because the bacteria Pseudomonas aureofaciens lack N2O reductive activity, the reaction stops at N2O, unlike the typical denitrification reaction that goes to N2. After several hours, the conversion is complete, and the N2O is extracted from the vial, separated from volatile organic vapor and water vapor by an automated -65 °C isopropanol-slush trap, a Nafion drier, a CO2 and water removal unit (Costech #021020 carbon dioxide absorbent with Mg(ClO4)2), and trapped in a small-volume trap immersed in liquid nitrogen with a modified Finnigan MAT (now Thermo Scientific) GasBench 2 introduction system. After the N2O is released, it is further purified by gas chromatography before introduction to the isotope-ratio mass spectrometer (IRMS). The IRMS is a Thermo Scientific Delta V Plus continuous flow IRMS (CF-IRMS). It has a universal triple collector, consisting of two wide cups with a narrow cup in the middle; it is capable of simultaneously measuring mass/charge (m/z) of the N2O molecule 44, 45, and 46. The ion beams from these m/z values are as follows: m/z = 44 = N2O = 14N14N16O; m/z = 45 = N2O = 14N15N16O or 14N14N17O; m/z = 46 = N2O = 14N14N18O. The 17O contributions to the m/z 44 and m/z 45 ion beams are accounted for before δ15N values are reported.

  12. Determination of the δ15N of nitrate in water; RSIL lab code 2899

    USGS Publications Warehouse

    Coplen, Tyler B.; Qi, Haiping; Revesz, Kinga; Casciotti, Karen; Hannon, Janet E.

    2007-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2899 is to determine the δ15N of nitrate (NO3-) in water. The δ15N of the dissolved NO3- is analyzed by conversion of the NO3- to nitrous oxide (N2O), which serves as the analyte for mass spectrometry. A culture of denitrifying bacteria is used in the enzymatic conversion of the NO3- to N2O, which follows the pathway shown in equation 1: NO3- → NO2- → NO → 1/2 N2O (1) Because the bacteria Pseudomonas aureofaciens lack N2O reductive activity, the reaction stops at N2O, unlike the typical denitrification reaction that goes to N2. After several hours, the conversion is complete, and the N2O is extracted from the vial, separated from volatile organic vapor and water vapor by an automated -65 °C isopropanol-slush trap, a Nafion drier, a CO2 and water removal unit (Costech #021020 carbon dioxide absorbent with Mg(ClO4)2), and trapped in a small-volume trap immersed in liquid nitrogen with a modified Finnigan MAT (now Thermo Scientific) GasBench 2 introduction system. After the N2O is released, it is further purified by gas chromatography before introduction to the isotope-ratio mass spectrometer (IRMS). The IRMS is a Thermo Scientific Delta V Plus continuous flow IRMS (CF-IRMS). It has a universal triple collector, consisting of two wide cups with a narrow cup in the middle; it is capable of simultaneously measuring mass/charge (m/z) of the N2O molecule 44, 45, and 46. The ion beams from these m/z values are as follows: m/z = 44 = N2O = 14N14N16O; m/z = 45 = N2O = 14N15N16O or 14N14N17O; m/z = 46 = N2O = 14N14N18O. The 17O contributions to the m/z 44 and m/z 45 ion beams are accounted for before δ15N values are reported.

  13. Optical Microscopy Characterization for Borehole U-15n#12 in Support of NCNS Source Physics Experiment

    SciTech Connect

    Wilson, Jennifer E.; Sussman, Aviva Joy

    2015-05-22

    Optical microscopy characterization of thin sections from corehole U-15n#12 is part of a larger material characterization effort for the Source Physics Experiment (SPE). The SPE program was conducted in Nevada with a series of explosive tests designed to study the generation and propagation of seismic waves inside Stock quartz monzonite. Optical microscopy analysis includes the following: 1) imaging of full thin sections (scans and mosaic maps); 2) high magnification imaging of petrographic texture (grain size, foliations, fractures, etc.); and 3) measurement of microfracture density.

  14. Structural study of ammonium metatungstate

    SciTech Connect

    Christian, Joel B. Whittingham, M. Stanley

    2008-08-15

    Several techniques have been used to study the structure of the Keggin-type polyoxometalate salt ammonium metatungstate (AMT)-(NH{sub 4}){sub 6}[H{sub 2}W{sub 12}O{sub 40}]*nH{sub 2}O, a potential fuel cell catalyst. The dehydrated salt is comprised of a mixture of crystallites of different unit cells in a centered eutactic cubic configuration, with an average unit cell of a{approx_equal}12.295. Varied orientations of the Keggin ions in the cubic arrangement create the differences, and orientational variation within each unit cell size represents an energy well. Progressive hydration of each crystallite leads to expansion of the lattice, with the degree of expansion depending on the locations of the water added in relation to the Keggin ion, which is influenced by cation location and hydrogen bonding. The structural hypothesis is supported by electron diffraction of single and multicrystal samples, by powder density measurements, X-ray powder diffraction studies, synchrotron powder X-ray diffraction, and a priori structural modeling studies. Based on the structure, projected active site densities are compared with nanostructured platinum catalysts for fuel cell application. - Graphical abstract: The structure of ammonium metatungstate powders are highly dependent on hydration and POM molecule rotation, with cation and hydrogen bonding forces directing a mixture of structures that have been studied with bulk and single-crystal methods. The illustration shows Monte Carlo simulated anion structural disorder for the fully dehydrated form of the title compound.

  15. Phase Diagram of Ammonium Nitrate

    NASA Astrophysics Data System (ADS)

    Dunuwille, Mihindra; Yoo, Choong-Shik

    2013-06-01

    Ammonium Nitrate (AN) has often been subjected to uses in improvised explosive devices, due to its wide availability as a fertilizer and its capability of becoming explosive with slight additions of organic and inorganic compounds. Yet, the origin of enhanced energetic properties of impure AN (or AN mixtures) is neither chemically unique nor well understood - resulting in rather catastrophic disasters in the past1 and thereby a significant burden on safety, in using ammonium nitrates even today. To remedy this situation, we have carried out an extensive study to investigate the phase stability of AN, in different chemical environments, at high pressure and temperature, using diamond anvil cells and micro-Raman spectroscopy. The present results confirm the recently proposed phase IV-to-IV' transition above 15 GPa2 and provide new constraints for the melting and phase diagram of AN to 40 GPa and 673 K. The present study has been supported by the U.S. DHS under Award Number 2008-ST-061-ED0001.

  16. Nitrate-Dependent Ferrous Iron Oxidation by Anaerobic Ammonium Oxidation (Anammox) Bacteria

    PubMed Central

    Oshiki, M.; Ishii, S.; Yoshida, K.; Fujii, N.; Ishiguro, M.; Satoh, H.

    2013-01-01

    We examined nitrate-dependent Fe2+ oxidation mediated by anaerobic ammonium oxidation (anammox) bacteria. Enrichment cultures of “Candidatus Brocadia sinica” anaerobically oxidized Fe2+ and reduced NO3− to nitrogen gas at rates of 3.7 ± 0.2 and 1.3 ± 0.1 (mean ± standard deviation [SD]) nmol mg protein−1 min−1, respectively (37°C and pH 7.3). This nitrate reduction rate is an order of magnitude lower than the anammox activity of “Ca. Brocadia sinica” (10 to 75 nmol NH4+ mg protein−1 min−1). A 15N tracer experiment demonstrated that coupling of nitrate-dependent Fe2+ oxidation and the anammox reaction was responsible for producing nitrogen gas from NO3− by “Ca. Brocadia sinica.” The activities of nitrate-dependent Fe2+ oxidation were dependent on temperature and pH, and the highest activities were seen at temperatures of 30 to 45°C and pHs ranging from 5.9 to 9.8. The mean half-saturation constant for NO3− ± SD of “Ca. Brocadia sinica” was determined to be 51 ± 21 μM. Nitrate-dependent Fe2+ oxidation was further demonstrated by another anammox bacterium, “Candidatus Scalindua sp.,” whose rates of Fe2+ oxidation and NO3− reduction were 4.7 ± 0.59 and 1.45 ± 0.05 nmol mg protein−1 min−1, respectively (20°C and pH 7.3). Co-occurrence of nitrate-dependent Fe2+ oxidation and the anammox reaction decreased the molar ratios of consumed NO2− to consumed NH4+ (ΔNO2−/ΔNH4+) and produced NO3− to consumed NH4+ (ΔNO3−/ΔNH4+). These reactions are preferable to the application of anammox processes for wastewater treatment. PMID:23624480

  17. MUSIC in triple-resonance experiments: amino acid type-selective (1)H-(15)N correlations

    PubMed

    Schubert; Smalla; Schmieder; Oschkinat

    1999-11-01

    Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective (1)H-(15)N correlations are presented. They are based on novel coherence transfer schemes, the MUSIC pulse sequence elements, that replace the initial INEPT transfer and are selective for XH(2) or XH(3) (X can be (15)N or (13)C). The desired amino acid type is thereby selected based on the topology of the side chain. Experiments for Gly (G-HSQC); Ala (A-HSQC); Thr, Val, Ile, and Ala (TAVI-HSQC); Thr and Ala (TA-HSQC), as well as Asn and Gln (N-HSQC and QN-HSQC), are described. The new experiments are recorded as two-dimensional experiments and therefore need only small amounts of spectrometer time. The performance of the experiments is demonstrated with the application to two protein domains. Copyright 1999 Academic Press. PMID:10527741

  18. Oxygen determination in materials by 18O(p,αγ)15N nuclear reaction

    NASA Astrophysics Data System (ADS)

    Kumar, Sanjiv; Sunitha, Y.; Reddy, G. L. N.; Sukumar, A. A.; Ramana, J. V.; Sarkar, A.; Verma, Rakesh

    2016-07-01

    The paper presents a proton induced γ-ray emission method based on 18O(p,αγ)15N nuclear reaction to determine bulk oxygen in materials. The determination involves the measurement of 5.27 MeV γ-rays emitted following the de-excitation of 15N nuclei. A description of the energetics of the reaction is given to provide an insight into the origin of 5.27 MeV γ-rays. In addition, thick target γ-ray yields and the limits of detection are measured to ascertain the analytical potential of the reaction. The thick-target γ-ray yields are measured with a high purity germanium detector and a bismuth germanate detector at 0° as well as 90° angles in 3.0-4.2 MeV proton energy region. The best limit of detection of about 1.3 at.% is achieved at 4.2 MeV proton energy for measurements at 0° as well 90° angles with the bismuth germanate detector while the uncertainty in quantitative analysis is <8%. The reaction has a probing depth of several tens of microns. Interferences can arise from fluorine due to the occurrence of 19F(p,αγ)16O reaction that emits 6-7 MeV γ-rays. The analytical potential of the methodology is demonstrated by determining oxygen in several oxide as well as non-oxide materials.

  19. The coral δ15N record of terrestrial nitrate loading varies with river catchment land use

    NASA Astrophysics Data System (ADS)

    Yamazaki, A.; Watanabe, T.; Tsunogai, U.; Hasegawa, H.; Yamano, H.

    2015-03-01

    We analysed the nitrogen isotopes in two coral cores (δ15Ncoral) from the mouth of the Todoroki River, Ishigaki Island, Japan, to examine whether the δ15Ncoral reflects the run-off of nitrate related to the land use in the river catchment. The two coral cores were used to examine the seasonal variation in δ15Ncoral for 14 years (CORE1; 1993-2007) and the annual variation of δ15Ncoral for 52 years (CORE2; 1958-2010). In CORE1, the 5-month running mean of δ15Ncoral was positively correlated with that of monthly precipitation, excluding all strong precipitation events (>150 mm d-1). In CORE2, the δ15Ncoral mean in the earlier period (1958-1980) was 1.0 ‰ greater than that in the later period (1981-2010). The annual averages of δ15Ncoral are positively correlated with the total precipitation in the rainy season (May-June) for both time periods. The difference in the δ15Ncoral between the earlier and later periods is probably caused by the land use changed from paddy fields with 15N-rich manure to sugar cane fields in the early 1980s. Although some uncertainties still remain regarding the precision of δ15N coral proxy records, this study emphasises the clear potential for their use in reconstructing terrestrial nitrate discharge records from corals.

  20. Covalent binding of aniline to humic substances. 2. 15N NMR studies of nucleophilic addition reactions

    USGS Publications Warehouse

    Thorn, K.A.; Pettigrew, P.J.; Goldenberg, W.S.; Weber, E.J.

    1996-01-01

    Aromatic amines are known to undergo covalent binding with humic substances in the environment. Although previous studies have examined reaction conditions and proposed mechanisms, there has been no direct spectroscopic evidence for the covalent binding of the amines to the functional groups in humic substances. In order to further elucidate the reaction mechanisms, the Suwannee River and IHSS soil fulvic and humic acids were reacted with 15N-labeled aniline at pH 6 and analyzed using 15N NMR spectrometry. Aniline underwent nucleophilic addition reactions with the quinone and other carbonyl groups in the samples and became incorporated in the form of anilinohydroquinone, anilinoquinone, anilide, imine, and heterocyclic nitrogen, the latter comprising 50% or more of the bound amine. The anilide and anilinohydroquinone nitrogens were determined to be susceptible to chemical exchange by ammonia. In the case of Suwannee River fulvic acid, reaction under anoxic conditions and pretreatment with sodium borohydride or hydroxylamine prior to reaction under oxic conditions resulted in a decrease in the proportion of anilinohydroquinone nitrogen incorporated. The relative decrease in the incorporation of anilinohydroquinone nitrogen with respect to anilinoquinone nitrogen under anoxic conditions suggested that inter- or intramolecular redox reactions accompanied the nucleophilic addition reactions.

  1. UV-visible and (1)H-(15)N NMR spectroscopic studies of colorimetric thiosemicarbazide anion sensors.

    PubMed

    Farrugia, Kristina N; Makuc, Damjan; Podborska, Agnieszka; Szaciłowski, Konrad; Plavec, Janez; Magri, David C

    2015-02-14

    Four model thiosemicarbazide anion chemosensors containing three N-H bonds, substituted with phenyl and/or 4-nitrophenyl units, were synthesised and studied for their anion binding abilities with hydroxide, fluoride, acetate, dihydrogen phosphate and chloride. The anion binding properties were studied in DMSO and 9 : 1 DMSO-H2O by UV-visible absorption and (1)H/(13)C/(15)N NMR spectroscopic techniques and corroborated with DFT studies. Significant changes were observed in the UV-visible absorption spectra with all anions, except for chloride, accompanied by dramatic colour changes visible to the naked eye. These changes were determined to be due to the deprotonation of the central N-H proton and not due to hydrogen bonding based on (1)H/(15)N NMR titration studies with acetate in DMSO-d6-0.5% water. Direct evidence for deprotonation was confirmed by the disappearance of the central thiourea proton and the formation of acetic acid. DFT and charge distribution calculations suggest that for all four compounds the central N-H proton is the most acidic. Hence, the anion chemosensors operate by a deprotonation mechanism of the central N-H proton rather than by hydrogen bonding as is often reported. PMID:25451865

  2. Attenuation of Nitrate-15N by Vegetated Buffers in an Irrigated Pasture System

    NASA Astrophysics Data System (ADS)

    Haughn, A. B.; Tate, K.; Kessel, C. V.

    2003-12-01

    Irrigated pastures are found within watersheds providing much of Western North America's surface drinking water supply. Vegetative buffers are often proposed to attenuate nutrient pollutants in runoff, but there is limited information on the mechanistic functioning of buffers adjacent to irrigated pastures. This study is intended to fill this gap in knowledge by examining specific vegetation, soil, and landscape characteristics controlling buffer efficiency and capacity. At the University of California Sierra Foothill Research and Extension Center, established flood-irrigated plots with three different buffer treatments are being used to characterize the attenuation of N, P, and sediment by buffers. Stable 15N isotope tracer was applied to quantify the fate of nitrate moving through the pasture and buffers. In the first 10 days following application of the 15N tracer, 2% of the tracer was lost as runoff, with more than half of the total loss occurring from plots with no buffers. Of the remaining tracer, 47% was taken up by grass in the zone of application, 3% was taken up by vegetation within the buffers (primarily in the first 4m of buffer), 20% was stored in the A horizon of the soil, and 28% was lost via leaching and/or gaseous losses. Results presented will include the effect of buffer length on nutrient attenuation and the relative importance of different N pools for nitrate retention. This research will allow land managers to maximize efficiency of riparian buffers adjacent to irrigated pasture, potentially increasing the adoption of vegetated buffers as a management tool.

  3. Automated Protein Turnover Calculations from 15N Partial Metabolic Labeling LC/MS Shotgun Proteomics Data

    PubMed Central

    Lyon, David; Castillejo, Maria Angeles; Staudinger, Christiana; Weckwerth, Wolfram; Wienkoop, Stefanie; Egelhofer, Volker

    2014-01-01

    Protein turnover is a well-controlled process in which polypeptides are constantly being degraded and subsequently replaced with newly synthesized copies. Extraction of composite spectral envelopes from complex LC/MS shotgun proteomics data can be a challenging task, due to the inherent complexity of biological samples. With partial metabolic labeling experiments this complexity increases as a result of the emergence of additional isotopic peaks. Automated spectral extraction and subsequent protein turnover calculations enable the analysis of gigabytes of data within minutes, a prerequisite for systems biology high throughput studies. Here we present a fully automated method for protein turnover calculations from shotgun proteomics data. The approach enables the analysis of complex shotgun LC/MS 15N partial metabolic labeling experiments. Spectral envelopes of 1419 peptides can be extracted within an hour. The method quantifies turnover by calculating the Relative Isotope Abundance (RIA), which is defined as the ratio between the intensity sum of all heavy (15N) to the intensity sum of all light (14N) and heavy peaks. To facilitate this process, we have developed a computer program based on our method, which is freely available to download at http://promex.pph.univie.ac.at/protover. PMID:24736476

  4. Contribution of 19F resonances on 18O( p, α)15N reaction rate

    NASA Astrophysics Data System (ADS)

    Benmeslem, Meriem; Chafa, Azzedine; Barhoumi, Slimane; Tribeche, Mouloud

    2014-08-01

    The 18O( p, α)15N reaction influences the isotopes production such as 19F, 18O, and 15N which can be used to test the models of stellar evolution. 19F is synthesized in both asymptotic giant branch (AGB) and metal-rich Wolf-Rayet (WR) stars. Using R-matrix theory we allow new values of resonances parameters in 19F. We show that the most important contribution to the differential and total cross section at low energies, comes from the levels in 19F situated at resonances energies E R =151, 680 and 840 keV with spin and parity 1/2+. The total width of the 680 keV resonance is badly known. So, we have focused on this broad resonance corresponding to the 8.65 MeV level in 19F. We delimit the temperature range in which each resonance contribution to the total reaction rate occurs by analyzing the ratio ( N A < σν> i / N A < σν>). This allowed us to show that the 680 and 840 keV broad resonances strongly dominate the reaction rate over the stellar temperature range T 9=0.02-0.06 and T 9=0.5-5. Finally, these results were compared to NACRE and Iliadis astrophysical compilations.

  5. Measurement and interpretation of 15N-1H residual dipolar couplings in larger proteins.

    PubMed

    Bhattacharya, Akash; Revington, Matthew; Zuiderweg, Erik R P

    2010-03-01

    A decade ago, Dr. L.E. Kay and co-workers described an ingenious HNCO-based triple-resonance experiment from which several protein backbone RDCs can be measured simultaneously (Yang et al. (1999) [1]). They implemented a J-scaling technique in the (15)N dimension of the 3D experiment to obtain the NH RDCs. We have used this idea to carry out J-scaling in a 2D (15)N-(1)H-TROSY experiment and have found it to be an excellent method to obtain NH RDCs for larger proteins upto 70 kDa, far superior to commonly used HSQC in-phase/anti-phase and HSQC/TROSY comparisons. Here, this method, dubbed "RDC-TROSY" is discussed in detail and the limits of its utility are assessed by simulations. Prominent in the latter analysis is the evaluation of the effect of amide proton flips on the "RDC-TROSY" linewidths. The details of the technical and computational implementations of these methods for the determination of domain orientations in 45-60 kDa Hsp70 chaperone protein constructs are described. PMID:20018538

  6. MUSIC in Triple-Resonance Experiments: Amino Acid Type-Selective 1H- 15N Correlations

    NASA Astrophysics Data System (ADS)

    Schubert, Mario; Smalla, Maika; Schmieder, Peter; Oschkinat, Hartmut

    1999-11-01

    Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective 1H-15N correlations are presented. They are based on novel coherence transfer schemes, the MUSIC pulse sequence elements, that replace the initial INEPT transfer and are selective for XH2 or XH3 (X can be 15N or 13C). The desired amino acid type is thereby selected based on the topology of the side chain. Experiments for Gly (G-HSQC); Ala (A-HSQC); Thr, Val, Ile, and Ala (TAVI-HSQC); Thr and Ala (TA-HSQC), as well as Asn and Gln (N-HSQC and QN-HSQC), are described. The new experiments are recorded as two-dimensional experiments and therefore need only small amounts of spectrometer time. The performance of the experiments is demonstrated with the application to two protein domains.

  7. MUSIC in triple-resonance experiments: amino acid type-selective (1)H-(15)N correlations

    PubMed

    Schubert; Smalla; Schmieder; Oschkinat

    1999-11-01

    Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective (1)H-(15)N correlations are presented. They are based on novel coherence transfer schemes, the MUSIC pulse sequence elements, that replace the initial INEPT transfer and are selective for XH(2) or XH(3) (X can be (15)N or (13)C). The desired amino acid type is thereby selected based on the topology of the side chain. Experiments for Gly (G-HSQC); Ala (A-HSQC); Thr, Val, Ile, and Ala (TAVI-HSQC); Thr and Ala (TA-HSQC), as well as Asn and Gln (N-HSQC and QN-HSQC), are described. The new experiments are recorded as two-dimensional experiments and therefore need only small amounts of spectrometer time. The performance of the experiments is demonstrated with the application to two protein domains. Copyright 1999 Academic Press.

  8. Stereospecific assignments of glycine in proteins by stereospecific deuteration and {sup 15}N labeling

    SciTech Connect

    Hansen, A.P.; Curley, R.W. Jr.; Panigot, M.J.; Fesik, S.W.

    1994-12-01

    Stereospecific assignments are important for accurately determining the three-dimensional structures of proteins through the use of multidimensional NMR techniques. It is especially important to stereospecifically assign the glycine {alpha}-protons in proteins because of the potential for different backbone conformations of this residue. These stereospecific assignments are critical for interpreting the {sup 3}J{sub NH,{alpha}H} coupling constants and NOEs involving the glycine {alpha}-protons that determine the conformation of this part of the protein. However, it is often difficult to unambiguously obtain the stereospecific assignments for glycine residues by using only NOE data. In this poster, we present a method for unambiguous, stereospecific assignment of the {alpha}-protons of glycine residues. This method involves synthesis of stereo-specifically deuterated and {sup 15}N-labeled Gly using a slightly modified procedure originally described by Woodard and coworkers for the stereoselective deuteration of glycine. The stereospecifically deuterated and {sup 15}N-labeled Gy has been incorporated into recombinant proteins expressed in both bacterial systems (FKBP) and mammalian cells (u-PA). Two- and three-dimensional isotope-filtered and isotope-edited NMR experiments were used to obtain the stereospecific assignments of the glycine {alpha}-protons for these proteins.

  9. Precursor discrimination of designer drug benzylpiperazine using δ13C and δ15N stable isotopes.

    PubMed

    Beckett, Nicola M; Grice, Darren I; Carter, James F; Cresswell, Sarah L

    2015-01-01

    Advances in analytical technology and emerging techniques have resulted in the increased exploitation of chemical and isotopic profiling for source linkage/discrimination of illicit drugs for forensic purposes. Although not routinely used for illicit drug investigations, such information has been obtained and its application demonstrated through the use of isotope ratio mass spectrometry (IRMS). There is a solid platform of research available relating to the isotopic analysis of methylenedioxymethamphetamine (MDMA) and methamphetamine (MA), however with the recently flourishing designer drug market it was of interest to examine the isotopic profiles of the popular 'party drug' benzylpiperazine hydrochloride (BZP·HCl). A preliminary analysis of δ13C and δ15N isotopic ratios in BZP·HCl products and corresponding synthetic intermediates (piperazine·HCl) synthesized in-house from three different precursor suppliers was conducted using IRMS. Analysis of the δ13C and δ15N isotopic data indicated that discrimination and correct grouping of all the intermediates and some of the product samples examined in this study were achievable.

  10. Plant and soil natural abundance delta (15)N: indicators of relative rates of nitrogen cycling in temperate forest ecosystems.

    PubMed

    Templer, Pamela H; Arthur, Mary A; Lovett, Gary M; Weathers, Kathleen C

    2007-08-01

    Watersheds within the Catskill Mountains, New York, receive among the highest rates of nitrogen (N) deposition in the northeastern United States and are beginning to show signs of N saturation. Despite similar amounts of N deposition across watersheds within the Catskill Mountains, rates of soil N cycling and N retention vary significantly among stands of different tree species. We examined the potential use of delta (15)N of plants and soils as an indicator of relative forest soil N cycling rates. We analyzed the delta (15)N of foliage, litterfall, bole wood, surface litter layer, fine roots and organic soil from single-species stands of American beech (Fagus grandifolia), eastern hemlock (Tsuga canadensis), red oak (Quercus rubra), and sugar maple (Acer saccharum). Fine root and organic soil delta (15)N values were highest within sugar maple stands, which correlated significantly with higher rates of net mineralization and nitrification. Results from this study suggest that fine root and organic soil delta (15)N can be used as an indicator of relative rates of soil N cycling. Although not statistically significant, delta (15)N was highest within foliage, wood and litterfall of beech stands, a tree species associated with intermediate levels of soil N cycling rates and forest N retention. Our results show that belowground delta (15)N values are a better indicator of relative rates of soil N cycling than are aboveground delta (15)N values.

  11. Diffusion technique for 15N and inorganic N analysis of low-N aqueous solutions and Kjeldahl digests.

    PubMed

    Chen, Rui Rui; Dittert, Klaus

    2008-06-01

    Diffusion of ammonia is a common sample preparation method for the stable isotope analysis of inorganic nitrogen in aqueous solution. Classical diffusion methods usually require 6-12 days of diffusion and often focus on (15)N/(14)N analysis only. More recent studies have discussed whether complete N recovery was necessary for the precise analysis of stable N isotope ratios. In this paper we present a newly revised diffusion technique that allows correct and simultaneous determination of total N and (15)N at% from aqueous solutions and Kjeldahl digests, with N concentrations down to sub-0.5-mg N L(-1) levels, and it is tested under different conditions of (15)N isotope labelling. With the modification described, the diffusion time was reduced to 72 h, while the ratios of measured and expected (15)N at% were greater than 99% and the simultaneous recovery of total N was >95%. Analysis of soil microbial biomass N and its (15)N/(14)N ratio is one of the most important applications of this diffusion technique. An experiment with soil extracts spiked with (15)N-labelled yeast showed that predigestion was necessary to prevent serious N loss during Kjeldahl digestion of aqueous samples (i.e. soil extracts). The whole method of soil microbial biomass N preparation for (15)N/(14)N analysis included chloroform fumigation, predigestion, Kjeldahl digestion and diffusion. An experiment with soil spiked with (15)N-labelled yeast was carried out to evaluate the method. Results showed a highly significant correlation of recovered and added N, with the same recovery rate (0.21) of both total N and (15)N. A k(N) value of 0.25 was obtained based on the data. In conclusion, the diffusion method works for soil extracts and microbial biomass N determination and hence could be useful in many types of soil/water studies.

  12. Glacial-interglacial dynamics of Antarctic firn columns: comparison between simulations and ice core air-?15N measurements

    NASA Astrophysics Data System (ADS)

    Capron, E.; Landais, A.; Buiron, D.; Cauquoin, A.; Chappellaz, J. A.; Debret, M.; Jouzel, J.; Leuenberger, M.; Martinerie, P.; Masson-Delmotte, V.; Mulvaney, R.; Parrenin, F.; Prié, F.

    2013-12-01

    Correct estimation of the firn lock-in depth is essential for correctly linking gas and ice chronologies in ice core studies. Here, two approaches to constrain the firn depth evolution in Antarctica are presented over the last deglaciation: outputs of a firn densification model, and measurements of δ15N of N2 in air trapped in ice core, assuming that δ15N is only affected by gravitational fractionation in the firn column. Since the firn densification process is largely governed by surface temperature and accumulation rate, we have investigated four ice cores drilled in coastal (Berkner Island, BI, and James Ross Island, JRI) and semi-coastal (TALDICE and EPICA Dronning Maud Land, EDML) Antarctic regions. Combined with available ice core air- δ15N measurements from the EPICA Dome C (EDC) site, the studied regions encompass a large range of surface accumulation rates and temperature conditions. Our δ15N profiles reveal a heterogeneous response of the firn structure to glacial-interglacial climatic changes. While firn densification simulations correctly predict TALDICE δ15N variations, they systematically fail to capture the large millennial-scale δ15N variations measured at BI and the δ15N glacial levels measured at JRI and EDML - a mismatch previously reported for central East Antarctic ice cores. New constraints of the EDML gas-ice depth offset during the Laschamp event (41 ka) and the last deglaciation do not favour the hypothesis of a large convective zone within the firn as the explanation of the glacial firn model- δ15N data mismatch for this site. While we could not conduct an in-depth study of the influence of impurities in snow for firnification from the existing datasets, our detailed comparison between the δ15N profiles and firn model simulations under different temperature and accumulation rate scenarios suggests that the role of accumulation rate may have been underestimated in the current description of firnification models.

  13. Glacial-interglacial dynamics of Antarctic firn columns: comparison between simulations and ice core air-δ15N measurements

    NASA Astrophysics Data System (ADS)

    Capron, E.; Landais, A.; Buiron, D.; Cauquoin, A.; Chappellaz, J.; Debret, M.; Jouzel, J.; Leuenberger, M.; Martinerie, P.; Masson-Delmotte, V.; Mulvaney, R.; Parrenin, F.; Prié, F.

    2013-05-01

    Correct estimation of the firn lock-in depth is essential for correctly linking gas and ice chronologies in ice core studies. Here, two approaches to constrain the firn depth evolution in Antarctica are presented over the last deglaciation: outputs of a firn densification model, and measurements of δ15N of N2 in air trapped in ice core, assuming that δ15N is only affected by gravitational fractionation in the firn column. Since the firn densification process is largely governed by surface temperature and accumulation rate, we have investigated four ice cores drilled in coastal (Berkner Island, BI, and James Ross Island, JRI) and semi-coastal (TALDICE and EPICA Dronning Maud Land, EDML) Antarctic regions. Combined with available ice core air-δ15N measurements from the EPICA Dome C (EDC) site, the studied regions encompass a large range of surface accumulation rates and temperature conditions. Our δ15N profiles reveal a heterogeneous response of the firn structure to glacial-interglacial climatic changes. While firn densification simulations correctly predict TALDICE δ15N variations, they systematically fail to capture the large millennial-scale δ15N variations measured at BI and the δ15N glacial levels measured at JRI and EDML - a mismatch previously reported for central East Antarctic ice cores. New constraints of the EDML gas-ice depth offset during the Laschamp event (~41 ka) and the last deglaciation do not favour the hypothesis of a large convective zone within the firn as the explanation of the glacial firn model-δ15N data mismatch for this site. While we could not conduct an in-depth study of the influence of impurities in snow for firnification from the existing datasets, our detailed comparison between the δ15N profiles and firn model simulations under different temperature and accumulation rate scenarios suggests that the role of accumulation rate may have been underestimated in the current description of firnification models.

  14. Productivity and sedimentary δ15N variability for the last 17,000 years along the northern Gulf of Alaska continental slope

    USGS Publications Warehouse

    Addison, Jason A.; Finney, Bruce P.; Dean, Walter E.; Davies, Maureen H.; Mix, Alan C.; Stoner, Joseph S.; Jaeger, John M.

    2012-01-01

    Biogenic opal, organic carbon, organic matter stable isotope, and trace metal data from a well-dated, high-resolution jumbo piston core (EW0408–85JC; 59° 33.3′N, 144° 9.21′W, 682 m water depth) recovered from the northern Gulf of Alaska continental slope reveal changes in productivity and nutrient utilization over the last 17,000 years. Maximum values of opal concentration (∼10%) occur during the deglacial Bølling-Allerød (B-A) interval and earliest Holocene (11.2 to 10.8 cal ka BP), moderate values (∼6%) occur during the Younger Dryas (13.0 to 11.2 cal ka BP) and Holocene, and minimum values (∼3.5%) occur during the Late Glacial Interval (LGI). When converted to opal mass accumulation rates, the highest values (∼5000 g cm−2 kyr−1) occur during the LGI prior to 16.7 cal ka BP, which points to a strong influence by LGI glacimarine sedimentation regimes. Similar patterns are also observed in total organic carbon and cadmium paleoproductivity proxies. Mid-Holocene peaks in the terrestrial organic matter fraction at 5.5, 4.7, 3.5, and 1.2 cal ka BP indicate periods of enhanced delivery of glaciomarine sediments by the Alaska Coastal Current. The B-A and earliest Holocene intervals are laminated, and enrichments of redox-sensitive elements suggest dysoxic-to-anoxic conditions in the water column. The laminations are also associated with mildly enriched sedimentary δ15N ratios, indicating a link between productivity, nitrogen cycle dynamics, and sedimentary anoxia. After applying a correction for terrestrial δ15N contributions based on end-member mixing models of terrestrial and marine organic matter, the resulting B-A marine δ15N (6.3 ± 0.4 ‰) ratios are consistent with either mild denitrification, or increased nitrate utilization. These findings can be explained by increased micronutrient (Fe) availability during episodes of rapid rising sea level that released iron from the previously subaerial coastal plain; iron input from enhanced

  15. Biogeochemical Indicators in High- and Low-Arctic Marine and Terrestrial Avian Community Changes: Comparative Isotopic (13C, 15N, and 34S) Studies in Alaska and Greenland

    NASA Astrophysics Data System (ADS)

    Causey, D.; Bargmann, N. A.; Burnham, K. K.; Burnham, J. L.; Padula, V. M.; Johnson, J. A.; Welker, J. M.

    2011-12-01

    Fulmar). Generally, High Arctic species have tissues that are more enriched in δ15N compared to their Low Arctic counterparts, but δ13C values are similar in both regions. These patterns follow observed regional differences in Arctic isoscapes, and are probably related to isotopic variations in food rather than trophic level differentiation. Both Low- and High-Arctic bird communities show decadal-period shifts in stable isotope profiles, based on prior published results and previously collected specimens. For example, Aleutian birds feeding in upper trophic levels have tissues are more enriched in both δ13C and δ15N compared to specimens collected in 2000-2001. We anticipate broadening the time depth of this initial study using museum archival and archeological material, as well as continuing studies in 2013 and 2014.

  16. Natural-abundance 15N NMR studies of Turkey ovomucoid third domain. Assignment of peptide 15N resonances to the residues at the reactive site region via proton-detected multiple-quantum coherence

    NASA Astrophysics Data System (ADS)

    Ortiz-Polo, Gilberto; Krishnamoorthi, R.; Markley, John L.; Live, David H.; Davis, Donald G.; Cowburn, David

    Heteronuclear two-dimensional 1H{ 15N} multiple-quantum (MQ) spectroscopy has been applied to a protein sample at natural abundance: ovomucoid third domain from turkey ( Meleagris gallopavo), a serine proteinase inhibitor of 56 amino acid residues. Peptide amide 1H NMR assignments obtained by two-dimensional 1H{ 1H} NMR methods (R. Krishnamoorthi and J. L. Markley, unpublished data) led to identification of the corresponding 1H{ 15N} MQ coherence cross peaks. From these, 15N NMR chemical shifts were determined for several specific backbone amide groups of amino acid residues located around the reactive site region of the inhibitor. The results suggest that amide 15N chemical shifts, which are readily obtained in this way, may serve as sensitive probes for conformational studies of proteins.

  17. 49 CFR 176.410 - Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 2 2012-10-01 2012-10-01 false Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures. 176.410 Section 176.410 Transportation Other Regulations Relating to... nitrate and ammonium nitrate mixtures. (a) This section prescribes requirements to be observed...

  18. 49 CFR 176.410 - Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 2 2011-10-01 2011-10-01 false Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures. 176.410 Section 176.410 Transportation Other Regulations Relating to... nitrate and ammonium nitrate mixtures. (a) This section prescribes requirements to be observed...

  19. 49 CFR 176.410 - Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 2 2014-10-01 2014-10-01 false Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures. 176.410 Section 176.410 Transportation Other Regulations Relating to... nitrate and ammonium nitrate mixtures. (a) This section prescribes requirements to be observed...

  20. 49 CFR 176.410 - Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures. 176.410 Section 176.410 Transportation Other Regulations Relating to... nitrate and ammonium nitrate mixtures. (a) This section prescribes requirements to be observed...

  1. 49 CFR 176.410 - Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 2 2013-10-01 2013-10-01 false Division 1.5 materials, ammonium nitrate and ammonium nitrate mixtures. 176.410 Section 176.410 Transportation Other Regulations Relating to... nitrate and ammonium nitrate mixtures. (a) This section prescribes requirements to be observed...

  2. Phase diagram of ammonium nitrate.

    PubMed

    Dunuwille, Mihindra; Yoo, Choong-Shik

    2013-12-01

    Ammonium Nitrate (AN) is a fertilizer, yet becomes an explosive upon a small addition of chemical impurities. The origin of enhanced chemical sensitivity in impure AN (or AN mixtures) is not well understood, posing significant safety issues in using AN even today. To remedy the situation, we have carried out an extensive study to investigate the phase stability of AN and its mixtures with hexane (ANFO-AN mixed with fuel oil) and Aluminum (Ammonal) at high pressures and temperatures, using diamond anvil cells (DAC) and micro-Raman spectroscopy. The results indicate that pure AN decomposes to N2, N2O, and H2O at the onset of the melt, whereas the mixtures, ANFO and Ammonal, decompose at substantially lower temperatures. The present results also confirm the recently proposed phase IV-IV' transition above 17 GPa and provide new constraints for the melting and phase diagram of AN to 40 GPa and 400°C.

  3. Phase diagram of ammonium nitrate

    NASA Astrophysics Data System (ADS)

    Dunuwille, Mihindra; Yoo, Choong-Shik

    2013-12-01

    Ammonium Nitrate (AN) is a fertilizer, yet becomes an explosive upon a small addition of chemical impurities. The origin of enhanced chemical sensitivity in impure AN (or AN mixtures) is not well understood, posing significant safety issues in using AN even today. To remedy the situation, we have carried out an extensive study to investigate the phase stability of AN and its mixtures with hexane (ANFO-AN mixed with fuel oil) and Aluminum (Ammonal) at high pressures and temperatures, using diamond anvil cells (DAC) and micro-Raman spectroscopy. The results indicate that pure AN decomposes to N2, N2O, and H2O at the onset of the melt, whereas the mixtures, ANFO and Ammonal, decompose at substantially lower temperatures. The present results also confirm the recently proposed phase IV-IV' transition above 17 GPa and provide new constraints for the melting and phase diagram of AN to 40 GPa and 400°C.

  4. Effects of climate on deer bone δ15N and δ13C: Lack of precipitation effects on δ15N for animals consuming low amounts of C 4 plants

    NASA Astrophysics Data System (ADS)

    Cormie, A. B.; Schwarcz, H. P.

    1996-11-01

    We have examined the relationship of bone collagen δ15N and δ13C to climatic variables, humidity, temperature, and amount of precipitation using fifty-nine specimens of North American white-tailed deer ( Odocoileus virginianus) from forty-six different locations. In previous studies of African mammals there was a significant correlation between bone collagen δ15N and local amount of precipitation. Results presented here similarly show an increase in δ15N with decreasing amount of precipitation but only for 25% of the animals, namely those consuming more than 10% C 4 plants. These animals also exhibited a significant correlation between δ13C and temperature which mirrors previous observations for grasses suggesting that these deer consume grasses during times of population and nutrient stress. In contrast, even in dry areas containing high proportions of C 4 grasses, the majority of the deer had consumed low amounts of C 4 plants and these deer did not have δ15N which correlate with amount of precipitation. Only when deer deviated from their normal feeding pattern by consuming C 4 plants or grasses did their δ15N correlate with amount of rainfall. For these animals, consumption of C 4 plants or grasses may signal conditions of water and nutrient stress. An increase in δ15N of bone collagen may result from combined effects from excretion of concentrated urine (to conserve water) and increased internal recycling of nitrogen (to conserve nitrogen).

  5. Symbiotic N2-Fixation Estimated by the 15N Tracer Technique and Growth of Pueraria phaseoloides (Roxb.) Benth. Inoculated with Bradyrhizobium Strain in Field Conditions

    PubMed Central

    Sarr, Papa Saliou; Okon, Judith Wase; Begoude, Didier Aime Boyogueno; Araki, Shigeru; Ambang, Zachée; Shibata, Makoto; Funakawa, Shinya

    2016-01-01

    This field experiment was established in Eastern Cameroon to examine the effect of selected rhizobial inoculation on N2-fixation and growth of Pueraria phaseoloides. Treatments consisted of noninoculated and Bradyrhizobium yuanmingense S3-4-inoculated Pueraria with three replications each. Ipomoea batatas as a non-N2-fixing reference was interspersed in each Pueraria plot. All the twelve plots received 2 gN/m2 of 15N ammonium sulfate 10% atom excess. At harvest, dry matter yields and the nitrogen derived from atmospheric N2-fixation (%Ndfa) of inoculated Pueraria were significantly (P < 0.05) higher (81% and 10.83%, resp.) than those of noninoculated Pueraria. The inoculation enhanced nodule dry weight 2.44-fold. Consequently, the harvested N significantly (P < 0.05) increased by 83% in inoculated Pueraria, resulting from the increase in N2-fixation and soil N uptake. A loss of 55 to 60% of the N fertilizer was reported, and 36 to 40% of it was immobilized in soil. Here, we demonstrated that both N2-fixing potential of P. phaseoloides and soil N uptake are improved through field inoculations using efficient bradyrhizobial species. In practice, the inoculation contributes to maximize N input in soils by the cover crop's biomass and represent a good strategy to improve soil fertility for subsequent cultivation. PMID:26904363

  6. Symbiotic N 2 -Fixation Estimated by the (15) N Tracer Technique and Growth of Pueraria phaseoloides (Roxb.) Benth. Inoculated with Bradyrhizobium Strain in Field Conditions.

    PubMed

    Sarr, Papa Saliou; Okon, Judith Wase; Begoude, Didier Aime Boyogueno; Araki, Shigeru; Ambang, Zachée; Shibata, Makoto; Funakawa, Shinya

    2016-01-01

    This field experiment was established in Eastern Cameroon to examine the effect of selected rhizobial inoculation on N2-fixation and growth of Pueraria phaseoloides. Treatments consisted of noninoculated and Bradyrhizobium yuanmingense S3-4-inoculated Pueraria with three replications each. Ipomoea batatas as a non-N2-fixing reference was interspersed in each Pueraria plot. All the twelve plots received 2 gN/m(2) of (15)N ammonium sulfate 10% atom excess. At harvest, dry matter yields and the nitrogen derived from atmospheric N2-fixation (%Ndfa) of inoculated Pueraria were significantly (P < 0.05) higher (81% and 10.83%, resp.) than those of noninoculated Pueraria. The inoculation enhanced nodule dry weight 2.44-fold. Consequently, the harvested N significantly (P < 0.05) increased by 83% in inoculated Pueraria, resulting from the increase in N2-fixation and soil N uptake. A loss of 55 to 60% of the N fertilizer was reported, and 36 to 40% of it was immobilized in soil. Here, we demonstrated that both N2-fixing potential of P. phaseoloides and soil N uptake are improved through field inoculations using efficient bradyrhizobial species. In practice, the inoculation contributes to maximize N input in soils by the cover crop's biomass and represent a good strategy to improve soil fertility for subsequent cultivation. PMID:26904363

  7. 15N tracer application to evaluate nitrogen dynamics of food webs in two subtropical small-scale aquaculture ponds under different managements.

    PubMed

    Pucher, Johannes; Mayrhofer, Richard; El-Matbouli, Mansour; Focken, Ulfert

    2014-01-01

    Small, semi-intensively managed aquaculture ponds contribute significantly to the food security of small-scale farmers around the world. However, little is known about nutrient flows within natural food webs in such ponds in which fish production depends on the productivity of natural food resources. (15)N was applied as ammonium at 1.1 and 0.4 % of total nitrogen in a traditionally managed flow-through pond and a semi-intensively managed stagnant pond belonging to small-scale farmers in Northern Vietnam and traced through the natural food resources over 7 days. Small-sized plankton (1-60 μ m) was the dominant pelagic biomass in both ponds with higher biomass in the stagnant pond. This plankton assimilated major portions of the applied tracer and showed a high sedimentation and turnover rate. High re-activation of settled nutrients into the pelagic food web was observed. The tracer was removed more quickly from the flow-through pond than from the stagnant pond. A steady nutrient supply could increase fish production. PMID:24995524

  8. 15N tracer application to evaluate nitrogen dynamics of food webs in two subtropical small-scale aquaculture ponds under different managements.

    PubMed

    Pucher, Johannes; Mayrhofer, Richard; El-Matbouli, Mansour; Focken, Ulfert

    2014-01-01

    Small, semi-intensively managed aquaculture ponds contribute significantly to the food security of small-scale farmers around the world. However, little is known about nutrient flows within natural food webs in such ponds in which fish production depends on the productivity of natural food resources. (15)N was applied as ammonium at 1.1 and 0.4 % of total nitrogen in a traditionally managed flow-through pond and a semi-intensively managed stagnant pond belonging to small-scale farmers in Northern Vietnam and traced through the natural food resources over 7 days. Small-sized plankton (1-60 μ m) was the dominant pelagic biomass in both ponds with higher biomass in the stagnant pond. This plankton assimilated major portions of the applied tracer and showed a high sedimentation and turnover rate. High re-activation of settled nutrients into the pelagic food web was observed. The tracer was removed more quickly from the flow-through pond than from the stagnant pond. A steady nutrient supply could increase fish production.

  9. Juvenile psittacine environmental enrichment.

    PubMed

    Simone-Freilicher, Elisabeth; Rupley, Agnes E

    2015-05-01

    Environmental enrichment is of great import to the emotional, intellectual, and physical development of the juvenile psittacine and their success in the human home environment. Five major types of enrichment include social, occupational, physical, sensory, and nutritional. Occupational enrichment includes exercise and psychological enrichment. Physical enrichment includes the cage and accessories and the external home environment. Sensory enrichment may be visual, auditory, tactile, olfactory, or taste oriented. Nutritional enrichment includes variations in appearance, type, and frequency of diet, and treats, novelty, and foraging. Two phases of the preadult period deserve special enrichment considerations: the development of autonomy and puberty.

  10. Nitrogen cycling between sediment and the shallow-water column in the transition zone of the Potomac River and Estuary. II. The role of wind-driven resuspension and adsorbed ammonium

    USGS Publications Warehouse

    Simon, N.S.

    1989-01-01

    During periods of sediment resuspension, desorption of ammonium from sediment solids can be the major pathway for enriching the water column with the ammonium that is produced by bacterial degradation of organic matter in the bottom material. This hyopthesis is based on a three-year study of diffusive flux in the transition zone of the Potomac River at a site 35 m from the Virginia shore where the average water-column depth is approximately 1 m over sandy sediment. A diffusion-controlled sampler was used to collect water samples at the interface between the water column and sediment and at several tens of centimeters into the sediment. Interstitial water concentration gradients showed that diffusive flux of ammonium from the sandy shallow-water sediments was approximately 1% of the diffusive flux of ammonium from the silty channel sediments in the same zone of the Potomac River. Organic nitrogen and bound or adsorbed ammonium were the predominant nitrogen forms in the sediment. Adsorbed ammonium concentrations ranged from nondetectable to 3??7 ??mol g-1 of sediment. Concentrations of adsorbed ammonium per gram of sediment were one to three orders of magnitude more than interstitial water ammonium concentrations. Desorption of ammonium from sediment solids appeared to be the controlling factor in the degree of water-column ammonium enrichment. In laboratory experiments that simulated sediment resuspension, 40-80% of the adsorbed ammonium predicted to desorb did so after approximately 30 min of mixing. Based on calculations for 1 m2 to a depth of 4 cm, one resuspenion event lasting minutes could mix more ammonium into the water column from desorption of ammonium from sediment solids than could be delivered to the water column by diffusive flux from shallow-water sediments in 10-1000 days and would be comparable to enrichment by ammonium diffusive flux for 5-50 days from channel sediments in the same river zone. ?? 1989.

  11. Biodegradation of rocket propellant waste, ammonium perchlorate

    NASA Technical Reports Server (NTRS)

    Naqvi, S. M. Z.; Latif, A.

    1975-01-01

    The short term effects of ammonium perchlorate on selected organisms were studied. A long term experiment was also designed to assess the changes incurred by ammonium perchlorate on the nitrogen and chloride contents of soil within a period of 3 years. In addition, an attempt was made to produce methane gas from anaerobic fermentation of the aquatic weed, Alternanthera philoxeroides.

  12. 76 FR 46907 - Ammonium Nitrate Security Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-03

    ... Federal Bureau of Investigation FR Federal Register HMR Hazardous Materials Regulations HMT Hazardous... ``Secure Handling of Ammonium Nitrate Program'' on October 29, 2008. See 73 FR 64280. The ANPRM solicited... interacting with state and local governments regarding ammonium nitrate security. ] See 73 FR 64280,...

  13. 21 CFR 573.170 - Ammonium formate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.170 Ammonium formate. The food additive, partially ammonium formate, may be safely used in...) To assure safe use of the additive, in addition to the other information required by the Federal...

  14. 21 CFR 573.170 - Ammonium formate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.170 Ammonium formate. The food additive, partially ammonium formate, may be safely used in...) To assure safe use of the additive, in addition to the other information required by the Federal...

  15. 21 CFR 573.170 - Ammonium formate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.170 Ammonium formate. The food additive, partially ammonium formate, may be safely used in...) To assure safe use of the additive, in addition to the other information required by the Federal...

  16. 21 CFR 573.170 - Ammonium formate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.170 Ammonium formate. The food additive, ammonium formate, may be safely used in the... safe use of the additive, in addition to the other information required by the Federal Food, Drug,...

  17. 21 CFR 558.340 - Maduramicin ammonium.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Maduramicin ammonium. 558.340 Section 558.340 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... for Use in Animal Feeds § 558.340 Maduramicin ammonium. (a) Approvals. Type A medicated articles:...

  18. Nitrogen use efficiency evaluation of aerobic rice under field capacity water potential using {sup 15}N isotopic tracer technique

    SciTech Connect

    Wahid, Ahmad Nazrul Abd; Rahim, Sahibin Abd; Rahim, Khairuddin Abdul; Harun, Abdul Rahim

    2015-09-25

    This study was carried out to evaluate the efficiency use of the nitrogen fertilizer on aerobic rice varieties MR219-4 and MR219-9 which were grown aerobically under field capacity water potential at the controlled environment area or shield house. Direct {sup 15}N isotope tracer method was used in this study, whereby the {sup 15}N isotope was utilized as a tracer for nitrogen nutrient uptake. {sup 15}N isotope presence in the samples is determined by using emission spectrometer analysis and percentage of total nitrogen is determined by using Kjeldahl method. {sup 15}N atom access value contained in the sample will be used in determining the effectiveness of the use of nitrogen in fertilizers through the specific calculation formulas. In this work, the data several data of nitrogen derived from fertilizer (Ndff), total nitrogen, nitrogen uptake and nitrogen use efficiency was obtained.

  19. Cosine Modulated HSQC: A Rapid Determination of 3JHNHα Scalar Couplings in 15N-labeled Proteins

    NASA Astrophysics Data System (ADS)

    Petit, Audrey; Vincent, Sébastien J. F.; Zwahlen, Catherine

    2002-06-01

    A two-dimensional HSQC-based NMR method, 15N-COSMO-HSQC, is presented for the rapid determination of homonuclear 3JHNHα couplings in 15N-labeled proteins in solution. Scalar couplings are extracted by comparing the intensity of two separate datasets recorded with and without decoupling of the 3JHNHα during a preparation period. The scalar couplings are introduced through a cosine modulation of the peak intensities. The experiment relies on a BIRD sandwich to selectively invert all amide protons H N and is very simple to implement. 3JHNHα couplings were determined using both the 15N-COSMO-HSQC and quantitative- J on 15N-labeled chemokine RANTES. The two experiments show well-correlated values.

  20. Nitrogen use efficiency evaluation of aerobic rice under field capacity water potential using 15N isotopic tracer technique

    NASA Astrophysics Data System (ADS)

    Wahid, Ahmad Nazrul Abd; Rahim, Sahibin Abd; Rahim, Khairuddin Abdul; Harun, Abdul Rahim

    2015-09-01

    This study was carried out to evaluate the efficiency use of the nitrogen fertilizer on aerobic rice varieties MR219-4 and MR219-9 which were grown aerobically under field capacity water potential at the controlled environment area or shield house. Direct 15N isotope tracer method was used in this study, whereby the 15N isotope was utilized as a tracer for nitrogen nutrient uptake. 15N isotope presence in the samples is determined by using emission spectrometer analysis and percentage of total nitrogen is determined by using Kjeldahl method. 15N atom access value contained in the sample will be used in determining the effectiveness of the use of nitrogen in fertilizers through the specific calculation formulas. In this work, the data several data of nitrogen derived from fertilizer (Ndff), total nitrogen, nitrogen uptake and nitrogen use efficiency was obtained.

  1. The 15N(α,γ)19F reaction and nucleosynthesis of 19F

    NASA Astrophysics Data System (ADS)

    Wilmes, S.; Wilmes, V.; Staudt, G.; Mohr, P.; Hammer, J. W.

    2002-12-01

    Several resonances in the 15N(α,γ)19F reaction have been investigated in the energy range between 0.6 MeV and 2.7 MeV. Resonance strengths and branching ratios have been determined. High sensitivity could be obtained by the combination of the Dynamitron high current accelerator, the windowless gas target system Rhinoceros, and actively shielded germanium detectors. Two levels of 19F could be observed for the first time in the (α,γ) channel, and several weak branchings below the detection limits of previous experiments were measured. Two observed resonances correspond to α-cluster states in 19F, which have been assigned unambiguously. The astrophysical reaction rate is derived from this set of resonance strengths.

  2. Nanoscale studies of cement chemistry with 15N resonance reaction analysis

    NASA Astrophysics Data System (ADS)

    Schweitzer, Jeffrey S.; Livingston, Richard A.; Rolfs, Claus; Becker, Hans-Werner; Kubsky, Stefan; Spillane, Timothy; Castellote, Marta; de Viedma, Paloma G.

    2005-12-01

    Analyses of materials with ion beams have proven to be a valuable technique for describing the spatial distributions of specific elements in host materials. We have applied this technique using the 15N(p, αγ)12C reaction to study the time dependence of the chemical reactions involved in the curing of cement. By using the Dynamitron Tandem accelerator at the Ruhr Universität, Bochum, Germany, we have been able to achieve a few nanometer spatial resolution at the surface of cement grains and to study the hydrogen distributions to a depth of about 2 μm. By applying a technique for stopping the chemical reactions at arbitrary times, the time dependence of the chemical reactions involving specific components of cement can be investigated. In addition, the effects of additives on the chemical reactions have been studied, as have materials that are components of concrete.

  3. 1H, 13C, and 15N resonance assignments of murine amelogenin, an enamel biomineralization protein.

    SciTech Connect

    Buchko, Garry W.; Bekhazi, Jacky G.; Cort, John R.; Valentine, Nancy B.; Snead, Malcolm L.; Shaw, Wendy J.

    2008-06-01

    Amelogenin is the predominant matrix protein in developing dental enamel. Making extensive use of residue-specific 15N-labeled amino acids samples, the majority of the main and side chain resonances for murine amelogenin were assigned in 2% aqueous acetic acid at pH 3.0. This research was performed at Pacific Northwest National Laboratory, operated by Battelle for the US-DOE. A large part of this research was performed at the W.R. Wiley Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by U.S. Department of Energy’s Office of Biological and Environmental Research (BER) program located at Pacific Northwest National Laboratory (PNNL).

  4. Determination of 15N/14N and 13C/12C in Solid and Aqueous Cyanides

    USGS Publications Warehouse

    Johnson, C.A.

    1996-01-01

    The stable isotopic compositions of nitrogen and carbon in cyanide compounds can be determined by combusting aliquots in sealed tubes to form N2 gas and CO2 gas and analyzing the gases by mass spectrometry. Free cyanide (CN-aq + HCNaq) in simple solutions can also be analyzed by first precipitating the cyanide as copper(II) ferrocyanide and then combusting the precipitate. Reproducibility is ??0.5??? or better for both ??15N and ??13C. If empirical corrections are made on the basis of carbon yields, the reproducibility of ??13C can be improved to ??0.2???. The analytical methods described herein are sufficiently accurate and precise to apply stable isotope techniques to problems of cyanide degradation in natural waters and industrial process solutions.

  5. Experimental and calculated 1H, 13C, 15N NMR spectra of famotidine

    NASA Astrophysics Data System (ADS)

    Barańska, M.; Czarniecki, K.; Proniewicz, L. M.

    2001-05-01

    Famotidine, 3-[[[2-[(aminoiminomethyl)amino]-4-thiazolyl]methyl]thio]- N-(aminosulfonyl), is a histamine H 2-receptor blocker that has been used mainly for the treatment of peptic ulcers and the Zollinger-Ellison syndrome. Its NMR spectra in different solvents were reported earlier; however, detailed interpretation has not been done thus far. In this work, experimental 1H, 13C and 15N NMR spectra of famotidine dissolved in DMSO-d 6 are shown. The assignment of observed chemical shifts is based on quantum chemical calculation at the Hartree-Fock/6-31G ∗ level. The geometry optimization of the famotidine molecule with two internal hydrogen bonds, i.e. [N(3)-H(23)⋯N(9) and N(3)⋯H(34)-N(20)], is done by using the B3LYP method with the 6-31G ∗ basis set.

  6. Bedrock Nitrogen and Hydrothermal Ammonium in Yellowstone National Park, WY, USA

    NASA Astrophysics Data System (ADS)

    Holloway, J. M.; Bohlke, J. K.; Nordstrom, D. K.

    2003-12-01

    High ammonium concentrations in some of the hot springs of Yellowstone National Park (up to 880 mg L-1 as N at Washburn Hot Springs) have been attributed to leaching of sedimentary rock by hydrothermal solutions. However, relatively little is known about the direct relationship between rock geochemistry in volcanic centers and nitrogen in thermal waters. For this study, a suite of core samples from US Geological Survey drill holes in Yellowstone National Park were characterized for nitrogen and carbon in different lithologies. These data were related to the aqueous geochemistry and δ 15N-NH4+ of thermal waters in different hot spring basins in the park to better understand the water-rock interactions. Core samples selected for study included tuff, water-reworked volcanic sediments, glacial sediment, lacustrine sedimentary rock, and marine sedimentary rock. Substantial amounts of nitrogen were present in all bedrock types, with the highest nitrogen concentrations measured in marine sedimentary rocks (430-830 mg N kg-1) from Y10, which is located at Mammoth Hot Springs. Although the underlying bedrock has elevated nitrogen concentrations, hydrothermal ammonium concentrations at Mammoth Hot Springs are relatively low (~1 mg L-1 as N). These solutions are buffered by carbonate (pH >8) and may have lost some N by volatilization as ammonia gas. Thermal waters in Norris Geyser Basin are acid to circumneutral with ammonium concentrations ranging from <0.03 to 80 mg L-1 as N. Nitrogen in tuffs (400-620 mg N kg-1) from drill holes Y9 and Y12 at Norris Geyser Basin may be present as a result of ammonium partitioning from solution to zeolites or other secondary minerals. Thermal waters sampled at Mammoth, Norris, and other geyser basins in the park varied widely in ammonium concentrations and isotopic compositions (δ 15N), from <0.3 to 450 mg L-1 as N and -5 to +25‰ , respectively. The isotope data are interpreted to reflect multiple processes, including leaching of

  7. Search for d3/2 single particle strength in 15N in Unbound Levels

    NASA Astrophysics Data System (ADS)

    Mertin, C. E.; Caussyn, D. D.; Crisp, A. M.; Keeley, N.; Kemper, K. W.; Momotyuk, O.; Roeder, B. T.; Volya, A.

    2013-10-01

    The population of states in the nucleus 15N provides the opportunity to investigate both single particle and cluster structures in the 1p and 2s1d shells. Single, two, three and four particle transfer reactions selectively excite states in 15N thus providing a way to explore current nuclear structure models. Narrow structures are observed in the various transfer reactions up to at least 20 MeV in excitation well above the neutron (10.8 MeV) and proton (10.2 MeV) separation energies. In the present work new results for the reaction 14N(d,p) are presented that explore possible single particle strengths up to 18 MeV in excitation. The beam energies used in the present work were between 10.5 and 16 MeV. An early work with a beam energy of 8 MeV clearly populated strong sharp levels at 10.07 and 11.23 MeV and the present work confirms their existence. In addition, very weak broader levels are populated at 12.13 and 12.5 MeV but no other structures are found experimentally at higher excitation energies. The results of shell model calculations that include the 1p and 2s1d shells will be presented. The centroid energies for the 1d5/2 and 2s1/2 single particle strength have been obtained through comparison with FRESCO calculations. This work was supported by the NSF, DOE and Florida State University.

  8. δ15N constraints on long-term nitrogen balances in temperate forests

    USGS Publications Warehouse

    Perakis, S.S.; Sinkhorn, E.R.; Compton, J.E.

    2011-01-01

    Biogeochemical theory emphasizes nitrogen (N) limitation and the many factors that can restrict N accumulation in temperate forests, yet lacks a working model of conditions that can promote naturally high N accumulation. We used a dynamic simulation model of ecosystem N and δ15N to evaluate which combination of N input and loss pathways could produce a range of high ecosystem N contents characteristic of forests in the Oregon Coast Range. Total ecosystem N at nine study sites ranged from 8,788 to 22,667 kg ha−1 and carbon (C) ranged from 188 to 460 Mg ha−1, with highest values near the coast. Ecosystem δ15N displayed a curvilinear relationship with ecosystem N content, and largely reflected mineral soil, which accounted for 96–98% of total ecosystem N. Model simulations of ecosystem N balances parameterized with field rates of N leaching required long-term average N inputs that exceed atmospheric deposition and asymbiotic and epiphytic N2-fixation, and that were consistent with cycles of post-fire N2-fixation by early-successional red alder. Soil water δ15NO3 − patterns suggested a shift in relative N losses from denitrification to nitrate leaching as N accumulated, and simulations identified nitrate leaching as the primary N loss pathway that constrains maximum N accumulation. Whereas current theory emphasizes constraints on biological N2-fixation and disturbance-mediated N losses as factors that limit N accumulation in temperate forests, our results suggest that wildfire can foster substantial long-term N accumulation in ecosystems that are colonized by symbiotic N2-fixing vegetation.

  9. Regional, seasonal and interspecific variation in 15N and 13C in sympatric mouse lemurs.

    PubMed

    Rakotondranary, S Jacques; Struck, Ulrich; Knoblauch, Christian; Ganzhorn, Jörg U

    2011-11-01

    Madagascar provides some of the rare examples where two or more primate species of the same genus and with seemingly identical niche requirements occur in sympatry. If congeneric primate species co-occur in other parts of the world, they differ in size in a way that is consistent with Hutchinson's rule for coexisting species, or they occupy different ecological niches. In some areas of Madagascar, mouse lemurs do not follow these "rules" and thus seem to violate one of the principles of community ecology. In order to understand the mechanisms that allow coexistence of sympatric congeneric species without obvious niche differentiation, we studied food composition of two identical sized omnivorous mouse lemur species, Microcebus griseorufus and M. murinus with the help of stable isotope analyses (δ(15)N and δ(13)C). The two species are closely related sister species. During the rich season, when food seems abundant, the two species do not differ in their nitrogen isotope composition, indicating that the two species occupy the same trophic level. But they differ in their δ(13)C values, indicating that M. griseorufus feeds more on C(4) and CAM (Crassulacean-acid-metabolism) plants than M. murinus. During the lean season, M. murinus has lower δ(15)N values, indicating that the two species feed at different trophic levels during times of food shortage. Hybrids between the two species showed intermediate food composition. The results reflect subtle differences in foraging or metabolic adaptations that are difficult to quantify by traditional observations but that represent possibilities to allow coexistence of species. PMID:21881908

  10. Backbone dynamics of the oligomerization domain of p53 determined from 15N NMR relaxation measurements.

    PubMed

    Clubb, R T; Omichinski, J G; Sakaguchi, K; Appella, E; Gronenborn, A M; Clore, G M

    1995-05-01

    The backbone dynamics of the tetrameric p53 oligomerization domain (residues 319-360) have been investigated by two-dimensional inverse detected heteronuclear 1H-15N NMR spectroscopy at 500 and 600 MHz. 15N T1, T2, and heteronuclear NOEs were measured for 39 of 40 non-proline backbone NH vectors at both field strengths. The overall correlation time for the tetramer, calculated from the T1/T2 ratios, was found to be 14.8 ns at 35 degrees C. The correlation times and amplitudes of the internal motions were extracted from the relaxation data using the model-free formalism (Lipari G, Szabo A, 1982, J Am Chem Soc 104:4546-4559). The internal dynamics of the structural core of the p53 oligomerization domain are uniform and fairly rigid, with residues 327-354 exhibiting an average generalized order parameter (S2) of 0.88 +/- 0.08. The N- and C-termini exhibit substantial mobility and are unstructured in the solution structure of p53. Residues located at the N- and C-termini, in the beta-sheet, in the turn between the alpha-helix and beta-sheet, and at the C-terminal end of the alpha-helix display two distinct internal motions that are faster than the overall correlation time. Fast internal motions (< or = 20 ps) are within the extreme narrowing limit and are of uniform amplitude. The slower motions (0.6-2.2 ns) are outside the extreme narrowing limit and vary in amplitude.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7663341

  11. Regional, seasonal and interspecific variation in 15N and 13C in sympatric mouse lemurs

    NASA Astrophysics Data System (ADS)

    Rakotondranary, S. Jacques; Struck, Ulrich; Knoblauch, Christian; Ganzhorn, Jörg U.

    2011-11-01

    Madagascar provides some of the rare examples where two or more primate species of the same genus and with seemingly identical niche requirements occur in sympatry. If congeneric primate species co-occur in other parts of the world, they differ in size in a way that is consistent with Hutchinson's rule for coexisting species, or they occupy different ecological niches. In some areas of Madagascar, mouse lemurs do not follow these "rules" and thus seem to violate one of the principles of community ecology. In order to understand the mechanisms that allow coexistence of sympatric congeneric species without obvious niche differentiation, we studied food composition of two identical sized omnivorous mouse lemur species, Microcebus griseorufus and M. murinus with the help of stable isotope analyses ( δ 15N and δ 13C). The two species are closely related sister species. During the rich season, when food seems abundant, the two species do not differ in their nitrogen isotope composition, indicating that the two species occupy the same trophic level. But they differ in their δ 13C values, indicating that M. griseorufus feeds more on C4 and CAM (Crassulacean-acid-metabolism) plants than M. murinus. During the lean season, M. murinus has lower δ 15N values, indicating that the two species feed at different trophic levels during times of food shortage. Hybrids between the two species showed intermediate food composition. The results reflect subtle differences in foraging or metabolic adaptations that are difficult to quantify by traditional observations but that represent possibilities to allow coexistence of species.

  12. 15N solid-state NMR provides a sensitive probe of oxidized flavin reactive sites.

    PubMed

    Koder, Ronald L; Walsh, Joseph D; Pometun, Maxim S; Dutton, P Leslie; Wittebort, Richard J; Miller, Anne-Frances

    2006-11-29

    Flavins are central to the reactivity of a wide variety of enzymes and electron transport proteins. There is great interest in understanding the basis for the different reactivities displayed by flavins in different protein contexts. We propose solid-state nuclear magnetic resonance (SS-NMR) as a tool for directly observing reactive positions of the flavin ring and thereby obtaining information on their frontier orbitals. We now report the SS-NMR signals of the redox-active nitrogens N1 and N5, as well as that of N3. The chemical shift tensor of N5 is over 720 ppm wide, in accordance with the predictions of theory and our calculations. The signal of N3 can be distinguished on the basis of coupling to 1H absent for N1 and N5, as well as the shift tensor span of only 170 ppm, consistent with N3's lower aromaticity and lack of a nonbonding lone pair. The isotropic shifts and spans of N5 and N1 reflect two opposite extremes of the chemical shift range for "pyridine-type" N's, consistent with their electrophilic and nucleophilic chemical reactivities, respectively. Upon flavin reduction, N5's chemical shift tensor contracts dramatically to a span of less than 110 ppm, and the isotropic chemical shift changes by approximately 300 ppm. Both are consistent with loss of N5's nonbonding lone pair and decreased aromaticity, and illustrate the responsiveness of the 15N chemical shift principal values to electronic structure. Thus. 15N chemical shift principal values promise to be valuable tools for understanding electronic differences that underlie variations in flavin reactivity, as well as the reactivities of other heterocyclic cofactors. PMID:17117871

  13. Species specific and environment induced variation of δ13C and δ15N in alpine plants

    PubMed Central

    Yang, Yang; Siegwolf, Rolf T. W.; Körner, Christian

    2015-01-01

    Stable carbon and nitrogen isotope signals in plant tissues integrate plant-environment interactions over long periods. In this study, we hypothesized that humid alpine life conditions are narrowing the scope for significant deviations from common carbon, water and nitrogen relations as captured by stable isotope signals. We explored the variation in δ13C and δ15N in 32 plant species from tissue type to ecosystem scale across a suite of locations at c. Two thousand five hundred meter elevation in the Swiss Alps. Foliar δ13C and δ15N varied among species by about 3–4‰ and 7–8‰ respectively. However, there was no overall difference in means of δ13C and δ15N for species sampled in different plant communities or when bulk plant dry matter harvests of different plant communities were compared. δ13C was found to be highly species specific, so that the ranking among species was mostly maintained across 11 habitats. However, δ15N varied significantly from place to place in all species (a range of 2.7‰) except in Fabaceae (Trifolium alpinum) and Juncaceae (Luzula lutea). There was also a substantial variation among individuals of the same species collected next to each other. No difference was found in foliar δ15N of non-legumes, which were either collected next to or away from the most common legume, T. alpinum. δ15N data place Cyperaceae and Juncaceae, just like Fabaceae, in a low discrimination category, well separated from other families. Soil δ15N was higher than in plants and increased with soil depth. The results indicate a high functional diversity in alpine plants that is similar to that reported for low elevation plants. We conclude that the surprisingly high variation in δ13C and δ15N signals in the studied high elevation plants is largely species specific (genetic) and insensitive to obvious environmental cues. PMID:26097487

  14. Comparison of five soil organic matter decomposition models using data from a 14C and 15N labeling field experiment

    NASA Astrophysics Data System (ADS)

    Pansu, Marc; Bottner, Pierre; Sarmiento, Lina; Metselaar, Klaas

    2004-12-01

    Five alternatives of the previously published MOMOS model (MOMOS-2 to -6) are tested to predict the dynamics of carbon (C) and nitrogen (N) in soil during the decomposition of plant necromass. 14C and 15N labeled wheat straw was incubated over 2 years in fallow soils of the high Andean Paramo of Venezuela. The following data were collected: soil moisture, total 14C and 15N and microbial biomass (MB)-14C and -15N, daily rainfall, air temperature and total radiation. Daily soil moisture was predicted using the SAHEL model. MOMOS-2 to -4 (type 1 models) use kinetic constants and flow partitioning parameters. MOMOS-2 can be simplified to MOMOS-3 and further to MOMOS-4, with no significant changes in the prediction accuracy and robustness for total-14C and -15N as well as for MB-14C and -15N. MOMOS-5 (type 2 models) uses only kinetic constants: three MB-inputs (from labile and stable plant material and from humified compounds) and two MB-outputs (mortality and respiration constants). MOMOS-5 did not significantly change the total-14C and -15N predictions but markedly improved the predictive quality and robustness of MB-14C and -15N predictions (with a dynamic different from the predictions by other models). Thus MOMOS-5 is proposed as an accurate and ecologically consistent description of decomposition processes. MOMOS-6 extends MOMOS-5 by including a stable humus compartment for long-term simulations of soil native C and N. The improvement of the predictions is not significant for this 2-year experiment, but MOMOS-6 enables prediction of a sequestration in the stable humus compartment of 2% of the initially added 14C and 5.4% of the added 15N.

  15. Evolution of Electrogenic Ammonium Transporters (AMTs).

    PubMed

    McDonald, Tami R; Ward, John M

    2016-01-01

    The ammonium transporter gene family consists of three main clades, AMT, MEP, and Rh. The evolutionary history of the AMT/MEP/Rh gene family is characterized by multiple horizontal gene transfer events, gene family expansion and contraction, and gene loss; thus the gene tree for this family of transporters is unlike the organismal tree. The genomes of angiosperms contain genes for both electrogenic and electroneutral ammonium transporters, but it is not clear how far back in the land plant lineage electrogenic ammonium transporters occur. Here, we place Marchantia polymorpha ammonium transporters in the AMT/MEP/Rh phylogeny and we show that AMTs from the liverwort M. polymorpha are electrogenic. This information suggests that electrogenic ammonium transport evolved at least as early as the divergence of bryophytes in the land plant lineage. PMID:27066024

  16. Evolution of Electrogenic Ammonium Transporters (AMTs)

    PubMed Central

    McDonald, Tami R.; Ward, John M.

    2016-01-01

    The ammonium transporter gene family consists of three main clades, AMT, MEP, and Rh. The evolutionary history of the AMT/MEP/Rh gene family is characterized by multiple horizontal gene transfer events, gene family expansion and contraction, and gene loss; thus the gene tree for this family of transporters is unlike the organismal tree. The genomes of angiosperms contain genes for both electrogenic and electroneutral ammonium transporters, but it is not clear how far back in the land plant lineage electrogenic ammonium transporters occur. Here, we place Marchantia polymorpha ammonium transporters in the AMT/MEP/Rh phylogeny and we show that AMTs from the liverwort M. polymorpha are electrogenic. This information suggests that electrogenic ammonium transport evolved at least as early as the divergence of bryophytes in the land plant lineage. PMID:27066024

  17. Lineshape-based polarimetry of dynamically-polarized 15N2O in solid-state mixtures

    PubMed Central

    Kuzma, N.N.; Håkansson, P.; Pourfathi, M.; Ghosh, R.K.; Kara, H.; Kadlecek, S.J.; Pileio, G.; Levitt, M.H.; Rizi, R.R.

    2013-01-01

    Dynamic nuclear polarization (DNP) of 15N2O, known for its long-lived singlet-state order at low magnetic field, is demonstrated in organic solvent/trityl mixtures at ~1.5 K and 5 T. Both 15N polarization and intermolecular dipolar broadening are strongly affected by the sample's thermal history, indicating spontaneous formation of N2O clusters. In situ 15N NMR reveals four distinct powder-pattern spectra, attributed to the chemical-shift anisotropy (CSA) tensors of the two 15N nuclei, further split by the intramolecular dipolar coupling between their magnetic moments. 15N polarization is estimated by fitting the free-induction decay (FID) signals to the analytical model of four single-quantum transitions. This analysis implies (10:2 ± 2:2)% polarization after 37 h of DNP, and provides a direct, instantaneous probe of the absolute 15N polarization, without a need for time-consuming referencing to a thermal-equilibrium NMR signal. PMID:23851025

  18. Using dual-bacterial denitrification to improve δ15N determinations of nitrates containing mass-independent 17O

    USGS Publications Warehouse

    Coplen, T.B.; Böhlke, J.K.; Casciotti, K.L.

    2004-01-01

    The bacterial denitrification method for isotopic analysis of nitrate using N2O generated from Pseudomonas aureofaciens may overestimate ??15N values by as much as 1-2??? for samples containing atmospheric nitrate because of mass-independent 17O variations in such samples. By analyzing such samples for ??15N and ??18O using the denitrifier Pseudomonas chlororaphis, one obtains nearly correct ??15N values because oxygen in N 2O generated by P. chlororaphis is primarily derived from H 2O. The difference between the apparent ??15N value determined with P. aureofaciens and that determined with P. chlororaphis, assuming mass-dependent oxygen isotopic fractionation, reflects the amount of mass-independent 17O in a nitrate sample. By interspersing nitrate isotopic reference materials having substantially different ?? 18O values with samples, one can normalize oxygen isotope ratios and determine the fractions of oxygen in N2O derived from the nitrate and from water with each denitrifier. This information can be used to improve ??15N values of nitrates having excess 17O. The same analyses also yield estimates of the magnitude of 17O excess in the nitrate (expressed as ??17O) that may be useful in some environmental studies. The 1-?? uncertainties of ??15N, ??18O and ??17O measurements are ??0.2, ??0.3 and ??5???, respectively. Copyright ?? 2004 John Wiley & Sons, Ltd.

  19. Investigating patterns of symbiotic nitrogen fixation during vegetation change from grassland to woodland using fine scale δ(15) N measurements.

    PubMed

    Soper, Fiona M; Boutton, Thomas W; Sparks, Jed P

    2015-01-01

    Biological nitrogen fixation (BNF) in woody plants is often investigated using foliar measurements of δ(15) N and is of particular interest in ecosystems experiencing increases in BNF due to woody plant encroachment. We sampled δ(15) N along the entire N uptake pathway including soil solution, xylem sap and foliage to (1) test assumptions inherent to the use of foliar δ(15) N as a proxy for BNF; (2) determine whether seasonal divergences occur between δ(15) Nxylem sap and δ(15) Nsoil inorganic N that could be used to infer variation in BNF; and (3) assess patterns of δ(15) N with tree age as indicators of shifting BNF or N cycling. Measurements of woody N-fixing Prosopis glandulosa and paired reference non-fixing Zanthoxylum fagara at three seasonal time points showed that δ(15) Nsoil inorganic N varied temporally and spatially between species. Fractionation between xylem and foliar δ(15) N was consistently opposite in direction between species and varied on average by 2.4‰. Accounting for these sources of variation caused percent nitrogen derived from fixation values for Prosopis to vary by up to ∼70%. Soil-xylem δ(15) N separation varied temporally and increased with Prosopis age, suggesting seasonal variation in N cycling and BNF and potential long-term increases in BNF not apparent through foliar sampling alone.

  20. 15N/14N variations in Cretaceous Atlantic sedimentary sequences: implication for past changes in marine nitrogen biogeochemistry

    USGS Publications Warehouse

    Rau, G.H.; Arthur, M.A.; Dean, W.E.

    1987-01-01

    At two locations in the Atlantic Ocean (DSDP Sites 367 and 530) early to middle Cretaceous organic-carbon-rich beds ("black shales") were found to have significantly lower ??15N values (lower 15N/14N ratios) than adjacent organic-carbon-poor beds (white limestones or green claystones). While these lithologies are of marine origin, the black strata in particular have ??15N values that are significantly lower than those previously found in the marine sediment record and most contemporary marine nitrogen pools. In contrast, black, organic-carbon-rich beds at a third site (DSDP Site 603) contain predominantly terrestrial organic matter and have C- and N-isotopic compositions similar to organic matter of modern terrestrial origin. The recurring 15N depletion in the marine-derived Cretaceous sequences prove that the nitrogen they contain is the end result of an episodic and atypical biogeochemistry. Existing isotopic and other data indicate that the low 15N relative abundance is the consequence of pelagic rather than post-depositional processes. Reduced ocean circulation, increased denitrification, and, hence, reduced euphotic zone nitrate availability may have led to Cretaceous phytoplankton assemblages that were periodically dominated by N2-fixing blue-green algae, a possible source of this sediment 15N-depletion. Lack of parallel isotopic shifts in Cretaceous terrestrially-derived nitrogen (Site 603) argues that the above change in nitrogen cycling during this period did not extend beyond the marine environment. ?? 1987.

  1. 1H-13C/1H-15N Heteronuclear Dipolar Recoupling by R-Symmetry Sequences Under Fast Magic Angle Spinning for Dynamics Analysis of Biological and Organic Solids

    PubMed Central

    Hou, Guangjin; Byeon, In-Ja L.; Ahn, Jinwoo; Gronenborn, Angela M.; Polenova, Tatyana

    2011-01-01

    Fast magic angle spinning (MAS) NMR spectroscopy is becoming increasingly important in structural and dynamics studies of biological systems and inorganic materials. Superior spectral resolution due to the efficient averaging of the dipolar couplings can be attained at MAS frequencies of 40 kHz and higher with appropriate decoupling techniques, while proton detection gives rise to significant sensitivity gains, therefore making fast MAS conditions advantageous across the board compared with the conventional slow- and moderate-MAS approaches. At the same time, many of the dipolar recoupling approaches that currently constitute the basis for structural and dynamics studies of solid materials and that are designed for MAS frequencies of 20 kHz and below, fail above 30 kHz. In this report, we present an approach for 1H-13C/1H-15N heteronuclear dipolar recoupling under fast MAS conditions using R-type symmetry sequences, which is suitable even for fully protonated systems. A series of rotor-synchronized R-type symmetry pulse schemes are explored for the determination of structure and dynamics in biological and organic systems. The investigations of the performance of the various RNnv-symmetry sequences at the MAS frequency of 40 kHz experimentally and by numerical simulations on [U-13C,15N]-alanine and [U-13C,15N]-N-acetyl-valine, revealed excellent performance for sequences with high symmetry number ratio (N/2n > 2.5). Further applications of this approach are presented for two proteins, sparsely 13C/uniformly 15N enriched CAP-Gly domain of dynactin and U-13C,15N-Tyr enriched C-terminal domain of HIV-1 CA protein. 2D and 3D R1632-based DIPSHIFT experiments carried out at the MAS frequency of 40 kHz, yielded site-specific 1H-13C/1H-15N heteronuclear dipolar coupling constants for CAP-Gly and CTD CA, reporting on the dynamic behavior of these proteins on time scales of nano- to microseconds. The R-symmetry based dipolar recoupling under fast MAS is expected to find

  2. 49 CFR 176.415 - Permit requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... nitrates, and certain ammonium nitrate fertilizers. 176.415 Section 176.415 Transportation Other... requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers. (a) Except as... Captain of the Port (COTP). (1) Ammonium nitrate UN1942, ammonium nitrate fertilizers containing more...

  3. 49 CFR 176.415 - Permit requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... nitrates, and certain ammonium nitrate fertilizers. 176.415 Section 176.415 Transportation Other... requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers. (a) Except as... Captain of the Port (COTP). (1) Ammonium nitrate UN1942, ammonium nitrate fertilizers containing more...

  4. 49 CFR 176.415 - Permit requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... nitrates, and certain ammonium nitrate fertilizers. 176.415 Section 176.415 Transportation Other... requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers. (a) Except as... Captain of the Port (COTP). (1) Ammonium nitrate UN1942, ammonium nitrate fertilizers containing more...

  5. 49 CFR 176.415 - Permit requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... nitrates, and certain ammonium nitrate fertilizers. 176.415 Section 176.415 Transportation Other... requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers. (a) Except as... Captain of the Port (COTP). (1) Ammonium nitrate UN1942, ammonium nitrate fertilizers containing more...

  6. 49 CFR 176.415 - Permit requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... nitrates, and certain ammonium nitrate fertilizers. 176.415 Section 176.415 Transportation Other... requirements for Division 1.5, ammonium nitrates, and certain ammonium nitrate fertilizers. (a) Except as... Captain of the Port (COTP). (1) Ammonium nitrate UN1942, ammonium nitrate fertilizers containing more...

  7. The nucleation kinetics of ammonium metavanadate precipitated by ammonium chloride

    NASA Astrophysics Data System (ADS)

    Du, Guangchao; Sun, Zhaohui; Xian, Yong; Jing, Han; Chen, Haijun; Yin, Danfeng

    2016-05-01

    The nucleation kinetics of ammonium metavanadate (NH4VO3) was investigated under conditions of the simulated process for precipitation of NH4VO3 from the vanadium-containing solution. Induction periods for the nucleation of NH4VO3 were experimentally determined as a function of supersaturation at temperatures from 30 to 45 °C. Using the classical nucleation theory, the interfacial tension between NH4VO3 and supersaturated solution, the nucleation rate and critical radius of nucleus for the homogeneous nucleation of NH4VO3 were estimated. With temperature increasing, the calculated interfacial tension gradually decreased from 29.78 mJ/m2 at 30 °C to 23.66 mJ/m2 at 45 °C. The nucleation rate was found to proportionally increase but the critical radius of nucleus exponentially decreased, with increase in supersaturation ratio at a constant temperature. The activation energy for NH4VO3 nucleation was obtained from the relationship between temperature and induction period, ranging from 79.17 kJ/mol at S=25 to 115.50 kJ/mol at S=15. FT-IR and Raman spectrum indicated that the crystals obtained in the precipitation process were NH4VO3.

  8. Characterization (δ13C, δ15N and TOC/TN) of marine sediments from restored seagrass (Zostera marina) meadows in coastal lagoons of Virginia

    NASA Astrophysics Data System (ADS)

    Egge, N.; Macko, S. A.

    2011-12-01

    Restoration of the Chesapeake Bay and its neighboring coastal areas has been a major focus of scientific research in the last ten years. The decline of the area is often linked with the explosive population growth and subsequent increases in agriculture and urbanization of the region. In the coastal bays of Virginia, large efforts have been made to restore seagrass (Zostera marina) meadows, which were devastated by a combination of events in the 1930s. As a result of the restoration efforts begun in 2001, seagrass has come to dominate portions of South Bay and Hog Island Bay. Sedimentary characterization (δ13C, δ15N, total organic carbon and total nitrogen) is used to assess sources of organic matter in marine environments. Push cores were used to collect samples from sites that have different records of seagrass cover. Sediments dominated by seagrass contain organic matter enriched in 13C relative to autochthonous algal sources. Within the sediment record there are two regions that show heightened 13C content-one near the surface and another at lower depths. Organic matter enriched in 13C near the surface is from current seagrass cover. Similar characteristics of sediment deeper in the core imply that the area was previously dominated comparable vegetation. Results may be used as a proxy to infer the spatial extent of the historic (prior to 1930) presence of seagrass and guide future restoration projects.

  9. MILLIMETER-WAVE OBSERVATIONS OF CN AND HNC AND THEIR {sup 15}N ISOTOPOLOGUES: A NEW EVALUATION OF THE {sup 14}N/{sup 15}N RATIO ACROSS THE GALAXY

    SciTech Connect

    Adande, G. R.; Ziurys, L. M.

    2012-01-10

    The N = 1 {yields} 0 transitions of CN and C{sup 15}N (X{sup 2}{Sigma}{sup +}), as well as the J = 1 {yields} 0 lines of HN{sup 13}C and H{sup 15}NC, have been observed toward 11 molecular clouds using the new 3 mm ALMA-type receiver of the 12 m telescope of the Arizona Radio Observatory. These sources span a wide range of distances from the Galactic center and are all regions of star formation. From these observations, {sup 14}N/{sup 15}N ratios have been determined using two independent methods. First, the measurements of C{sup 14}N and C{sup 15}N were directly compared to establish this ratio, correcting for high opacities when needed, as indicated by the nitrogen hyperfine intensities. Second, the ratio was calculated from the quantity [HN{sup 13}C]/[H{sup 15}NC], determined from the HNC data, and then scaled by {sup 12}C/{sup 13}C ratios previously established, i.e., the so-called double isotope method. Values from both methods are in reasonable agreement, and fall in the range {approx}120-400, somewhat lower than previous {sup 14}N/{sup 15}N ratios derived from HCN. The ratios exhibit a distinct positive gradient with distance from the Galactic center, following the relationship{sup 14}N/{sup 15}N = 21.1 (5.2) kpc{sup -1} D{sub GC} + 123.8 (37.1). This gradient is consistent with predictions of Galactic chemical evolution models in which {sup 15}N has a secondary origin in novae, while primary and secondary sources exist for {sup 14}N. The local interstellar medium value was found to be {sup 4}N/{sup 15}N = 290 {+-} 40, in agreement with the ratio found in nearby diffuse clouds and close to the value of 272 found in Earth's atmosphere.

  10. Characterization of the N2O isotopic composition (15N, 18O and N2O isotopomers) emitted from incubated Amazon forest soils. Implications for the global N2O isotope budget

    NASA Astrophysics Data System (ADS)

    Pérez, T.; García, D.; Trumbore, S.; Tyler, S.; de Camargo, P.; Moreira, M.; Piccolo, M.; Park, S.; Boering, K.; Cerri, C.

    2003-04-01

    Tropical rain forest soils are the largest natural source of N2O to the atmosphere. Uncertainty in the signature of this source limits the utility of isotopes in constraining the global N2O budget. Differentiating the relative contribution of nitrification and denitrification to the emitted N2O using stable isotopes has been difficult due to the lack of enrichment factors values for each process measured in situ. We have devised a method for measuring enrichment factors using soil incubation experiments. We selected three Amazon rain forest soils: (1) Clay and (2) Sandy from Santarem, Pará State, and (3) Sandy from Nova Vida Farm, Rondonia State, Brazil. The enrichment factor values for nitrification and denitrification are: -97.8±4.2 and -9.9±3.8 per mil for clay Santarem soil, -86.8±4.3 and -45.2±4.5 per mil for sandy Santarem soil and-112.6±3.8 and -10.4±3.5 per mil for Nova Vida Farm soils, respectively. Our results show that enrichment factors for both processes differ with soil texture and location. The enrichment factors for nitrification are significantly smaller than the range reported in the literature (-66 to -42 per mil). Also, the enrichment factors for the Santarem soils (clay and sandy) differ significantly implying that soil texture (which will affect the soil air filled pore space at a given water content) is influencing the bacteria isotopic discrimination. However, the enrichment factors for the Santarem clay sand Nova Vida sandy soils do not differ by much. This suggests that the enrichment factors not only can be affected by texture but also by the microbial fauna present in these soils. We also determined the measurement of the N2O positional dependence. N2O is a linear molecule with two nitrogen atoms. The 15N isotope can be located in either the central nitrogen (alpha position) or in the terminal nitrogen (beta position). The isotopomer site preference (15N alpha - 15N beta) can be used to differentiate processes of production and

  11. A stable isotope ( δ13C, δ15N) model for the North Water food web: implications for evaluating trophodynamics and the flow of energy and contaminants

    NASA Astrophysics Data System (ADS)

    Hobson, Keith A.; Fisk, Aaron; Karnovsky, Nina; Holst, Meike; Gagnon, Jean-Marc; Fortier, Martin

    The North Water Polynya is an area of high biological activity that supports large numbers of higher trophic-level organisms such as seabirds and marine mammals. An overall objective of the Upper Trophic-Level Group of the International North Water Polynya Study (NOW) was to evaluate carbon and contaminant flux through these high trophic-level (TL) consumers. Crucial to an evaluation of the role of such consumers, however, was the establishment of primary trophic linkages within the North Water food web. We used δ15N values of food web components from particulate organic matter (POM) through polar bears ( Ursus maritimus) to create a trophic-level model based on the assumptions that Calanus hyperboreus occupies TL 2.0 and there is a 2.4‰ trophic enrichment in 15N between birds and their diets, and a 3.8‰ trophic enrichment for all other components. This model placed the planktivorous dovekie ( Alle alle) at TL 3.3, ringed seal ( Phoca hispida) at TL 4.5, and polar bear at TL 5.5. The copepods C. hyperboreus, Chiridius glacialis and Euchaeta glacialis formed a trophic continuum (TL 2.0-3.0) from primary herbivore through omnivore to primary carnivore. Invertebrates were generally sorted according to planktonic, benthic and epibenthic feeding groups. Seabirds formed three trophic groups, with dovekie occupying the lowest, black-legged kittiwake ( Rissa tridactyla), northern fulmar ( Fulmarus glacialis), thick-billed murre ( Uria aalge), and ivory gull ( Pagophilia eburnea) intermediate (TL 3.9-4.0), and glaucous gull ( Larus hyperboreus) the highest (TL 4.6) trophic positions. Among marine mammals, walrus ( Odobenus rosmarus) occupied the lowest (TL 3.2) and bearded seal ( Erignathus barbatus), ringed seal, beluga whale ( Delphinapterus leucas), and narwhal ( Monodon monoceros) intermediate positions (TL 4.1-4.6). In addition to arctic cod ( Boreogadus saida), we suggest that lower trophic-level prey, in particular the amphipod Themisto libellula, contribute

  12. 40 CFR 721.4095 - Quaternary ammonium alkyltherpropyl trialkylamine halides.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Quaternary ammonium alkyltherpropyl... Specific Chemical Substances § 721.4095 Quaternary ammonium alkyltherpropyl trialkylamine halides. (a... generically as quaternary ammonium alkyltherpropyl trialkylamine halides (PMNs...