Distinguishing Nitrous Oxide Production from Nitrification and Denitrification on the Basis of Isotopomer Abundances

Sutka, R. L.; Ostrom, Nathaniel E.; Ostrom, Peggy H.; Breznak, John A.; Gandhi, Hasand; Pitt, Adam; Li, F.

doi:10.1128/aem.72.1.638-644.2006

Cited by 460 publications

(741 citation statements)

References 39 publications

(44 reference statements)

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“…The two enzymes are structurally very different and show significantly different isotope fractionations on bulk (Martin and Casciotti, 2016). Sutka and Ostrom (2006) conclude that a difference in the nitrite reductase does not have an effect on the SP during denitrification. This conclusion is based on measurements of two denitrifiers (P. chlororaphis, ATCC 43928; and P. aureofaciens, ATCC 13985) possessing cd1-type nitrite reductase and Cu-containing nitrite reductase, respectively.…”

Section: Discussionmentioning

confidence: 81%

“…Applying different incubation techniques on soils with different properties showed SP during production of N 2 O between −3 and 9 ‰ and between −0.9 and −8.2 ‰ during reduction (Well and Flessa, 2009a, b;Köster et al, 2013a;Lewicka-Szczebak et al, 2014. During production of N 2 O in bacterial culture experiments (involving P. chlororaphis, ATCC 43928; P. aureofaciens, ATCC 13985; and P. denitrificans, ATCC 17741), Thompson et al (2004), Toyoda et al (2005), Sutka and Ostrom (2006), and Ostrom and Ostrom (2011) found SP values of −27.4 and 1.3 ‰ and bulk values of between −42.6 and 36.7 ‰. The wide range of bulk presented both in this study and by others confirms the difficulties in the use of bulk in N 2 O evolution analysis.…”

Section: Comparison To Previous Studiesmentioning

confidence: 99%

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Continuous measurements of nitrous oxide isotopomers during incubation experiments

et al. 2018

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Abstract. Nitrous oxide (N 2 O) is an important and strong greenhouse gas in the atmosphere. It is produced by microbes during nitrification and denitrification in terrestrial and aquatic ecosystems. The main sinks for N 2 O are turnover by denitrification and photolysis and photo-oxidation in the stratosphere. In the linear N=N=O molecule 15 N substitution is possible in two distinct positions: central and terminal. The respective molecules, 14 N 15 N 16 O and 15 N 14 N 16 O, are called isotopomers. It has been demonstrated that N 2 O produced by nitrifying or denitrifying microbes exhibits a different relative abundance of the isotopomers. Therefore, measurements of the site preference (difference in the abundance of the two isotopomers) in N 2 O can be used to determine the source of N 2 O, i.e., nitrification or denitrification. Recent instrument development allows for continuous positiondependent δ 15 N measurements at N 2 O concentrations relevant for studies of atmospheric chemistry. We present results from continuous incubation experiments with denitrifying bacteria, Pseudomonas fluorescens (producing and reducing N 2 O) and Pseudomonas chlororaphis (only producing N 2 O). The continuous measurements of N 2 O isotopomers reveals the transient isotope exchange among KNO 3 , N 2 O, and N 2 . We find bulk isotopic fractionation of −5.01 ‰ ± 1.20 for P. chlororaphis, in line with previous results for production from denitrification. For P. fluorescens, the bulk isotopic fractionation during production of N 2 O is −52.21 ‰ ± 9.28 and 8.77 ‰ ± 4.49 during N 2 O reduction.The site preference (SP) isotopic fractionation for P. chlororaphis is −3.42 ‰ ± 1.69. For P. fluorescens, the calculations result in SP isotopic fractionation values of 5.73 ‰ ± 5.26 during production of N 2 O and 2.41 ‰ ± 3.04 during reduction of N 2 O. In summary, we implemented continuous measurements of N 2 O isotopomers during incubation of denitrifying bacteria and believe that similar experiments will lead to a better understanding of denitrifying bacteria and N 2 O turnover in soils and sediments and ultimately hands-on knowledge on the biotic mechanisms behind greenhouse gas exchange of the globe.

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Section: Discussionmentioning

confidence: 81%

Section: Comparison To Previous Studiesmentioning

confidence: 99%

Continuous measurements of nitrous oxide isotopomers during incubation experiments

et al. 2018

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“…Characteristic SP values of 33‰ and 0‰ for NH 2 OH oxidation and NO 2 -reduction (nitrifier denitrification and heterotrophic bacterial denitrification), respectively, which were estimated in specific pure cultures, were used for estimation of the contribution to each process (Sutka et al, 2006). Approximate contributions of NH 2 OH oxidation and NO 2 -reduction to N 2 O production were estimated by assuming that each process is linearly proportional to the SP value using the following equation:…”

Section: Isotopomer Analysismentioning

confidence: 99%

“…Analyses of the intermolecular distributions of 15 N in N 2 O (isotopomers) are regarded as useful parameters to infer the predominant N 2 O production pathway (Wunderlin et al, 2013;Sutka et al, 2006;Toyoda et al, 2005;. Isotopomer ratios (site-specific N isotope ratios in asymmetric molecules of NNO) give us qualitative information on N 2 O production and consumption pathways.…”

Section: Introductionmentioning

confidence: 99%

Source identification of nitrous oxide on autotrophic partial nitrification in a granular sludge reactor

et al. 2013

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Emission of nitrous oxide (N 2 O) during biological wastewater treatment is of growing concern since N 2 O is a major stratospheric ozone-depleting substance and an important greenhouse gas. The emission of N 2 O from a lab-scale granular sequencing batch reactor (SBR) for partial nitrification (PN) treating synthetic wastewater without organic carbon was therefore determined in this study, because PN process is known to produce more N 2 O than conventional nitrification processes. The average N 2 O emission rate from the SBR was 0.32 ± 0.17 mg-N L -1 h -1 , corresponding to the average emission of N 2 O of 0.8 ± 0.4% of the incoming nitrogen load (1.5 ± 0.8% of the converted NH 4 + ). Analysis of dynamic concentration profiles during one cycle of the SBR operation demonstrated that N 2 O concentration in off-gas was the highest just after starting aeration whereas N 2 O concentration in effluent was gradually increased in the initial 40 min of the aeration period and was decreased thereafter. Isotopomer analysis was conducted to identify the main N 2 O production pathway in the reactor during one cycle. The hydroxylamine (NH 2 OH) oxidation pathway accounted for 65% of the total N 2 O production in the initial phase during one cycle, whereas contribution of the NO 2 -reduction pathway to N 2 O production was comparable with that of the NH 2 OH oxidation pathway in the latter phase. In addition, 3 spatial distributions of bacteria and their activities in single microbial granules taken from the reactor were determined with microsensors and by in situ hybridization. Partial nitrification occurred mainly in the oxic surface layer of the granules and ammonia-oxidizing bacteria were abundant in this layer. N 2 O production was also found mainly in the oxic surface layer. Based on these results, although N 2 O was produced mainly via NH 2 OH oxidation pathway in the autotrophic partial nitrification reactor, N 2 O production mechanisms were complex and could involve multiple N 2 O production pathways.

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“…Site preference has emerged as a potential conservative tracer for microbial N 2 O production because 1) it is independent of the isotopologue composition of the substrates and 2) it does not exhibit changes during the course of production. Distinct site preference values for N 2 O production due to bacterial denitrification, including nitrifier denitrification, relative to hydroxylamine oxidation and fungal denitrification provide a fundamental basis that facilitates the resolution of production pathways in the natural environment (Sutka et al, 2006.…”

Section: Introductionmentioning

confidence: 99%

Isotopic signatures of N2O produced by ammonia-oxidizing archaea from soils

Well²,

et al. 2013

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N 2 O gas is involved in global warming and ozone depletion. The major sources of N 2 O are soil microbial processes. Anthropogenic inputs into the nitrogen cycle have exacerbated these microbial processes, including nitrification. Ammonia-oxidizing archaea (AOA) are major members of the pool of soil ammonia-oxidizing microorganisms. This study investigated the isotopic signatures of N 2 O produced by soil AOA and associated N 2 O production processes. All five AOA strains (I.1a, I.1a-associated and I.1b clades of Thaumarchaeota) from soil produced N 2 O and their yields were comparable to those of ammonia-oxidizing bacteria (AOB). The levels of site preference (SP), d 15 N bulk and d 18 O -N 2 O of soil AOA strains were 13-30%, À 13 to À 35% and 22-36%, respectively, and strains MY1-3 and other soil AOA strains had distinct isotopic signatures. A 15 N-NH 4 þ -labeling experiment indicated that N 2 O originated from two different production pathways (that is, ammonia oxidation and nitrifier denitrification), which suggests that the isotopic signatures of N 2 O from AOA may be attributable to the relative contributions of these two processes. The highest N 2 O production yield and lowest site preference of acidophilic strain CS may be related to enhanced nitrifier denitrification for detoxifying nitrite. Previously, it was not possible to detect N 2 O from soil AOA because of similarities between its isotopic signatures and those from AOB. Given the predominance of AOA over AOB in most soils, a significant proportion of the total N 2 O emissions from soil nitrification may be attributable to AOA.

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Distinguishing Nitrous Oxide Production from Nitrification and Denitrification on the Basis of Isotopomer Abundances

Cited by 460 publications

References 39 publications

Continuous measurements of nitrous oxide isotopomers during incubation experiments

Continuous measurements of nitrous oxide isotopomers during incubation experiments

Source identification of nitrous oxide on autotrophic partial nitrification in a granular sludge reactor

Isotopic signatures of N2O produced by ammonia-oxidizing archaea from soils

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