Sulfur-driven autotrophic denitrification of nitric oxide for efficient nitrous oxide recovery

Nitrous oxide (N₂O) was previously deemed as a potent greenhouse gas but is actually an untapped energy source, which can accumulate during the microbial denitrification of nitric oxide (NO). Compared with the organic electron donor required in heterotrophic denitrification, elemental sulfur (S⁰) is a promising electron donor alternative due to its cheap cost and low biomass yield in sulfur-driven autotrophic denitrification. However, no effort has been made to test N₂O recovery from sulfur-driven denitrification of NO so far. Therefore, in this study, batch and continuous experiments were carried out to investigate the NO removal performance and N₂O recovery potential via sulfur-driven NO-based denitrification under various Fe(II)EDTA-NO concentrations. Efficient energy recovery was achieved, as up to 35.5%–40.9% of NO was converted to N₂O under various NO concentrations. N₂O recovery from Fe(II)EDTA-NO could be enhanced by the low bioavailability of sulfur and the acid environment caused by sulfur oxidation. The NO reductase (NOR) and N₂O reductase (N₂OR) were inhibited distinctively at relatively low NO levels, leading to efficient N₂O accumulation, but were suppressed irreversibly at NO level beyond 15 mM in continuous experiments. Such results indicated that the regulation of NO at a relatively low level would benefit the system stability and NO removal capacity during long-term system operation. The continuous operation of the sulfur-driven Fe(II)EDTA-NO-based denitrification reduced the overall microbial diversity but enriched several key microbial community. Thauera, Thermomonas, and Arenimonas that are able to carry out sulfur-driven autotrophic denitrification became the dominant organisms with their relative abundance increased from 25.8% to 68.3%, collectively.     

The impact of organic matter on nitric oxide formation by Nitrosomonas europaea

Chemolithoautotrophically growing cells of Nitrosomonas europaea quantitatively oxidized ammonia to nitrite under aerobic conditions with no loss of inorganic nitrogen. Significant inorganic nitrogen losses occurred when cells were growing mixotrophically with ammonium, pyruvate, yeast extract and peptone. Under oxygen limitation the nitrogen losses were even higher. In the absence of oxygen pyruvate was metabolized slowly while nitrite was consumed concomitantly. Nitrogen losses were due to the production of nitric oxide and nitrous oxide. In mixed cultures of Nitrosomonas and Nitrobacter, strong inhibition of nitrite oxidation was reproducibly measured. NO and ammonium were not inhibitory to Nitrobacter. First evidence is given that hydroxylamine, the intermediate of the Nitrosomonas monooxygenase-reaction, is formed. 0.2 to 1.7 M NH₂OH were produced by mixotrophically growing cells of Nitrosomonas and Nitrosovibrio. Hydroxylamine was both a selective inhibitory agent to Nitrobacter cells and a strong reductant which reduced nitrite to NO and N₂O. It is discussed whether chemodenitrification or denitrification is the most abundant process for NO and N₂O production of Nitrosomonas.     

Nitrous oxide and dinitrogen emissions from urine-affected soil under controlled conditions

A ¹⁵N labelling technique was used to measure N₂O and N₂ emissions from an undisturbed grassland soil treated with cow urine and held at 30 cm water tension and 20°C in a laboratory. Large emissions of dinitrogen were detected immediately following urine application to pasture. These coincided with a rapid and large increase in soil water-soluble carbon levels, some of this increase being attributed to solubilization of soil organic matter by high pH and ammonia concentrations. Emissions of nitrous oxide generally increased with time in contrast to dinitrogen fluxes which decreased as time progressed. Estimated losses of N₂O and N₂ over a 30 day period were between 1 to 5% and 30 to 65% of the urine N applied plus N mineralized from soil organic matter, respectively. Most of the N₂ and N₂O originated from denitrification with nitrification-denitrification being of minor significance as a source of N₂O. Comparisons of the ¹⁵N enrichments in the soil mineral N pools and the evolved N₂O suggested that much of the N₂O was produced in the 5–8 cm zone of the soil. It is concluded that established grassland soils contain large amounts of readily-oxidizable organic carbon which may be used by soil denitrifying organisms when nitrate is non-limiting and soil redox potential is lowered due to high rates of biological activity and high soil moisture contents. ei]{gnR}{fnMerckx}     

Impact of COD/N ratio on nitrous oxide emission from microcosm wetlands and their performance in removing nitrogen from wastewater

Constructed wetlands (CWs) are considered to be important sources of nitrous oxide (N₂O). In order to investigate the effect of influent COD/N ratio on N₂O emission and control excess emission from nitrogen removal, free water surface microcosm wetlands were used and fed with different influent. In addition, the transformation of nitrogen was examined for better understanding of the mechanism of N₂O production under different operating COD/N ratios. It was found that N₂O emission and the performance of microcosm wetlands were significantly affected by COD/N ratio of wastewater influent. Strong relationships exist between N₂O production rate and nitrite (r = 0.421, p < 0.01). During denitrification process, DO concentration crucially influences N₂O production rate. An optimal influent COD/N ratio was obtained by adjusting external carbon sources for most effective N₂O emission control and best performance of the CWs in nitrogen removal from wastewater. It is concluded that under the operating condition of COD/N ratio = 5, total N₂O emission is minimum and the microcosm wetland is most effective in wastewater nitrogen removal.     

Effect of carbon source and competition for electrons on nitrous oxide reduction in a mixed denitrifying microbial community

The competition for electrons has been recently demonstrated to affect the reduction rates of the nitrogen oxides in a methanol enriched denitrifying community. The aim of this study was to test if electron competition also occurred when other substrates were used for denitrification and if that could have an effect on the potential nitrous oxide (N₂O) production and subsequent consumption. A denitrifying culture was developed in a sequencing batch reactor using nitrate as electron acceptor and a combination of acetate, ethanol and methanol as carbon sources. Four sets of batch tests were conducted using acetate, ethanol, methanol and a combination of the three carbon sources respectively. For each set the effect of nitrate, nitrite and nitrous oxide on each other reduction rates when present individually or in combination was assessed. Results show that reduction rates are affected by the type of substrate added, probably due to different microbial populations specialized with consuming a particular substrate. Also, N₂O reduction rate is the most reduced under the different electron competition scenarios tested, which results in N₂O accumulation in some cases. The effect of substrate limitation on N₂O reduction was also assessed.     

Production of nitric oxide in Nitrosomonas europaea by reduction of nitrite

Nitrosomonas europaea and Nitrosovibrio sp. produced NO and N₂O during nitrification of ammonium. Less then 15% of the produced NO was due to chemical decomposition of nitrite. Production of NO and especially of N₂O increased when the bacteria were incubated under anaerobic conditions at decreasing flow rates of air, or at increasing cell densities. Low concentrations of chlorite (10 M) inhibited the production of NO and N₂, but not of nitrite indicating that NO and N₂O were not produced during the oxidative conversion of ammonium to nitrite. NO and N₂O were produced during reduction of nitrite with hydrazine as electron donor in almost stoichiometric quantities indicating that reduction of nitrite was the main source of NO and N₂O.     

Modeling nitrogen removal for a denitrification biofilter

Nitrous oxide (N₂O) is a major greenhouse gas, heavily contributing to global warming. N₂O is emitted from various sources such as wastewater treatment plants, during the nitrification and denitrification steps. ASM models, which are commonly used in wastewater treatment, usually consider denitrification as a one-step process (NO₃ ⁻ directly reduced to N₂) and are as such unable to provide values for intermediate products of the reaction like N₂O. In this study, a slightly modified ASM1 model was implemented in the GPS-X™ software to simulate the concentration of such intermediate products (NO₂ ⁻, NO and N₂O) and to estimate the amounts of gaseous N₂O emitted by the denitrification stage (12 biofilters) of the Seine-Centre WWTP (SIAAP, Paris). Simulations running on a 1-year period have shown good agreements with measured effluent data for nitrate and nitrite. The calculated mean value for emitted N₂O is 4.95 kgN–N₂O/day, which stands in the typical range of estimated experimental values of 4–31 kgN–N₂O/day. Nitrous oxide emissions are usually not measured on WWTPs and so, as obtained results show, there is a certain potential for using models that quantify those emissions using traditionally measured influent data.     

Role of maize stover incorporation on nitrogen oxide emissions in a non-irrigated Mediterranean barley field

Aims

Agricultural soils in semiarid Mediterranean areas are characterized by low organic matter contents and low fertility levels. Application of crop residues and/or manures as amendments is a cost-effective and sustainable alternative to overcome this problem. However, these management practices may induce important changes in the nitrogen oxide emissions from these agroecosystems, with additional impacts on carbon dioxide emissions. In this context, a field experiment was carried out with a barley (Hordeum vulgare L.) crop under Mediterranean conditions to evaluate the effect of combining maize (Zea mays L.) residues and N fertilizer inputs (organic and/or mineral) on these emissions.

Methods

Crop yield and N uptake, soil mineral N concentrations, dissolved organic carbon (DOC), denitrification capacity, N₂O, NO and CO₂ fluxes were measured during the growing season.

Results

The incorporation of maize stover increased N₂O emissions during the experimental period by c. 105 %. Conversely, NO emissions were significantly reduced in the plots amended with crop residues. The partial substitution of urea by pig slurry reduced net N₂O emissions by 46 and 39 %, with and without the incorporation of crop residues respectively. Net emissions of NO were reduced 38 and 17 % for the same treatments. Molar DOC:NO ₃ ^? ratio was found to be a robust predictor of N₂O and NO fluxes.

Conclusions

The main effect of the interaction between crop residue and N fertilizer application occurred in the medium term (4–6 month after application), enhancing N₂O emissions and decreasing NO emissions as consequence of residue incorporation. The substitution of urea by pig slurry can be considered a good management strategy since N₂O and NO emissions were reduced by the use of the organic residue.     

Relative Rates of Nitric Oxide and Nitrous Oxide Production by Nitrifiers, Denitrifiers, and Nitrate Respirers

Biogenic emissions of nitric and nitrous oxides have important impacts on the photochemistry and chemistry of the atmosphere. Although biogenic production appears to be the overwhelming source of N₂O, the magnitude of the biogenic emission of NO is very uncertain. In soils, possible sources of NO and N₂O include nitrification by autotrophic and heterotrophic nitrifiers, denitrification by nitrifiers and denitrifiers, nitrate respiration by fermenters, and chemodenitrification. The availability of oxygen determines to a large extent the relative activities of these various groups of organisms. To better understand this influence, we investigated the effect of the partial pressure of oxygen (pO₂) on the production of NO and N₂O by a wide variety of common soil nitrifying, denitrifying, and nitrate-respiring bacteria under laboratory conditions. The production of NO per cell was highest by autotrophic nitrifiers and was independent of pO₂ in the range tested (0.5 to 10%), whereas N₂O production was inversely proportional to pO₂. Nitrous oxide production was highest in the denitrifier Pseudomonas fluorescens, but only under anaerobic conditions. The molar ratio of NO/N₂O produced was usually greater than unity for nitrifiers and much less than unity for denitrifiers. Chemodenitrification was the major source of both the NO and N₂O produced by the nitrate respirer Serratia marcescens. Chemodenitrification was also a possible source of NO and N₂O in nitrifier cultures but only when high concentrations of nitrite had accumulated or were added to the medium. Although most of the denitrifiers produced NO and N₂O only under anaerobic conditions, chemostat cultures of Alcaligenes faecalis continued to emit these gases even when the cultures were sparged with air. Based upon these results, we predict that aerobic soils are primary sources of NO and that N₂O is produced only when there is sufficient soil moisture to provide the anaerobic microsites necessary for denitrification by either denitrifiers or nitrifiers.     

Mechanism of nitrite reduction to nitrous oxide in a photodenitrifier,Rhodopseudomonas sphaeroide forma sp. denitrificans

The mechanism of anaerobic reduction of NO₂^? to N₂O in a photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans, was investigated. With ascorbate-reduced phenazine methosulfate (PMS) as the electron donor, the nitrite reductase of this photodenitrifier reduced NO₂^? to NO and a trace amount of N₂O. With dithionite-reduced benzyl viologen as the electron donor, the major product of NO₂^? reduction was NH₂OH, and a trace amount of N₂O was also produced. The nitrate reductase itself had no NO reductase activity with ascorbate-reduced PMS. It was concluded that the essential product of NO₂^? reduction by the purified nitrite reductase is NO. Chromatophore membranes stoichiometrically produced N₂O from NO₂^? with any electron donor, such as dithionite-redduced benzyl viologen, ascorbate-reduced PMS or NADH/FMN. The membranes also contrained activity of NO reduction of N₂O with either ascorbate-reduced PMS or duroquinol. The NO reductase activity with duroquinol was inhibited by antimycin A. Stoichiometric production of N₂O from N₂^? also was observed in the reconstituted NO₂^? reduction system which contained the cytochrome bc₁ complex, cytochrome c₂, the nitrite reductase and duroquinol as the electron donor. The preparation of the cytochrome bc₁ complex itself contianed NO reductase activity. From these results the mechanism of NO₂^? reduction to N₂O in this photodenitrifier was determined as the nitrite reductase reducing NO₂^? to NO with electrons from the cytochrome bc₁ complex, and NO subsequently being reduced, without release, to N₂O with electrons from the cytochrome bc₁ complex by the NO reductase, which is closely associated with the complex.     

Denitrification by Actinomycetes and Purification of Dissimilatory Nitrite Reductase and Azurin from Streptomyces thioluteus

Many actinomycete strains are able to convert nitrate or nitrite to nitrous oxide (N₂O). As a representative of actinomycete denitrification systems, the system of Streptomyces thioluteus was investigated in detail. S. thioluteus attained distinct cell growth upon anaerobic incubation with nitrate or nitrite with concomitant and stoichiometric conversion of nitrate or nitrite to N₂O, suggesting that the denitrification acts as anaerobic respiration. Furthermore, a copper-containing, dissimilatory nitrite reductase (CuNir) and its physiological electron donor, azurin, were isolated. This is the first report to show that denitrification generally occurs among actinomycetes.     

Soil environmental conditions rather than denitrifier abundance and diversity drive potential denitrification after changes in land uses

Land‐use practices aiming at increasing agro‐ecosystem sustainability, e.g. no‐till systems and use of temporary grasslands, have been developed in cropping areas, but their environmental benefits could be counterbalanced by increased N₂O emissions produced, in particular during denitrification. Modelling denitrification in this context is thus of major importance. However, to what extent can changes in denitrification be predicted by representing the denitrifying community as a black box, i.e. without an adequate representation of the biological characteristics (abundance and composition) of this community, remains unclear. We analysed the effect of changes in land uses on denitrifiers for two different agricultural systems: (i) crop/grassland conversion and (ii) cessation/application of tillage. We surveyed potential denitrification (PD), the abundance and genetic structure of denitrifiers (nitrite reducers), and soil environmental conditions. N₂O emissions were also measured during periods of several days on control plots. Time‐integrated N₂O emissions and PD were well correlated among all control plots. Changes in PD were partly due to changes in denitrifier abundance but were not related to changes in the structure of the denitrifier community. Using multiple regression analysis, we showed that changes in PD were more related to changes in soil environmental conditions than in denitrifier abundance. Soil organic carbon explained 81% of the variance observed for PD at the crop/temporary grassland site, whereas soil organic carbon, water‐filled pore space and nitrate explained 92% of PD variance at the till/no‐till site, without any residual effect of denitrifier abundance. Soil environmental conditions influenced PD by modifying the specific activity of denitrifiers, and to a lesser extent by promoting a build‐up of denitrifiers. Our results show that an accurate simulation of carbon, oxygen and nitrate availability to denitrifiers is more important than an accurate simulation of denitrifier abundance and community structure to adequately understand and predict changes in PD in response to land‐use changes.     

Excessive use of nitrogen in Chinese agriculture results in high N2O/(N2O+N2) product ratio of denitrification,primarily due to acidification of the soils

China is the world's largest producer and consumer of fertilizer N, and decades of overuse has caused nitrate leaching and possibly soil acidification. We hypothesized that this would enhance the soils' propensity to emit N₂O from denitrification by reducing the expression of the enzyme N₂O reductase. We investigated this by standardized oxic/anoxic incubations of soils from five long‐term fertilization experiments in different regions of China. After adjusting the nitrate concentration to 2 mM, we measured oxic respiration (R), potential denitrification (D), substrate‐induced denitrification, and the denitrification product stoichiometry (NO, N₂O, N₂). Soils with a history of high fertilizer N levels had high N₂O/(N₂O+N₂) ratios, but only in those field experiments where soil pH had been lowered by N fertilization. By comparing all soils, we found a strong negative correlation between pH and the N₂O/(N₂O+N₂) product ratio (r² = 0.759, P < 0.001). In contrast, the potential denitrification (D) was found to be a linear function of oxic respiration (R), and the ratio D/R was largely unaffected by soil pH. The immediate effect of liming acidified soils was lowered N₂O/(N₂O+N₂) ratios. The results provide evidence that soil pH has a marginal direct effect on potential denitrification, but that it is the master variable controlling the percentage of denitrified N emitted as N₂O. It has been known for long that low pH may result in high N₂O/(N₂O+N₂) product ratios of denitrification, but our documentation of a pervasive pH‐control of this ratio across soil types and management practices is new. The results are in good agreement with new understanding of how pH may interfere with the expression of N₂O reductase. We argue that the management of soil pH should be high on the agenda for mitigating N₂O emissions in the future, particularly for countries where ongoing intensification of plant production is likely to acidify the soils.     

Deciphering the oxygen isotope composition of nitrous oxide produced by nitrification

The ability to use δ¹⁸O values of nitrous oxide (N₂O) to apportion environmental emissions is currently hindered by a poor understanding of the controls on δ¹⁸O–N₂O from nitrification (hydroxylamine oxidation to N₂O and nitrite reduction to N₂O). In this study fertilized agricultural soils and unfertilized temperate forest soils were aerobically incubated with different ¹⁸O/¹⁶O waters, and conceptual and mathematical models were developed to systematically explain the δ¹⁸O–N₂O formed by nitrification. Modeling exercises used a set of defined input parameters to emulate the measured soil δ¹⁸O–N₂O data (Monte Carlo approach). The Monte Carlo simulations implied that abiotic oxygen (O) exchange between nitrite (NO₂^?) and H₂O is important in all soils, but that biological, enzyme‐controlled O‐exchange does not occur during the reduction of NO₂^? to N₂O (nitrifier‐denitrification). Similarly, the results of the model simulations indicated that N₂O consumption is not characteristic of aerobic N₂O formation. The results of this study and a synthesis of the published literature data indicate that δ¹⁸O–N₂O formed in aerobic environments is constrained between +13‰ and +35‰ relative to Vienna Standard Mean Ocean Water (VSMOW). N₂O formed via hydroxylamine oxidation and nitrifier‐denitrification cannot be separated using δ¹⁸O unless ¹⁸O tracers are employed. The natural range of nitrifier δ¹⁸O–N₂O is discussed and explained in terms of our conceptual model, and the major and minor controls that define aerobically produced δ¹⁸O–N₂O are identified. Despite the highly complex nature of δ¹⁸O–N₂O produced by nitrification this δ¹⁸O range is narrow. As a result, in many situations δ¹⁸O values may be used in conjunction with δ¹⁵N–N₂O data to apportion nitrifier‐ and denitrifier‐derived N₂O. However, when biological O‐exchange during denitrification is high and N₂O consumption is low, there may be too much overlap in δ¹⁸O values to distinguish N₂O formed by these pathways.     

A meta‐analysis of fertilizer‐induced soil NO and combined NO+N2O emissions

Soils are among the important sources of atmospheric nitric oxide (NO) and nitrous oxide (N₂O), acting as a critical role in atmospheric chemistry. Updated data derived from 114 peer‐reviewed publications with 520 field measurements were synthesized using meta‐analysis procedure to examine the N fertilizer‐induced soil NO and the combined NO+N₂O emissions across global soils. Besides factors identified in earlier reviews, additional factors responsible for NO fluxes were fertilizer type, soil C/N ratio, crop residue incorporation, tillage, atmospheric carbon dioxide concentration, drought and biomass burning. When averaged across all measurements, soil NO‐N fluxes were estimated to be 4.06 kg ha^?1 yr^?1, with the greatest (9.75 kg ha^?1 yr^?1) in vegetable croplands and the lowest (0.11 kg ha^?1 yr^?1) in rice paddies. Soil NO emissions were more enhanced by synthetic N fertilizer (+38%), relative to organic (+20%) or mixed N (+18%) sources. Compared with synthetic N fertilizer alone, synthetic N fertilizer combined with nitrification inhibitors substantially reduced soil NO emissions by 81%. The global mean direct emission factors of N fertilizer for NO (EF_NO) and combined NO+N₂O (EF_c) were estimated to be 1.16% and 2.58%, with 95% confidence intervals of 0.71–1.61% and 1.81–3.35%, respectively. Forests had the greatest EF_NO (2.39%). Within the croplands, the EF_NO (1.71%) and EF_c (4.13%) were the greatest in vegetable cropping fields. Among different chemical N fertilizer varieties, ammonium nitrate had the greatest EF_NO (2.93%) and EF_c (5.97%). Some options such as organic instead of synthetic N fertilizer, decreasing N fertilizer input rate, nitrification inhibitor and low irrigation frequency could be adopted to mitigate soil NO emissions. More field measurements over multiyears are highly needed to minimize the estimate uncertainties and mitigate soil NO emissions, particularly in forests and vegetable croplands.     

Nitrous oxide fluxes in estuarine environments: response to global change

Nitrous oxide is a powerful, long‐lived greenhouse gas, but we know little about the role of estuarine areas in the global N₂O budget. This review summarizes 56 studies of N₂O fluxes and associated biogeochemical controlling factors in estuarine open waters, salt marshes, mangroves, and intertidal sediments. The majority of in situ N₂O production occurs as a result of sediment denitrification, although the water column contributes N₂O through nitrification in suspended particles. The most important factors controlling N₂O fluxes seem to be dissolved inorganic nitrogen (DIN) and oxygen availability, which in turn are affected by tidal cycles, groundwater inputs, and macrophyte density. The heterogeneity of coastal environments leads to a high variability in observations, but on average estuarine open water, intertidal and vegetated environments are sites of a small positive N₂O flux to the atmosphere (range 0.15–0.91; median 0.31; Tg N₂O‐N yr^?1). Global changes in macrophyte distribution and anthropogenic nitrogen loading are expected to increase N₂O emissions from estuaries. We estimate that a doubling of current median NO₃^? concentrations would increase the global estuary water–air N₂O flux by about 0.45 Tg N₂O‐N yr^?1 or about 190%. A loss of 50% of mangrove habitat, being converted to unvegetated intertidal area, would result in a net decrease in N₂O emissions of 0.002 Tg N₂O‐N yr^?1. In contrast, conversion of 50% of salt marsh to unvegetated area would result in a net increase of 0.001 Tg N₂O‐N yr^?1. Decreased oxygen concentrations may inhibit production of N₂O by nitrification; however, sediment denitrification and the associated ratio of N₂O:N₂ is expected to increase.     

Dominance of bacterial ammonium oxidizers and fungal denitrifiers in the complex nitrogen cycle pathways related to nitrous oxide emission

Organic compounds and mineral nitrogen (N) usually increase nitrous oxide (N₂O) emissions. Vinasse, a by‐product of bio‐ethanol production that is rich in carbon, nitrogen, and potassium, is recycled in sugarcane fields as a bio‐fertilizer. Vinasse can contribute significantly to N₂O emissions when applied with N in sugarcane plantations, a common practice. However, the biological processes involved in N₂O emissions under this management practice are unknown. This study investigated the roles of nitrification and denitrification in N₂O emissions from straw‐covered soils amended with different vinasses (CV: concentrated and V: nonconcentrated) before or at the same time as mineral fertilizers at different time points of the sugarcane cycle in two seasons. N₂O emissions were evaluated for 90 days, the period that occurs most of the N₂O emission from fertilizers; the microbial genes encoding enzymes involved in N₂O production (archaeal and bacterial amoA, fungal and bacterial nirK, and bacterial nirS and nosZ), total bacteria, and total fungi were quantified by real‐time PCR. The application of CV and V in conjunction with mineral N resulted in higher N₂O emissions than the application of N fertilizer alone. The strategy of vinasse application 30 days before mineral N reduced N₂O emissions by 65% for CV, but not for V. Independent of rainy or dry season, the microbial processes were nitrification by ammonia‐oxidizing bacteria (AOB) and archaea and denitrification by bacteria and fungi. The contributions of each process differed and depended on soil moisture, soil pH, and N sources. We concluded that amoA‐AOB was the most important gene related to N₂O emissions, which indicates that nitrification by AOB is the main microbial‐driven process linked to N₂O emissions in tropical soil. Interestingly, fungal nirK was also significantly correlated with N₂O emissions, suggesting that denitrification by fungi contributes to N₂O emission in soils receiving straw and vinasse application.     

Residue incorporation depth is a controlling factor of earthworm‐induced nitrous oxide emissions

Earthworms can increase nitrous oxide (N₂O) emissions, particularly in no‐tillage systems where earthworms are abundant. Here, we study the effect of residue incorporation depth on earthworm‐induced N₂O emissions. We hypothesized that cumulative N₂O emissions decrease with residue incorporation depth, because (i) increased water filled pore space (WFPS) in deeper soil layers leads to higher denitrification rates as well as more complete denitrification; and (ii) the longer upward diffusion path increases N₂O reduction to N₂. Two 84‐day laboratory mesocosm experiments were conducted. First, we manually incorporated maize (Zea mays L.) residue at different soil depths (incorporation experiment). Second, ¹³C‐enriched maize residue was applied to the soil surface and anecic species Lumbricus terrestris (L.) and epigeic species Lumbricus rubellus (Hoffmeister) were confined to different soil depths (earthworm experiment). Residue incorporation depth affected cumulative N₂O emissions in both experiments (P < 0.001). In the incorporation experiment, N₂O emissions decreased from 4.91 mg N₂O–N kg^?1 soil (surface application) to 2.71 mg N₂O–N kg^?1 soil (40–50 cm incorporation). In the earthworm experiment, N₂O emissions from L. terrestris decreased from 3.87 mg N₂O–N kg^?1 soil (confined to 0–10 cm) to 2.01 mg N₂O–N kg^?1 soil (confined to 0–30 cm). Both experimental setups resulted in dissimilar WFPS profiles that affected N₂O dynamics. We also found significant differences in residue C recovery in soil organic matter between L. terrestris (28–41%) and L. rubellus (56%). We conclude that (i) N₂O emissions decrease with residue incorporation depth, although this effect was complicated by dissimilar WFPS profiles; and (ii) larger residue C incorporation by L. rubellus than L. terrestris indicates that earthworm species differ in their C stabilization potential. Our findings underline the importance of studying earthworm diversity in the context of greenhouse gas emissions from agro‐ecosystems.     

Effect of N-fixing and non N-fixing trees and crops on NO and N2O emissions from Senegalese soils

Aim Agroforestry systems incorporating N‐fixing trees have been shown to be socially beneficial and are thought to be environmentally friendly, both enriching and stabilizing soil. However, the effect of such systems on the emissions of the important greenhouse gas nitrous oxide (N₂O) and the tropospheric ozone precursor nitric oxide (NO) is largely unknown. Location Soil was collected from the research plots of Institut Sénégalais de Recherches Agricoles at Bandia and Bambey, Senegal, West Africa, and from neighbouring farmers’ fields. Trace gas flux measurements and chemical analysis of the soil were carried out at the Centre for Ecology and Hydrology (CEH), Edinburgh, UK. Methods Nitric oxide (NO) and nitrous oxide (N₂O) emissions were measured following simulated rainfall events (10 and 20 mm equivalents) from repacked soil cores collected under two tree species (Acacia raddiana) and Eucalyptus camaldulensis) in each of two provenance trails. In addition, soil samples were collected in local fields growing peanut (Arachis hypogaea) and Sorghum (Sorghum vulgare), close to the species trials in Bambey. NO was measured using a flow through system and was analysed by chemiluminescence. Nitrous oxide was measured from the repacked soil core headspace and was analysed by electron capture gas chromatography. Soil mineral N was extracted with KCl and analysed by colorimetric methods on separate soil columns. Results Light rainfall, which increased the gravimetric soil moisture content to 20%, stimulated an increase in NO emission but there was no detectable N₂O emission. A heavy rainfall event, which increased the gravimetric soil moisture to 30%, stimulated N₂O emission with a subsequent peak in NO emissions when the soils became drier. Soil collected under the N‐fixing tree species emitted significantly more N₂O than soil collected under the N‐fixing crop species (P < 0.01). NO and N₂O emissions significantly correlated with soil available N (NH₄ and NO₃) (P < 0.05). Main conclusions Rainfall intensity, supply of mineral N from organic matter and N fixation were the prime drivers of NO and N₂O emissions from seasonally dry tropical soils. The improved soil fertility underneath the trees provided a larger pool of mineral N and yielded larger rates of NO and N₂O emissions.