Dataset Information

Dimethyl pyrazol-based nitrification inhibitors effect on nitrifying and denitrifying bacteria to mitigate N2O emission.

ABSTRACT: Nitrous oxide (N2O) emissions have been increasing as a result of intensive nitrogen (N) fertilisation. Soil nitrification and denitrification are the main sources of N2O, and the use of ammonium-based fertilisers combined with nitrification inhibitors (NIs) could be useful in mitigating N2O emissions from agricultural systems. In this work we looked at the N2O mitigation capacity of two dimethylpyrazol-based NIs, 3,4-dimethylpyrazole phosphate (DMPP) and 2-(N-3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture (DMPSA), on soil nitrifying and denitrifying microbial populations under two contrasting soil water contents (40% and 80% soil water filled pore space; WFPS). Our results show that DMPP and DMPSA are equally efficient at reducing N2O emissions under 40% WFPS conditions by inhibiting bacterial ammonia oxidation. In contrast, at 80% WFPS DMPSA was less efficient than DMPP at reducing N2O emissions. Interestingly, at 80% WFPS, where lowered oxygen availability limits nitrification, both DMPP and DMPSA not only inhibited nitrification but also stimulated N2O reduction to molecular nitrogen (N2) via nitrous oxide reductase activity (Nos activity). Therefore, in this work we observed that DMP-based NIs stimulated the reduction of N2O to N2 by nitrous oxide reductase during the denitrification process.

SUBMITTER: Torralbo F

PROVIDER: S-EPMC5653738 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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Dimethyl pyrazol-based nitrification inhibitors effect on nitrifying and denitrifying bacteria to mitigate N2O emission.

Torralbo Fernando F Menéndez Sergio S Barrena Iskander I Estavillo José M JM Marino Daniel D González-Murua Carmen C

Scientific reports 20171023 1

Nitrous oxide (N2O) emissions have been increasing as a result of intensive nitrogen (N) fertilisation. Soil nitrification and denitrification are the main sources of N2O, and the use of ammonium-based fertilisers combined with nitrification inhibitors (NIs) could be useful in mitigating N2O emissions from agricultural systems. In this work we looked at the N2O mitigation capacity of two dimethylpyrazol-based NIs, 3,4-dimethylpyrazole phosphate (DMPP) ...[more]

PMID: 29062007

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Project description:Denitrification is a critical component of soil nitrogen (N) cycling, including its role in the production and loss of nitrous oxide (N2O) from the soil system. However, restoration effects on the contribution of denitrification to soil N2O emissions, the abundance and diversity of denitrifying bacteria, and relationships among N2O emissions, soil properties, and denitrifying bacterial community composition remains poorly known. This is particularly true for fragile semiarid ecosystems. In order to address this knowledge gap, we utilized 42-year chronosequence of Robinia pseudoacacia plantations in the Chinese hilly gullied Loess Plateau. Soil potential N2O emission rates were measured using anaerobic incubation experiments. Quantitative polymerase chain reaction (Q-PCR) and Illumina MiSeq high-throughput sequencing were used to reveal the abundance and community composition of denitrifying bacteria. In this study, the afforestation practices following farmland abandonment had a strong negative effect on soil potential N2O emission rates during the first 33 years. However, potential N2O emission rates steadily increased in 42 years of restoration, leading to enhanced potential risk of greenhouse gas emissions. Furthermore, active afforestation increased the abundance of denitrifying functional genes, and enhanced microbial biomass. Actinobacteria and Proteobacteria were the dominant denitrifying bacterial phyla in the 0 to 33-years old sites, while the 42-years sites were dominated by Planctomycetes and Actinobacteria, implying that the restoration performed at these sites promoted soil microbial succession. Finally, correlation analyses revealed that soil organic carbon concentrations had the strongest relationship with potential N2O emission rates, followed by the abundance of the nosZ functional gene, bulk density, and the abundance of Bradyrhizobium and Variovorax across restoration stages. Taken together, our data suggest above-ground restoration of plant communities results in microbial community succession, improved soil quality, and significantly altered N2O emissions.

Project description:The Achères wastewater treatment plant, located just downstream of Paris, discharges its effluents into the lower Seine River. The effluents contain large numbers of heterotrophic bacteria, organic matter, and ammonium and are a source of nitrifying bacteria. As a result, degradation of organic matter by heterotrophic bacteria and subsequent oxygen depletion occur immediately downstream of the effluent outlet, whereas nitrifying bacteria apparently need to build up a significant biomass before ammonium oxidation significantly depletes the oxygen. We quantified the potential total nitrifying activity and the potential activities of the ammonia- and nitrite-oxidizing communities along the Seine River. In the summer, the maximum nitrifying activity occurs in the upper freshwater estuary, approximately 200 km downstream of Achères. The quantities of nitrifying bacteria, based on amoA gene copy numbers, and of Nitrobacter organisms, based on 16S rRNA gene copy numbers, were correlated with the potential nitrifying activities. The species composition of ammonia-oxidizing bacteria was investigated at two sites: the Triel station just downstream from Achères (km 84) and the Seine freshwater estuary at the Duclair station (km 278). By means of PCR primers targeting the amoA gene, a gene library was created. Phylogenetic analysis revealed that the majority of the analyzed clones at both sites were affiliated with the genus NITROSOMONAS: The Nitrosomonas oligotropha- and Nitrosomonas urea-related clones represented nearly 81% of the community of ammonia-oxidizing bacteria at Triel and 60% at Duclair. Two other ammonia-oxidizing clusters of the beta subclass of the Proteobacteria, i.e., Nitrosomonas europaea- and Nitrosospira-like bacteria, were found in smaller numbers. The major change in the ammonia-oxidizing community between the two stations along the Seine River-upper estuary continuum was the replacement of the N. oligotropha- and N. urea-related bacteria by the Nitrosospira-affiliated bacteria. Although the diversities of the ammonia oxidizers appear to be similar for the two sites, only half of the restriction patterns are common to both sites, which could be explained by the differences in ammonium concentrations, which are much lower in the upper estuary than in the river at the effluent outlet. These results imply a significant immigration and/or selection of the ammonia-oxidizing bacterial population along the continuum of the Seine River from Paris to the estuary.

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

Project description:Grassland cultivation can mobilize large pools of N in the soil, with the potential for N leaching and N2O emissions. Spraying with the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) before cultivation was simulated by use of soil columns in which the residue distribution corresponded to plowing or rotovation to study the effects of soil-residue contact on N transformations. DMPP was sprayed on aboveground parts of ryegrass and white clover plants before incorporation. During a 42-day incubation, soil mineral N dynamics, potential ammonia oxidation (PAO), denitrifying enzyme activity (DEA), nitrifier and denitrifier populations, and N2O emissions were investigated. The soil NO3- pool was enriched with 15N to trace sources of N2O. Ammonium was rapidly released from decomposing residues, and PAO was stimulated in soil near residues. DMPP effectively reduced NH4+ transformation irrespective of residue distribution. Ammonia-oxidizing archaea (AOA) and bacteria (AOB) were both present, but only the AOB amoA transcript abundance correlated with PAO. DMPP inhibited the transcription of AOB amoA genes. Denitrifier genes and transcripts (nirK, nirS, and clades I and II of nosZ) were recovered, and a correlation was found between nirS mRNA and DEA. DMPP showed no adverse effects on the abundance or activity of denitrifiers. The 15N enrichment of N2O showed that denitrification was responsible for 80 to 90% of emissions. With support from a control experiment without NO3- amendment, it was concluded that DMPP will generally reduce the potential for leaching of residue-derived N, whereas the effect of DMPP on N2O emissions will be significant only when soil NO3- availability is limiting. IMPORTANCE:Residue incorporation following grassland cultivation can lead to mobilization of large pools of N and potentially to significant N losses via leaching and N2O emissions. This study proposed a mitigation strategy of applying 3,4-dimethylpyrazole phosphate (DMPP) prior to grassland cultivation and investigated its efficacy in a laboratory incubation study. DMPP inhibited the growth and activity of ammonia-oxidizing bacteria but had no adverse effects on ammonia-oxidizing archaea and denitrifiers. DMPP can effectively reduce the potential for leaching of NO3- derived from residue decomposition, while the effect on reducing N2O emissions will be significant only when soil NO3- availability is limiting. Our findings provide insight into how DMPP affects soil nitrifier and denitrifier populations and have direct implications for improving N use efficiency and reducing environmental impacts during grassland cultivation.

Project description:Nitrification is an essential process for nutrient removal from wastewater and an important emission source of nitrous oxide (N2O), which is a powerful greenhouse gas and a dominant ozone-depleting substance. In this study, nitrification and N2O emissions were tested in two weakly acidic (pH 6.3 to 6.8) reactors: one with dissolved oxygen (DO) at over 2.0 mg/liter and the other with DO at approximately 0.5 mg/liter. Efficient nitrification was achieved in both reactors. Compared to that in the high-DO reactor, N2O emission in the low-DO reactor decreased slightly, by 20%, and had insignificant correlation with the fluctuations of DO (P = 0.935) and nitrite (P = 0.713), indicating that N2O might not be produced mainly via nitrifier denitrification. Based on quantitative PCR (qPCR), quantitative fluorescent in situ hybridization (qFISH), and functional gene amplicon and metagenome sequencing, it was found that complete ammonia oxidizers (comammox), i.e., Nitrospira organisms, significantly outnumbered canonical ammonia-oxidizing bacteria (AOB) in both weakly acidic reactors, especially in the low-DO reactor with the comammox/AOB amoA gene ratio increasing from 6.6 to 17.1. Therefore, it was speculated that the enriched comammox was the primary cause for the slightly decreased N2O emission under long-term low DO in the weakly acidic reactor. This study demonstrated that the comammox Nitrospira can survive well under the weakly acidic and low-DO conditions, implying that achieving efficient nitrification with low N2O emission as well as low energy and alkalinity consumption is feasible for wastewater treatment.IMPORTANCE Nitrification in wastewater treatment is an important process for eutrophication control and an emission source for the greenhouse gas N2O. The nitrifying process is usually operated at a slightly alkaline pH and high DO (>2 mg/liter) to ensure efficient nitrification. However, it consumes a large amount of energy and chemicals, especially for wastewater without sufficient alkalinity. This paper demonstrates that comammox can adapt well to the weakly acidic and low-DO bioreactors, with a result of efficient nitrification and low N2O emission. These findings indicate that comammox organisms are significant for sustainable wastewater treatment, which provides an opportunity to achieve efficient nitrification with low N2O production as well as low energy and chemical consumption simultaneously.

Dataset Information

Dimethyl pyrazol-based nitrification inhibitors effect on nitrifying and denitrifying bacteria to mitigate N2O emission.

Publications

Dimethyl pyrazol-based nitrification inhibitors effect on nitrifying and denitrifying bacteria to mitigate N<sub>2</sub>O emission.

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