Inhibition Of Denitrification Research Articles

Inhibition of denitrification and enhancement of microbial interactions in the AGS system by high concentrations of quinoline

Quinoline represents a highly toxic and structurally stable nitrogen-containing heterocyclic compound in coking wastewater, posing a potential threat to human beings and the ecological environment. In this study, we investigated the impact of gradually elevating quinoline concentration on pollutant removal efficiency, sludge characteristics, microbial community and their interactions in the aerobic granular sludge (AGS) system. The results demonstrated that AGS was capable of effectively degrading quinoline, with a final removal rate of 90 mg/L quinoline reaching 98.54 ± 0.28%. Notably, the denitrification process was significantly impeded in the presence of 90 mg/L quinoline, with the Phase D effluent displaying a notably high NO3--N concentration of 37.09 ± 21.81 mg/L, primarily attributed to the reduced abundance of norank_f_A4b bacteria. As the quinoline concentration increased, the sludge particle size diminished from 3.46 to 2.60 mm, while the settling performance deteriorated significantly, escalating from 31.29 ± 1.63 mL/g to 62.32 ± 2.87 mL/g. Meanwhile, the protein (PN) content in EPS gradually increased (from 19.87 ± 0.88 mg/g MLVSS to 51.22 ± 3.21 mg/g MLVSS), while the polysaccharide (PS) content fluctuated. Quinoline profoundly modified microbial community composition and structure, with deterministic processes dominating community assembly. Network analysis indicated intensified and complex microbial interactions at 90 mg/L quinoline, characterized by significantly higher positive correlations. In addition, rare taxa (RT) dominated the network nodes, with 74 of 93 key species belonging to RT, highlighting their pivotal roles in sustaining system functions and strengthening microbial connections. This study provides new insights into the effects of quinoline on microbial community structure and interactions in AGS system.

Nutrient removal performance and the nitrogen-sulfur conversion pathways in sulfur-iron based biofilter under acidic/alkaline conditions

The nitrate and phosphate removal in sulfur based (Vp) and sulfur-iron based autotrophic denitrification systems (Sp) with sponge iron as the iron source was investigated under different pH (6–9). In this study, Sp (>98 %) and Vp (>97 %) had higher nitrate removal efficiency at pH 7–8 and the highest phosphate removal efficiency (94.9 % ± 3.57 %) at pH 6. The flow cytometry detection revealed that the cell integrity was damaged due to the formation of iron crusts at pH 6, which resulted in irreversible inhibition in Sp. Electron transfer and nitrogen transformation showed that compared with the sustained inhibition of denitrification at pH 6, the inhibition at pH 9 occurred mainly in the initial 0–2 h due to the enhanced flow of electrons toward the formation of biological element sulfur (Sbio0). Coupled sponge iron promoted the adsorption and reduction of NO2−/N2O and reduced N2O emissions. Metatranscriptomics analysis showed that significant upregulation of key gene expression for nitrogen and sulfur metabolism in Sp, with complex regulatory mechanisms enhancing system stability and performance. Acidic conditions reduced the abundance of denitrification and sulfur oxidation functional genes. However, alkaline conditions (pH of 9) mainly caused the decreased abundance of denitrification functional genes, which was favorable for accumulation of Sbio0. This study provides a theoretical basis for the stable operation and regulation of the sulfur-iron based autotrophic denitrification process.

Single and Combined Effects of Phenanthrene and Silver Nanoparticles on Denitrification Processes in Coastal Marine Sediments.

The increasing production and utilization of polycyclic aromatic hydrocarbons (PAHs) and commercial silver nanoparticles (AgNPs) have raised concerns about their potential environmental release, with coastal sediments as a substantial sink. To better understanding the effects of these contaminants on denitrification processes in coastal marine sediments, a short-term exposure simulation experiment was conducted. We investigated the effects of single and combined contamination of phenanthrene (Phe) and AgNPs on denitrification processes in a coastal marine sediment. Results showed that all contaminated treatment groups had different degrees of inhibitory effect on denitrification activity, denitrifying enzyme activity, total bacteria count and denitrifying genes. The inhibitory effect sequence of each treatment group was combined treatment > AgNPs treatment > Phe treatment. Moreover, the inhibitory effects of denitrifying genes were much larger than that of total bacteria count, indicating that the pollutants had specific toxic effects on denitrifying bacteria. The sequence of sensitivity of three reduction process to pollutants was N2O > NO2- > NO3-. All contaminated treatment groups could increase NO3-, NO2- and N2O accumulation. Furthermore, according to the linear relationship between functional gene or reductase and denitrification process, we also found that the abundance of denitrifying genes could better predict the influence of Phe and AgNPs on sediment denitrification than the denitrifying bacterial diversity. In addition, at the genus level, the community structure of nirS- and nosZ-type denitrifying bacteria changed dramatically, while changes at the phylum level were comparatively less pronounced. Single and combined contamination of Phe and AgNPs could reduce the dominance of Pseudomonas, which may lead to a potential slow-down in the degradation of Phe and inhibition of denitrification, especially the combined contamination. Overall, our study revealed that combined contamination of Phe and AgNPs could lead to an increase in NO3-, NO2- and N2O accumulation in coastal sediment, which poses a risk of eutrophication in coastal areas, exacerbates the greenhouse effect and has adverse effects on global climate change.

Complete elimination of methane formation in stored livestock manure using plasma technology

Plasma-based nitrogen fixation has recently been shown to be applicable in the domain of manure management, as it has the ability to reduce ammoniacal nitrogen losses and increase the nitrogen content of organic wastes, with air and electricity as the only input. In addition, the plasma treatment confers antimicrobial properties, which we hypothesize to be transferable to methanogenic archaea and hence prevent methane formation during manure storage – a major contributor to global anthropogenic greenhouse gas emissions. In this work we compared the methane formation from cow manure to the methane formation in nitrogen enriched cow manure, kept in two outdoor storage tanks for 70–80 summer days over three consecutive years. In all instances, the methane formation was eliminated completely. To investigate the cause of inhibition, a controlled incubation experiment was conducted to show that neither the acidification nor the addition of nitrate or nitrite, alone or in combination, could explain the inhibition of methanogenesis and denitrification that occurred in plasma treated cow manure at moderate pH.

Denitrification regulates spatiotemporal pattern of N2O emission in an interconnected urban river-lake network

Urban rivers are hotspots of N2O production and emission. Interconnected river-lake networks are constructed to improve the water quality and hydrodynamic conditions of urban rivers in many cities of China. However, the impact of the river-lake connectivity project on N2O production and emission remains unclear. This study investigated dissolved N2O and emission of the river-lake network in Wuhan City, China from March 2021 to December 2021. The results showed that river-lake connection greatly decreased riverine Nitrogen (N) concentration and increased dissolved oxygen (DO) concentration compare to traditional urban rivers. N2O emissions from the urban river interconnected with lakes (LUR: 67.3 ± 92.6 μmol/m2/d) were much lower than those from the traditional urban rivers (UR: 467.3 ± 1075.7 μmol/m2/d) and agricultural rivers (AR: 20.4 ± 15.3μmol/m2/d). Regression tree analysis suggested that the N2O concentrations were extremely high when hypoxia exists (DO < 1.6 mg/L), and TDN was the primary factor regulating N2O concentrations when hypoxia does not occur. Thus, we ascribe the low N2O emission in the LUR and AR to the lower N contents and higher DO concentrations. The microbial process of N2O production and consumption were quantitatively estimated by isotopic models. The mean proportion of denitrification derived N2O (fbD) was 63.5 %, 55.6 %, 42.3 % and 42.7 % in the UR, LUR, lakes and AR, suggested denitrification dominated N2O production in the urban rivers, but nitrification dominated N2O production in the lakes and AR. The positive correlation between logN2O and fbD suggested that denitrification is the key process to regulate the N2O production and emission. The abundance of denitrification genes (nirS and nirK) was much higher than that of nitrification genes (amoA and amoB), also evidenced that denitrification was the main N2O source. Therefore, river-lake interconnected projects changed the nutrients level and hypoxic condition, leading to the inhibition of denitrification and nitrification, and ultimately resulting in a decrease of N2O production and emission. These results advance the knowledge on the microbial processes that regulate N2O emissions in inland waters and illustrate the integrated management of water quality and N2O emission.

Biological inhibition of denitrification (BDI): an early plant strategy for Fallopia × bohemica seedling development.

The successful plant Fallopia × bohemica presents interesting capacities for control of the soil nitrogen cycle at the adult stage, termed biological inhibition of denitrification (BDI). The BDI strategy allows the plant, via the production of secondary metabolites (procyanidins), to compete with the denitrifying microbial community and to divert nitrate from the soil for its benefit. In this study, we analysed whether seedlings of F. × bohemica can implement BDI at the seedling stage. We also determined whether soil nitrogen availability influences the implementation of BDI and seedling growth. We sowed achenes of F. × bohemica in soils representing a nitrogen gradient (six treatments) and harvested seedlings after 20 or 40 days of growth. The denitrification and related microbial communities (i.e. functional gene abundances of nirK and nirS), soil parameters (nitrate content and humidity) and plant performance (biomass, growth and root morphology) were determined. On soil without addition of nitrogen, BDI was observed after 20 days of growth, whereas a stimulation of denitrification was found after 40 days. The increase of soil N content had few effects on the activity and structure of the soil denitrifying community and on the plant biomasses or the relative growth rates. Correlations between plant and microbial parameters were observed after 20 days of growth, reflecting early and strong chemical interactions between plants and the denitrifying community, which decreased with plant growth after 40 days. This study shows that an early BDI enhances the efficiency of nitrogen acquisition in the first weeks of growth, allowing for a conservative root strategy after 40 days. This switch to a conservative strategy involved resource storage, an altered allocation to above- and below-ground parts and an investment in fine roots. It now seems clear that this storage strategy starts at a very young age with early establishment of BDI, giving this clonal plant exceptional capacities for storage and multiplication.

Effects of inorganic ions on autotrophic denitrification by Thiobacillus denitrificans and on heterotrophic denitrification by an enrichment culture

Salinity of nitrate-laden wastewaters, such as those produced by metal industries, tanneries, and wet flue gas cleaning systems may affect their treatment by denitrification. Salt inhibition of denitrification has been reported, while impacts of individual ions remain poorly understood whilst being relevant for wastewaters where often the concentration of a single ion rather than the salts varies. The aim of this study was to determine the inhibition by inorganic ions (Na+, Cl−, SO42− and K+) commonly present in saline wastewaters on denitrification and reveal its potential for the treatment of such waste streams, like those produced by NOx-SOx removal scrubbers. The inhibitory effects were investigated for both heterotrophic (enrichment culture) and autotrophic (T. denitrificans) denitrification in batch assays, by using NaCl, Na2SO4, KCl and K2SO4 salts at increasing concentrations. The half inhibition concentrations (IC50) of Na+ (as NaCl), Na+ (as Na2SO4) and Cl− (as KCl) were: 4.3 ± 0.3, 7.9 ± 0.5 and 5.2 ± 0.3 g/L for heterotrophic, and 1–2.5, 2.5–5 and 4.1 ± 0.3 g/L for autotrophic denitrification, respectively. Heterotrophic denitrification was completely inhibited at 20 g/L Na+ (as NaCl), 30 g/L Na+ (as Na2SO4) and 30 g/L Cl− (as KCl), while autotrophic at 8 g/L Na+ (as NaCl), 10 g/L Na+ (as Na2SO4) and 15 g/L Cl− (as KCl). In both cases, Cl− addition had the most important role in decreasing denitrification rate, while Na+ at 1 g/L stimulated autotrophic denitrification but rapidly inhibited the rate at higher concentrations. Nitrite reduction was less inhibited by the ions than nitrate reduction and both the osmotic pressure and the toxicity of the single ions played key roles in the overall inhibition of denitrification. Eventually, both autotrophic and heterotrophic denitrification showed potential for the treatment of a saline wastewater from a NOx-SO2 removal scrubber from a pulp mill.

Effects of native plants on nitrogen cycling microorganisms in soil

Plants modify the nitrogen (N) cycle in soil through plant N uptake, root exudation, and nitrification inhibition resulting from root exudates and changes to the physicochemical properties of soil. Plants with specific traits can be selected to manage N fluxes in the soil following the land application of N-rich wastes thereby reducing losses of N from these systems into the atmosphere (via nitrous oxide) and waterbodies (via nitrate). Previous work has shown that some New Zealand native myrtaceous species can reduce such losses more than other species, but the underlying mechanisms are unknown. It was hypothesized that lower N losses may result from the inhibition of nitrification and denitrification. We aimed to determine the effect of New Zealand native plant species on the abundance of nitrifying and denitrifying microorganisms in the soil using a pot experiment with five native plant species (Carex secta, Coprosma robusta, Kunzea robusta, Leptospermum scoparium, and Metrosideros umbellata) and exotic pasture (Lolium perenne). Half of the pots received fertilisation with N (urea), phosphorus, potassium, and sulphur, while the remainder were unfertilised controls. We quantified the abundance of bacteria, archaea, and the functional genes encoding nitrite reductase (nirK, nirS), nitrous oxide reductase (nosZ), and bacterial and archaeal ammonia monooxygenase (amoA) in the rhizosphere of these plants. Results indicated that plant species had a significant effect on the abundance of nosZ and bacterial amoA. In fertilised soil, the abundance of bacterial amoA was lower under mono- than dicotyledonous species. Similarly, the chemical properties of the soil differed between these groups. Monocotyledonous species took up more N and had lower concentrations of mineral N in the rhizosphere. This indicates that the increased competition for N likely reduced the abundance of amoA and therefore nitrification and nitrate losses. The results support the utilisation of species selection to reduce N losses. In particular, monocotyledonous species may be planted in high-fertility environments to mitigate N contamination of ground- and surface water. Future work should determine the mechanisms of plant specific interaction with ammonia-oxidising bacteria, although plant uptake can explain some of the observed differences.

Effects of air blower and pump failures on the performance of A2O processes for wastewater treatment

The unexpected failure of equipment such as pumps and fans in wastewater treatment plants can reduce wastewater treatment efficiency, leading to direct leakage of untreated wastewater into the environment. It is hence important to predict the possible consequences of equipment failure to minimize the leakage of harmful substances. This study examines the impacts of equipment shutdown on the performance and recovery time of a laboratory-scale anaerobic/anoxic/aerobic system with regard to reactor conditions and water quality. Two days after the air blowers are stopped, the concentrations of the soluble chemical oxygen demand, NH4–N, and PO4–P in the effluent of the settling tank increase to 122 mg/L, 23.8 mg/L, and 46.6 mg/L, respectively. These concentrations return to their initial values after 12, 24, and 48 h of restarting the air blowers. The concentrations of PO4–P and NO3–N in the effluent increase to 58 mg/L and 20 mg/L, respectively, about 24 h after stopping the return activated sludge and mixed liquor recirculation pumps, owing to the release of phosphates in the settling tank and inhibition of denitrification.

An integrated full-scale system for the treatment of real ship domestic wastewater: Shore-test experiments

With the increasing number of ocean-sailing ships, the pollution of the oceans' water caused by the discharge of ship wastewater has become one of the serious concerns for the whole world. Most of existing ship wastewater treatment studies were carried out in bench/pilot-scale and cannot meet the latest discharge standards of International Maritime Organization (IMO) and China. To overcome those barriers and prepare for the onboard test, we evaluated the hydraulic retention time (HRT) variations (10.5-12 h) for treating low COD/TN ship wastewater by a full-scale oxic-anoxic micro-sludge membrane biofilm reactor (O-AMSMBR) system. The full-scale O-AMSMBR achieved over 82 % removal of chemical oxygen demand (COD) and 76 % removal of total nitrogen (TN) from a ship wastewater with continuous operation at a HRT of 10.5 h. Although anoxic membrane biofilm partly compensated for the deterioration of TN removal in performance of the aerobic sludge, the effluent TN concentrations still cannot meet IMO discharge standards at HRT as low as 9 h due to the inhibition of simultaneous nitrification and denitrification (SND) by low HRT and C/N. Wavelet Neural Networks (WNN) was used to explore the removal mechanism under HRT variations. Simulation results show that HRT andinfluent pollutant’s concentration are the two most sensitive factors for COD and TN removal in daily operation so special attention should be paid to the dramatic change of influent flow rate in ship sewage treatment operation. In particular, techno-economic analysis was conducted in this study to provides a baseline for developing economic options for ship wastewater treatment in practice.

Does the aging behavior of microplastics affect the process of denitrification by the difference of copper ion adsorption?

Riparian sediment is a hot zone for denitrification that can withhold copper and microplastics (MPs) from outside. It has been proven that MPs affect denitrification and the existing forms of copper in the environment. However, the impact of copper on sediment denitrification under exposure to MPs remains unclear. This study revealed the response of sediment denitrification to copper availability under the adsorption of MPs and the complexation of MP-derived dissolved organic matter (DOM). These results showed that MP accumulation inhibited denitrification. However, aged MPs increased the activity of nitrite reductase (12.64%), nitrogen dioxide reductase (37.68%), and electron transport (28.93%) compared with pristine MPs. The aging behavior of MPs alleviated 28.18% nitrite accumulation and 16.41–118.35% nitrous oxide emissions. Thus, the aging behavior of MPs alleviated the inhibition of denitrification. Notably, we resolved the copper ion adsorption and complexation by MPs, MP-derived DOM contributed to the denitrification process, and we found that the key nitrogen removal factors were affected by KL, KM, and K2. These results fill a gap in our understanding of biochemical synthesis of MPs during denitrification. Furthermore, it can be used to build a predictive understanding of the long-term effects of MPs on the sediment nitrogen cycle.

Altering sliver nanoparticles-induced inhibition to bacterial denitrification via visible light by regulating silver transformation and adaptive mechanism under anaerobic conditions

Silver nanoparticles (Ag NPs) are toxic to bacteria in the environment, and visible light has been widely reported to aggravate their inhibitory impacts, especially under aerobic conditions. Bacterial denitrification is crucial for the nitrogen cycle, which mainly depends on the anaerobic environment. It is still unclear whether visible light alters the impacts of Ag NPs on bacterial denitrification under anaerobic conditions. This work found that Ag NP-induced inhibition of the anaerobic denitrification of Paracoccus denitrificans was alleviated by visible light. Compared with the reactor with 5 mg/L Ag NPs, visible light alleviated the negative impacts on denitrifying bacteria, and thus improved denitrification efficiency by 264.8 %. The mechanism studies indicated that visible light provoked the agglomeration of Ag NPs and the formation of insoluble and nontoxic Ag oxides (Ag2O and Ag2S) on the surface of particles under anaerobic conditions, resulting in the decreased dissolution of Ag NPs. Furthermore, the released Ag+ could be photoreduced under visible light to drastically reduce the concentration of free Ag+. The secretion of extracellular polymeric substances (especially proteins) could be stimulated by visible light to adapt to Ag NPs exposure. In addition, visible light promoted bacterial growth and the activities of critical enzymes that participated in denitrification (nitrate reductase and nitrite reductase). Hence, by altering Ag NPs transformation and bacterial adaptation, the inhibition of bacterial denitrification by Ag NPs was alleviated via visible light under anaerobic conditions.

Development of Bio-Electrochemical Reactor for Groundwater Denitrification: Effect of Electric Current and Water Hardness

Nitrate-nitrogen (NO3-N) contaminating groundwater is an environmental issue in many areas, and is difficult to treat by simple processes. A bio-electrochemical reactor (BER) using copper wire and graphite plate was developed to purify the NO3-N-contaminated groundwater. The low (of 10 mA) and high (of 20 mA) electric currents were applied to the BERs, and various influent hardness levels from 20 to 80 mg/L as CaCO3 due to groundwater characteristics were supplied to clarify the total nitrogen removal efficiency and NO3-N removal mechanisms. In the BER-10, the bio-electrochemical reactions caused 85% of total nitrogen to be removed through heterotrophic and autohydrogenotrophic denitrification in the suspended sludge and biofilm. However, the chemical deposit occurring at the cathode from water hardness affected the decreasing denitrification performance; 12.6% of Mg and 8.8% of Ca elements were observed in the biofilm. The enhancement of electrochemical reactions in the BER-20 caused integrating electrochemical and bio-electrochemical reactions; the NO3-N was electrochemically reduced to NO2-N, and it was further biologically reduced to N2. A better total nitrogen removal of 95% was found; although, a larger deposit of Mg (22.8%) and Ca (10.8%) was observed. The relatively low dissolved H2 in the BER-20 confirmed that the deposit affected the decreasing gaseous H2 transfer and inhibition of autohydrogenotrophic denitrification in the suspended sludge. According to the microbial analysis, both heterotrophic and autohydrogenotrophic denitrification were obtained in the suspended sludge of both BERs; Nocadia (26.8%) was the most abundant genus in the BER-10, whereas Flavobacterium (27.1%) and Nocadia (25.0%) were the dominant genera in the BER-20.

The individual and combined effects of polystyrene and silver nanoparticles on nitrogen transformation and bacterial communities in an agricultural soil

The effects of emerging contaminants micro/nanoplastics (MPs/NPs) and silver nanoparticles (Ag NPs) on health have attracted universal concern throughout the world. However, it is unclear on the combined effects of MPs/NPs and Ag NPs on the biogeochemistry cycle such as nitrogen transformation and functional microorganism in the soil. In the present study, we conducted a 45-day soil microcosm experiment with polystyrene (PS) MPs/NPs and Ag NPs to investigate their combined impact on nitrogen cycling and the bacterial community. The results showed that MPs or NPs exerted limited effects on nitrogen transformation in the soil. The combined effects of PS MPs/NPs and Ag NPs were mainly caused by the presence of Ag NPs. However, PS NPs alleviated the inhibition of anammox and denitrification induced by Ag NPs via upregulating anammox-related genes and elevating nitrate and nitrite reductase activities. PS MPs + Ag NPs treatment significantly reduced bacterial diversity. PS MPs/NPs + Ag NPs increased the relative abundances of denitrifying Cupriavidus by 0.32% and 0.06% but decreased nitrogen-fixing functional microorganisms of Microvirga (by 2.05% and 2.24%), Bacillus (by 0.16% and 0.22%), and Herbaspirillum (by 0.14% and 0.07%) at the genus level compared with Ag NPs alone. The significant downregulation of nitrogen-fixing genes (K02586, K02588, and K02591) was observed in PS MPs/NPs + Ag NPs treatment compared to Ag NPs in the nitrogen metabolism pathway. Moreover, g-Lysobacter and g-Aquimonas were identified as biomarkers in PS MPs + Ag NPs and PS NPs + Ag NPs by LEfSe analysis. Our study sheds the light that changes of functional microorganism abundances contributed to the alteration of nitrogen transformation. Taking the particle size of plastics into account will be helpful to accurately assess the combined ecological risks of plastics and nanomaterial contaminants.

Mitigation of CyanoHABs Using Phoslock® to Reduce Water Column Phosphorus and Nutrient Release from Sediment.

Cyanobacterial blooms can be stimulated by excessive phosphorus (P) input, especially when diazotrophs are the dominant species. A series of mesocosm experiments were conducted in a lake dominated by a cyanobacteria bloom to study the effects of Phoslock®, a phosphorus adsorbent. The results showed that the addition of Phoslock® lowered the soluble reactive phosphate (SRP) concentrations in water due to efficient adsorption and mitigated the blooms. Once settled on the sediments, Phoslock® serves as a barrier to reduce P diffusion from sediments into the overlying waters. In short-term (1 day) incubation experiments, Phoslock® diminished or reversed SRP effluxes from bottom sediments. At the same time, the upward movement of the oxic–anoxic interface through the sediment column slightly enhanced NH4+ release and depressed N2 release, suggesting the inhibition of nitrification and denitrification. In a long-term (28 days) experiment, Phoslock® hindered the P release, reduced the cyanobacterial abundance, and alleviated the bloom-driven enhancements in the pH and oxygen. These results suggest that, through suppression of internal nutrient effluxes, Phoslock® can be used as an effective control technology to reduce cyanobacteria blooms common to many freshwater systems.

Novel Alcaligenes ammonioxydans sp. nov. from wastewater treatment sludge oxidizes ammonia to N2 with a previously unknown pathway.

Heterotrophic nitrifiers are able to oxidize and remove ammonia from nitrogen-rich wastewaters but the genetic elements of heterotrophic ammonia oxidation are poorly understood. Here, we isolated and identified a novel heterotrophic nitrifier, Alcaligenes ammonioxydans sp. nov. strain HO-1, oxidizing ammonia to hydroxylamine and ending in the production of N2 gas. Genome analysis revealed that strain HO-1 encoded a complete denitrification pathway but lacks any genes coding for homologous to known ammonia monooxygenases or hydroxylamine oxidoreductases. Our results demonstrated strain HO-1 denitrified nitrite (not nitrate) to N2 and N2 O at anaerobic and aerobic conditions respectively. Further experiments demonstrated that inhibition of aerobic denitrification did not stop ammonia oxidation and N2 production. A gene cluster (dnfT1RT2ABCD) was cloned from strain HO-1 and enabled E. coli accumulated hydroxylamine. Sub-cloning showed that genetic cluster dnfAB or dnfABC already enabled E. coli cells to produce hydroxylamine and further to 15 N2 from (15 NH4 )2 SO4 . Transcriptome analysis revealed these three genes dnfA, dnfB and dnfC were significantly upregulated in response to ammonia stimulation. Taken together, we concluded that strain HO-1 has a novel dnf genetic cluster for ammonia oxidation and this dnf genetic cluster encoded a previously unknown pathway of direct ammonia oxidation (Dirammox) to N2 .

The inhibitory efficacy of procyanidin on soil denitrification varies with N fertilizer type applied

Denitrification is a major process of the nitrogen (N) cycle by converting nitrate (NO3−) back to gaseous nitrogen (N2), which leads to massive losses of N, including fertilizer N, from agricultural systems. One mitigation strategy for these N losses involves denitrification inhibition by plant-derived biological denitrification inhibitors (BDIs). Procyanidin was recently identified as a new class of BDI in root extracts from Fallopia spp. However, the efficacy of this compound on soil denitrification under different N fertilizer sources is not well understood. Here, a 14-day microcosm experiment was conducted using three nitrate-based fertilizers (NH4NO3, KNO3, and Ca(NO3)2) to investigate the impact of procyanidin on soil denitrification and associated microbial pathways. Results showed that procyanidin inhibited denitrification activity regardless of the source of N fertilizer applied, but the inhibitory efficacy of procyanidin varied with N fertilizer types. Addition of procyanidin had greater denitrification inhibition in the soils applied with NH4NO3 than with other types of N fertilizer. Moreover, nitrate reductase activity was significantly suppressed by procyanidin addition across all three N fertilizers tested. Quantification of denitrifying functional genes (nirS, nirK, and nosZ) demonstrated that procyanidin inhibited the activity and growth of nirS- and nirK-type denitrifiers, but stimulated the growth of nosZI-containing denitrifiers. These findings indicate that the inhibition of soil denitrification by procyanidin was mainly a result of the suppression of nitrate reductase activity and nirS- and nirK-type denitrifiers abundance. The use of procyanidin together with N fertilizers, especially NH4NO3, can be an effective way to reduce the N losses by denitrification.

Metagenomic insights into the explanation of biofilter performance distinction induced by dissolved oxygen increment

In this study, a set of novel double-layer-packed sequencing batch biofilm reactors (SBBRs) were constructed to treat secondary effluent with a low carbon-to-nitrogen ratio (C/N = 3.0). All the reactors were operated under identical conditions except dissolved oxygen (DO) in the aerobic section, which was controlled at seven different levels (0.5, 1.0, 1.5, 2.0, 2.5, 3.5 and 4.5 mg/L). It was found that the aerobic section contributed most to performance at an optimal DO of 2.0 mg/L. The change in DO accounted for the varied bacterial communities, which resulted in different performances. Both bacterial community structure and potential functions displayed close associations with performance implying nutrients were mainly removed by simultaneous nitrification-endogenous denitrification and phosphorus removal (SNEDPR) process. Moreover, the proliferation of nonfunctional nitrifiers/denitrifiers led to the inhibition of denitrification resulting in a lower removal of total dissolved nitrogen and phosphorus.

Biological inhibition of denitrification (BDI) in the field: Effect on plant growth in two different soils

Intensive agriculture uses increasingly large amounts of nitrogen fertilizers to increase yields because nitrogen is one of the limiting factor of plant growth. Plants are in direct competition with denitrifying bacteria for nitrate, but biological denitrification inhibition (BDI) is a strategy developed by some plants in which procyanidins are produced that inhibit denitrification. This phenomenon increases the available nitrate in the soil. Previous results showed that the addition of procyanidins to lettuce fields allowed effective BDI and increased soil nitrate and plant growth. However, agriculture is performed in different soils, and the action of procyanidins could depend on the type of soil. In this study, we tested the effect of procyanidin amendment on lettuce growth in two types of soils: loamy sand and sandy clay loam. Our results show that in both soils, the addition of procyanidins causes inhibition of denitrification, an increase in available nitrate, counter-selection of denitrification communities and a gain in plant mass without modification of the soil structure. This study highlights a sustainable agricultural method effective in a variety of soils.

Elucidating the microbiological characteristics of cyromazine affecting the nitrogen cycle during aerobic composting of pig manure

Cyromazine as insect growth inhibitor have been frequently detected in the environment, which show a potential threat to environment and soil health. Nitrogen is an essential component of all living organisms and the main nutrient limiting life on our planet. In this study, quantitative polymerase chain reaction (qPCR) and sequencing of nitrifying and denitrifying bacteria were conducted to investigate the dynamic effects of cyromazine on nitrogen conversion during laboratory-based composting. Results showed that the presence of cyromazine significantly reduced the abundance of amoA gene during the thermophilic phase of composting (p < 0.01), resulting in lower oxidation of NH4+-N. The archaea amoA gene was more resistant to cyromazine. The nirK gene was more abundant than the nirS gene during composting and was significantly reduced only under high concentrations of cyromazine (p < 0.01). The high dose of cyromazine (15 mg/kg) severely damaged the nitrogen fixation capacity of compost products. Cyromazine exhibited an inhibition effect on richness (ACE, Chao) of nitrifying and denitrifying microorganisms during the thermophilic period, while increased the diversity (shannon) at all stages of composting. Pseudomonas_formosensis was the core denitrifiers that harbored nosZ gene, Nitrosomonas_eutropha and Nitrosospira_sp_Nl5 were the dominant nitrifier that harbored amoA gene, and these species have a negative response to cyromazine. Network analysis indicated that the dominant bacteria harboring amoA and nosZ genes were hubs of nitrogen oxidation and reduction processes. Structural equation modeling revealed that NO2−-N conversion played a crucial role in driving denitrification, and increase of NH4+-N content was attributed to the inhibition of nitrification and denitrification during composting caused by cyromazine.