Hydroxylamine Oxidoreductase Research Articles

Effect of mineral additives and ammonia-oxidizing bacteria additions on physicochemical properties, nitrogen retention and microbial dynamics during vermicomposting of chicken manure and green waste

Vermicomposting is an effective technology to transform organic wastes into organic fertilizers. However, a large amount of nitrogen will be lost through ammonia (NH3) volatilization during verimicomposting. Herein, we investigated the effects of mineral additives (diatomite, zeolite) and exogenous ammonia-oxidizing bacteria (AOB) (Nitrosomonas europaea, Nitrosospira multiformis) added individually or combinedly on the physicochemical changes, the conversion of nitrogen, and the resident microbial community succession during chicken manure and green waste co-vermicomposting. The results showed that the combined addition of AOB and mineral additives significantly improved vermicompost quality and maturity in terms of humus, carbon/nitrogen ratio, humic acid/fulvic acid ratio and germination index. Compared with the control, jointly adding AOB and mineral additives reduced NH3 emissions by 65.72∼71.05 %, accelerated the conversion of NH4+-N to NO3−-N, increased total nitrogen content by 23.22∼25.21 %. The combination of AOB and mineral additives significantly increased the activities of ammonia monooxygenase, hydroxylamine oxidase, and nitrate oxidoreductase, and enhanced the amount of AOB community. Additionally, the combined addition of AOB and mineral additives increased bacterial richness and diversity, accompanied by increased relative abundances of Luteimonas, Cellvibrio, Saccharimonadales, and Pseudomonas. These microbial shifts were correlated with enhanced lignocellulose degradation, organic phosphate mineralization, and promotion of plant growth. Moreover, vermiompost-based growing media with mineral additives and AOB can significantly promote the growth of purple cabbage as compared to the vermicompost without additives. Overall, the synergistic use of AOB and mineral additives (diatomite, zeolite) was recommended as a favorable approach for reducing nitrogen losses and improving the efficiency of chicken manure and green waste vermicomposting. This study offers a novel approach to mediate nitrogen transformation processes and reduce nitrogen loss.

Rapid achievement of partial nitrification process by adopting the combined strategy of anoxic starvation and free ammonia inhibition

ABSTRACT Partial nitrification (PN) is a prerequisite step for the short-cut nitrogen removal process, which is crucial to provide stable nitrite accumulation for subsequent units. The present study innovatively proposed a new strategy for the rapid establishment of PN by adopting short-term anoxic starvation combined with high free ammonia inhibition. The sludge obtained from the secondary sedimentation tank of a municipal wastewater treatment plant was starved for 7 days under anoxic conditions, and then wastewater with high ammonia nitrogen (400 mg L−1) was introduced. Within 17 days, stable nitrite accumulation was achieved in the sequencing batch reactor, and the nitrite accumulation rate reached more than 95.0%. The activity of ammonia monooxygenase enzyme increased from 0.0364 ± 0.0074 to 0.1275 ± 0.0021 μg NO2 −-N·mg−1 protein min−1, while that of hydroxylamine oxidoreductase enzyme increased from 1.5350 ± 0.0208 to 6.3852 ± 0.0400 EU g−1 SS. The relative abundance of Nitrosomonas increased from 0.10% to 25.90%, while that of Nitrospira consistently remained below 0.04%. And the relative abundance of short-cut denitrifying bacteria, including Truepera, OLB8, and OLB13 all increased. The results proved that the short-term anoxic starvation combined with high free ammonia inhibition was an effective strategy for rapid establishment of PN.

Long-term effect of phenol, quinoline, and pyridine on nitrite accumulation in the nitrification process: performance, microbial community, metagenomics and molecular docking analysis

Phenol, quinoline, and pyridine, commonly found in industrial wastewater, disrupt the nitrification process, leading to nitrite accumulation. This study explores the potential mechanisms through which these biotoxic organic compounds affect nitrite accumulation, using metagenomic and molecular docking analyses. Despite increasing concentrations of these compounds from 40 to 160 mg/L, ammonia nitrogen removal was not hindered, and stable nitrite accumulation rates exceeding 90 % were maintained. Additionally, these compounds inhibited nitrite-oxidizing bacteria (NOB) and enriched ammonia-oxidizing bacteria (AOB) in situ. As the concentration of these compounds rose, protein (PN) and polysaccharide (PS) concentrations also increased, along with a higher PN/PS ratio. Metagenomic analysis further revealed an increase in hao relative abundance, while microbial community analysis showed increased Nitrosomonas abundance, which contributed to nitrite accumulation stability. Molecular docking indicated that these compounds have lower binding energy with hydroxylamine oxidoreductase (HAO) and nitrate reductase (NAR), theoretically supporting the observed sustained nitrite accumulation.

Discovery of Candidatus Nitrosomaritimum as a New Genus of Ammonia-Oxidizing Archaea Widespread in Anoxic Saltmarsh Intertidal Aquifers.

Ammonia-oxidizing archaea (AOA) are widely distributed in marine and terrestrial habitats, contributing significantly to global nitrogen and carbon cycles. However, their genomic diversity, ecological niches, and metabolic potentials in the anoxic intertidal aquifers remain poorly understood. Here, we discovered and named a novel AOA genus, Candidatus Nitrosomaritimum, from the intertidal aquifers of Yancheng Wetland, showing close metagenomic abundance to the previously acknowledged dominant Nitrosopumilus AOA. Further construction of ammonia monooxygenase-based phylogeny demonstrated the widespread distribution of Nitrosomaritimum AOA in global estuarine-coastal niches and marine sediment. Niche differentiation among sublineages of this new genus in anoxic intertidal aquifers is driven by salinity and dissolved oxygen gradients. Comparative genomics revealed that Candidatus Nitrosomaritimum has the genetic capacity to utilize urea and possesses high-affinity phosphate transporter systems (phnCDE) for surviving phosphorus-limited conditions. Additionally, it contains putative nosZ genes encoding nitrous-oxide (N2O) reductase for reducing N2O to nitrogen gas. Furthermore, we gained first genomic insights into the archaeal phylum Hydrothermarchaeota populations residing in intertidal aquifers and revealed their potential hydroxylamine-detoxification mutualism with AOA through utilizing the AOA-released extracellular hydroxylamine using hydroxylamine oxidoreductase. Together, this study unravels the overlooked role of priorly unknown but abundant AOA lineages of the newly discovered genus Candidatus Nitrosomaritimum in biological nitrogen transformation and their potential for nitrogen pollution mitigation in coastal environments.

Ultrasound-induced structural changes in partial nitrification sludge: Unravelling the mechanism for improved nitrogen removal

Low-intensity ultrasound, as a form of biological enhancement technology, holds significant importance in the field of biological nitrogen removal. This study utilized low-intensity ultrasound (200 W, 6 min) to enhance partial nitrification and investigated its impact on sludge structure, as well as the internal relationship between structure and properties. The results demonstrated that ultrasound induced a higher concentration of nitrite in the effluent (40.16 > 24.48 mg/L), accompanied by a 67.76% increase in the activity of ammonia monooxygenase (AMO) and a 41.12% increase in the activity of hydroxylamine oxidoreductase (HAO), benefiting the partial nitrification. Based on the extended Derjaguin–Landau–Verwey–Overbeek (XDLVO) theoretical analysis, ultrasonic treatment enhanced the electrostatic interaction energy (WR) between sludge flocs, raising the total interaction energy from 46.26 kT to 185.54 kT, thereby causing sludge dispersion. This structural alteration was primarily attributed to the fact that the tightly bonded extracellular polymer (TB-EPS) after ultrasound was found to increase hydrophilicity and negative charge, weakening the adsorption between sludge cells. In summary, this study elucidated that the change in sludge structure caused by ultrasonic treatment has the potential to enhance the nitrogen removal performance by partial nitrification.

Norfloxacin affects inorganic nitrogen compound transformation in tailwater containing Corbicula fluminea

The Asian clam, Corbicula fluminea, commonly used in engineered wetlands receiving tailwater, affects nitrogen compound transformation in water. This study investigates how a commonly observed antibiotic in tailwater, norfloxacin, impact nitrogen compound transformation in tailwater containing C. fluminea. The clam was exposed to artificial tailwater with norfloxacin (0, 0.2, 20, and 2000 μg/L) for 15 days. Water properties, C. fluminea ecotoxicity responses, microorganism composition and nitrification- or denitrification-related enzyme activities were measured. Results revealed norfloxacin-induced increases and reductions in tailwater NH4+ and NO2− concentrations, respectively, along with antioxidant system inhibition, organ histopathological damage and disruption of water filtering and digestion system. Microorganism composition, especially biodiversity indices, varied with medium (clam organs and exposure water) and norfloxacin concentrations. Norfloxacin reduced NO2− content by lowering the ratio between microbial nitrifying enzyme (decreased hydroxylamine oxidoreductase and nitrite oxidoreductase activity) and denitrifying enzyme (increased nitrate reductase and nitrite reductase activity) in tailwater. Elevated NH4+ content resulted from upregulated ammonification and inhibited nitrification of microorganisms in tailwater, as well as increased ammonia emission from C. fluminea due to organ damage and metabolic disruption of the digestion system. Overall, this study offers insights into using benthic organisms to treat tailwater with antibiotic residues, especially regarding nitrogen treatment.

Insight into mitigation mechanisms of N2O emission by biochar during agricultural waste composting

The effects and mitigation mechanisms of biochar added at different composting stages on N2O emission were investigated. Four treatments were set as follows: CK: control, BB10%: +10 % biochar at beginning of composting, BB5%&T5%: +5% biochar at beginning and + 5 % biochar after thermophilic stage of composting, BT10%: +10 % after thermophilic stage of composting. Results showed that treatment BB10%, BB5%&T5%, and BT10% reduced total N2O emissions by 55 %, 37 %, and 36 %, respectively. N2O emission was closely related to most physicochemical properties, while it was only related to amoA gene and hydroxylamine oxidoreductase. Different addition strategies of biochar changed the contributions of physicochemical properties, functional genes and enzymes to N2O emission. Organic matter and C/N contributed 23.7 % and 27.6 % of variations in functional gene abundances (P < 0.05), respectively. pH and C/N (P < 0.05) contributed 37.3 % and 17.3 % of variations in functional enzyme activities. These findings provided valuable insights into mitigating N2O emissions during composting.

Biochar Combined with Garbage Enzyme Enhances Nitrogen Conservation during Sewage Sludge Composting: Evidence from Microbial Community and Enzyme Activities Related to Ammoniation

Nitrogen loss is an unavoidable problem during composting processes, and the ammonia oxidation process significantly affects nitrogen transformation. The objective of this study was to evaluate nitrogen transformation when garbage enzyme (GE), biochar (BC), pelelith (PL) and combinations thereof were added during sewage sludge composting. Meanwhile, the succession of ammonia-oxidizing bacteria (AOB) and archaea (AOA) were also explored via quantitative polymerase chain reaction and high-throughput sequencing. The results showed that GE + BC and GE + PL treatments decreased ammonia (NH3) formation by 23.8% and 8.3%, and that of nitrous oxide (N2O) by 25.7% and 26.3% relative to the control, respectively. Simultaneously, the GE, GE + BC, and GE + PL treatments boosted the succession of AOA and AOB, and increased the activities of ammonia monooxygenase (AMO) and hydroxylamine oxidoreductase (HAO) activities and the gene copies of AOA and AOB. The AMO activities, NH4-N, NO3-N, and C/N, significantly affect AOA and AOB community structures. The network analysis predicted that the AMO and HAO were secreted mainly by the unclassified_Archaea and norank_Crenarchaeota, whereas it also showed that the GE + BC improved microbial associations with AOA, enzymatic activity, and environmental factors. Thus, the addition of garbage enzyme and biochar appears to be a promising mitigation strategy to reduce nitrogen losses during the composting process.

The different responses of AOA and AOB communities to irrigation systems in the semi-arid region of Northeast China.

Ammonia oxidation is the rate-limiting step in nitrification and the key step in the nitrogen (N) cycle. Most soil nutrients and biological indicators are extremely sensitive to irrigation systems, from the perspective of improving soil fertility and soil ecological environment, the evaluation of different irrigation systems and suitability of selection, promote crop production and soil quality, study the influence of the soil microenvironment contribute to accurate evaluation of irrigation farmland soil health. Based on the amoA gene, the abundance and community diversity of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) and their responses to soil physicochemical factors and enzyme activities were studied in semi-arid areas of Northeast China. The study consisted of three irrigation systems: flood irrigation (FP), shallow buried drip irrigation (DI), and mulched drip irrigation (MF). The results showed that DI and MF significantly increased the contents of alkaline hydrolyzed nitrogen (AN), nitrate nitrogen (NO3--N), soil moisture, and the activities of ammonia monooxygenase (AMO) and hydroxylamine oxidase (HAO). Compared with FP, DI significantly increased the abundance of soil AOA and AOB, while MF significantly increased the abundance of soil AOB. Irrigation systems significantly affected the community composition of ammonia-oxidizing microorganisms (AOM). Also, AN and soil moisture had the greatest influence on the community composition of AOA and AOB, respectively. The AOB community had better stability and stress resistance. Moreover, the symbiotic network of AOB in the three irrigation systems was more complex than that of AOA. Compared with FP, the AOA community under treatment DI had higher complexity and stability, maintaining the versatility and sustainability of the ecosystem, while the AOB community under treatment MF had higher transfer efficiency in terms of matter and energy. In conclusion, DI and MF were more conducive to the propagation of soil AOM in the semi-arid area of Northeast China, which can provide a scientific basis for rational irrigation and N regulation from the perspective of microbiology.

Inhibitory effect of ammonium conversion by deep nitrogen application effectively reduces gaseous nitrogen losses

Increasing the depth of nitrogen (N) fertilizer application is an effective agronomic management measure for mitigating NH3 volatilization and N2O emissions. However, the information about reasons for the reduction of gaseous N loss and the responses of soil N transformation processes to deep N fertilization is limited. This field experiment monitored NH3 volatilization, N2O emission, and soil NH4+/NO3- dynamics under three N application depths (8, 16, and 24 cm) in the typical soil of Loess Plateau (Cumulic Anthrosol) during 2021–2022 and 2022–2023. The N transformation rate under different soil depths was determined by laboratory 15N tracing experiments, and driving contributions of soil physicochemical properties and biological characteristics to gaseous N emissions were considered comprehensively. Our field results verified that increasing N fertilization depth effectively reduces NH3 volatilization and N2O emission by 20.7–55.6% and 17.9–45.8%; Simultaneously, deep N fertilization delayed the peak time of NH3 or N2O and retained a higher concentration of NH4+ in deep soil (37.3–46.4 mg kg−1). The 15N tracing experiments revealed that the lower soil enzymatic activity and microbial abundance in deep soil significantly decreased N mineralization, NH4+ consumption, and nitrification rates by 30.6–44.1%, 37.1–44.9%, and 22.5–31.1% (P < 0.05), compared with the shallow soil. Random forest analysis explored that the predictive factors of NH3 volatilization were soil urease activity, NH4+ consumption rate, soil NH4+, and TOC (P < 0.01). Interestingly, N2O emission was also mainly affected by soil urease activity, hydroxylamine oxidoreductase (HAO) activity, soil NH4+, and NO3- (P < 0.01); Through partial least squares path models (PLS-PMs), deep N fertilization mainly reduced risk of gaseous N loss by changing the environmental conditions required for N transformation and affecting the N mineralization and nitrification processes to limit NH4+ generation and consumption. This study provides a theoretical foundation for the demonstration and promotion of deep fertilization in the Loess Plateau.

Performance of microbial inoculation and tricalcium phosphate on nitrogen retention and conversion: Core microorganisms and enzyme activity during kitchen waste composting

The substantial release of NH3 during composting leads to nitrogen (N) losses and poses environmental hazards. Additives can mitigate nitrogen loss by adsorbing NH3/NH4, adjusting pH, and enhancing nitrification, thereby improving compost quality. Herein, we assessed the effects of combining bacterial inoculants (BI) (1.5%) with tricalcium phosphate (CA) (2.5%) on N retention, organic N conversion, bacterial biomass, functional genes, network patterns, and enzyme activity during kitchen waste (KW) composting. Results revealed that adding of 1.5%/2.5% (BI + CA) significantly (p < 0.05) improved ecological parameters, including pH (7.82), electrical conductivity (3.49 mS/cm), and N retention during composting. The bacterial network properties of CA (265 node) and BI + CA (341 node) exhibited a substantial niche overlap compared to CK (210 node). Additionally, treatments increased organic N and total N (TN) content while reducing NH4+-N by 65.42% (CA) and 77.56% (BI + CA) compared to the control (33%). The treatments, particularly BI + CA, significantly (p < 0.05) increased amino acid N, hydrolyzable unknown N (HUN), and amide N, while amino sugar N decreased due to bacterial consumption. Network analysis revealed that the combination expanded the core bacterial nodes and edges involved in organic N transformation. Key genes facilitating nitrogen mediation included nitrate reductase (nasC and nirA), nitrogenase (nifK and nifD), and hydroxylamine oxidase (hao). The structural equation model suggested that combined application (CA) and microbial inoculants enhance enzyme activity and bacterial interactions during composting, thereby improving nitrogen conversion and increasing the nutrient content of compost products.

Comammox and unknown ammonia oxidizers contribute to nitrite accumulation in an integrated A-B stage process that incorporates side-stream EBPR (S2EBPR)

A novel integrated pilot-scale A-stage high rate activated sludge, B-stage short-cut biological nitrogen removal and side-stream enhanced biological phosphorus removal (A/B-shortcut N-S2EBPR) process for treating municipal wastewater was demonstrated with the aim to achieve simultaneous and carbon- and energy-efficient N and P removal. In this studied period, an average of 7.62 ± 2.17 mg-N/L nitrite accumulation was achieved through atypical partial nitrification without canonical known NOB out-selection. Network analysis confirms the central hub of microbial community as Nitrospira, which was one to two orders of magnitude higher than canonical aerobic oxidizing bacteria (AOB) in a B-stage nitrification tank. The contribution of comammox Nitrospira as AOB was evidenced by the increased amoB/nxr ratio and higher ammonia oxidation activity. Furthermore, oligotyping analysis of Nitrospira revealed two dominant sub-clusters (microdiveristy) within the Nitrospira. The relative abundance of oligotype II, which is phylogenetically close to Nitrospira_midas_s_31566, exhibited a positive correlation with nitrite accumulation in the same operational period, suggesting its role as comammox Nitrospira. Additionally, the phylogenetic investigation suggested that heterotrophic organisms from the family Comamonadacea and the order Rhodocyclaceae embedding ammonia monooxygenase and hydroxylamine oxidase may function as heterotrophic nitrifiers. This is the first study that elucidated the impact of integrating the S2EBPR on nitrifying populations with implications on short-cut N removal. The unique conditions in the side-stream reactor, such as low ORP, favorable VFA concentrations and composition, seemed to exert different selective forces on nitrifying populations from those in conventional biological nutrient removal processes. The results provide new insights for integrating EBPR with short-cut N removal process for mainstream wastewater treatment.

An effective strategy for rapid nitrite accumulation related to quorum sensing and the impact of quorum quenching enzyme

Nitrite accumulation is crucial in nitrogen removal processes that treat wastewater with a low carbon-to-nitrogen (C/N) ratio. This study investigates the effectiveness of effluent reflux in accelerating nitrite accumulation, considering quorum sensing, and explores the impact of quorum quenching enzymes. The results demonstrate that effluent reflux successfully accelerates the nitrite accumulation rate to 92.2% and increases the abundance of Nitrosomonas from 0.2% to 19.5%, as well as hydroxylamine oxidoreductase (HAO) from 0.285 Eu g−1 SS to 0.475 Eu g−1 SS. The quenching enzyme suppresses the growth of aerobic ammonia-oxidizing bacteria, leading to a decrease in Nitrosomonas abundance to 4.2% and HAO to 0 Eu g−1 SS. However, it enhances denitrification, which consumes nitrite and results in the deterioration of nitrite accumulation. Consequently, the total nitrogen (TN) removal efficiency increases from 15.5% to 43.2%, and the abundance of Hyphomicrobium increases to 33.7% accordingly. Quorum sensing proves to be an effective strategy for regulating ammonia removal and nitrite accumulation, and effluent reflux has a positive effect on enhancing quorum sensing.

Juglone, a plant-derived 1,4-naphthoquinone, binds to hydroxylamine oxidoreductase and inhibits the electron transfer to cytochrome c554.

Nitrification, the microbial conversion of ammonia to nitrate via nitrite, plays a pivotal role in the global nitrogen cycle. However, the excessive use of ammonium-based fertilizers in agriculture has disrupted this cycle, leading to groundwater pollution and greenhouse gas emissions. In this study, we have demonstrated the inhibitory effects of plant-derived juglone and related 1,4-naphthoquinones on the nitrification process in Nitrosomonas europaea. Notably, the inhibition mechanism is elucidated in which 1,4-naphthoquinones interact with hydroxylamine oxidoreductase, disrupting the electron transfer to cytochrome c554, a physiological electron acceptor. These findings support the notion that phytochemicals can impede nitrification by interfering with the essential electron transfer process in ammonia oxidation. The findings presented in this article offer valuable insights for the development of strategies aimed at the management of nitrification, reduction of fertilizer utilization, and mitigation of greenhouse gas emissions.

The Effect of Foam-Recycle on Ammonium Removal by Aerobic Denitrification Using Alcaligenes faecalis No. 4

Aerobic denitrifier Alcaligenes faecalis No. 4 removes ammonium-nitrogen to nitrogen gas via denitrification in a single aerobic condition. In our previous studies, factors such as ammonium removal rate, denitrification ratio, and cell growth were tested in various conditions. The removal pathway from ammonium to nitrogen gas still needs to be determined in detail. To clarify this pathway of Alcaligenes faecalis No. 4, we in this study investigated the effects of several factors on ammonium removal, such as foam-recycle, initial pH, initial ammonium concentration, and airflow rate. Denitrification ratio was improved by up to 23% through foam-recycle. The improvement of the denitrification ratio was resulted by the higher enzyme activity of hydroxylamine oxidoreductase (HAO) in the produced foam, which was about 28 times higher than that in the culture broth (i.e., without foam-recycle). The stripped ammonia was significantly high (above pH 9). The initial ammonium concentration and airflow rate also influenced the denitrification ratio.

Evaluation of long-term operational and production performance of biofloc technology (BFT) system within various particle size

Biofloc technology (BFT) is a promising technique, but there has been ongoing debate between biofloc's properties and particle size. This study investigated the relationship between long-term particle size variation and the production performance of BFT aquaculture systems. External shearing force introduction could keep bioflocs at a smaller size level chronically, and 97% of smaller bioflocs were below 181.97 μm which was obviously lower than the normal size group (Control). No significant difference in the daily water treatment capacity for bioflocs within different particle size was detected (p > 0.05), but the activity of Soil-Dehydrogenase (S-DHA), Ammonia Monooxygenase (AMO), Hydroxylamine Oxidase (HAO) and Nitrite Oxidase (NOR) were all higher in smaller bioflocs. Smaller bioflocs were better enriched heavy metal elements and less damage to the gill lamellae was also detected in smaller size group. The survival rate and final mean weight of tilapia (Oreochromis niloticus) cultured in the BFT system based on smaller size bioflocs were significantly higher than the control (p < 0.05). The microbial compositions of varying particle sizes bioflocs demonstrated remarkable distinctions, and higher abundance of particle-attached bacteria (e.g., Flavobacteriaceae, Rhodobacteraceae) was detected in normal size bioflocs. Overall, this study comprehensively evaluated the long-term application of various particle size bioflocs and provided a practical basis for the sustainable BFT development.

Comammox dominate soil nitrification under different N fertilization regimes in semi-arid areas of Northeast China

The newly identified complete ammonia oxidation (Comammox), which is capable of oxidizing ammonia directly to nitrate, has complemented the knowledge of nitrification in the global nitrogen (N) cycle. Knowledge of the community compositions and contributions to nitrification of autotrophic ammonia-oxidizing archaea (AOA) and bacteria (AOB) and Comammox remain void. In this study, the abundance, community compositions, and contributions to nitrification of AOA, AOB, and Comammox were observed in five N gradients (0, 90, 150, 210, and 270 kg ha−1) in a semi-arid area of Northeast China. The results indicated that, compared with low N application rates, higher N application rates significantly improved the soil ammonia monooxygenase (AMO) and the hydroxylamine oxidase (HAO) activities while total nitrogen (TN) was noted to be the main factor driving AMO and HAO activities. Soil potential nitrification rate (PNR) significantly increased with the increase in N application rate, and soil PNR was positively correlated with TN, nitrate nitrogen (NO3−-N), soil organic matter (SOM), and ammonium nitrogen (NH4+-N). The structural equation model (SEM) showed that TN was the main factor driving the community composition of Comammox; SOM had a greatest effect on the community composition of AOA while NH4+-N had a greatest effect on the community composition of AOB. The soil PNR had a direct effect on the yield. Overall, the findings of this study highlighted that N application was more conducive to the reproduction of soil nitrifying microorganisms while Comammox was the dominant contributor to nitrification under N fertilization regimes in the semi-arid area of Northeast China.

Efficient nitrogen removal from municipal wastewater by an autotrophic-heterotrophic coupled anammox system: The up-regulation of key functional genes

The metabolic pathways based on key functional genes were innovatively revealed in the autotrophic-heterotrophic coupled anammox system for real municipal wastewater treatment. The nitrogen removal performance of the system was stabilized at 88.40 ± 3.39 % during the treatment of real municipal wastewater. The relative abundances of the nitrification functional genes ammonia oxidase (amoA/B/C), hydroxylamine oxidoreductase (hao), and nitrite oxidoreductases (nxrA/B) were increased by 1.2–2.4 times, and these three nitrification functional genes were mostly contributed by Nitrospira that dominated the efficient nitrification of the system. The relative abundance of anammox bacteria Candidatus Brocadia augmented from 0.35 % to 0.75 %, accompanied with the increased expression of hydrazine synthase (hzs) and hydrazine dehydrogenase (hdh), resulting in the major role of anammox (81.24 %) for nitrogen removal. The expression enhancement of the functional genes nitrite reductase (narG/H, napA/B) that promoted partial denitrification (PD) of the system weakened the adverse effects of the sharp decline in the population of PD microbe Thauera (from 5.7 % to 2.2 %). The metabolic module analysis indicated that the carbon metabolism pathways of the system mainly included CO2 fixation and organic carbon metabolism, and the stable enrichment of autotrophic bacteria ensured stable CO2 fixation. Furthermore, the enhanced expression of the glucokinases (glk, GCK, HK, ppgk) and the abundant pyruvate kinase (PK) achieved stable hydrolysis ability of organic carbon metabolism function of the system. This study offers research basics to practical application of the mainstream anammox process.

Sludge granulation in PN/A enhances nitrogen removal from mainstream anaerobically pretreated wastewater

Treating anaerobically pretreated wastewater using partial nitritation/anammox (PN/A) process faces severe challenges because of the complex syntrophic and competitive relationship among various bacteria. Results of this study suggested a continuous low dissolved oxygen (DO) concentration failed to sustain NH4+ removal (<80 %), whereas moderate DO concentrations with high aerobic periods suppressed anammox reaction. Through implementing a moderate DO concentration with low aerobic periods (MDO-LA), NH4+ and total nitrogen removal efficiency reached 91.5 ± 5.5 % and 71.3 ± 2.8 % respectively. The specific activities of ammonium-oxidizing bacteria (AOB) and anaerobic ammonium-oxidizing bacteria (AnAOB) reached 0.942 ± 0.030 and 0.277 ± 0.010 g nitrogen per gram mixed liquor volatile suspended solids, respectively, mainly because MDO-LA favored Thiothrix (filamentous bacteria) wash-out and promoted Nitrosomonas growth. Moreover, sludge granules covered by a thin exterior rim with abundant AOB were formed, favoring Ca. Brocadia growth (5.4 % to 13.2 %) and mass transfer between AOB and AnAOB, which consequently increased the expression of genes coding hydroxylamine oxidase and hydrazine synthase. Overall, achievements in this study provide a promising operating strategy for PN/A treating anaerobically pretreated wastewater.

Mixed legume-grass seeding and nitrogen fertilizer input enhance forage yield and nutritional quality by improving the soil enzyme activities in Sichuan, China.

Information regarding relationships between forage yield and soil enzymes of legume-grass mixtures under nitrogen (N) fertilization can guide the decision-making during sustainable forage production. The objective was to evaluate the responses of forage yield, nutritional quality, soil nutrients, and soil enzyme activities of different cropping systems under various N inputs. Alfalfa (Medicago sativa L.), white clover (Trifolium repens L.), orchardgrass (Dactylis glomerata L.), and tall fescue (Festuca arundinacea Schreb.) were grown in monocultures and mixtures (A1: alfalfa, orchardgrass, and tall fescue; A2: alfalfa, white clover, orchardgrass, and tall fescue) under three N inputs (N1: 150kg ha-1; N2, 300kg ha-1; and N3: 450kg ha-1) in a split plot arrangement. The results highlight that A1 mixture under N2 input had a greater forage yield of 13.88t ha-1 year-1 than the other N inputs, whereas A2 mixture under N3 input had a greater forage of 14.39t ha-1 year-1 than N1 input, but it was not substantially greater than N2 input (13.80t ha-1 year-1). The crude protein (CP) content of grass monocultures and mixtures significantly (P < 0.05) increased with an increase in the rate of N input, and A1 and A2 mixtures under N3 input had a greater CP content of 18.91% and 18.94% dry matter, respectively, than those of grass monocultures under various N inputs. The A1 mixture under N2 and N3 inputs had a substantially greater (P < 0.05) ammonium N content of 16.01 and 16.75 mg kg-1, respectively, whereas A2 mixture under N3 had a greater nitrate N content of 4.20 mg kg-1 than the other cropping systems under various N inputs. The A1 and A2 mixtures under N2 input had a substantial higher (P < 0.05) urease enzyme activity of 0.39 and 0.39 mg g-1 24 h-1 and hydroxylamine oxidoreductase enzyme activity of 0.45 and 0.46 mg g-1 5 h-1, respectively, than the other cropping systems under various N inputs. Taken together, growing legume-grass mixtures under N2 input is cost-effective, sustainable, and eco-friendly, which provide greater forage yield and improved nutritional quality by the better utilization of resources.