Ammonia Monooxygenase Activity Research Articles

Effects of exogenous thermophilic bacteria and ripening agent on greenhouse gas emissions, enzyme activity and microbial community during straw composting

This research examined the impact of exogenous thermophilic bacteria and ripening agents on greenhouse gas (GHG) emission, enzyme activity, and microbial community during composting. The use of ripening agents alone resulted in a 30.9 % reduction in CO2 emissions, while the use of ripening agents and thermophilic bacteria resulted in a 50.8 % reduction in N2O emissions. Pearson’s analysis showed that organic matter and nitrate nitrogen were the key parameters affecting GHG emissions. There was an inverse correlation between CO2 and CH4 releases and methane monooxygenase α subunit and N2O reductase activity (P<0.05). Additionally, N2O emissions were positively related to β-1, 4-N-acetylglucosaminidase, and ammonia monooxygenase activity (P<0.05). Deinococcota, Chloroflexi, and Bacteroidota are closely related to CO2 and N2O emissions. Overall, adding thermophilic bacteria represents an effective strategy to mitigate GHG emissions during composting.

Ammonia monooxygenase-mediated cometabolic biotransformation of volatile 4-chlorophenol in nitrifying counter-diffused biofilms: A combined molecular dynamics simulation, DFT calculation and experimental study

Ammonia monooxygenase (AMO)-mediated cometabolism of organic pollutants has been widely observed in biological nitrogen removal process. However, its molecular mechanism remains unclear, hindering its practical application. Furthermore, conventional nitrification systems encounter significant challenges such as air pollution and the loss of ammonia-oxidizing bacteria, when dealing with wastewater containing volatile organic pollutants. This study developed a nitrifying membrane-aerated biofilm reactor (MABR) to enhance the biodegradation of volatile 4-chlorophenol (4-CP). Results showed that 4-CP was primarily removed via Nitrosomonas nitrosa-mediated cometabolism in the presence of NH4+-N, supported by the increased nicotinamide adenine dinucleotide (NADH) and adenosine triphosphate (ATP) content, AMO activity and the related genes abundance. Hydroquinone, detected for the first time and produced via oxidative dechlorination, as well as 4-chlorocatechol was primary transformation products of 4-CP. Nitrosomonas nitrosa AMO structural model was constructed for the first time using homology modeling. Molecular dynamics simulation suggested that the ortho-carbon in the benzene ring of 4-CP was more prone to metabolismcompared to the ipso-carbon. Density functional theory calculation revealed that 4-CP was metabolized by AMO via H-abstraction-OH-rebound reaction, with a significantly higher rebound barrier at the ipso-carbon (16.37 kcal·mol-1) as compared to the ortho-carbon (6.7 kcal·mol-1). This study fills the knowledge gap on the molecular mechanism of AMO-mediated cometabolism of organic pollutants, providing practical and theoretical foundations for improving volatile organic pollutants removal through nitrifying MABR.

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.

Impacts of exogenous C6-HSL on the enhanced cometabolic removal of sulfadiazine by enriched nitrifying sludge

The antibiotics in the wastewater could be effectively removed by the cometabolism of nitrifying sludge mediated by ammonia monooxygenase (AMO). The effects of quorum sensing (QS) system regulated by N-acyl homoserine lactones (AHLs) on the performance and functional microorganisms in wastewater treatment have also been increasingly reported, yet few studies report the impacts of AHLs on the cometabolic removal of antibiotics by nitrifying bacteria. In this study, N-hexanoyl-L-homoserine lactone (C6-HSL) and sulfadiazine (SDZ) were selected, and a series of batch tests were implemented to study the cometabolic degradation of SDZ by enriched nitrifying sludge and the responses of the sludge under different concentrations of exogenous C6-HSL (0.1, 0.5 and 1 μM). The results indicated C6-HSL enhanced the cometabolic removal of SDZ by nitrifying sludge, with the highest removal efficiency of 84.60 % achieved with 1 μM C6-HSL. The extracellular polymeric substances (EPS) increased by 50.8 %–139.5 %，and a positive correlation between the increase of protein content in EPS and the concentration of C6-HSL was established. C6-HSL also brought an increase in cellular adenosine triphosphate content, which increased by 51.8 % with 1 μM C6-HSL. Moreover, C6-HSL up-regulated the amoA gene and enhanced AMO activity, which led to the sufficient degradation of SDZ.

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.

Italian ryegrass (Lolium multiflorum L.)-rice (Oryza sativa L.) rotation promotes the nitrogen cycle in the rice rhizosphere through dominant ammonia-oxidizing bacteria

The dominant ammonium bacteria established in different crop rotations are directly related to the conversion and utilization of nitrogen (N) in paddy soil. This study investigated the ammonia oxidation process in the rice rhizosphere of Italian ryegrass (Lolium multiflorum L.)-rice (IRR), celery (Apium graveolens)-rice (CLR) and winter fallow-rice (WFR) rotations in South China through the amoA functional gene analysis based on Miseq high-throughput sequencing platform and qPCR methods. The abundance of amoA gene in IRR was increased by 116.67 % (P < 0.05) and 42.76 % (P > 0.05) compared with CLR and WFR, respectively. Nitrosomonas became the dominant genus in the rhizosphere of IRR, with the highest relative abundance (59.30 %). The contents of soil organic matter, total N and nitrate nitrogen (NO3−-N) in the rice rhizosphere soil in IRR were significantly higher than other two treatments (P < 0.05) by 12.47–14.31 %, 16.88–25.00 %, and 97.48–159.60 %, respectively. Although lower microbial diversity was observed in IRR, the soil fluorescein diacetate hydrolysis enzyme activities and ammonia monooxygenase activities in the rice rhizosphere were higher in IRR than those in CLR and WFR (P < 0.05). Also, the soluble sugar and crude protein of milled rice were highest in IRR among the three rotation treatments (P < 0.05). The ammonia oxidation efficiency of ammonium-oxidizing bacteria was largely dependent on key taxa, such as Nitrosomonas (dominant genus in IRR), rather than the dynamics of AOB community structure. The key amoA-nitrifier taxa of the AOB community developed in IRR are capable of promoting rice production by improving soil N cycle efficiency and reducing the loss of N. Our findings illustrate the regulatory mechanism of IRR on the rice rhizosphere AOB community and reveal the important role of winter ryegrass in promoting soil nitrogen cycling during nutrient transformation of agricultural ecosystems in paddy fields.

Effects of UV-B Radiation on the Chemical Composition of Azolla and Its Decomposition after Returning to the Field and Nitrogen Transformation in Soil

In the present research, the effects of UV-B radiation (5.00 kJ·m−2) on the chemical composition of Azolla were investigated, and the decomposition of Azolla residues after UV-B radiation, the nitrogen form, enzyme activity, and bacterial community in paddy soil were analyzed. Compared to the natural light treatment, the total nitrogen content of Azolla was significantly increased by 17.0% under UV-B radiation treatment. Compared to returned Azolla grown under natural light, the decomposition rate of cellulose, lignin, and total nitrogen of returned Azolla grown under UV-B radiation significantly increased, which led to an increase in the activities of nitrogen transformation enzymes, including neutral protease, ammonia monooxygenase, nitrogenase, nitrate reductase, and nitrite reductase, and the contents of different nitrogen forms (NH4+-N, NO3−-N, soluble organic nitrogen, and microbial biomass nitrogen) in paddy soil, while N2O emission flux was significantly reduced by 20–30%. The dominant bacteria in soil supplied with Azolla grown under natural light were Firmicutes, Clostridia, Clostridiales, and Lachnospiraceae. However, returning Azolla grown under UV-B radiation to the soil significantly changed the bacterial community structure in soil, resulting in a decrease in the number of ammonifying bacteria, nitrifying bacteria, and nitrogen-fixing bacteria and an increase in the number of denitrifying bacteria, inducing changes in the dominant bacteria to Methanomicrobiales, Methanoregulaceae, and Methanoregula. According to the structural equation model, returning Azolla to the field would reduce N2O emissions by increasing Azolla lignin decomposition and ammonia monooxygenase activity, reducing the number of nitrifying bacteria and reducing nitrite reductase activity in soil. Thus, UV-B radiation can directly change the phytochemical components and their decomposition in soil, thus indirectly affecting the bacterial community structure, enzyme activity, and nitrogen transformation, which play important ecological roles in regulating the nutrient transformation of terrestrial ecosystems.

Understanding the cometabolic degradation of sulfadiazine by an enriched ammonia oxidizing bacteria culture from both extracellular and intracellular perspectives

Antibiotics are widely used drugs in the world and pose serious threats to ecosystems and human health. Although it has been reported that ammonia oxidizing bacteria (AOB) can cometabolize antibiotics, little has been reported on how AOB would respond to the exposure of antibiotics on extracellular and enzymatic levels, as well as the impact of antibiotics on the bioactivity of AOB. Therefore, in this study, a typical antibiotic, sulfadiazine (SDZ), was selected, and a series short-term batch tests using enriched AOB sludge were conducted to investigate the intracellular and extracellular responses of AOB along the cometabolic degradation process of SDZ. The results showed the cometabolic degradation of AOB made the main contribution to SDZ removal. When the enriched AOB sludge was exposed to SDZ, ammonium oxidation rate, ammonia monooxygenase activity, adenosine triphosphate concentration and dehydrogenases activity were negatively affected. The amoA gene abundance increased 1.5 folds within 24 h, which may enhance the uptake and utilization of substrates and maintain stable metabolic activity. In the tests with and without ammonium, the concentration of total EPS increased from 264.9 to 231.1 mg/gVSS to 607.7 and 538.2 mg/gVSS, respectively, under the exposure to SDZ, which was mainly contributed by the increase of proteins in tightly bound extracellular polymeric substances (EPS) and polysacharides in tightly bound EPS and soluble microbial products. The proportion of tryptophan-like protein and humic acid-like organics in EPS also increased. Moreover, SDZ stress stimulated the secretion of three quorum sensing signal molecules, C4-HSL (from 140.3 to 164.9 ng/L), 3OC6-HSL (from 17.8 to 42.4 ng/L) and C8-HSL (from 35.8 to 95.9 ng/L) in the enriched AOB sludge. Among them, C8-HSL may be a key signal molecule that promoted the secretion of EPS. The findings of this study could shed more light on the cometabolic degradation of antibiotics by AOB.

Biochar-mediated remediation impacts on nitrogen cycling bacteria and ammonia monooxygenase activity in crude oil polluted soil

This study adopted an ecosystem services approach to pollution management by investigating the impact of biochar-mediated remediation on soil nitrogen, abundance of nitrogen cycling bacteria and the activity of ammonia monooxygenase (AMO) enzyme in petroleum-polluted soil using two biochar types applied at two treatment levels with monitoring over 15 weeks. The corn cob-derived biochar (CDB), generally, had a stronger restorative effect on soil ammonium nitrogen, nitrate and total organic nitrogen concentrations than the bone-derived biochar (BDB). Both biochar types had a more robust impact on restoration of Nitrosomonas, Nitrobacter and Azotobacter abundance (with the re-establishment of pre-pollution levels) than on Rhizobium and Pseudomonas aeruginosa. Biochar amendment restored the activity of AMO enzyme in the soil by week 15. The CDB (72.4% – 73.7%) showed more effective total petroleum hydrocarbon (TPH) elimination capacity than the BDB (51.1% – 57.7%). Biochar amendments exhibited great potential for restoration of nitrogen cycling while facilitating remediation of petroleum-polluted soils.

Dissolved oxygen benefits N-decanoyl-homoserine lactone regulated biological nitrogen removal system to resist acute ZnO nanoparticle exposure

The beneficial effects of N-decanoyl-homoserine lactone (C10-HSL), one of the typical N-acyl-homoserine lactones on biological nitrogen removal (BNR) system to resist the acute exposure of zinc oxide nanoparticles (ZnO NPs) has attracted extensive attentions. Nevertheless, the potential impact of dissolved oxygen (DO) concentration on the regulatory capacity of C10-HSL in the BNR system has yet to be investigated. This study conducted a systematic investigation of the impact of DO concentration on the C10-HSL-regulated BNR system against short-term ZnO NP exposure. Based on the findings, sufficient DO played a crucial role to improve the BNR system's resistance capacity to ZnO NPs. Under the micro-aerobic condition (0.5 mg/L DO), the BNR system was more sensitive to ZnO NPs. The ZnO NPs induced increased intracellular reactive oxygen species (ROS) accumulation, reduced antioxidant enzyme activities, and decreased specific ammonia oxidation rates in the BNR system. Furthermore, the exogenous C10-HSL had a positive effect on the BNR system's resistance to ZnO NP-induced stress, primarily by decreasing ZnO NPs-induced ROS generation and improving ammonia monooxygenase activities, especially under low DO concentrations. The findings contributed to the theoretical foundation for regulation strategy development of wastewater treatment plants under NP shock threat.

Reclamation intensifies the positive effects of warming on N2O emission in an alpine meadow.

Climatic warming can alter grassland nitrous oxide (N2O) emissions due to soil property alterations. However, how the reclamation affect grassland N2O flux under warming conditions remains unclear in alpine meadow ecosystems. We conducted a long-term manipulative warming experiment in a natural alpine meadow and a cultivated grassland on the Qinghai-Tibetan Plateau to explore the separate and interactive effects of warming and reclamation on the soil N2O emission flux. N2O fluxes were measured under four treatments including control (CK), warming (W), reclamation (R) and warming under reclamation (WR) from August 2018 to July 2019. We measured the content of soil C, N nutrients and 5 enzymatic activities in 2018 and 2019. Correlation analysis and structural equation modeling were used to clarify how soil N availability and soil enzyme activities affect N2O emission. Our results indicated that compared to the ambient conditions for the growing and non-growing seasons, soil N2O flux was significantly increased 59.1% and 152.0% by warming and 28.4% and 142.4% by reclamation, respectively. Compared with W, WR significantly increased N2O flux by 18.9% and 81.1% during the growing and non-growing seasons, respectively. Soil moisture was negatively correlated to enzymatic activity and N2O flux. Both warming and reclamation promoted soil nitrification by increasing related enzymatic activities that acted to increase the N2O flux. Reclamation resulted in a greater sensitivity of the activity of ammonia monooxygenase and hydroxylamine oxidoreductase to warming, thus enhancing the effects of warming on increasing the N2O flux. Our research indicated that reclamation can additionally increase the effects of warming on N2O emissions for alpine meadows. Therefore, excessive expansion of arable land should be avoided, and new reclamation sites should be planned scientifically, as warming is expected to intensify in the future.

Effect of biochar and nitrogen fertilizers on maize seedling growth and enzyme activity relating to nitrification

AbstractMaximizing nitrogen use efficiency (NUE) involves synchronizing the interplay between nitrogen preferential crops and the nitrogen transformation pathways of soil. Biochar may benefit specific N‐preference crops in relatively unsuitable soil environments; however, experimental data are lacking. This study tested eight treatments, consisting of four nitrogen treatments (N0 = control; N1 = NH4Cl; N2 = NaNO3; and N3 = 1:1 ratio of NH4+ and NO3−) each with biochar applied at 0% or 2% (w/w). The results show that biochar and/or nitrogen application enhanced maize seedling biomass and NO3−‐based fertilizer resulted in higher seedling biomass than NH4+‐based fertilizer. With the application of biochar and NH4+‐based fertilizer, maize seedling biomass increased and soil NH4+‐N content was significantly reduced compared with NH4Cl sole application. Correlation analysis and redundancy analysis revealed that SOC content and inorganic nitrogen content were the main factors influencing maize growth and N absorption. Biochar with or without nitrogen fertilizer (except N1 treatment) significantly increased β‐1,4‐glucosidase (BG) activity. Co‐application treatments also resulted in higher vector length, an indicator of C limitation—the increment might add to the risk of microbial C limitation. The activity of ammonia monooxygenase (AMO), a key enzyme in nitrification, decreased with the co‐application of biochar and nitrogen, suggesting the alteration of nitrogen transformation.

Agronomic performances of biodegradable and non-biodegradable plastic film mulching on a maize cropping system in the semi-arid Loess Plateau, China

Biodegradable plastic film mulch (PFM) is considered an alternative to non-biodegradable PFM to mitigate the negative impacts of residual film. However, the agronomic performance of biodegradable PFM in comparison to non-biodegradable PFM still needs to be tested. In this study, we evaluated the effects of biodegradable and non-biodegradable PFM on soil physicochemical properties, microbial community, and enzyme activities, as well as maize growth performance. Biodegradable and non-biodegradable PFM both increased soil temperature, water content, N content, and microbial biomass and maize yield by up to 30%, but decreased soil enzyme activities as compared to no mulching (control, CK). Most soil physicochemical properties, microbial community, and enzyme activities were similar under non-biodegradable and biodegradable PFM at the early stages of maize growth. However, at the late stages, soil temperature, water content, mineral N, NO3--N, ammonia monooxygenase (AMO) activity, and total phospholipid fatty acids (PLFAs) decreased under biodegradable PFM owing to film fragmentation. White PFM increased soil temperature, water content, and total PLFAs at the early stages of maize growth but decreased soil mineral N and total PLFAs at the late stages, as compared to black PFM. As soil temperature and N availability were the major factors affecting soil microbial community, microbial activity decreased after the fragmentation of biodegradable PFM, owing to the decreased soil temperature, water content, and mineral N. Notably, biodegradable PFM could decrease NO3--N accumulation in topsoil by decreasing N transformation due to the lower microbial and N-related enzyme (e.g., AMO) activities, compared with non-biodegradable PFM, which may avoid negative environmental impacts, such as NO3--N leaching or gas emission after harvest. Maize yield, height, aboveground biomass, and N uptake under biodegradable PFM were similar to those under non-biodegradable PFM during maize growth, implying that biodegradable PFM has no negative impact on crop growth and yield. In general, biodegradable PFM was equivalent to non-biodegradable PFM in terms of maize yield increase and N uptake, but was environmentally friendly. Therefore, biodegradable PFM can be used as an alternative to non-biodegradable PFM in semi-arid areas for sustainable agricultural practices.

Impacts of exogenous quorum sensing signal molecule-acylated homoserine lactones (AHLs) with different addition modes on Anammox process

Anammox was proved having the quorum sensing ability, and several acylated homoserine lactones (AHLs) signal molecules were detected in the system. In this study, the impact of exogenous N-dodecanoyl homoserine lactone (C12-HSL) with different addition modes on the nitrogen removal, key enzymes’ activity, and microbial revolution were investigated in Anammox system. Results showed that once-addition of C12-HSL had no obvious impact on Anammox. Daily-addition with 40 nM slightly improved the TN removal from 71.1 % to 74.5 %, while 80 and 200 nM significantly decreased it to 62.7 % and 61.8 %, respectively. The enzyme activity of ammonia monooxygenase increased from 0.015 to 0.068, nitrite reductase increased from 0.25 to 1.23, and nitrate reductase increased from 0.05 to 0.11 μg NO2–-N mg−1 Protein min−1. Arenimonas abundance showed positive correlation with TN removal while Candidatus Kuenenia was continuously suppressed. C12-HSL was beneficial for partial nitrification, and it could be adopted for regulating the nitrite production.

Regulating performance of CANON process via adding external quorum sensing signal molecules in membrane bioreactor

In this study, the regulation effect of the external quorum sensing signals, N-dodecanoyl homoserine lactone (C12-HSL) on CANON process were investigated in a membrane bioreactor. C12-HSL significantly enhanced the aerobic ammonia-oxidizing bacteria and improved the ammonia monooxygenase activity to 0.134 from 0.076 μg NO2−-N mg−1 protein min−1, while suppressed anaerobic ammonia-oxidizing bacteria and limited the TN removal to 0.07 from 0.22 kg m−3 d-1. Key enzymes synthesis were enhanced during the operation without C12-HSL addition, enabling the resistance of CANON system to high C12-HSL. As a result, the hydroxylamine oxidoreductase and nitrite reductase activity reached 35.9 EU g−1 SS and 1.28 μg NO2−-N mg−1 protein min−1, respectively; Nitrosomonas and Candidatus Kuenenia, with the abundance as 12.5 % and 22.9 %, cooperatively contributed to the TN removal, which maintained at 0.19 kg m−3 d-1. C12-HSL was profitable for aerobic ammonia oxidation, which could be adopted for regulating the nitrite production rate.

The response and anti-stress mechanisms of nitrifying sludge under long-term exposure to CdSe quantum dots

The wide application of CdSe quantum dots (QDs) increases its stress risk to sewage treatment systems. This study evaluated the response of nitrification performance, floc characteristics and microbial community of nitrifying sludge under long-term exposure to CdSe QDs. Results showed CdSe QDs (≥1 mg/L) would decrease the activity of ammonia monooxygenase (AMO). Under the stress of 30 mg/L CdSe QDs, the activity of AMO was reduced by 66%, while the activities of hydroxylamine oxidase and nitrite oxidoreductase were enhanced by 19.1% and 26%, respectively. Thus, the final nitrification effects were not adversely affected, and the production rates of NO2−-N and NO3−-N were accelerated. Additionally, CdSe QDs improved biomass concentration in sludge and maintained the stability of sludge settleability. High throughput sequencing analysis showed that CdSe QDs evidently reduced the abundance and diversity of microbial community in nitrifying sludge. The abundances of amino acid metabolism and lipid metabolism were enriched. Moreover, CdSe QDs decreased the fluorescence intensity of tryptophan-like protein from 2,326 to 1,179 a.u. in loosely bound extracellular polymeric substances (EPSs) and from 3,792 to 3,117 a.u. in tightly bound EPSs. To relieve CdSe QD stress, the polysaccharide content increased from 0.31 to 0.61 mg/g MLSS and intracellular antioxidant defense was activated. With CdSe QD level increasing to 30 mg/L, the total antioxygenic capacity and the activities of catalase were enhanced up to 411% and 143.2%, respectively. Thereby, CdSe QDs had little adverse effects on cell membrane integrity, microbial metabolism and the abundance of Nitrospirae.

Nitrogen occurrence forms and bacterial community in sediment influenced by Bellamya purificata bioturbation

Endogenous water pollution and excessive nitrogen in ponds have always been prominent problems threatening aquaculture. To solve this problem, a 70-day indoor simulation experiment was conducted. The snail Bellamya purificata was stocked at four different densities: 0, 15, 30, and 60 individuals/tank, represented as CON (control), LD (low density), MD (medium density), and HD (high density), respectively, to investigate the effects of B. purificata on the nitrogen occurrence forms and bacterial communities in the sediment. At the end of the experiment, the nitrate concentration was significantly higher, while the total nitrogen content was significantly lower in the MD group than in the other three groups. Ammonia monooxygenase activity was significantly lower in the CON group than in the other three groups, and hydroxylamine oxidase activity was significantly higher in the HD group than in the other three groups. The CON and MD groups showed the highest and lowest values in nitrate reductase activity, respectively. The hydroxylamine reductase activity decreased significantly with increasing density. Through 16S ribosomal RNA (rRNA) high-throughput sequencing, significantly affected bacterial communities by B. purificata were found. Alpha diversity results showed that, a significantly lower Shannon index was observed in the MD group than in the other three groups. The LD and MD groups showed the highest and lowest Chao1 index values, respectively. Phyla Nitrospinae and family Nitrosomonadaceae were significantly enriched in the HD and MD groups, respectively. Redundancy analysis (RDA) indicated a significant correlation between differential bacterial taxa and TN content. Predicted functional analysis based on FAPROTAX (Functional annotation of prokaryotic taxa) database showed that functional groups aerobic ammonia oxidation and aerobic nitrite oxidation were significantly enriched in the MD group. Overall, B. purificata significantly altered the bacterial community composition, increased hydroxylamine oxidase and ammonia monooxygenase activities, enhanced the bacterial nitrification process, and promoted the transformation of total nitrogen to nitrate. Moreover, B. purificata stocked at 30 and 60 individuals/tank appeared to have a significant promotion effect on the bacterial community and nitrogen occurrence forms in the sediment. Hence, co-culturing B. purificata may be a feasible and effective ecological restoration method to alleviate excess nitrogen and reduce water eutrophication.

Rapid start-up of partial nitrification reactor by exogenous AHLs and Vanillin combined with intermittent aeration

Quorum sensing (QS) and quorum quenching (QQ) are common phenomena in microbial systems and play an important role in the nitrification process. However, rapidly start up partial nitrification regulated by N-acyl-homoserine lactones (AHLs)-mediated QS or QQ has not been reported. Hence, we chose N-butyryl homoserine lactone (C4-HSL) and N-hexanoyl homoserine lactone (C6-HSL) as the representative AHLs, and Vanillin as the representative quorum sensing inhibitor (QSI) combined intermittent aeration to investigate their effects on the start-up process of partial nitrification. The start-up speed in the group with C4-HSL or C6-HSL addition was 1.42 or 1.26 times faster than that without addition, respectively. Meanwhile, the ammonium removal efficiency with C4-HSL or C6-HSL addition was increased by 13.87 % and 17.30 % than that of the control group, respectively. And, partial nitrification could maintain for a certain period without AHLs further addition. The increase of Nitrosomonas abundance and ammonia monooxygenase (AMO) activity, and the decrease of Nitrobacter abundance and nitrite oxidoreductase (NXR) activity were the reasons for the rapid start-up of partial nitrification in the AHLs groups. Vanillin addition reduced AMO and hydroxylamine oxidoreductase (HAO) activity, and increased Nitrobacter abundance and NXR activity, thus these were not conducive to achieving partial nitrification. Denitrifying bacteria (Hydrogenophaga, Thauera and Aquimonas) abundance increased in the Vanillin group. QS-related bacteria and gene abundance were elevated in the AHLs group, and reduced in the Vanillin group. Function prediction demonstrated that AHLs promoted the nitrogen cycle while Vanillin enhanced the carbon cycle. This exploration might provide a new technical insight into the rapid start-up of partial nitrification based on QS control.

Synergistically mitigating nitric oxide emission by co-applications of biochar and nitrification inhibitor in a tropical agricultural soil

Agricultural soils are the hotspots of nitric oxide (NO) emissions, which are related to atmospheric pollution and greenhouse effect. Biochar application has been recommended as an important countermeasure, however, its mitigation efficiency is limited as biochar, under certain conditions, can stimulate soil nitrification. Therefore, biochar co-applied with nitrification inhibitor could optimize the mitigation potential of biochar. Herein, a laboratory-scale experiment was conducted to investigate the effects of co-application of biochar and nitrification inhibitor on NO emission, nitrogen cycling function and bacterial community in a tropical vegetable soil. Results showed that a single application of biochar or nitrification inhibitor significantly decreased NO emissions, and this mitigation effectiveness was amplified by their co-applications. Soil NO2−−N intensity, along with abundances of AOB-amoA and nirK were significantly and positively correlated with cumulative NO emissions. The stimulated activity of ammonia monooxygenase and growths of AOB and total comammox Nitrospira by biochar were weakened by nitrification inhibitor, implying decreased nitrification-driven NO production. The nitric oxide reductase activity and related qnorB abundance in nitrification inhibitor-added soils were increased by biochar, indicating promoted NO consumption during denitrification. The nirK abundance and NO2−−N intensity were decreased more by co-applications of biochar or nitrification inhibitor. Moreover, both biochar and nitrification inhibitor changed bacterial β-diversity, and their co-application synergistically enriched Armatimonadetes and Verrucomicrobia abundances and decreased WPS-2 abundance. This study highlights that co-applications of biochar and nitrification inhibitor can make their respective advantages complementary to each other, thereby achieving a larger mitigation of NO emissions from agricultural soils in tropical regions.

Response mechanism of microorganisms to the inhibition of endogenous pollution release by calcium peroxide

To further explore the response mechanism of microorganisms to the synchronous control of nitrogen and phosphorus release from sediments by CaO2, the spatiotemporal changes in the physical, chemical and biological indicators of the overlying water, interstitial water and sediments in each reactor were measured in the experiment. The experiment results showed that CaO2 could increase the ammonia monooxygenase activity, nitrite oxidase activity and Nitrospira abundance in the sediment near its dosing position, and enhanced the activities of nitrate reductase and nitrite reductase at a certain distance from the dosing position, thereby promoting nitrogen removal in sediments through the alternating process of nitrification and denitrification. At the same time, the increase of alkaline phosphatase activity and Saccharimonadales abundance in the test groups accelerated the hydrolysis of organic phosphorus, and the P immobilization in sediments was realized through the subsequent precipitation reaction of Ca2+ and PO43− under alkaline conditions. In addition, the enhanced activities of dehydrogenase and catalase ensured that CaO2 would not cause great killing effect on microorganisms when improving the hypoxic conditions and inhibiting endogenous release. As a result, the dissolved product of CaO2 such as Ca(OH)2 and H2O2 reduced the nutrients concentration and killed the algae, which kept the algae density and chlorophyll a concentration at a low level throughout the test groups. Therefore, this study systematically clarified the microbial mechanism of CaO2 synchronously controlling the release of nitrogen and phosphorus from sediments, which provided a new idea for the remediation of endogenous pollution in the water system.