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Nitrogen removal and sludge reduction in a symbiotic activated sludge system between anaerobic archaea and bacteria.

The possible symbiosis between bacteria and anaerobic archaea was investigated in intermittent aeration (I/A) systems. Archaea solution added to I/A reactor might play an important role in biological activities as well as in improvement of mineralization of organic matter. I/A reactor with archaea solution (I/A-arch) could increase both nitrification and denitrification rate and also reduce the sludge yield remarkably. These results indicate the possibility of the symbiotic activated sludge system with anaerobic archaea by controlling the DO level in the aeration tank. In this study, DO was controlled by intermittent aeration schemes and a successful symbiotic activated sludge system was achieved to reach the following conclusions. 1) SOUR of I/A-arch system was 2.9 mg-O2/g-VSS x min. SOUR and nitrification rate of the sludge from I/A-arch was higher than those from the I/A and A/S reactors. 2) Removal efficiencies of organic matter (TCOD(Cr)) in I/A-arch, I/A and conventional activated sludge (A/S) reactors were 93, 90 and 87%, respectively. 3) Nitrification occurred successfully in each reactor, while denitrification rate was much higher in the I/A-arch reactor. Efficiencies of TN removal in A/I-arch, I/A and A/S reactors were 75, 63 and 33%, respectively. 4) Observed yield coefficients of I/A-arch, I/A and A/S reactors were 0.28, 0.41 and 0.37 g-VSS/g-COD.

Anaerobiosis↗

Nitrogen removal in a fluidized bed bioreactor by using mixed culture under oxygen-limited conditions.

Nitrogen removal involving nitrification and denitrification was investigated in a fluidized bed bioreactor by using mixed culture sludge under oxygen-limited conditions. Methane was used as a sole carbon source for denitrification. In this study, optimal nitrification and denitrification rates were examined by varying methane and oxygen gas dissolution flow rates, 90 ml/min, 400 ml/min and 650 ml/min, in each. Simultaneously nitrification and denitrification was achieved. The total nitrogen removal rate was 15-mg N/g VSS. d, 21-mg N/g VSS. d and 26.4-mg N/g VSS. d at gas dissolution flow rate 90 ml/min, 400 ml/min and 650 ml/min, respectively. No significant accumulation of nitrite was found in this experiment. Nitrogen removal rates depend on gas dissolution flow rates. DO concentration was at 0.5-2 mg/L.

Bioreactors↗

[Effects of elemental sulphur and dicyandiamide on mitigating NO3- -N leaching loss from vegetable soil].

In a pot experiment with allium as test plant and NH4HCO3 as nitrogen source, this paper studied the effects of element sulphur (S0) and dicyandiamide (DCD) on mitigating the NO3- -N leaching loss from soil and on soil inorganic nitrogen (NO3- -N and NH4+ -N) content. The results showed that within the 12 weeks of the experiment, the cumulative leaching loss of soil NO3- -N in treatments S0 + DCD and S0 was 83%-86% and 83% lower, while that of soil NH4+ -N was 16.8-21.0 mg x pot(-1) and 20.4-25.0 mg x pot(-1) higher than CK, respectively, and the cumulative loss of soil (NH4+ + NO3-)-N was 60% lower. By the end of the experiment, soil inorganic nitrogen content in treatments S0 + DCD and S0 was 79.9%-85.4% and 74.9-82.6% higher than CK, respectively. The cumulative leaching loss of inorganic N in treatment S0 + DCD was 4.6%-14.4% and 15.4%-30.1% lower, and the soil inorganic nitrogen content by the end of the experiment was 6.1% and 16.8%-36.0% higher than that of treatments S0 and DCD, respectively. Similar results were obtained when S0 was replaced by Na2S2O3, but not by Na2SO4. The fact that the application of S0 could obviously decrease the NO3- -N leaching loss from soil could be contributed to the inhibitory effects of S2O3(2-) and S4O6(2-) originated from S0 oxidation in soil on the nitrification of NH4+ -N. S0 could retard the decomposition of DCD due to the effect of its oxidized products S2O3(2-) and S4O6(2-), and thus, extend the inhibitory effect of DCD on NH4+ -N nitrification. It is suggested that S0 combined with DCD could be used as an effective nitrification inhibitor to control the NO3- -N leaching loss from vegetable soils.

Fertilizers↗

In situ net mineralization rates in a heterogeneous mixed deciduous forest receiving elevated n deposition.

In situ net mineralization was studied at 6 locations (E, Eb, Ec, F1, F2, F3) of a heterogeneous mixed deciduous forest ("De Gulkeputten") with oak (Quercus robur L., Quercus rubra) and birch (Betula pendula) as dominant species. Net nitrogen mineralization was determined by means of a sequential in situ incubation experiment using intact soil cores. For all incubations, the net mineralization rates of the organic (F+H) layer varied between -0.4 and 2.0 g N m(-2) month(-1), while the net nitrification rates varied between -0.6 and 0.7 g NO3- -N m(-2) month(-1). The net mineralization and nitrification rates of the mineral (0-30 cm) layer ranged from 4.5 g N m(-2) month(-1)to 8.8 g N m(-2) month(-1)and from -1.0 to 4.9 g NO3- -N m(-2) month(-1) respectively. In general, net mineralization rates increased from August 1998 to October 1998. Net mineralization rates were positively correlated with the gravimetrical moisture content and mineralization and nitrification rates were mutually positively correlated.

Betula↗

Effectiveness of oxidation-reduction potential and pH as monitoring and control parameters for nitrogen removal in swine wastewater treatment by sequencing batch reactors.

Two bench-scale sequencing batch reactors (SBRs) were operated in a fixed hydraulic retention time study to investigate the effectiveness of oxidation-reduction potential (ORP), pH and dissolved oxygen as parameters for indicating denitrification followed by nitrification in SBRs for swine wastewater treatment. The ORP and pH profiles were monitored and evaluated under different denitrification and nitrification conditions with and without a supplemental carbon source. With a low C/N ratio, and using a suitable C/N ratio adjustment control, ORP and pH could be used as monitoring and control parameters in both the anoxic and oxic phases for practical swine wastewater treatment. High-level accumulation of nitrate was observed without any C/N ratio adjustment. In this case, ORP and pH were not useful for monitoring denitrification followed by nitrification in SBRs. According to our research, with regard to N removal, it would be better to use pH as a parameter during the oxic phase and ORP as a parameter during the anoxic phase. Using a suitable adjustment of a ON ratio in the influent by adding swine slurry, a high total nitrogen removal efficiency of up to 95.5% was reached. It was found that, in this case, the use of ORP and pH as parameters for real-time control processes was possible in swine wastewater treatment.

Journal Article↗

A comparison between the theory and reality of full-scale step-feed nutrient removal systems.

Capacity enhancement and volume reduction benefits of step-feeding fully aerobic bioreactors has been well documented. Application of step-feed technology to biological nutrient removal (BNR) systems, particularly those removing nitrogen alone or both nitrogen and phosphorus, is relatively new to the industry. In recent years, a number of full-scale step-feed facilities have been brought into service. This paper reviews nine full-scale step-feed biological nutrient removal systems--both nitrogen removal alone, and nitrogen and phosphorus removal. The objective is to compare the theoretical benefits of such systems with their actual operation. The predicted benefits of reduced bioreactor volume or increased process capacity, reduced energy usage, more robust nitrification performance, and the flexibility to tune (or de-tune) nitrification efficiency were verified in full-scale systems. Equations are also presented that may be used in the prediction of step-feed benefits. There are two primary drivers for considering a step-feed biological reactor system: 1. Reduced bioreactor volume for a defined capacity or performance or increased process capacity given a fixed bioreactor volume. 2. More robust nitrification performance. Full-scale operation of these step-feed nutrient removal systems provides a real world basis for the claimed benefits of step-feed operation. These systems have uniformly shown additional capacity. A number of them have also exhibited more robust performance, especially during storms. Where possible, side-by-side comparisons of full-scale step-feed systems with non-step-feed systems have exhibited greater process reliability and flexibility.

Bioreactors↗

[Effect of nitrite on aerobic phosphate uptake by phosphate accumulating organisms].

Effect of nitrite at various concentration levels on aerobic phosphate uptake was investigated through a series of batch experiments. Furthermore, the effect of nitrite accumulated in the process of nitrification on aerobic phosphate uptake was studied in saline wastewater treatment. The results show that NO2(-) -N concentration of 4 mg/L inhibits aerobic phosphate uptake by phosphate uptake by phosphate accumulating organisms (PAO) and phosphate uptake rate decreases 8%. Exposure to higher nitrite concentration levels inhibits aerobic phosphate uptake severely. At NO2(-) -N concentration of 15 mg/L, phosphate uptake rate decreases 61%. The toxic effect of nitrite is presumed to be linked with free nitrous acid (FNA). Significant inhibition on aerobic phosphate uptake appeared at 0.000 2 mg/L of FNA concentration. The inhibiting effect of nitrite is found to occur only when nitrite is present. The ability of accumulating phosphate resumes afternitrite is no longer present. During the process of nitrification of nitrification of saline wastewater, the inhibition of nitrite on aerobic phosphate uptake is slight at the initial 1 - 2 h due to low nitrite accumulation. With the build-up of NO2(-) -N(up to about 8 to 9 mg/L), the inhibiting effect of nitrite increases gradually. It is found that higher ammonium concentration causes lower pH value and higher FNA concentration, which could decrease the amount of phosphate uptake.

Bacteria, Aerobic↗

[Influence of N-serve on mineral nitrogen forms and microorganisms of the soil (author's transl)].

In a model experiment with soil, 2 stages of nitrogen (without additional N and with 200 kg N/ha in the form of ammonia) and 3 N-Serve stages (without inhibitor and with 0.25 per cent and 1 per cent of inhibitor, relative to 200 kg of N/ha) were followed up over a period of 3 and 6 weeks, respectively, with respect to the dynamics of the nitrification process, the saprophytic soil microorganisms, and the autotrophic nitrifiers. In a second experiment, the ammonia form of nitrogen was substituted by the nitrate and nitrite form, respectively. No serious influence on the heterotrophic soil micro-organisms could be detected in any experimental variant; N-Serve was slightly stimulating, the autotrophic nitrifiers were noticeably depressed by N-Serve. The nitrification process could clearly be proved after 3 weeks already; by N-Serve it was remarkably delayed. Nitrite that had been added to the soil was scarcely transformed into nitrate, whereas it was somewhat more ammonified. The high loss of nitrogen in the nitrite variant cannot be caused by denitrification alone. When nitrate was used, the nitrification was not delayed.

Ammonia↗

[The effect of biocides on the microflora of soils and their degradation. 4. The influence of substances inhibiting the straw decomposition on the nitrogen transformation (author's transl)].

1. Active ingredients retarding the carbon-transformation of straw (TCP, Thiuram, TBTO) also retard the nitrification. Their active period depends on the concentration and on the presence of organic matter. High doses of urea can inhibit the nitrification as well. 2. The activity of urease and the ammonification could not be influenced by adequate doses of the tested active ingredients. 3. The species of bacterium Azotobacter fixing the nitrogen from the air were inhibited by TCP, urea and D-chloramphenicol and promoted by Thiuram and WTZ II 277 after a short period of inhibition. 4. The fungi are very important in the biological nitrogen fixation after manuring straw and in prevention of nitrogen leaching in the autumn. 5. The sufficient concentration of the active ingredients to inhibit the nitrification are lower than the fungicidal concentrations. The soil bacteria could even not be restrained by doses of 5000 ppm. The bacilli were more retarded in presence of organic matters than in a not manured soil.

Ammonia↗

[Biotransformation of nitrogen-containing compounds in the process of the continuous final purification of effluents from swine-breeding farms by immobilized cells].

After their two-step purification in aeration tanks, effluents from a swine-breeding farm were subjected to final purification in a flow bioreactor with immobilized cells derived from activated sludge. The dynamics of nitrogen compounds were studied at the reactor inlet and outlet at three flow rates. The use of a simple mathematical model of the nitrogen balance allowed the rates of nitrogen assimilation-mineralization two nitrification stages, and nitrogen fixation-denitrification to be estimated. The first stage of nitrification was found to proceed intensely and produce high nitrite concentrations (up to 100 mg N/l, accounting for about half of the total N at the inlet). The second nitrification stage was inhibited, and nitrate was only produced after 3-4 weeks of continuous operation. The strategy of the final purification of effluents from nitrogen compounds is discussed on the basis of the known regulatory mechanisms of microbial activities in activated sludge.

Animal Husbandry↗

Effect of high concentrations of nitrogen on mixed populations of nitrifying bacteria isolated from industrial nitrogenous wastewaters.

The effect of high concentrations of different forms of nitrogen (NH4+, NO2-, NO3- and urea N) on nitrification by mixed populations of nitrosobacters isolated from nitrogen fertilizer plant wastewaters and nitrobacters isolated from effluents from a biological bed treating these wastewaters was determined. The inhibitory activity (within the concentration values for industrial wastewaters) of only the reaction products was observed, i.e. nitrites for nitrification phase I and nitrates for nitrification phase II. A mixed population of phase II nitrifying bacteria is highly resistant to high concentrations of ammonia nitrogen (50% inhibition at 3,000 mg N/l).

Ammonia↗

Phenacetin inhibited but acetaminophen stabilized partial nitrification/anammox system: Studies on microbial metabolism and resistance genes in biofilm and plastisphere.

Partial nitrification (PN) inhibitors, such as phenacetin (PNCT) and acetaminophen (APAP), ensure a stable nitrite supply for anaerobic ammonium oxidation (anammox). But the unknown impact of inhibitors on anammox limit the application of inhibitors. In addition to the biofilm carriers used in biological nitrogen removal systems, microplastics (MPs) (a type of emerging contaminants) are the common substrate for microbial colonization, even enriched resistance genes (RGs). This research compared the effects of 0.5, 1 and 5 mg/L PNCT or APAP on partial nitrification-anammox (PN/A) biofilm and plastisphere. 1 mg/L PNCT inhibited the nitrogen removal functional bacteria (Nitrosomonas, Candidatus Kuenenia, Candidatus Brocadia and Nitrospira), resulting in the sharp deteriorated performance of PN/A system. 5 mg/L PNCT inhibited multiple metabolism pathways, resulting in the absence of electrons and energy supply of microorganisms. 0.5-1 mg/L APAP maintained the stable operation of PN/A system. Nitrospira abundances declined from 2.8% to 1.1% after 0.5 mg/L APAP exposure. But 5 mg/L APAP inhibited the abundance of amoA and the production of extracellular polymeric substances, which caused the slight fluctuation of PN/A performance. PN inhibitors did not cause the sharp increase of most RGs in biofilm and water. However, MPs exhibited the huge capacity of enriching RGs, which should be removed. This study proposed that 0.5 mg/L of APAP was suitable for the PN/A system to control dosage for practical application.

Biofilms↗

Potential survival strategies of novel comammox and nitrite-oxidizing Nitrospira synthesizing osmoprotectants in a wastewater microbiome treating high-ammonia brackish landfill leachate.

BACKGROUND: In the late stages of landfill operation, leachate becomes brackish and contains high concentrations of ammonia with limited organic carbon. At leachate treatment facilities, it is typically subjected to nitrification followed by denitrification, with methanol supplied as an external electron donor. This unique environment may harbor novel microorganisms, including nitrifiers. Although a variety of microorganisms are involved in nitrification, their substrate specificity and salinity tolerance remain insufficiently understood. In this study, a genome-centric metagenome analysis was conducted on the microbiome from a leachate treatment facility at a closed landfill. RESULTS: A total of 68 metagenome-assembled genomes (MAGs) were reconstructed, including 64 putative novel species. Among these, two Nitrospira MAGs were recovered: a novel complete ammonia-oxidizing bacterium (comammox), Nitrospira LAS72 (88.72% completeness, 2.10% contamination), and canonical nitrite-oxidizing Nitrospira LAS18 (99.98% completeness, 2.29% contamination). Comparative genomic analysis with 260 publicly available Nitrospira genomes revealed that LAS18 represents a new sub-lineage within lineage VII of the Nitrospira genus. Two ammonia-oxidizing archaea (AOA), Candidatus Nitrosocosmicus LAS21 and Nitrosarchaeum LAS73, were also identified, while canonical ammonia-oxidizing bacteria were not detected. Given the brackish conditions (1.23% salinity) and the methanol-fed operation of the treatment facility, the genomic potential for osmotic stress adaptation and methanol metabolism was investigated. Comammox Nitrospira LAS72 harbors biosynthetic pathways for several compatible solutes (osmoprotectants), including glycine betaine, proline, trehalose, and L-glutamate. Moreover, comammox Nitrospira LAS72 possesses genetic potential for oxidizing formaldehyde, suggesting that it may exploit these methanol-derived intermediates as energy sources. These features indicate that LAS72 may withstand osmotic fluctuations through the production of various osmoprotectants and thrive under the unique conditions of a methanol-fed environment. CONCLUSIONS: The discovery of novel comammox Nitrospira and canonical Nitrospira forming a new sub-lineage within lineage VII of the Nitrospira genus in an ammonia-rich brackish environment provides the first genomic evidence for evolutionary adaptation among nitrifiers to saline, methanol-fed environments. These findings enhance our understanding of the ecological and evolutionary dynamics shaping nitrifier communities in complex treatment ecosystems. Video Abstract.

Ammonia↗

On-line estimation of the maximum specific growth rate of nitrifiers in activated sludge systems.

The on-line estimation of the maximum specific growth rate of autotrophic biomass is addressed in this article. A general nitrification process model, which is valid for any realistic flow pattern, is used to develop the estimation algorithm. Depending on the measurements available, two estimation equations are derived. While both require measuring the nitrification activity of the activated sludge, one requires the additional measurement of the nitrifiable nitrogen concentrations at the two ends of the bioreactor, and the other requires the nitrate nitrogen concentrations at the same locations. The algorithm also requires some stoichiometric and kinetic parameters. However, sensitivity analysis shows that the estimate is insensitive to the parameters other than the autotrophic decay rate. Compared to the existing algorithms, the algorithm developed in this article does not rely on the assumption of ideal flow pattern in the plant and does not require an error-prone estimate of the autotrophic biomass concentration. Experimental and simulation studies show that the algorithm performs well and is robust to influent variations and accidental sludge losses.

Algorithms↗

Application of Toxkit microbiotests for toxicity assessment in soil and compost.

The potential of Toxkit microbiotests to detect and analyze pollution in agricultural soil and the quality of compost was studied. The toxicity tests used included seed germination biotests using cress salad (Lepidum sativum L.), tomato (Lycopersicum esculentum L.), and cucumber (Cucumis sativus L.), and the Toxkit microbiotests included those with microalgae (Selenastrum capricornutum), protozoa (Tetrahymena thermophila), crustaceans (Daphnia magna, Thamnocephalus platyurus, and Heterocypris incongruens), and rotifers (Brachionus calyciflorus). Experiments on compost were undertaken in a modified solid-state fermentation system (SSF) and under field conditions (in a windrow). To promote the composting process, two strains of Trichoderma (Trichoderma lignorum and Trichoderma viride), as well as a nitrification association that regulated the nitrogen-ammonification and nitrification processes were applied.

Animals↗

Impact of five selected xenobiotics on isolated ammonium oxidizers and on nitrifying activated sludge.

Sewage treatment plants (STPs) are usual receptors of xenobiotic compounds that have to be cotreated with municipal wastewaters before being discharged to the water environment. The presence of organic contaminants, such as surfactants, polycyclic aromatic hydrocarbons (PAHs), phthalates, and their primary degradation products in the influents of STPs may inhibit irreversibly sensitive biological processes, such as nitrification. The first step of nitrification, i.e., the oxidation of ammonium to nitrite (nitritification), is particularly sensitive. Inhibition of this step under uncontrolled conditions may completely inhibit biological nitrogen removal. The aim of this work was to study the possible inhibitory effect of five selected xenobiotics on (a) a mixed culture of ammonium-oxidizing bacteria isolated from activated sludge and (b) nitrifying activated sludge directly. The xenobiotics that were tested include nonylphenols (NP), nonylphenolethoxylates (NPEO), linear alkylbenzene sulfonates (LAS), di(2-ethylhexyl) phthalate (DEHP), as a representative phthalate ester, and the PAH phenanthrene. Remarkable inhibitory effects for all tested compounds were observed in this study even at xenobiotic concentrations as low as 1 mg/L. The observed inhibition of xenobiotics on nitrifying activated sludge was less pronounced, because of the masking effect exerted by the sludge flocs, but was still significant for many of the tested substances at concentrations up to 10 mg/L.

Bacteria↗

A screening technique for assessing effects of pesticides on population and activities of non-target soil microorganisms.

A simplified technique for assessing the effect of pesticides on non-target soil microorganisms was developed. Changes in microbial population determined by the dilution plate method, in nitrification by the soil perfusion technique, and in oxygen consumption using a differential respirometer are the only measurements required. A comparison of the effects observed for three pesticides, dieldrin, chlorpyrifos and Vorlex, in three soil types with those produced by an antibiotic, a fungicide, a nitrification inhibitor and steam pasteurization of the soil clearly demonstrate the effectiveness of the simple technique.

Chlorpyrifos↗

The evolution of nitrogen cycling.

Based upon arguments concerning properties of the environment and the energetics of nitrogen transformation reactions, new hypotheses regarding their evolution are presented. These hypotheses are supported by new calculations and observations germane to understanding the evolution of the nitrogen cycle. From calculations of shock production by meteor impact, we suggest that impact produced fixed nitrogen could have resulted in the entire reservoir of Earth's N2 being converted into fixed nitrogen at the end of accretion. We have significantly improved upon previous calculations of the abiotic fixation rate on the early earth and find a rate of fixation by lightning of approximately 1-3 x 10(16) molecules NO/J, which is 2 to 3 times greater than previous estimates. This strengthens the suggestion, corroborated by the predominance of a single nitrogenase enzyme, that biological nitrogen fixation may have been a late evolutionary development, after the development of an aerobic atmosphere. In addition, we show for the first time that HNO, predicted to be the main product of atmospheric photochemical reactions involving NO on the primitive Earth by photochemical models, would eventually become NO2- and NO3- after reaching the Earth's surface. Based upon microbe-environment interactions on an ecological as well as a biochemical scale we suggest that denitrification arose prior to aerobic respiration and that nitrification arose after the advent of an aerobic atmosphere. We hypothesize the following evolutionary sequence for the biological transformation of nitrogen compounds: Ammonification --> Denitrification --> Nitrification --> Nitrogen fixation.

Ammonia↗