Biological nitrogen removal with nitrification and denitrification via nitrite pathway

Presently, the wastewater treatment practices can be significantly improved through the introduction of new microbial treatment technologies. To meet increasingly stringent discharge standards, new applications and control strategies for the sustainable removal of ammonium from wastewater have to be implemented. Partial nitrification to nitrite was reported to be technically feasible and economically favorable, especially when wastewater with high ammonium concentrations or low C/N ratios is treated. For successful implementation of the technology, the critical point is how to maintain partial nitrification of ammonium to nitrite. Partial nitrification can be obtained by selectively inhibiting nitrite oxidizing bacteria through appropriate regulation of the system’s DO concentration, microbial SRT, pH, temperature, substrate concentration and load, operational and aeration pattern, and inhibitor. The review addressed the microbiology, its consequences for their application, the current status regarding application, and the future developments.     

限氧自养硝化-反硝化生物脱氮新技术

限氧自养硝化—反硝化是部分硝化与厌氧氨氧化相耦联的生物脱氮反应过程,通过严格控制溶解氧在0．1～0．3mg·L^-1,实现硝化反应控制在亚硝酸阶段,然后以硝化阶段剩余的NH4^+作为电子供体,在厌氧条件下实现反硝化,该反应过程是完全的自养硝化—反硝化过程,具有能耗低、脱氮效率高、反应系统占地面积小等优点,适用于处理COD／NH4^+—N低的废水,是一种非常有应用前景的生物脱氮技术,文中详细介绍了限氧自养硝化—反硝化生物脱氮反应过程的研究进展,讨论了其微生物学机理及应用前景。     

Simultaneous nitrification and denitrification using an autotrophic membrane-immobilized biofilm reactor

AIMS: To develop a laboratory-scale autotrophic membrane-immobilized biofilm reactor to remove nitrogen from drinking water. METHODS AND RESULTS: A polyvinyl alcohol (PVA) immobilized biofilm, attached to the surface of a silicone tube, was used as the basis of a bioreactor for simultaneous nitrification and denitrification of water. The bioreactor was aerated with air to supply oxygen for nitrification. Pure hydrogen was supplied to the silicone tube and diffused through the membrane wall to feed the biofilm for autotrophic denitrification. The bioreactor was effective for the simultaneous nitrification and denitrification of water after a short period of acclimation, while the biofilm exhibited good resistance to the inhibition of denitrification by dissolved oxygen; the denitrification rate decreased by only 8% as the dissolved oxygen increased from 2 mg l(-1) to saturation. CONCLUSIONS: By using PVA crosslinked with sodium nitrate to entrap nitrifying and denitrifying sludge on the surface of a silicone tube, a novel bioreactor for simultaneous nitrification and denitrification was developed. In addition to performing as an immobilizing agent to strengthen the biofilm, PVA protected the denitrifying microorganisms to reduce the inhibition by dissolved oxygen under aerobic condition. Therefore, nitrification and denitrification occurred simultaneously within the biofilm. Furthermore, the immobilization technique shortened the acclimation period of the bioreactor. SIGNIFICANCE AND IMPACT OF THE STUDY: The described space saving and simple to operate bioreactor for nitrogen removal performed autotrophic denitrification to solve the problem of residual carbon in heterotrophic denitrification, and thus is suitable for removing nitrogen from drinking water.     

Efficient nitrogen removal in a pilot system based on upflow multi-layer bioreactor for treatment of strong nitrogenous swine wastewater

In order to prevent the toxic effect caused by high strength ammonium in a swine wastewater treatment system, a patented upflow multi-layer bioreactor (UMBR) as a pre-anoxic tank was applied to a pilot-scale plant with a treatment capacity of 5 m³/d. This plant was operated for 4 months at a high IR ratio in the range of 10–17, in order to alleviate the toxic effects caused by high strength ammonium. A computational fluid dynamic (CFD) analysis was also conducted to design and configure the rotating distributors in the UMBR. At an IR ratio of about 17, the influent NH₄⁺-N (1169 mg N/L) was diluted to less than 80 mg N/L at the head of the UMBR, and then was completely nitrified (about 98.3%) in the aeration tank, without any inhibition caused by high strength ammonium. The nitrate at a concentration of about 58.2 mg N/L recycled from the aeration tank was completely denitrified in UMBR #1, which was operated at an actual hydraulic retention time (HRT) of 3.5 h.     

Characteristics of the bioreactor landfill system using an anaerobic-aerobic process for nitrogen removal

A sequential upflow anaerobic sludge blanket (UASB) and air-lift loop sludge blanket (ALSB) treatment was introduced into leachate recirculation to remove organic matter and ammonia from leachate in a lab-scale bioreactor landfill. The results showed that the sequential anaerobic-aerobic process might remove above 90% of COD and near to 100% of NH4+ -N from leachate under the optimum organic loading rate (OLR). The total COD removal efficiency was over 98% as the OLR increased to 6.8-7.7 g/l d, but the effluent COD concentration increased to 2.9-4.8 g/l in the UASB reactor, which inhibited the activity of nitrifying bacteria in the subsequent ALSB reactor. The NO3- -N concentration in recycled leachate reached 270 mg/l after treatment by the sequential anaerobic-aerobic process, but the landfill reactor could efficiently denitrify the nitrate. After 56 days operation, the leachate TN and NH4+ -N concentrations decreased to less than 200 mg/l in the bioreactor landfill system. The COD concentration was about 200 mg/l with less than 8 mg/l BOD in recycled leachate at the late stage. In addition, it was found that nitrate in recycled leachate had a negative effect on waste decomposition.     

Novel autotrophic nitrogen removal system using gel entrapment technology

A pilot plant involving a nitritation-anammox process was operated for treating digester supernatant. In the preceding nitritation process, ammonium-oxidizing bacteria were immobilized in gel carriers, and the growth of nitrite-oxidizing bacteria was suppressed by heat-shock treatment. For the following anammox process, in order to maintain the anammox biomass in the reactor, a novel process using anammox bacteria entrapped in gel carriers was also developed. The nitritation performance was stable, and the average nitrogen loading and nitritation rates were 3.0 and 1.7 kg N m⁻³ d⁻¹, respectively. In the nitritation process, nitrate production was completely suppressed. For the anammox process, the startup time was about two months. Stable nitrogen removal was achieved, and an average nitrogen conversion rate of 5.0 kg N m⁻³ d⁻¹ was obtained. Since the anammox bacteria were entrapped in gel carriers, stable nitrogen removal performance was attained even at an influent suspended solids concentration of 1500 mg L⁻¹.     

Efficient COD removal and nitrification in an upflow microaerobic sludge blanket reactor for domestic wastewater

The treatment performance of an upflow microaerobic sludge blanket reactor (UMSB) for synthetic domestic wastewater was investigated at two dissolved oxygen (DO) levels, 0.3–0.5 and 0.7–0.9 mg l⁻¹, focusing on nitrification performance. The higher DO level induced complete nitrification of ammonia nitrogen (NH₃–N), achieving chemical oxygen demand and NH₃–N removals of 97 and 92%, respectively. There were consistently significantly higher nitrate nitrogen (NO₃–N) and nitrite nitrogen (NO₂–N) levels in the effluent, with ~66% of newly-produced oxidised nitrogen as NO₂–N. Despite the high nitrification efficiency, only about 23% of the removed NH₃–N amount from the influent was ultimately transformed into oxidised nitrogen due to the simultaneous nitrification-denitrification. Sludge blanket development and granulation occurred simultaneously in the UMSB.     

Post-treatment of fish canning effluents by sequential nitrification and autotrophic denitrification processes

In this research study a nitrifying/autotrophic denitrifying system was used for the post-treatment of an effluent coming from an anaerobic digester treating the wastewater produced in a fish canning industry. The nitrifying reactor achieved 100% of ammonia oxidation into nitrate. The effluent from this unit was fed to the autotrophic denitrifying reactor which treated a maximum sulphide loading rate (SLR) of 200 mg S^2?/L d with removal percentages of 100% and 30% for sulphide and nitrate, respectively. The low nitrate removal efficiency is attributed to sulphide limitations.The operational costs of this system were estimated as 0.92 €/kg N_removed, lower than those for conventional nitrification/denitrification processes. For nitrogen removal the SHARON/anammox processes is the cheapest option. However the combination of nitrification and autotrophic denitrification (using elemental sulphur) processes would present a better operational stability compared to the SHARON/anammox system.     

Using the combined bioelectrochemical and sulfur autotrophic denitrification system for groundwater denitrification

A combined bioelectrochemical and sulfur autotrophic denitrification system (CBSAD) was evaluated to treat a groundwater with nitrate contamination (20.9-22.0mgNO(3)(-)-N/L). The reactor was operated continuously for several months with groundwater to maximize treatment efficiency under different hydraulic retention times (HRT) and electric currents. The denitrification rate of sulfur autotrophic part followed a half-order kinetics model. Moreover, the removal efficiency of bioelectrochemical part depended on the electric current. The reactor could be operated efficiently at the HRT ranged from 4.2 to 2.1h (corresponding nitrogen volume-loading rates varied from 0.12 to 0.24 kg N/m(3)d; and optimum current ranged from 30 to 1000 mA), and the NO(3)(-)-N removal rate ranged from 95% to 100% without NO(3)(-)-N accumulation. The pH of effluent was satisfactorily adjusted by bioelectrochemical part, and the sulfate concentration of effluent was lower than 250 mg/L, meeting the drinking water standard of China EPA.     

Closed water recirculating system for fish rearing equipped with bioreactor capable of simultaneous nitrification and denitrification.

Five crucian carp, Carassius auratus langsdorfiicarps had been reared in a closed water recirculating system. The system was equipped with the compact bioreactor using the plate gels capable of both nitrification and denitrification in a single unit. Ammonia and nitrite concentrations in the rearing water had been maintained below 0.05 mg-N/L, and nitrate concentration also controlled between 2 and 8 mg-N/L with the bioreactor. As concerns nitrogen budget in the closed system, 95.0% of nitrogen income from feed was lost as nitrogen gas from the closed system. All fish was alive for 91 days without any unusual behavior. Thus, the bioreactor performed both nitrification and denitrification abilities enough to rear the five fish for 91 days. The bioreactor using the plate gels would be effective to simplify the closed system both physically and operationally, since it can remove the ammonia excreted from fish as nitrogen gas by a single step.     

Application of low-intensity ultrasound to enhance simultaneous nitrification/iron-based autotrophic denitrification

Biotechnology Letters - Intermittent ultrasound with an intensity of 0.2 W/ml was applied during simultaneous nitrification/iron-based autotrophic denitrification to evaluate its impacts...     

生物反应器填埋场系统渗滤液的脱氮性能

利用填埋场垂直分布的好氧-缺氧-厌氧的独特生态环境,并采用填埋垃圾上层间歇曝气充氧的方式,研究了生物反应器填埋场系统渗滤液的脱N性能.结果表明,填埋垃圾上层间歇曝气充氧,促进了填埋垃圾层硝化细菌和反硝化细菌的生长,且可使反硝化细菌的数量比普通的填埋垃圾层高4～13个数量级,硝化细菌的最大数量可达到10⁹个·g^-1;营建了填埋场内硝化、反硝化等脱N反应的生物环境,有利于回流渗滤液含N化合物的去除.试验结束时,其渗滤液NH₄⁺-N和TN浓度分别为186和289 mg·L^-1,仅为对照的18%和26％.此外,填埋垃圾上层间歇曝气充氧也有利于填埋垃圾的降解,提高垃圾的稳定化效果.     

Combined removal of sulfur compounds and nitrate by autotrophic denitrification in bioaugmented activated sludge system

An autotrophic denitrification process using reduced sulfur compounds (thiosulfate and sulfide) as electron donor in an activated sludge system is proposed as an efficient and cost effective alternative to conventional heterotrophic denitrification for inorganic (or with low C/N ratio) wastewaters and for simultaneous removal of sulfide or thiosulfate and nitrate. A suspended culture of sulfur-utilizing denitrifying bacteria was fast and efficiently established by bio-augmentation of activated sludge with Thiobacillus denitrificans. The stoichiometry of the process and the key factors, i.e. N/S ratio, that enable combined sulfide and nitrogen removal, were determined. An optimum N/S ratio of 1 (100% nitrate removal without nitrite formation and low thiosulfate concentrations in the effluent) has been obtained during reactor operation with thiosulfate at a nitrate loading rate (NLR) of 17.18 mmol N L(-1) d(-1). Complete nitrate and sulfide removal was achieved during reactor operation with sulfide at a NLR of 7.96 mmol N L(-1) d(-1) and at N/S ratio between 0.8 and 0.9, with oxidation of sulfide to sulfate. Complete nitrate removal while working at nitrate limiting conditions could be achieved by sulfide oxidation with low amounts of oxygen present in the influent, which kept the sulfide concentration below inhibitory levels.     

Modeling nitrogen removal for a denitrification biofilter

Nitrous oxide (N₂O) is a major greenhouse gas, heavily contributing to global warming. N₂O is emitted from various sources such as wastewater treatment plants, during the nitrification and denitrification steps. ASM models, which are commonly used in wastewater treatment, usually consider denitrification as a one-step process (NO₃ ⁻ directly reduced to N₂) and are as such unable to provide values for intermediate products of the reaction like N₂O. In this study, a slightly modified ASM1 model was implemented in the GPS-X™ software to simulate the concentration of such intermediate products (NO₂ ⁻, NO and N₂O) and to estimate the amounts of gaseous N₂O emitted by the denitrification stage (12 biofilters) of the Seine-Centre WWTP (SIAAP, Paris). Simulations running on a 1-year period have shown good agreements with measured effluent data for nitrate and nitrite. The calculated mean value for emitted N₂O is 4.95 kgN–N₂O/day, which stands in the typical range of estimated experimental values of 4–31 kgN–N₂O/day. Nitrous oxide emissions are usually not measured on WWTPs and so, as obtained results show, there is a certain potential for using models that quantify those emissions using traditionally measured influent data.     

Nitrogen-removal bioreactor capable of simultaneous nitrification and denitrification for application to industrial wastewater treatment

A bioreactor system with 30 packed gel envelopes was installed in a thermal power plant for the removal of nitrogen from ammonia-containing desulfurization wastewater. Each envelope consisted of double-sided plate gels containing Nitrosomonas europaea and Paracoccus denitrificans cells with an internal space in between for injecting an electron donor. The envelope can remove ammonia from wastewater in a single step. When the wastewater was continuously treated with the bioreactor system, it removed 95.0% of the total nitrogen in the inlet, and the total nitrogen concentration in the outlet was below 9.0 mg L⁻¹. The maximum nitrogen removal rate was 6.0 g day⁻¹ per square meter of the gel area. The maximum utilization efficiency of the injected ethanol for denitrification was 98.4%, and the total organic carbon concentration in the outflow was maintained at a low level. Since the bioreactor system could use the electron donor effectively, it was not necessary to use an additional aerobic tank to remove the electron donor and a settling tank to segregate the surplus sludge containing bacteria from wastewater. Our concept of using packed gel envelopes would be highly effective for constructing a simple and efficient nitrogen removal system capable of simultaneous nitrification and denitrification.     

Effect of sludge age on simultaneous nitrification and denitrification in membrane bioreactor

This study evaluated the effect of sludge age on simultaneous nitrification and denitrification in a membrane bioreactor treating black water. A membrane bioreactor with no separate anoxic volume was operated at a sludge age of 20 days under low dissolved oxygen concentration of 0.1-0.2 mg/L. Its performance was compared with the period when the sludge age was adjusted to 60 days. Floc size distribution, apparent viscosity, and nitrogen removal differed significantly, together with different biomass concentrations: nitrification was reduced to 40% while denitrification was almost complete. Modelling indicated that both nitrification and denitrification kinetics varied as a function of the sludge age. Calibrated values of half saturation coefficients were reduced when the sludge age was lowered to 20 days. Model simulation confirmed the validity of variable process kinetics for nitrogen removal, specifically set by the selected sludge age.     

A novel nonwoven hybrid bioreactor (NWHBR) for enhancing simultaneous nitrification and denitrification

This study proposed a nonwoven hybrid bioreactor (NWHBR) in which the nonwoven fabric played dual roles as a biofilm carrier and membrane‐like separation of the flocculent sludge in the reactor. The results of long‐term monitoring demonstrated that the NWHBR could achieve simultaneous nitrification and denitrification (SND), with nearly complete ammonium removal and 80% removal of total nitrogen. The biofilm attached to the nonwoven fabric removed 27% of the chemical oxygen demand (COD) and 36% of the nitrate in the reactor, an enhanced elimination of nutrients that was attributed to the increased mass transfer within the biofilm due to permeate drag. The results of batch experiments showed that the flocculent sludge played a more dominant role in nitrification and denitrification (79% and 61%, respectively) than the biofilm (21% and 36%, respectively). The batch experiments also revealed that the enforced mass transfer, with an effluent recirculation rate of 4.3 L/m²h (which was the same as the flux during the reactor's long‐term operation), improved the denitrification rate by 58% (i.e., from 9.0 to 14.2 mg‐NO‐N/h). Pyrosequencing of the 16S rRNA gene amplification revealed a high microbial diversity in both the flocculent sludge and biofilm, with Proteobacteria, Bacteroidetes and Chloroflexi as the dominant groups. A phylogenetic (P) test indicated that the NWHBR contained phylogenetically distinct microbial communities: those in the biofilm differed from those in the flocculent sludge. However, the communities on the exterior and interior of the biofilm were more similar to each other. Due to its good SND performance, low physical back‐washing frequency and low air‐to‐water ratio, the NWHBR represents an attractive alternative for the wider application of either low‐cost membrane bioreactors or biofilm reactors. Biotechnol. Bioeng. 2013; 110: 1903–1912. © 2013 Wiley Periodicals, Inc.     

Influence of hydrological connectivity of riverine wetlands on nitrogen removal via denitrification

Wetland ecosystems in agricultural areas often become progressively more isolated from main water bodies. Stagnation favors the accumulation of organic matter as the supply of electron acceptors with water renewal is limited. In this context it is expected that nitrogen recycling prevails over nitrogen dissipation. To test this hypothesis, denitrification rates, fluxes of dissolved oxygen (SOD), inorganic carbon (DIC) and nitrogen and sediment features were measured in winter and summer 2007 on 22 shallow riverine wetlands in the Po River Plain (Northern Italy). Fluxes were determined from incubations of intact cores by measurement of concentration changes or isotope pairing in the case of denitrification. Sampled sites were eutrophic to hypertrophic; 10 were connected and 12 were isolated from the adjacent rivers, resulting in large differences in nitrate concentrations in the water column (from <5 to 1,133 μM). Benthic metabolism and denitrification rates were investigated by two overarching factors: season and hydrological connectivity. SOD and DIC fluxes resulted in respiratory quotients greater than one at most sampling sites. Sediment respiration was coupled to both ammonium efflux, which increased from winter to summer, and nitrate consumption, with higher rates in river-connected wetlands. Denitrification rates measured in river-connected wetlands (35–1,888 μmol N m^?2 h^?1) were up to two orders of magnitude higher than rates measured in isolated wetlands (2–231 μmol N m^?2 h^?1), suggesting a strong regulation of the process by nitrate availability. These rates were also significantly higher in summer (9–1,888 μmol N m^?2 h^?1) than in winter (2–365 μmol N m^?2 h^?1). Denitrification supported by water column nitrate (D_W) accounted for 60–100% of total denitrification (Dtot); denitrification coupled to nitrification (D_N) was probably controlled by limited oxygen availability within sediments. Denitrification efficiency, calculated as the ratio between N removal via denitrification and N regeneration, and the relative role of denitrification for organic matter oxidation, were high in connected wetlands but not in isolated sites. This study confirms the importance of restoring hydraulic connectivity of riverine wetlands for the maintenance of important biogeochemical functions such as nitrogen removal via denitrification.     

Potential rates of nitrification and denitrification in an oligotrophic freshwater sediment system

Potential rates of nitrification and denitrification were measured in an oligotrophic sediment system. Nitrification potential was estimated using the CO oxidation technique, and potential denitrification was measured by the acetylene blockage technique. The sediments demonstrated both nitrifying and denitrifying activity. E_h, O₂, and organic C profiles showed two distinct types of sediment. One type was low in organic C, had high O₂ and E_h, and had rates of denitrification 1,000 times lower than the other which had high organic C, low O₂, and low E_h. Potential nitrification and denitrification rates were negatively correlated with E_h. This suggests that environmental heterogeneity in denitrifier and nitrifier populations in oligotrophic sediment systems may be assessed using E_h before sampling protocols for nitrification or denitrification rates are established. There was no correlation between denitrification and nitrification rates or between either of these processes and NH₄ ⁺ or NO₃ ^– concentrations. The maximum rate of denitrification was 0.969 nmole N cm^–3 hour^–1, and the maximum rate of nitrification was 23.6 nmole cm^–3 hour^–1, suggesting nitrification does not limit denitrification in these oligotrophic sediments. Some sediment cores had mean concentrations of 6.0 mg O₂/liter and still showed both nitrification and denitrification activity.     

Control of carbon and ammonium ratio for simultaneous nitrification and denitrification in a sequencing batch bioreactor

This study shows how the carbon and nitrogen (C/N) ratio controls the simultaneous occurrence of nitrification and denitrification in a sequencing batch reactor (SBR). Data demonstrated that a low C/N ratio resulted in a rapid carbon deficit, causing an unbalanced simultaneous nitrification–denitrification (SND) process in SBR. When the initial COD/NH₄⁺-N ratio was adjusted to 11.1, the SND-based SBR achieved complete removal of NH₄-N and COD without leaving any NO₂⁻-N in the effluent. The nitrogen removal efficiency decreases gradually with increasing ammonium-loading rate to the SND–SBR system. Altogether, data showed that appropriate controls of carbon and nitrogen input are required to achieve an efficient SND–SBR. An established SND technology can save operation time and energy, and might replace the traditional two-stage biological nitrification and denitrification process.