Nitrogen removal from sludge reject water by a two-stage oxygen-limited autotrophic nitrification denitrification process.

Nitrogen removal from sludge reject water was obtained by oxygen-limited partial nitritation resulting in nitrite accumulation in a first stage, followed by autotrophic denitrification of nitrite with ammonium as electron donor (similar to anaerobic ammonium oxidation) in a second stage. Two membrane-assisted bioreactors (MBRs) were used in series to operate with high sludge ages and subsequent high volumetric loading rates, achieving 1.45 kg N m(-3) day(-1) for the partial nitritation MBR and 1.1 kg N m(-3) day(-1) for the anaerobic ammonium oxidation MBR. Biomass retention in the nitritation stage ensured flexibility towards loading rate and operating temperature. Nitrite oxidisers were out-competed at low oxygen and high free ammonia concentration. Biomass retention in the second MBR prevented wash-out of the slowly growing bacteria. Nitrite and ammonium were converted to dinitrogen gas in a reaction ratio of 1.05, thereby maintaining nitrite limitation to assure process stability. The anoxic consortium catalysing the autotrophic denitrification process consisted of Nitrosomonas-like aerobic ammonium oxidizers and anaerobic ammonium oxidizing bacteria closely related to Kuenenia stuttgartiensis. The overall removal efficiency of the combined process was 82% of the incoming ammonium according to a total nitrogen removal rate of 0.55 kg N m(-3) day(-1), without adding extra carbon source.     

Anammox brings WWTP closer to energy autarky due to increased biogas production and reduced aeration energy for N-removal.

Fifty years ago when only BOD was removed at municipal WWTPs primary clarifiers were designed with 2-3 hours hydraulic retention time (HRT). This changed with the introduction of nitrogen removal in activated sludge treatment that needed more BOD for denitrification. The HRT of primary clarification was reduced to less than one hour for dry weather flow with the consequence that secondary sludge had to be separately thickened and biogas production was reduced. Only recently the ammonia rich digester liquid (15-20% of the inlet ammonia load) could be treated with the very economic autotrophic nitritation/anammox process requiring half of the aeration energy and no organic carbon source compared to nitrification and heterotrophic denitrification. With the introduction of this new innovative digester liquid treatment the situation reverts, allowing us to increase HRT of the primary clarifier to improve biogas production and reduce aeration energy for BOD removal and nitrification at similar overall N-removal.     

Granular biomass capable of partial nitritation and anammox

A novel and efficient way of removing nitrogen from wastewater poor in biodegradable organic carbon, is the combination of partial nitritation and anoxic ammonium oxidation (anammox), as in the one-stage oxygen-limited autotrophic nitrification/denitrification (OLAND) process. Since anoxic ammonium-oxidizing bacteria grow very slowly, maximum biomass retention in the reactor is required. In this study, a lab-scale sequencing batch reactor (SBR) was used to develop granular, rapidly settling biomass. With SBR cycles of one hour and a minimum biomass settling velocity of 0.7 m/h, OLAND granules were formed in 1.5 months and the nitrogen removal rate increased from 50 to 450 mg N L(-1) d(-1) in 2 months. The granules had a mean diameter of 1.8 mm and their aerobic and anoxic ammonium-oxidizing activities were well equilibrated to perform the OLAND reaction. Fluorescent in-situ hybridization (FISH) demonstrated the presence of both beta-proteobacterial aerobic ammonium oxidizers and planctomycetes (among which anoxic ammonium oxidizers) in the granules. The presented results show the applicability of rapidly settling granular biomass for one-stage partial nitritation and anammox.     

Nitrogen removal rates at a technical-scale pilot plant with the one-stage partial nitritation/Anammox process.

Traditional nitrification/denitrification is not suitable for nitrogen removal when wastewater contains high concentrations of ammonium nitrogen and low concentrations of biodegradable carbon. Recently, a deammonification process was developed and proposed as a new technology for treatment of such streams. This process relies on a stable interaction between aerobic bacteria Nitrosomonas, that accomplish partial nitritation and anaerobic bacteria Planctomycetales, which conduct the Anammox reaction. Simultaneous performance of these two processes can lead to a complete autotrophic nitrogen removal in one single reactor. The experiments where nitrogen was removed in one reactor were performed at a technical-scale moving-bed pilot plant, filled with Kaldnes rings and supplied with supernatant after dewatering of digested sludge. It was found that a nitrogen removal rate obtained at the pilot plant was 1.9 g m(-2) d(-1). Parallel to the pilot plant run, a series of batch tests were carried out under anoxic and aerobic conditions. Within the batch tests, where the pilot plant's conditions were simulated, removal rates reached up to 3 g N m(-2)d(-1). Moreover, the batch tests with inhibition of Nitrosomonas showed that only the Anammox bacteria (not anoxic removal by Nitrosomonas) are responsible for nitrogen removal.     

Nitrogen loss in a nitrifying rotating contactor treating ammonium rich leachate without organic carbon

Extended loss (up to 70%) of nitrogen is observed in a nitrifying rotating biological contactor (RBC) treating ammonium - rich leachate of a hazardous waste landfill. Due to pretreatment (flocculation, BOD-removal, activated carbon) DOC was less than 20 mg/l so heterotrophic denitrification can be excluded. The nitrification rate reaches 3-4 g NH₄-N m⁻² d⁻¹ at a pH of 7-7.3 in the first two of three RBC compartments. An increasing partial pressure of oxygen and ammonium concentration favor nitrogen removal over ammonium oxidation. The reduction of nitrite produced in the aerobic biofilm layer close to the surface might therefore be coupled with ammonium oxidation and takes place in the deeper or temporarily anoxic layer of the biofilm.     

Anammox bacteria enrichment and characterization from municipal activated sludge

A sustainable option for nitrogen removal is the anaerobic ammonium-oxidizing (anammox) process in which ammonium is oxidized to nitrogen gas with nitrite as electron acceptor. Application of this process, however, is limited by the availability of anammox biomass. In this study, two Brocadia-like anammox phylotypes were successfully enriched, detected and identified from an activated sludge taken from a domestic wastewater treatment plant (Minas Gerais, Brazil) employing a Sequencing Batch Reactor (SBR). The dominant phylotype was closely related to 'Candidatus Brocadia sinica', but one clone seemed to represent a novel species for which we propose the name 'Candidatus Brocadia brasiliensis'. Based on Fluorescence in situ hybridization (FISH) analysis, this enrichment led to a relative population size of 52.7% (±15.6) anammox bacteria after 6 months of cultivation. The cultivation process can be divided into three phases: phase 1 (approximately 25 days) was characterized by heterotrophic denitrification metabolism, phase 2 was the propagation phase and phase 3 (from the 87th day onwards), in which significant anammox activity was detected. A long-term performance of the SBR showed a near perfect removal of nitrite based on the influent NO(2)(-)-N concentration of 61-95 mg L(-1). The average ammonia removal efficiency was 90% with the influent NH(4)(+)-N concentration of 55-82 mg L(-1). Therefore, anammox cultivation and enrichment from activated sludge was possible under a controlled environment within 3 months.     

Anaerobic ammonium oxidation of ammonium-rich waste streams in fixed-bed reactors.

The feasibility of anaerobic ammonium oxidation (Anammox) in fixed-bed reactors was evaluated on laboratory and pilot scales. Using synthetic wastewater, the specific nitrogen removal rate was increased from 0.05-0.1 kgNm(-3)(reactor)d(-1) to 0.35-0.38 kgNm(-3)(reactor)d(-1) within a year (T= 22-27 degrees C) in all applications. However, the anammox activity was seriously and repeatedly inhibited at prolonged high nitrite concentrations (e.g. six days at 30-50 gNO2-Nm(-3)) and recovery was always a lengthy process. But even at a moderate nitrite concentration (11+/-10 gNO2-Nm(-3)), the observed specific growth rate was only 0.018 d(-1) at 26.4+/-0.8 degrees C, which corresponds to approximately 0.025 d(-1) at 30 degrees C (doubling time: 28 days). In a second experimental period for another 250 days, one of the laboratory reactors was fed with partially nitrified sludge liquors from a domestic wastewater treatment plant (WWTP). In this case, the specific elimination rate was as high as 3.5 kgNm(-3)(reactor)d(-1) at 26-27 degrees C. Independently of the feed, the average nitrogen elimination rate lay between 80-85% in all applications. An appropriate hydraulic design is essential to prevent clogging and local nitrite inhibition in fixed-bed reactors.     

Nitrite accumulation followed by denitrification using sequencing batch reactor.

Biological ammonia-nitrogen removal utilizes two distinct processes, nitrification and denitrification. In nitrification, ammonia oxidizes to nitrite then to nitrate. In this study, elimination of nitrite oxidation to nitrate step was attempted in order to directly remove nitrite to nitrogen gas by denitrification. For this study the supernatant from an anaerobic digester was used as an ammonia source and a sequencing batch reactor (SBR) was employed. Emphasis was given to the evaluation of the operational factors affecting nitrite accumulation and the elucidation of kinetics for biological nitrification and denitrification. Accumulation of nitrite in the nitrification process was achieved by suppressing the growth of Nitrobacter, a nitrite oxidizer, by loading high concentration ammonia supernatant immediately after all ammonia in the previous loading was oxidized to nitrite. Nitrite oxidation was taking place as the solid retention time (SRT) was increased from 2.5 days to 3.0 days in a continuously aerated SBR mode with daily feeding. However, nitrite accumulation was achieved even at longer SRT of 5 days when the aeration and non-aeration periods were appropriately combined and the non-aeration period can be used for denitrification of the accumulated nitrite with a carbon source supplied.     

A two-sludge system for simultaneous biological C, N and P removal via the nitrite pathway

Nitrogen removal via the nitrite pathway results in significant savings in both aeration costs and COD requirements for denitrification when compared to the conventional biological nitrogen removal process. Implementation of the nitrite pathway for simultaneous C/N/P removal in a single sludge system has a major drawback: the aeration phase disfavours denitrifying phosphorus removal. A possible configuration to overcome this issue is the utilisation of a two-sludge system where autotrophic and heterotrophic populations are physically separated. This paper experimentally demonstrates the feasibility of a nitrite-based two-sludge system with sequencing batch reactors (SBR) for the treatment of urban wastewater: a heterotrophic SBR with denitrifying PAOs for P removal and an aerobic SBR for N removal. Partial nitrification was attained in the autotrophic SBR so that shortcut biological nitrogen removal was achieved by using the anoxic dephosphatation activity of DPAOs. Finally, the effect of operating this system without pH control was studied using different influent pH values (pH = 6.8, 7.5 and 8.2) and, despite some efficiency lost due to the pH fluctuations, the system was able to remove most of the C, N and P present in the wastewater.     

Autotrophic nitrogen removal from anaerobic supernatant of Florence's WWTP digesters.

In municipal WWTP with anaerobic sludge digestion, 10-20% of total nitrogen load comes from the return supernatant produced by the final sludge dewatering. In recent years a completely autotrophic nitrogen removal process based on Anammox biomass has been tested in a few European countries, in order to treat anaerobic supernatant and to increase the COD/N ratio in municipal wastewater. This work reports the experimental results of the SHARON-ANAMMOX process application to anaerobic supernatant taken from the urban Florentine area wastewater treatment plant (S. Colombano WWTP). A nitritation labscale chemostat (7.4 L) has been started-up seeded with the S. Colombano WWTP nitrifying activated sludge. During the experimental period, nitrite oxidising bacteria wash-out was steadily achieved with a retention time ranging from 1 to 1.5 d at 35 degrees C. The Anammox inoculum sludge was taken from a pilot plant at EAWAG (Zurich). Anammox biomass has been enriched at 33 degrees C with anaerobic supernatant diluted with sodium nitrite solution until reaching a maximum specific nitrogen removal rate of 0.065 kgN kg(-1) VSS d(-1), which was 11 times higher than the one found in inoculum sludge (0.005 kgN kg(-1) VSS d(-1). In a lab-scale SBR reactor (4 L), coupled with nitritation bioreactor, specific nitrogen removal rate (doubling time equal to 26 d at 35 degrees C and at nitrite-limiting condition) reached the value of 0.22 kgN kg(-1) VSS d(-1), which was approximately 44 times larger than the rate measured in the inoculum Anammox sludge.     

Energy saving processes for nitrogen removal in organic wastewater from food processing industries in Thailand.

Nitrogen removal from organic wastewater is becoming a demand in developed communities. The use of nitrite as intermediate in the treatment of wastewater has been largely ignored, but is actually a relevant energy saving process compared to conventional nitrification/denitrification using nitrate as intermediate. Full-scale results and pilot-scale results using this process are presented. The process needs some additional process considerations and process control to be utilized. Especially under tropical conditions the nitritation process will round easily, and it must be expected that many AS treatment plants in the food industry already produce NO2-N. This uncontrolled nitrogen conversion can be the main cause for sludge bulking problems. It is expected that sludge bulking problems in many cases can be solved just by changing the process control in order to run a more consequent nitritation. Theoretically this process will decrease the oxygen consumption for oxidation by 25% and the use of carbon source for the reduction will be decreased by 40% compared to the conventional process.     

ANAMMOX and partial denitritation in anaerobic nitrogen removal from piggery waste.

The anaerobic ammonium removal from a piggery waste with high strength (56 g COD/L and 5 g T-N/L) was investigated using a lab-scale upflow anaerobic sludge bed reactor at a mesophilic condition. Based on the nitrogen and carbon balance in the process, the contribution of autotrophic and heterotrophic organisms was also evaluated in terms of the influent NO2-N/NH4-N ratio (1:0.8 and 1:1.2 for Phase 1 and Phase 2, respectively). The result of this research demonstrates that the anaerobic ammonium removal from the piggery waste, using the UASB reactor, can be performed successfully. Furthermore, it appears that by using granular sludge as the seed biomass, the ANAMMOX reaction can start more quickly. Average nitrogen conversion was 0.59 kg T-N/m3 reactor-day (0.06 kg T-N/kg VSS/day) and 0.66 kg T-N/m3 reactor-day (0.08 kg T-N/kg VSS/day) for Phase 1 and Phase 2. The NO2-N/NH4-N removal ratio by the ANAMMOX was 1.48 and 1.79 for Phase 1 and Phase 2. The higher nitrite contents (about 50%) in the substrate resulted in higher nitrite nitrogen removal by the partial denitritation, as well as the ANAMMOX reaction, implying higher potential of partial denitritation. However, the result reveals that the ANAMMOX reaction was influenced less by the degree of partial denitritation, and the ANAMMOX bacteria did not compete with denitritation bacteria. The colour of the biomass at the bottom of the reactor changed from dark gray to dark red, which was accompanied by an increase in cytochrome content. At the end of the experiment, red-coloured granular sludge with diameter of 1-2 mm at the lower part of the reactor was also observed.     

Chemical denitrification for nitrogen removal from landfill leachate.

A new system that removes nitrogen from landfill leachate and other waste waters with similar properties has been proposed with nitritation (i.e. oxidation of ammonium to nitrite) of half of the influent ammonium followed by chemical denitrification with a reaction between equal amounts of ammonium and nitrite to form nitrogen gas. Chemical denitrification occurs at high concentrations and the reactions were studied in combination with a concentration step. Studied concentration methods were freezing/thawing and evaporation/drying. Chemical denitrification is well-known in inorganic chemistry and has been observed in natural systems. Studies in laboratory were focused on chemical denitrification and showed that nearly complete removal of soluble nitrogen can be obtained in evaporation/drying of water solutions or leachate with equal amounts of ammonium and nitrite. Freezing/drying was less efficient with a removal of about 50-60% at high initial concentrations. Chemical denitrification is much influenced by concentration, pH-value, temperature and some compounds in leachate have an inhibiting effect on the reaction. Factors as safety (ammonium nitrite as a salt is explosive above 60 degrees C) and possible side-reactions as formation of ammonia and nitrogen oxides must be carefully evaluated before use in full-scale. Conductivity is a suitable parameter to follow-up the chemical denitrification process.     

Factors affecting the activity of anammox bacteria during start up in the continuous culture reactor.

Factors affecting cultivation of extremely slow-growing bacteria (anaerobic ammonium oxidiser, doubling time 11 days) were investigated by using upflow anaerobic sludge blanket (UASB) reactors which can maintain high solid retention time. The effects of concentrations of DO, free ammonia (FA), and nitrite on activation of anammox activity were tested during the start-up period. The reactor was inoculated with granular sludge collected from a full-scale UASB reactor used for treating brewery wastewater, and sludge from a piggery wastewater treatment plant and rotating biological contactor treating sewage. Results of continuous operation showed that concentrations of DO, free ammonia (FA) and nitrite in the reactors played a key role in stimulating the anammox activity during start-up period. It is crucial to keep DO below 0.2 ppm, FA below 2 mg/L and nitrite nitrogen below 35 mg/L to cultivate anammox cells in the continuous bioreactor. When the levels of DO, FA and nitrite in the influent were controlled at less than the inhibition levels, the anammox activity increased gradually in the anaerobic condition. Addition of hydrogen sulphide into the reactor enhanced anammox activity in the continuous culture. Through the SEM, TEM and FISH analysis, anammox bacteria were detected in the granular sludge after 3 months of continuous operation.     

Development of a super high-rate Anammox reactor and in situ analysis of biofilm structure and function.

The anaerobic ammonium oxidation (Anammox) process is a new efficient and cost effective method of ammonium removal from wastewater. Under strictly anoxic condition, ammonium is directly oxidised with nitrite as electron acceptor to dinitrogen gas. However, it is extremely difficult to cultivate Anammox bacteria due to their low growth rate. This suggests that a rapid and efficient start-up of Anammox process is the key to practical applications. To screen appropriate seeding sludge with high Anammox potential, a real-time quantitative PCR assay with newly designed primers has been developed. Thereafter, the seeding sludge with high abundance of Anammox bacteria (1.7 x 10(8) copies/mg-dry weight) was selected and inoculated into an upflow anaerobic biofilters (UABs). The UABs were operated for more than 1 year and the highest nitrogen removal rate of 24.0 kg-N m-3 day(-1) was attained. In addition, the ecophysiology of Anammox bacteria (spatial distribution and in situ activity) in biofilms was analysed by combining a full-cycle 16S rRNA approach and microelectrodes. The microelectrode measurement clearly revealed that a successive vertical zonation of the partial nitrification (NH4+ to NO2-), Anammox reaction and denitrification was developed in the biofilm in the UAB. This result agreed with the spatial distribution of corresponding bacterial populations in the biofilm. We linked the micro-scale information (i.e. single cell and/or biofilm levels) with the macro-scale information (i.e. the reactor level) to understand the details of Anammox reaction occurring in the UABs.     

Combined partial nitritation/Anammox system for treatment of digester supernatant.

One-year (2004) comprehensive investigations in a semi-industrial pilot plant (5 m3) were carried out with the aim of assessing the influence of operational parameters on the partial nitritation/Anammox system performance. In the system designed as a moving-bed biofilm reactor, the influent nitrogen load to the Anammox reactor was progressively increased and a stable Anammox bacterial culture was obtained. Interaction between subsequent aerobic and anaerobic conditions in the partial nitritation and Anammox reactors, respectively, granted conditions to remove nitrogen through the nitrite route. It implies that the oxygen supply can be limited to a high extent. A control strategy for the partial nitritation step relied on concomitant adjustment of the air supply with a variable influent nitrogen load, which can be monitored by both pH and conductivity measurements. In the Anammox reactor, an influent nitrite-to-ammonium ratio plays a vital role in obtaining efficient nitrogen removal. During the 1-year experimental period, the Anammox reactor was operated steadily and average nitrogen removal efficiency was 84% with 97% as the maximum value.     

微生物在湿地氮循环系统的效应分析

从湿地植物根区微环境研究出发,分析了湿地系统氮循环中微生物效应及其湿地植物为微生物生长提供的微生态系统功能。指出在湿地系统氮循环中的微生物过程(包括亚硝化、硝化、反硝化和异化作用等)是氮降解的主要过程,植物对营养物质的直接去除所占比例很小,但植物的生理特性从不同途径影响不同氮化合物的微生物转化。厌氧氨氧化过程是一种氮素转化的新途径,能同时将两种氮污染物(氨氮和亚硝酸氮)转化成气态氮,从而高效地减缓氮污染问题。好氧/厌氧界面在湿地植物根区微环境中的广泛存在及有关研究成果,说明了湿地氮循环系统中厌氧氨氧化过程存在的可能性。     

Partial nitrification of sludge reject water by means of aerobic granulation

Granular sludge formation was performed in a laboratory scale Sequencing Batch Reactor (SBR) fed with supernatant of anaerobic digestion of sewage sludge. This effluent was concentrated progressively in order to enhance biomass capacity without inhibiting it. During the first part of the study, ammonium nitrogen was converted to nitrate, so conventional nitrification took place. When a nitrogen load of 0.8 g N L(-1) d(-1) was treated, the effluent concentration of nitrite started to increase while the nitrate concentration decreased until it disappeared. So, partial nitrification was achieved. At the end of this study, a nitrogen load of 1.1 g N L(-1) d(-1) was treated obtaining an effluent with 50% ammonium and 50% nitrite. The volatile suspended solids concentration in the reactor reached 10 g VSS L(-1). At this point the granule morphology was quite round and no filamentous bacteria was observed. The Feret's diameter was in the range between 1 and 6 mm with an average value of 4.5 mm. Roundness value was all the time higher than 0.7. Granule density increased during the experimental period, obtaining a final value of 7.0 g L(-1).     

The presence of ammonium facilitates nitrite reduction under PHB driven simultaneous nitrification and denitrification.

For economic and efficient nitrogen removal from wastewater treatment plants via simultaneous nitrification and denitrification the nitrification process should stop at the level of nitrite such that nitrite rather than nitrate becomes the substrate for denitrification. This study aims to contribute to the understanding of the conditions that are necessary to improve nitrite reduction over nitrite oxidation. Laboratory sequencing batch reactors (SBRs) were operated with synthetic wastewater containing acetate as COD and ammonium as the nitrogen source. Computer controlled operation of the reactors allowed reproducible simultaneous nitrification and denitrification (SND). The oxygen supply was kept precisely at a low level of 0.5 mgL(-1) and bacterial PHB was the only electron donor available for denitrification. During SND little nitrite or nitrate accumulated (< 20% total N), indicating that the reducing processes were almost as fast as the production of nitrite and nitrate from nitrification. Nitrite spiking tests were performed to investigate the fate of nitrite under different oxidation (0.1-1.5 mgL(-1) of dissolved oxygen) and reduction conditions. High levels of reducing power were provided by allowing the cells to build up to 2.5 mM of PHB. Nitrite added was preferentially oxidised to nitrate rather than reduced even when dissolved oxygen was low and reducing power (PHB) was excessively high. However, the presence of ammonium enabled significant reduction of nitrite under low oxygen conditions. This is consistent with previous observations in SBR where aerobic nitrite and nitrate reduction occurred only as long as ammonium was present. As soon as ammonium was depleted, the rate of denitrification decreased significantly. The significance of the observed strongly stimulating effect of ammonium on nitrite reduction under SND conditions is discussed and potential consequences for SBR operation are suggested.