Aerobic granulation and nitrogen removal with the effluent of internal circulation reactor in start-up of a pilot-scale sequencing batch reactor

Aerobic granular sludge was successfully cultivated with the effluent of internal circulation (IC) reactor in a pilot-scale sequencing batch reactor (SBR) using activated sludge as seeding sludge. N removal was investigated in the start-up of aerobic granulation process. Initially, the phenomenon of partial nitrification was observed and nitrite accumulation rates (NO₂ ^?-N/NO _x ^? -N) were between 84.6 and 99.1?%. It was potentially caused by ammonium oxidizing bacteria (AOB) in the seeding activated sludge, high external environmental temperature (~32?°C) and free ammonia (FA) concentration. After 50?days’ running, the aerobic granules-based bioreactor demonstrated perfect performance in simultaneous removal of organic matter and ammonia nitrogen, and average removal efficiencies were maintained above 93 and 96?%, respectively. The maximum nitrogen removal efficiency of 83.1?% was achieved after the formation of aerobic granules. The average diameter of mature aerobic granular sludge mostly ranged from 0.5 to 1.0?mm. Furthermore, one typical cyclic test indicated that pH and DO profiles could be used as effective parameters for biological reactions occurring in the aerobic/anoxic process. The obtained results could provide further information on the cultivation of aerobic granular sludge with practical wastewater, especially with regard to nitrogen-rich industrial wastewater.     

Effects of loading rate and aeration on nitrogen removal and N<Subscript>2</Subscript>O emissions in intermittently aerated sequencing batch reactors treating slaughterhouse wastewater at 11 °C

This study aimed to find optimal operation conditions for nitrogen removal from high strength slaughterhouse wastewater at 11 °C using the intermittently aerated sequencing batch reactors (IASBRs) so as to provide an engineering control strategy for the IASBR technology. Two operational parameters were examined: (1) loading rates and (2) aeration rates. Both the two parameters affected variation of DO concentrations in the IASBR operation cycles. It was found that to achieve efficient nitrogen removal via partial nitrification–denitrification (PND), “DO elbow” point must appear at the end of the last aeration period. There was a correlation between the ammonium oxidizing bacteria (AOB)/nitrite oxidizing bacteria (NOB) ratio and the average DO concentrations in the last aeration periods; when the average DO concentrations in the last aeration periods were lower than 4.86 mg/L, AOB became the dominant nitrifier population, which benefited nitrogen removal via PND. Both the nitrogen loading rate and the aeration rate influenced the population sizes of AOB and NOB. To accomplish efficient nitrogen removal via PND, the optimum aeration rate (A, L air/min) applied can be predicted according to the average organic loading rates based on mathematical equations developed in this study. The research shows that the amount of N₂O generation in the aeration period was reduced with increasing the aeration rate; however, the highest N₂O generation in the non-aeration period was observed at the optimum aeration rates.     

Dinitrogen oxide production by a mixed culture of nitrifying bacteria during ammonia shock loading and aeration failure

A number of experiments was conducted in order to establish if N₂O in the exhaust gas from an aerobic consortium of nitrifiers could be used as an indicator for monitoring the nitrification process. Laboratory-scale experiments with an activated sludge system showed a strong correlation between ammonia shock loads and both the concentration of N₂O and the rate of increase of N₂O in the exhaust gas for shock loads less than 1.60 mg ammonical nitrogen (NH₃-N) per g total suspended solids (TSS). For greater ammonia shock loads, correlation was found between build-up of nitrite in the aeration tank and the concentration of N₂O in the exhaust gas from the tank. When subjecting the system to aeration failure, a similar pattern was seen, with a correlation between nitrite build-up in the aeration tank and increases in the concentration of N₂O in the exhaust gas. The results from this work suggest that the changes in N₂O concentration in the exhaust gas from a nitrifying process may be a useful parameter for monitoring such processes. Received 15 October 2001/ Accepted in revised form 05 June 2002     

Effect of oxic and anoxic conditions on nitrous oxide emissions from nitrification and denitrification processes

A lab-scale sequencing batch reactor fed with real municipal wastewater was used to study nitrous oxide (N(2)O) emissions from simulated wastewater treatment processes. The experiments were performed under four different controlled conditions as follows: (1) fully aerobic, (2) anoxic-aerobic with high dissolved oxygen (DO) concentration, (3) anoxic-aerobic with low DO concentration, and 4) intermittent aeration. The results indicated that N(2)O production can occur from both incomplete nitrification and incomplete denitrification. N(2)O production from denitrification was observed in both aerobic and anoxic phases. However, N(2)O production from aerobic conditions occurred only when both low DO concentrations and high nitrite concentration existed simultaneously. The magnitude of N(2) O produced via anoxic denitrification was lower than via oxic denitrification and required the presence of nitrite. Changes in DO, ammonium, and nitrite concentrations influenced the magnitude of N(2)O production through denitrification. The results also suggested that N(2)O can be produced from incomplete denitrification and then released to the atmosphere during aeration phase due to air stripping. Therefore, biological nitrogen removal systems should be optimized to promote complete nitrification and denitrification to minimize N(2)O emissions.     

Comparison of biological removal via nitrite with real-time control using aerobic granular sludge and flocculent activated sludge

The process of nitrification–denitrification via nitrite for nitrogen removal under real-time control mode was tested in two laboratory-scale sequencing batch reactors (SBRs) with flocculent activated sludge (R1) and aerobic granular sludge (R2) to compare operational performance and real-time control strategies. The results showed that the average ammonia nitrogen, total inorganic nitrogen (TIN), and chemical oxygen demand (COD) removal during aeration phase were 97.6%, 57.0%, and 90.1% in R2 compared with 98.6%, 48.7%, and 88.1% in R1. The TIN removed in both SBRs was partially due to the presence of simultaneous nitrification–denitrification via nitrite, especially in R2. The specific nitrification and denitrification rates in R2 were 0.0416 mgNH₄⁺–N/gSS-min and 0.1889 mgNO_X⁻–N/gSS-min, which were 1.48 times and 1.35 times that of R1. The higher rates for COD removal, nitrification, and denitrification were achieved in R2 than R1 with similar influent quality. Dissolved oxygen (DO), pH, and oxidization reduction potential, corresponding to nutrient variations, were used as diagnostic parameters to control the organic carbon degradation and nitrification–denitrification via nitrite processes in both SBRs. The online control strategy of granular SBR was similar to that of the SBR with flocculent activated sludge. However, a unique U-type pattern on the DO curve in granular SBR was different from SBR with flocculent activated sludge in aerobic phase.     

Dynamic simulation of a WWTP operated at low dissolved oxygen condition by integrating activated sludge model and a floc model

While an aeration tank in an activated sludge process is often operated with high dissolved oxygen (DO) concentration to ensure organic degradation and nitrification, it may be operated at low DO concentration to reduce energy consumption and achieve desired denitrification. The ASM1 (Activated Sludge Model No. 1) can be used to describe the activated sludge process if the nitrification and denitrification occur either during different phases or in different tanks, but it may encounter problems in simulating the denitrification phenomenon caused by low DO concentration in the aeration tank. In the present work, we developed a model integrating the ASM1 kinetics and a biofloc model to account for the actual anoxic and aerobic rates. Oxygen was assumed the only substrate of both bio-kinetically and flux limiting in the flocs and its dispersion coefficient was estimated as 1.2 × 10⁻⁴ m² day⁻¹ by using a set of measured effluent qualities of a full-scale wastewater treatment plant (WWTP) operating at low DO concentration (∼0.80 mg L⁻¹) for 60 days. Simulation studies predicted the optimal DO level of 0.36 mg L⁻¹ which would lead to minimum total nitrogen of 15.7 mg N L⁻¹ and also showed the insignificance of the addition of carbon source for nitrogen removal for the operation under study. The developed model may be helpful for process engineers to predict the plant behaviors under various configurations or operating strategies.     

Simultaneous estimation of sludge biological activity and influent nitrogen load using ORP and DO dynamics

This paper proposes a new optimization strategy to estimate nitrifiable nitrogen concentration in wastewater, nitrification rate, denitrification rate and/or COD available for denitrification of an activated sludge process submitted to intermittent aeration. The approach uses the oxydo-reduction potential and dissolved oxygen measurements only. The parameter identification is based on a Simplex optimization of a cost function related to the error between an experimental cycle (an aerobic period followed by an anoxic one) and a simulation of a reduced model derived from ASM1. Results show very good prediction of experimental oxygen, ammonium and nitrate profiles. The estimation of nitrifiable nitrogen and removal rates has been validated both on simulated data obtained from COST action 624 benchmark and on experimental data.     

Feasibility of nitrification/denitrification in a sequencing batch biofilm reactor with liquid circulation applied to post-treatment

An investigation was performed on the biological removal of ammonium nitrogen from synthetic wastewater by the simultaneous nitrification/denitrification (SND) process, using a sequencing batch biofilm reactor (SBBR). System behavior was analyzed as to the effects of sludge type used as inoculum (autotrophic/heterotrophic), wastewater feed strategy (batch/fed-batch) and aeration strategy (continuous/intermittent). The presence of an autotrophic aerobic sludge showed to be essential for nitrification startup, despite publications stating the existence of heterotrophic organisms capable of nitrifying organic and inorganic nitrogen compounds at low dissolved oxygen concentrations. As to feed strategy, batch operation (synthetic wastewater containing 100 mg COD/L and 50 mg N-NH(4)(+)/L) followed by fed-batch (synthetic wastewater with 100 mg COD/L) during a whole cycle seemed to be the most adequate, mainly during the denitrification phase. Regarding aeration strategy, an intermittent mode, with dissolved oxygen concentration of 2.0mg/L in the aeration phase, showed the best results. Under these optimal conditions, 97% of influent ammonium nitrogen (80% of total nitrogen) was removed at a rate of 86.5 mg N-NH(4)(+)/Ld. In the treated effluent only 0.2 mg N-NO(2)(-)/L,4.6 mg N-NO(3)(-)/L and 1.0 mg N-NH(4)(+)/L remained, demonstrating the potential viability of this process in post-treatment of wastewaters containing ammonium nitrogen.     

Mechanism leading to N<Subscript>2</Subscript>O production in wastewater treating biofilm systems

Wastewater treatment plants are known to be important point sources for nitrous oxide (N₂O) in the anthropogenic N cycle. Biofilm based treatment systems have gained increasing popularity in the treatment of wastewater, but the mechanisms and controls of N₂O formation are not fully understood. Here, we review functional groups of microorganism involved in nitrogen (N) transformations during wastewater treatment, with emphasis on potential mechanism of N₂O production in biofilms. Biofilms used in wastewater treatment typically harbour aerobic and anaerobic zones, mediating close interactions between different groups of N transforming organisms. Current models of mass transfer and biomass interactions in biofilms are discussed to illustrate the complex regulation of N₂O production. Ammonia oxidizing bacteria (AOB) are the prime source for N₂O in aerobic zones, while heterotrophic denitrifiers dominate N₂O production in anoxic zones. Nitrosative stress ensuing from accumulation of NO₂ ^? during partial nitrification or denitrification seems to be one of the most critical factors for enhanced N₂O formation. In AOB, N₂O production is coupled to nitrifier denitrification triggered by nitrosative stress, low O₂ tension or low pH. Chemical N₂O production from AOB intermediates (NH₂OH, HNO, NO) released during high NH₃ turnover seems to be limited to surface-near AOB clusters, since diffusive mass transport resistance for O₂ slows down NH₃ oxidation rates in deeper biofilm layers. The proportion of N₂O among gaseous intermediates (NO, N₂O, N₂) in heterotrophic denitrification increases when NO or nitrous acid (HNO₂) accumulates because of increasing NO₂ ^?, or when transient oxygen intrusion impairs complete denitrification. Limited electron donor availability due to mass transport limitation of organic substrates into anoxic biofilm zones is another important factor supporting high N₂O/N₂ ratios in heterotrophic denitrifiers. Biofilms accommodating Anammox bacteria release less N₂O, because Anammox bacteria have no known N₂O producing metabolism and reduce NO₂ ^? to N₂, thereby lowering nitrosative stress to AOB and heterotrophs.     

Modeling of activated sludge wastewater treatment processes

The performance of an activated sludge wastewater treatment process consisting of an aeration tank and a secondary settler has been studied. A tanks-in-series model with backflow was used for mathematical modeling of the activated sludge wastewater treatment process. Non-linear algebraic equations obtained from the material balances of MLSS (mixed liquor suspended solids or activated sludge), BOD (biological oxygen demand) and DO (dissolved oxygen) for the aeration tank and the settler and from the behavior of the settler were solved simultaneously using the modified Newton-Raphson technique. The concentration profiles of MLSS, BOD and DO in the aeration tank were obtained. The simulation results were examined from the viewpoints of mixing in the aeration tank and flow in the secondary settling tank. The relationships between the overall performance of the activated sludge process and the operating and design parameters such as hydraulic residence time, influent BOD, recycle ratio and waste sludge ratio were obtained.     

Impact of COD/N ratio on nitrous oxide emission from microcosm wetlands and their performance in removing nitrogen from wastewater

Constructed wetlands (CWs) are considered to be important sources of nitrous oxide (N₂O). In order to investigate the effect of influent COD/N ratio on N₂O emission and control excess emission from nitrogen removal, free water surface microcosm wetlands were used and fed with different influent. In addition, the transformation of nitrogen was examined for better understanding of the mechanism of N₂O production under different operating COD/N ratios. It was found that N₂O emission and the performance of microcosm wetlands were significantly affected by COD/N ratio of wastewater influent. Strong relationships exist between N₂O production rate and nitrite (r = 0.421, p < 0.01). During denitrification process, DO concentration crucially influences N₂O production rate. An optimal influent COD/N ratio was obtained by adjusting external carbon sources for most effective N₂O emission control and best performance of the CWs in nitrogen removal from wastewater. It is concluded that under the operating condition of COD/N ratio = 5, total N₂O emission is minimum and the microcosm wetland is most effective in wastewater nitrogen removal.     

Isolation and characterization of a novel heterotrophic nitrifying and aerobic denitrifying bacterium Pseudomonas stutzeri KTB for bioremediation of wastewater

A novel heterotrophic nitrifying and aerobic denitrifying bacterium, KTB, was isolated from activated sludge flocci collected from a biological aerated filter according to the modified Takaya method and identified as Pseudomonas stutzeri by 16S rDNA gene sequence analysis. When shaking-cultured in the presence of 4.331 mmol/L of nitrate, 4.511 mmol/L of nitrite and 4.438 mmol/L of ammonium, the strain grew fast, with μ_max being 0.42, 0.45, and 0.56/h, and displayed high nitrogen removal efficiency, with nitrogen removal rate being 0.239, 0.362, and 0.361 mmol/L/h and nitrogen removal ratio being 99.1, 100.0, and 100.0% in 18 h, respectively. The removal mainly occurred in the logarithmic phase. Nitrite accumulation did not affect denitrification performance. Nitrate concentration was below the detectable limit during the whole growth cycle when ammonium was used as sole nitrogen source. It tolerated high DO level and exhibited excellent aggregation ability. A possible pathway involved in the nitrogen removal process, which demonstrated a full nitrification and denitrification route, was speculated. The strain might be a great candidate for biological removal of nitrogen compounds from wastewater.     

Long-term continuous partial nitritation-anammox reactor aeration optimization at different nitrogen loading rates for the treatment of ammonium rich digestate lagoon supernatant

Dissolved oxygen (DO) is an important parameter for partial nitritation-anammox process but previously not evaluated for the treatment of digested biosolid thickening lagoon supernatant. Using intermittent aeration we investigated nitrogen removal from such supernatant in an integrated fixed film activated sludge (IFAS) process operated under a variety of hydraulic retention times (1.2–2.5 days). The overall nitrogen removal rate (NRR) was significantly increased (P < 0.01) from 0.26 ± 0.01 kg N m⁻³ d^-1 at HRT of 2.5 days to 0.50 ± 0.01 kg N m^-3 d^-1 at HRT of 1.2 day. Higher nitrogen loading rates needed higher DO concentrations in order to cope with the increased oxygen demand by ammonium-oxidizing bacteria (AOB). Enhancing the DO concentration from 0.18 mg L^-1 to 0.35 mg L^-1 improved AOB activity. However, when the bulk liquid DO was in the range of 0.28−0.35 mg L^-1, anammox activity inhibition was observed associated with a significant free nitrous acid (FNA) accumulation (21.70 ± 4.10 μg L^-1). Batch studies confirmed the inhibition of anammox activity under high DO conditions (0.28−0.35 mg L^-1). Aeration strategies, other than increasing the DO set points, should be investigated in order to be able to work at high nitrogen loading rates without compromising anammox activity.     

Nitrogen removal from slaughterhouse wastewater in a sequencing batch reactor under controlled low DO conditions

The aim of this study was to examine nitrogen removal from slaughterhouse wastewater in a laboratory-scale sequencing batch reactor (SBR) operated at low dissolved oxygen (DO) levels under two aeration strategies: intermittent aeration (IA) and continuous aeration (CA). Under the IA strategy, during the aeration periods, the maximum DO was controlled at 10% saturation; under the CA strategy, in the first hour of the react phase, the DO was maintained at 10% saturation, and then it was kept at 2–3% saturation in the remaining react phase. Total nitrogen removals of up to 95 and 91% were achieved under the IA and CA aeration strategies, respectively. It is proposed that in situ measurement of oxygen utilization rates can be used to control the operation of SBRs for nitrogen removal.     

Biological treatment of high strength wastewater by fed-batch operation

Biological treatment systems for high strength wastewaters are usually operated in continuous mode such as activated sludge systems. When operated at steady-state, continuous systems result in constant effluent standards. However, in the presence of shock loadings and/or toxic compounds in feed wastewater, system performance drops quite significantly as a result of partial loss of microbial activity. In fed-batch operation, wastewater is fed to the aeration tank with a flow rate determined by effluent standards. In this type of operation, wastewater can be fed to biological oxidation unit intermittently or continuously with a low flow rate without any effluent removal. Feed flow rate is adjusted by measuring COD concentration in the effluent. As a result of intermittent addition of wastewater high COD concentrations and toxic compounds are diluted in large volume of aeration tank and inhibition effects of those compounds are reduced. As a result, biological oxidation of these compounds take place at a much higher rate. In order to show the aforementioned advantage of fed-batch operation, a high strength synthetic wastewater consisting of diluted molasses, urea, KH₂PO₄ and MgSO₄ was treated in an biological aeration tank by fed-batch operation. Organisms used were an active and dominant culture of Zooglea ramigera commonly encountered in activated sludge operations. COD removal kinetics was found to be first order and the rate constant was determined.     

Minimizing nitrous oxide in biological nutrient removal from municipal wastewater by controlling copper ion concentrations

In this study, nitrous oxide (N₂O) production during biological nutrient removal (BNR) from municipal wastewater was reported to be remarkably reduced by controlling copper ion (Cu²⁺) concentration. Firstly, it was observed that the addition of Cu²⁺ (10–100 μg/L) reduced N₂O generation by 54.5–73.2 % and improved total nitrogen removal when synthetic wastewater was treated in an anaerobic–aerobic (with low dissolved oxygen) BNR process. Then, the roles of Cu²⁺ were investigated. The activities of nitrite and nitrous oxide reductases were increased by Cu²⁺ addition, which accelerated the bio-reductions of both nitrite to nitric oxide (NO ₂ ^? ?→?NO) and nitrous oxide to nitrogen gas (N₂O?→?N₂). The quantitative real-time polymerase chain reaction assay indicated that Cu²⁺ addition increased the number of N₂O reducing denitrifiers. Further investigation showed that more polyhydoxyalkanoates were utilized in the Cu²⁺-added system for denitrification. Finally, the feasibility of reducing N₂O generation by controlling Cu²⁺ was examined in two other BNR processes treating real municipal wastewater. As the Cu²⁺ in municipal wastewater is usually below 10 μg/L, according to this study, the supplement of influent Cu²⁺ to a concentration of 10–100 μg/L is beneficial to reduce N₂O emission and improve nitrogen removal when sludge concentration in the BNR system is around 3,200 mg/L.     

Identifying sources of nitrous oxide emission in anoxic/aerobic sequencing batch reactors (A/O SBRs) acclimated in different aeration rates

Both long term and batch experiments were carried out to identify the sources of the N₂O emission in anoxic/aerobic sequencing batch reactors (A/O SBRs) under different aeration rates. The obtained results showed that aeration rate has an important effect on the N₂O emission of A/O SBR and most of the N₂O was emitted during the aerobic phase. During the anoxic phase, nitrate ammonification was the major source of N₂O emission while denitrification performed as a sink of N₂O, in all three bioreactors. The N₂O emission mechanisms during the aerobic phase differed with the aeration rate. At low and high aeration rates (Run 1 and Run 3), both coupled-denitrification and nitrifier denitrification were ascribed to be the source of N₂O emission. At mild aeration rate (Run 2), nitrifier denitrification by Nitrosomonas-like ammonia oxidizing-bacterial (AOB) was responsible for N₂O emission while coupled-denitrification turned out to be a sink of N₂O because of the presence of inner anaerobic region in sludge flocs.     

Aerobic and two-stage anaerobic-aerobic sludge digestion with pure oxygen and air aeration

The degradability of excess activated sludge from a wastewater treatment plant was studied. The objective was establishing the degree of degradation using either air or pure oxygen at different temperatures. Sludge treated with pure oxygen was degraded at temperatures from 22 degrees C to 50 degrees C while samples treated with air were degraded between 32 degrees C and 65 degrees C. Using air, sludge is efficiently degraded at 37 degrees C and at 50-55 degrees C. With oxygen, sludge was most effectively degraded at 38 degrees C or at 25-30 degrees C. Two-stage anaerobic-aerobic processes were studied. The first anaerobic stage was always operated for 5 days HRT, and the second stage involved aeration with pure oxygen and an HRT between 5 and 10 days. Under these conditions, there is 53.5% VSS removal and 55.4% COD degradation at 15 days HRT - 5 days anaerobic, 10 days aerobic. Sludge digested with pure oxygen at 25 degrees C in a batch reactor converted 48% of sludge total Kjeldahl nitrogen to nitrate. Addition of an aerobic stage with pure oxygen aeration to the anaerobic digestion enhances ammonium nitrogen removal. In a two-stage anaerobic-aerobic sludge digestion process within 8 days HRT of the aerobic stage, the removal of ammonium nitrogen was 85%.     

Microbial composition of the activated sludge of Moscow wastewater treatment plants

The contribution of the major technologically important microbial groups (ammonium- and nitrite-oxidizing, phosphate-accumulating, foam-inducing, and anammox bacteria, as well as planctomycetes and methanogenic archaea) was characterized for the aeration tanks of the Moscow wastewater treatment facilities. FISH investigation revealed that aerobic sludge were eubacterial communities; the metabolically active archaea contributed insignificantly. Stage II nitrifying microorganisms and planctomycetes were significant constituents of the bacterial component of activated sludges, with Nitrobacter spp. being the dominant nitrifiers. No metabolically active anammox bacteria were revealed in the sludge from aeration tanks. The sludge from the aeration tanks using different wastewater treatment technologies were found to have differing characteristics. Abundance of the nitrifying and phosphate-accumulating bacteria in the sludge generally correlated with microbial activity in microcosms and with efficiency of nitrogen and phosphorus removal from wastewater. The highest microbial numbers and activity were found in the sludge of the tanks operating according to the technologies developed in the universities of Hannover and Cape Town. The activated sludge from the Novokur’yanovo facilities, where abundant growth of filamentous bacteria resulted in foam formation, exhibited the lowest activity. The group of foaming bacteria included Gordonia spp. and Acinetobacter spp utilizing petroleum and motor oils, Sphaerotilus spp. utilizing unsaturated fatty acids, and Candidatus ‘Microthrix parvicella’. Thus, the data on abundance and composition of metabolically active microorganisms obtained by FISH may be used for the technological control of wastewater treatment.