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.     

Biological uptake of influent organic matter as an electron donor for denitrification by activated sludge

To preserve organic materials in the sludge required for denitrification, a study was made with a contact process. The contactor, when operated with a short retention time (30 min), gave incomplete metabolism of organic matter. With 64% of the influent soluble COD (SCOD) was adsorbed to activated sludge within 30 min. The specific mass of organic matter uptaken was 55 mg SCOD/g Mixed Liquor Suspended Solids that enhanced the denitrification efficiency up to 63% in the following denitrification step.     

Removal of ammonium via simultaneous nitrification-denitrification nitrite-shortcut in a single packed-bed batch reactor

A polyurethane packed-bed-biofilm sequential batch reactor was fed with synthetic substrate simulating the composition of UASB reactor effluents. Two distinct ammonia nitrogen concentrations (125 and 250 mg l(-1)) were supplied during two sequential long-term experiments of 160 days each (320 total). Cycles of 24h under intermittent aeration for periods of 1h were applied, and ethanol was added as a carbon source at the beginning of each anoxic period. Nitrite was the main oxidized nitrogen compound which accumulated only during the aerated phases of the batch cycle. A consistent decrease of nitrite concentration started always immediately after the interruption of oxygen supply and addition of the electron donor. Removal to below detection limits of all nitrogen soluble forms was always observed at the end of the 24h cycles for both initial concentrations. Polyurethane packed-bed matrices and ethanol amendments conferred high process stability. Microbial investigation by cloning suggested that nitrification was carried out by Nitrosomonas-like species whereas denitrification was mediated by unclassified species commonly observed in denitrifying environments. The packed-bed batch bioreactor favored the simultaneous colonization of distinct microbial groups within the immobilized microbial biomass. The biofilm was capable of actively oxidizing ammonium and denitrification at high ratios in intermittent intervals within 24h cycles.     

Simultaneous occurrence of denitrification and nitrate ammonification in sediments of the French Mediterranean Coast

Dissimilatory nitrate reductions in coastal marine sediment of Carteau Cove (French Mediterranean Coast) were studied between April 1993 and July 1994. Simultaneous determination of denitrification and dissimilatory nitrate reduction to ammonium was achieved by using a combination of acetylene blockage and 15N techniques. After short incubations (maximum 5 h), a part of 15N labelled nitrate added to the sediment was recovered as ammonium without incorporation in organic matter. The result indicate that a fraction of nitrate was reduced to ammonium by a dissimilatory mechanism instead of denitrifying. Denitrifying and nitrate ammonifying activities ranged from 0 to 19.8 μmol l-1 d-1 and from 2.3 to 83.2 μmol l-1 d-1, respectively. Denitrification rates were highest in early spring whereas nitrate ammonification were highest in fall. The recovery of nitrate reduced as N2O-N plus ammonium was between 40 and 100%, the highest nitrogen losses were recorded in July. Depending on the station and time of year denitrification accounted for between 0 and 43% of the total nitrate reduction whereas dissimilatory nitrate reduction to ammonium (DNRA) accounted for between 18 and 100%. The reduction rate data suggest that the pathway of nitrate reduction to ammonium may be important in coastal sediments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nitrification-denitrification via nitrite accumulation for nitrogen removal from wastewaters

The biological nitrification-denitrification process is used extensively for removal of ammonia nitrogen from wastewaters. Saves in aeration, organic matter (for denitrification) and surplus sludge are achievable if nitrite accumulation is possible in the nitrification step. In this paper, operational parameters were studied for each process for maximum nitrite accumulation in the nitrification step and nitrite adaptation in the denitrification step. Nitrite accumulation during nitrification can be controlled by the dissolved oxygen (DO) concentration, presenting a maximum of 65% at around 0.7 mg DO/L. Denitrification can be adapted to nitrite and the process is stable if nitrite in the reactor is keep low. The performance of a continuous stirred tank reactor (CSTR) and an up flow sludge blanket reactor (USB) were compared. Once the operational parameters were established, a CSTR for nitrification and an USB reactor for denitrification were operated in series for 25 days. The process was stable and a steady state was maintained for 20 days, and 93.5% of overall nitrogen removal was achieved in the nitrification-denitrification via the nitrite process.     

Application of biochemical oxygen demand (BOD) biosensor for optimization of biological carbon and nitrogen removal from synthetic wastewater in a sequencing batch reactor system

A bench scale reactor using a sequencing batch reactor process was used to evaluate the applicability of biosensors for the process optimization of biological carbon and nitrogen removal. A commercial biochemical oxygen demand (BOD) biosensor with a novel microbial membrane was used to determine the duration of each phase by measuring samples in real time in an SBR cycle with filling/anoxic-anaerobic/aerobic/sludge wasting/settling/withdrawal periods. Possible strategies to increase the efficiency for the biological removal of carbon and nitrogen from synthetic wastewater have been developed. The results show that application of a BOD biosensor enables estimation of organic carbon, in real time, allowing the optimization or reduction the SBR cycle time. Some typical consumption patterns for organic carbon in the non-aeration phase of a typical SBR operation were identified. The rate of decrease of BOD measured using a sensor BOD, was the highest in the initial glucose breakdown period and during denitrification. It then slowed down until a 'quiescent period' was observed, which may be considered as the commencement of the aeration period. Monitoring the BOD curve with a BOD biosensor allowed the reduction of the SBR cycle time, which leads to an increase in the removal efficiency. By reducing the cycle time from 8 to 4 h cycle, the removal efficiencies of nitrate, glucose, and phosphorus in a given time interval, were increased to nearly double, while the removal of nitrogen ammonium was increased by one-third.     

Factors Controlling Anaerobic Ammonium Oxidation with Nitrite in Marine Sediments

Factors controlling the anaerobic oxidation of ammonium with nitrate and nitrite were explored in a marine sediment from the Skagerrak in the Baltic-North Sea transition. In anoxic incubations with the addition of nitrite, approximately 65% of the nitrogen gas formation was due to anaerobic ammonium oxidation with nitrite, with the remainder being produced by denitrification. Anaerobic ammonium oxidation with nitrite exhibited a biological temperature response, with a rate optimum at 15°C and a maximum temperature of 37°C. The biological nature of the process and a 1:1 stoichiometry for the reaction between nitrite and ammonium indicated that the transformations might be attributed to the anammox process. Attempts to find other anaerobic ammonium-oxidizing processes in this sediment failed. The apparent K_m of nitrite consumption was less than 3 μM, and the relative importance of ammonium oxidation with nitrite and denitrification for the production of nitrogen gas was independent of nitrite concentration. Thus, the quantitative importance of ammonium oxidation with nitrite in the jar incubations at elevated nitrite concentrations probably represents the in situ situation. With the addition of nitrate, the production of nitrite from nitrate was four times faster than its consumption and therefore did not limit the rate of ammonium oxidation. Accordingly, the rate of this process was the same whether nitrate or nitrite was added as electron acceptor. The addition of organic matter did not stimulate denitrification, possibly because it was outcompeted by manganese reduction or because transport limitation was removed due to homogenization of the sediment.     

A model and its implications for denitrification in soil environment

We analyzed the mechanisms of a soil nitrogen (N) sub-model, which is a subroutine of the Crop-Environment Resources Synthesis (CERES)-maize; a model which was originally designed to simulate crop yield and has been calibrated and validated in Taiwan. Some experiments designed specifically for testing the N sub-model proved the capability of the model in reflecting field observations. With the mechanisms, we could write computer programs for calculating the relative sensitivities of major parameters in the model, and for simulating different treatments of organic matter. The purposes were to find how they affected N transformations, especially the processes of denitrification, which are considered to be responsible for N losses in upland soils and are an important environmental issue related to human disturbance of the N cycle. The results implied that soil water content and temperature were, respectively, the first and second dominant factors. They were much more sensitive than any other parameters, such as the decomposition rate coefficients, soil pH and bulk density. Decomposition of organic matter could slow down if organic matter with different carbon/nitrogen (C/N) ratios were treated in fractions. This treatment could also decrease the process of denitrification unless the organic matter was extremely large in quantity and has a high C/N ratio.     

Assessment of a bioaugmentation strategy with polyphosphate accumulating organisms in a nitrification/denitrification sequencing batch reactor

Different alternative configurations and strategies for the simultaneous biological removal of organic matter and nutrients (N and P) in wastewater have been proposed in the literature. This work demonstrates a new successful strategy to bring in enhanced biological phosphorus removal (EBPR) to a conventional nitrification/denitrification system by means of bioaugmentation with an enriched culture of phosphorus accumulating organisms (PAO). This strategy was tested in a sequencing batch reactor (SBR), where an 8 h configuration with 3 h anoxic, 4.5 h aerobic and 25 min of settling confirmed that nitrification, denitrification and PAO activity could be maintained for a minimum of 60 days of operation after the bioaugmentation step. The successful bioaugmentation strategy opens new possibilities for retrofitting full-scale WWTP originally designed for only nitrification/denitrification. These systems could remove P simultaneously to COD and N if they were bioaugmented with waste purge of an anaerobic/aerobic SBR operated in parallel treating part of the influent wastewater.     

Biological treatment of ammonium-rich wastewater by partial nitritation and subsequent anaerobic ammonium oxidation (anammox) in a pilot plant

In wastewater treatment plants with anaerobic sludge digestion, 15-20% of the nitrogen load is recirculated to the main stream with the return liquors from dewatering. Separate treatment of this ammonium-rich digester supernatant would significantly reduce the nitrogen load of the activated sludge system. Some years ago, a novel biological process was discovered in which ammonium is converted to nitrogen gas under anoxic conditions with nitrite as the electron acceptor (anaerobic ammonium oxidation, anammox). Compared to conventional nitrification and denitrification, the aeration and carbon-source demand is reduced by over 50 and 100%, respectively. The combination of partial nitritation to produce nitrite in a first step and subsequent anaerobic ammonium oxidation in a second reactor was successfully tested on a pilot scale (3.6 m(3)) for over half a year. This report focuses on the feasibility of nitrogen removal from digester effluents from two different wastewater treatment plants (WWTPs) with the combined partial nitritation/anammox process. Nitritation was performed in a continuously stirred tank reactor (V=2.0 m(3)) without sludge retention. Some 58% of the ammonium in the supernatant was converted to nitrite. At 30 degrees C the maximum dilution rate D(x) was 0.85 d(-1), resulting in nitrite production of 0.35 kg NO(2)-N m(-3)(reactor) d(-1). The nitrate production was marginal. The anaerobic ammonium oxidation was carried out in a sequencing batch reactor (SBR, V=1.6 m(3)) with a nitrogen elimination rate of 2.4 kg N m(-3)(reactor) d(-1) during the nitrite-containing periods of the SBR cycle. Over 90% of the inlet nitrogen load to the anammox reactor was removed and the sludge production was negligible. The nitritation efficiency of the first reactor limited the overall maximum rate of nitrogen elimination.     

Kinetics of nitrogen removal from sanitary landfill leachate

Leachates from municipal landfill ‘Lublinek‘ in Lodz city (Poland) were treated in two-sludge sequence batch reactor systems (type ND). The external carbon source—sodium acetate was used in denitrification. Two versions were tested—with and without supernatant recirculation from D-SBR to N-SBR. The 99% removal of inorganic nitrogen compounds was achieved in both versions. Due to recirculation, the buffering (by means of sodium bicarbonate) was not necessary. Kinetic models for nitrification and denitrification processes were proposed. Kinetic parameters of the models were estimated by the optimization method. The experimental results of both nitrification steps (oxidation of ammonia to nitrites and nitrites to nitrates) were best fitted with the interactive limitation model of nitrogen compounds and oxygen. The denitrification process was limited only by the concentration of nitrogen compounds.     

Fluidized-bed denitrification for mine waters. Part II: effects of Ni and Co

The dispersion of nitrogenous compounds and heavy metals into the environment is frequent during mining activities. The effects of nickel (Ni) and cobalt (Co) on denitrification of simulated mine waters were investigated in batch bioassays and fluidized-bed reactors (FBRs). At pH 7, batch tests revealed that Co did not exhibit inhibition on denitrification even at 86.6 mg/L. Ni showed to be inhibitory at 50 and 100 mg/L by decreasing nitrate removal efficiencies of 18 and 65 %, respectively. In two FBRs, operated at 7–8 and 22 °C, 5.5 mg/L Ni did not affect nitrate and nitrite removals because of FBR potential of diluting soluble Ni feed concentration. On the contrary, the effluent pH clearly decreased in both FBR1 and FBR2 because of nickel sulfide precipitation and Ni inhibition of the last two steps of denitrification. When Ni injection was stopped, the process recovered more slowly at 22 than 7–8 °C. This is the first study reporting the effect of Ni on denitrification in biological FBRs.     

Selection of organic substrates as potential reactive materials for use in a denitrification permeable reactive barrier (PRB)

The aim of the present study was to select a suitable natural organic substrate as a potential carbon source for use in a denitrification permeable reactive barrier (PRB). A number of seven organic substrates were first tested in batch tests. The materials attained varying degrees of success at promoting denitrification. Some of the organic substrates performed very well, achieving complete nitrate removal (>98%), while others were considered unsuitable for a variety of reasons, including: insufficient nitrate or nitrogen removal, excessive release of leachable nitrogen from the substrate or excessive reduction of nitrate to ammonium rather than removing it as gaseous N2. The top performing substrate in terms of denitrification extent (>98%) and rate (0.067 mgNO3(-)-N dm(-3)d(-1)g(sub)(-1)) was then selected for two bench-scale column experiments in an attempt to simulate the PRB. The inlet concentration was 50 mg dm(-3) NO3(-)-N and the columns operated at two different flow rates: 0.3 cm3 min(-1) (Column 1) and 1.1cm3 min(-1) (Column 2). The two columns showed different general patterns, making it clear that the flow rate was a key factor at the nitrate removal. Nitrate was completely removed (>96%) by the passage through Column 1, while only partially removed in Column 2 (66%). The results indicated that the selected organic substrate (Softwood) was applicable for further use as a filling material for a PRB.     

Production of N(2) through anaerobic ammonium oxidation coupled to nitrate reduction in marine sediments

In the global nitrogen cycle, bacterial denitrification is recognized as the only quantitatively important process that converts fixed nitrogen to atmospheric nitrogen gas, N(2), thereby influencing many aspects of ecosystem function and global biogeochemistry. However, we have found that a process novel to the marine nitrogen cycle, anaerobic oxidation of ammonium coupled to nitrate reduction, contributes substantially to N(2) production in marine sediments. Incubations with (15)N-labeled nitrate or ammonium demonstrated that during this process, N(2) is formed through one-to-one pairing of nitrogen from nitrate and ammonium, which clearly separates the process from denitrification. Nitrite, which accumulated transiently, was likely the oxidant for ammonium, and the process is thus similar to the anammox process known from wastewater bioreactors. Anaerobic ammonium oxidation accounted for 24 and 67% of the total N(2) production at two typical continental shelf sites, whereas it was detectable but insignificant relative to denitrification in a eutrophic coastal bay. However, rates of anaerobic ammonium oxidation were higher in the coastal sediment than at the deepest site and the variability in the relative contribution to N(2) production between sites was related to large differences in rates of denitrification. Thus, the relative importance of anaerobic ammonium oxidation and denitrification in N(2) production appears to be regulated by the availability of their reduced substrates. By shunting nitrogen directly from ammonium to N(2), anaerobic ammonium oxidation promotes the removal of fixed nitrogen in the oceans. The process can explain ammonium deficiencies in anoxic waters and sediments, and it may contribute significantly to oceanic nitrogen budgets.     

Effects of operating parameters on performances of nitrification and denitrification processes

Nitrification and denitrification of synthetic wastewater was studied by using two reactors in series. An activated sludge unit was used for nitrification followed by a downflow biofilter (packed column) for denitrification. A glucose solution was fed to the denitrification column to supply carbon source. Effects of important process variables such as sludge age, hydraulic residence time and feed ammonium concentration on system's performance were investigated. Effluent ammonium-nitrogen (NH4-N) concentration decreased with increasing sludge age and hydraulic residence time and remained constant for sludge age and hydraulic residence times greater than 12 d and 15 h, respectively. Feed ammonium-nitrogen concentration above 200 mg/l resulted in significant levels of NH4-N in the effluent at Š_c = 15 d and Š_H = 12 h in nitrification. Performance of denitrification stage was not satisfactory for feed NO₃-N concentrations above 150 mg N/l resulting in significant effluent NO₃-N levels at hydraulic residence time of Š_H = 6 h.     

Factors controlling anaerobic ammonium oxidation with nitrite in marine sediments

Factors controlling the anaerobic oxidation of ammonium with nitrate and nitrite were explored in a marine sediment from the Skagerrak in the Baltic-North Sea transition. In anoxic incubations with the addition of nitrite, approximately 65% of the nitrogen gas formation was due to anaerobic ammonium oxidation with nitrite, with the remainder being produced by denitrification. Anaerobic ammonium oxidation with nitrite exhibited a biological temperature response, with a rate optimum at 15 degrees C and a maximum temperature of 37 degrees C. The biological nature of the process and a 1:1 stoichiometry for the reaction between nitrite and ammonium indicated that the transformations might be attributed to the anammox process. Attempts to find other anaerobic ammonium-oxidizing processes in this sediment failed. The apparent K(m) of nitrite consumption was less than 3 microM, and the relative importance of ammonium oxidation with nitrite and denitrification for the production of nitrogen gas was independent of nitrite concentration. Thus, the quantitative importance of ammonium oxidation with nitrite in the jar incubations at elevated nitrite concentrations probably represents the in situ situation. With the addition of nitrate, the production of nitrite from nitrate was four times faster than its consumption and therefore did not limit the rate of ammonium oxidation. Accordingly, the rate of this process was the same whether nitrate or nitrite was added as electron acceptor. The addition of organic matter did not stimulate denitrification, possibly because it was outcompeted by manganese reduction or because transport limitation was removed due to homogenization of the sediment.     

Effect of operational cycle time length on nitrogen removal in an alternating oxidation ditch system

This paper refers to nitrogen removal optimization of an alternating oxidation ditch system through the use of a mathematical model and pilot testing. The pilot system where measurements have been made has a total volume of 120 m³ and consists of two ditches operating in four phases during one cycle and performs carbon oxidation, nitrification, denitrification and settling. The mathematical model consists of one-dimensional mass balance (convection–dispersion) equations based on the IAWPRC ASM 1 model. After the calibration and verification of the model, simulation system performance was made. Optimization is achieved by testing operational cycles and phases with different time lengths. The limits of EU directive 91/271 for nitrogen removal have been used for comparison. The findings show that operational cycles with smaller time lengths can achieve higher nitrogen removals and that an “equilibrium” between phase time percentages in the whole cycle, for a given inflow, must be achieved.     

Production of N2 through Anaerobic Ammonium Oxidation Coupled to Nitrate Reduction in Marine Sediments

In the global nitrogen cycle, bacterial denitrification is recognized as the only quantitatively important process that converts fixed nitrogen to atmospheric nitrogen gas, N₂, thereby influencing many aspects of ecosystem function and global biogeochemistry. However, we have found that a process novel to the marine nitrogen cycle, anaerobic oxidation of ammonium coupled to nitrate reduction, contributes substantially to N₂ production in marine sediments. Incubations with ¹⁵N-labeled nitrate or ammonium demonstrated that during this process, N₂ is formed through one-to-one pairing of nitrogen from nitrate and ammonium, which clearly separates the process from denitrification. Nitrite, which accumulated transiently, was likely the oxidant for ammonium, and the process is thus similar to the anammox process known from wastewater bioreactors. Anaerobic ammonium oxidation accounted for 24 and 67% of the total N₂ production at two typical continental shelf sites, whereas it was detectable but insignificant relative to denitrification in a eutrophic coastal bay. However, rates of anaerobic ammonium oxidation were higher in the coastal sediment than at the deepest site and the variability in the relative contribution to N₂ production between sites was related to large differences in rates of denitrification. Thus, the relative importance of anaerobic ammonium oxidation and denitrification in N₂ production appears to be regulated by the availability of their reduced substrates. By shunting nitrogen directly from ammonium to N₂, anaerobic ammonium oxidation promotes the removal of fixed nitrogen in the oceans. The process can explain ammonium deficiencies in anoxic waters and sediments, and it may contribute significantly to oceanic nitrogen budgets.     

Model-based design and integration of a two-step biopharmaceutical production process

This paper presents the design of a two-step process in which the first step is PEGylation of a protein, and the second step is chromatographic purification of the target mono-PEGylated protein from the unreacted and the di-PEGylated impurities. The difference between optimizing each process step separately and optimizing them simultaneously is studied. It was found that by optimizing the steps simultaneously up to a 100 % increase in productivity could be obtained without reduction in yield. Optimizing both steps at the same time makes it possible for the optimization method to take into account that the di-PEGylated protein is much easier to separate than the non-PEGylated protein. The easier separation makes it possible to get a higher yield and productivity at the same time. The effect of recycling was also studied and the yield could be increased by 30 % by recycling the unreacted protein. However, if maximum productivity is required, batch mode is preferable.