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Feasibility study on the removal of nitrous compounds with a hollow-fiber membrane biofilm reactor.

The objective of this study was to develop an integrated nitrogen treatment system using autotrophic organisms. A treatment system consists of an aerobic hollow-fiber membrane biofilm reactor (HfMBR) and anaerobic HfMBR. In the aerobic HfMBR, a mixture gas of air and O2 was supplied through the fibers for nitrification. Denitrification occurred in the anaerobic HfMBR using H2 as the electron donor. The treatment system was continuously operated for 190 days. NH4-N removal efficiencies ranging from 95% to 97% were achieved at NH4-N concentrations of influent ranging from 50 to 100 mg N/L. When glucose was added to the influent, the simultaneous nitrification and denitrification occurred in the aerobic HfMBR, and nitrogen removal rates were changed according to the COD/NH4-N ratio of influent. In the anaerobic HfMBR, autotrophic denitrification using H2 occurred and the removal rates achieved in this study were 23-58 mg N/m2 d. In this study, the achieved removal efficiency was lower than other study findings; however, the result suggested that this hybrid HfMBR system can be used effectively for nitrogen removal in oligotrophic water.

Aerobiosis↗

How to treat raw sewage with constructed wetlands: an overview of the French systems.

The development of vertical flow constructed wetlands treating raw wastewater in France has proved to be very successful over the last 20 years. In view of this a survey was carried out on more than 80 plants in order to study their performance and correct the design if necessary. This study shows that such systems perform well in terms of respecting the goals of both low level outlet COD and SS and nitrification. Pollutant removal performance in relation to the loads handled and the specific characteristics of the plants were investigated. Nitrification is shown to be the most sensitive process in such systems and performance in relation to sizing is discussed. Such systems, if well designed, can achieve an outlet level of 60 mg L(-1) in COD, 15 mg L(-1) in SS and 8 mg L(-1) in TKN with an area of 2 - 2.5 m2.PE(-1). The sludge deposit on the first stage must be removed after about 10-15 years.

Biodegradation, Environmental↗

Removal of nitrogen in different wetland filter materials: use of stable nitrogen isotopes to determine factors controlling denitrification and DNRA.

Laboratory incubations with varying O2 and NO3 concentrations were performed with a range of filter materials used in constructed wetlands (CWs). The study included material sampled from functioning CWs as well as raw materials subjected to laboratory pre-incubation. 15N-tracer techniques were used to assess the rates of denitrification versus dissimilatory nitrate reduction to ammonium (DNRA), and the relative role of nitrification versus denitrification in producing N2O. The N2O/(N2 + N2O) product ratio was assessed for the different materials. Sand, shell sand, and peat sustained high rates of denitrification. Raw light-weight aggregates (LWA) had a very low rate, while in LWA sampled from a functioning CW, the rate was similar to the one found in the other materials. The N2O/(N2 + N2O) ratio was very low for sand, shell sand and LWA from functioning CWs, but very high for raw LWA. The ratio was intermediate but variable for peat. The N2O produced by nitrification accounted for a significant percentage of the N2O accumulated during the incubation, but was dependent on the initial oxygen concentration. DNRA was significant only for shell sand taken from a functioning CW, suggesting that the establishment of active DNRA is a slower process than the establishment of a denitrifying flora.

Biodegradation, Environmental↗

Improving nitrogen reduction in waste stabilisation ponds.

This paper reviews the performance of two waste stablisation ponds (WSP) systems in the South Island of New Zealand that have been upgraded to multiple ponds-in-series to improve effluent quality. Results of monitoring are provided which show that it is possible to achieve relatively low ammonia (approximately 1 g/m3) and total nitrogen (approximately 10 g/m3) effluent concentrations through the use of nitrification filter beds (rock trickling filters) and sand filters. Evidence suggests that the nitrification and denitrification processes in the extra biofilm surface area provided by the rock filters or rock bank protection is primarily responsible for the improved effluent quality. The paper also compares the WSP results with effluent quality predicted by published formulae. It is concluded that these formulae do not reliably predict the performance of WSP systems and the development of universally applicable design guidelines would be useful.

Ammonia↗

Performance analysis of anoxic selector in upgrading activated sludge process in tropical climate.

The paper describes and analyses the performance of anoxic selectors in upgrading activated sludge process in a municipal wastewater treatment plant under tropical climate, where poor sludge settleability is a problem due to elevated temperature. Site monitoring and laboratory experiment were conducted to study the denitrification, sludge settleability, kinetics, mass balance, pH and alkalinity variation in the system. The sludge settleability measured in Sludge Volume Index (SVI) was observed to improve with the increasing degree of denitrification in the anoxic selector. Under well-developed stable state, an average SVI value of 136 ml/g was achieved, which was significantly lower than the value of 250 ml/g before the application of anoxic selector. The specific reaction rates for denitrification and nitrification at 30 degrees C were 0.06 mg NO3-N/mg MLSS day and 0.08 mg NH4-N/mg MLSS day, respectively. The overall efficiencies of nitrification and denitrification were 86 percent and 55.4 percent, respectively, with an alkalinity recovery ratio of 15.4 percent. 32 percent of total COD removed was used up as electron donor in the denitrification process. However, due to absence of the internal Mixed Liquor Recirculation (MLR), a higher degree of denitrification occurred in the secondary sedimentation tank than in the anoxic zone. Issues for further studies are also discussed.

Ammonia↗

Achieving and maintaining biological nitrogen removal via nitrite under normal conditions.

The principal aim of this paper is to develop an approach to realize stable biological nitrogen removal via nitrite under normal conditions. Validation of the new method was established on laboratory-scale experiments applying the sequencing batch reactor (SBR) activated sludge process to domestic wastewater with low C/N ratio. The addition of sodium chloride (NaCl) to influent was established to achieve nitrite build-up. The high nitrite accumulation, depending on the salinity in influent and the application duration of salt, was obtained in SBRs treating saline wastewater. The maintenance results indicated that the real-time SBRs can maintain stable nitrite accumulation, but conversion from shorter nitrification-denitrification to full nitrification-denitrification was observed after some operation cycles in the other SBR with fixed-time control. The presented method is valuable to offer a solution to realize and to maintain nitrogen removal via nitrite under normal conditions.

Bioreactors↗

Estimations of municipal point source pollution in the context of river basin management.

Integrated presentation of total emissions on catchment scale is prerequisite for many tasks in integrated management of point and diffuse sources of pollution. This paper will focus on emissions of nutrients from municipal point sources. Based on calculations of discharges of N, P from households into wastewater and on the detailed evaluation of data from 76 municipal wastewater treatments plants, this paper presents ranges of specific loads of inhabitants and population equivalents in the raw wastewater. In addition data of these treatment plants have been evaluated in respect of the treatment efficiency for nitrogen and phosphorus (average reduction rates) dependent on the design characteristic (with or without nitrification, denitrification or enhanced phosphorus removal). The results of the investigation show that the specific N and P loads from households in Austria lie within the range 1.6-2.0 g P/(inhabitant.d) and 11- 13 g N/(inhabitant.d). The specific contribution of industries to municipal wastewater varies between 0.3 and 2.0 gP/(pe.d) and 0 and 13 g N/(pe.d) with average values of 1.3 g P/(pe.d) and 6.5 g N/(population equivalent (pe)/d). As average values for municipal wastewater (contributions from household and industry) this leads to specific influent loads of 1.5 g P/(pe.d) and 8.8 g N/(pe.d). Average treatment efficiencies of treatment plants are for instance 50% nitrogen removal in treatment plants with nitrification and 80% in treatment plants with nitrification/denitrification. For phosphorus a removal of about 85% can be expected where the treatment plant was designed for enhanced phosphorus removal. Finally a method for load estimations based on standard values as mentioned above was tested for the estimation of emission from municipal point sources of selected regions.

Austria↗

Membrane bioreactor versus conventional activated sludge system: population dynamics of nitrifiers.

Although membrane bioreactors have attracted increasing attention in recent years, little research has been undertaken on the influence of the membrane separation on the microbial community composition. This paper compares the startup behaviour and the performance of the subsequent eight months of a membrane bioreactor with a conventional activated sludge pilot plant. Both plants were operated in parallel at the same sludge age and treated the same domestic wastewater. The identification of the nitrifying community composition using fluorescent in situ hybridization revealed only minor differences between the two reactors for both ammonia-oxidizing bacteria and nitrite-oxidizing bacteria. Accordingly, both systems exhibited the same maximum nitrification rates. Confocal laser scanning microscopy showed that the aggregates formed by nitrifying bacteria were located mostly in the inner part of the flocs and were overgrown by heterotrophic bacteria. It is concluded that the membrane separation itself does affect neither the nitrifying community composition nor the nitrification performance. However, impacts on kinetic parameters are emphasized.

Ammonia↗

Applicability of an extant batch respirometric assay in describing dynamics of ammonia and nitrite oxidation in a nitrifying bioreactor.

Several techniques have been proposed for biokinetic estimation of nitrification. Recently, an extant respirometric assay has been presented that yields kinetic parameters for both nitrification steps with minimal physiological change to the microorganisms during the assay. Herein, the ability of biokinetic parameter estimates from the extant respirometric assay to adequately describe concurrently obtained NH4+-N and NO(2-)-N substrate depletion profiles is evaluated. Based on our results, in general, the substrate depletion profiles resulted in a higher estimate of the maximum specific growth rate coefficient, micro(max) for both NH4+-N to NO(2-)-N oxidation and NO(2-)-N to NO(3-)-N oxidation compared to estimates from the extant respirograms. The trends in the kinetic parameter estimates from the different biokinetic estimation techniques are paralleled in the nature of substrate depletion profiles obtained from best-fit parameters. Based on a visual inspection, in general, best-fit parameters from optimally designed complete respirograms provided a better description of the substrate depletion profiles than estimates from isolated respirograms. Nevertheless, the sum of the squared errors for the best-fit respirometry based parameters was outside the 95% joint confidence interval computed for the best-fit substrate depletion based parameters. Notwithstanding the difference in kinetic parameter estimates determined in this study, the different biokinetic estimation techniques still are close to estimates reported in literature. Additional parameter identifiability and sensitivity analysis of parameters from substrate depletion assays revealed high precision of parameters and high parameter correlation. Although biokinetic estimation via automated extant respirometry is far more facile than via manual substrate depletion measurements, additional sensitivity analyses are needed to test the impact of differences in the resulting parameter values on continuous reactor performance.

Ammonia↗

[Main reactions in anaerobic ammonium oxidation reactor under organic carbon condition].

The main reactions occurred was studied in five ASBRs (1-5) which had run at steady ANAMMOX. The organic carbon condition was kept up by means of the addition of glucose in the wastewater. The results reveal that aerobic nitrifing bacteria, denitnifying bacteria and ANAMMOX bacteria could coexist, and the aerobic nitrification reaction, the ANAMMOX reaction and the denitrification reaction could occur according to certain order in the reactors. The optimal ANAMMOX performance was achieved in reactor 1 which C/NH4+ -N was 1.7. On hour 41, COD removal, NH4+ -N removal and NO2- -N removal in reactor 1 were 100%, 81.7% and 74.4%, respectively.

Anaerobiosis↗

Hypoosmolarity inhibits ammonia oxidation by terrestrial and freshwater Nitrosopumilaceae members.

Salinity strongly influences the physiology and distribution of nitrifying microorganisms, yet the effects of low salinity remain understudied. This study investigates the impact of hypoosmolarity on different groups of ammonia oxidizers in soil and freshwater reservoirs, as well as in pure culture isolates. In soil microcosms amended with ammonium, at low salinity levels (~120 μS/cm), comparable to values commonly found in pristine terrestrial and freshwater environments, the abundance of ammonia-oxidizing bacteria (AOB), dominated by Nitrosomonas oligotropha, significantly increased. In contrast, the growth of ammonia-oxidizing archaea (AOA), dominated by "Candidatus Nitrosotenuis" of the Nitrosopumilaceae family, was stimulated by high salinity (~760 μS/cm). In ammonium-fed freshwater microcosms, the abundance of AOB, dominated by N. oligotropha, significantly increased under both low (~170 μS/cm) and high salinity (~850 μS/cm) conditions. In the presence of allylthiourea (50 μM), used to inhibit bacterial ammonia oxidation, AOA were sensitive to low salinity in both soil and freshwater microcosms. Consistently, culture-dependent studies revealed marked growth inhibition of terrestrial AOA, especially members of Nitrosopumilaceae, under hypoosmolarity, unlike AOB and complete ammonia oxidizer (comammox) strains. Genomic analyses, along with transcriptomic studies, suggested that the sensitivity of AOA to hypoosmolarity stress was possibly due to a lack of osmoregulatory transport systems and their S-layer cell wall structure. Overall, this study indicates hypoosmolarity as an important factor shaping the ecological niches and distribution of ammonia oxidizers, as well as nitrification activities, in terrestrial and freshwater environments that are increasingly affected by intensified water cycles due to global change.

Ammonia↗

Nitrogen removal reactor using packed gel envelopes containing Nitrosomonas europaea and Paracoccus denitrificans.

Packed gel envelopes were constructed as simple, compact reactors for removing nitrogen from wastewater. Each packed gel envelope consisted of two plate gels with a spacer in between. Nitrosomonas europaea and Paracoccus denitrificans were co-immobilized in the plate gels, and ethanol, serving as an electron donor for denitrification, was injected into the internal spaces of the envelopes. The external surfaces of the envelopes were in contact with ammonia-containing wastewater; the N. europaea present in the gels oxidized the ammonia to nitrite aerobically. On the other hand, the internal surfaces of the envelopes were in contact with the ethanol solution, which P. denitrificans used to reduce the nitrite to nitrogen gas anaerobically. In this way, the reactor using the packed gel envelopes removed ammonia from wastewater in a single step. When artificial wastewater containing 200 mg-N/L was treated using the reactor using eight envelopes, the ammonia was removed by the reactor without accumulating nitrite or ethanol. This simple system exhibited high rates of nitrification (ammonia to nitrite; 1.9 kg-N/day for 1m(3) of reactor volume) and nitrogen removal (ammonia to nitrogen gas; 1.6 kg-N/day). It is presumed that these high rates were achieved as a consequence of cooperation between the N. europaea and P. denitrificans present in the gels and the efficient uptake and exhaust of gases leading to the smooth conversion of ammonia to nitrogen gas.

Ammonia↗

Sensitivity analysis of a biofilm model describing a one-stage completely autotrophic nitrogen removal (CANON) process.

A mathematical model for nitrification and anaerobic ammonium oxidation (ANAMMOX) processes in a single biofilm reactor (CANON) was developed. This model describes completely autotrophic conversion of ammonium to dinitrogen gas. Aerobic ammonium and nitrite oxidation were modeled together with ANAMMOX. The sensitivity of kinetic constants and biofilm and process parameters to the process performance was evaluated, and the total effluent concentrations were, in general, found to be insensitive to affinity constants. Increasing the amount of biomass by either increasing biofilm thickness and density or decreasing porosity had no significant influence on the total effluent concentrations, provided that a minimum total biomass was present in the reactor. The ANAMMOX process always occurred in the depth of the biofilm provided that the oxygen concentration was limiting. The optimal dissolved oxygen concentration level at which the maximum nitrogen removal occurred is related to a certain ammonium surface load on the biofilm. An ammonium surface load of 2 g N/m2. d, associated with a dissolved oxygen concentration level of 1.3 g O2/m3 in the bulk liquid and with a minimum biofilm depth of 1 mm seems a proper design condition for the one-stage ammonium removal process. Under this condition, the ammonium removal efficiency is 94% (82% for the total nitrogen removal efficiency) (30 degrees C). Better ammonium removal could be achieved with an increase in the dissolved oxygen concentration level, but this would strongly limit the ANAMMOX process and decrease total nitrogen removal. It can be concluded that a one-stage process is probably not optimal if a good nitrogen effluent is required. A two-stage process like the combined SHARON and ANAMMOX process would be advised for complete nitrogen removal.

Anaerobiosis↗

Development of a novel titration and off-gas analysis (TOGA) sensor for study of biological processes in wastewater treatment systems.

The development of the new TOGA (titration and off-gas analysis) sensor for the detailed study of biological processes in wastewater treatment systems is outlined. The main innovation of the sensor is the amalgamation of titrimetric and off-gas measurement techniques. The resulting measured signals are: hydrogen ion production rate (HPR), oxygen transfer rate (OTR), nitrogen transfer rate (NTR), and carbon dioxide transfer rate (CTR). While OTR and NTR are applicable to aerobic and anoxic conditions, respectively, HPR and CTR are useful signals under all of the conditions found in biological wastewater treatment systems, namely, aerobic, anoxic and anaerobic. The sensor is therefore a powerful tool for studying the key biological processes under all these conditions. A major benefit from the integration of the titrimetric and off-gas analysis methods is that the acid/base buffering systems, in particular the bicarbonate system, are properly accounted for. Experimental data resulting from the TOGA sensor in aerobic, anoxic, and anaerobic conditions demonstrates the strength of the new sensor. In the aerobic environment, carbon oxidation (using acetate as an example carbon source) and nitrification are studied. Both the carbon and ammonia removal rates measured by the sensor compare very well with those obtained from off-line chemical analysis. Further, the aerobic acetate removal process is examined at a fundamental level using the metabolic pathway and stoichiometry established in the literature, whereby the rate of formation of storage products is identified. Under anoxic conditions, the denitrification process is monitored and, again, the measured rate of nitrogen gas transfer (NTR) matches well with the removal of the oxidised nitrogen compounds (measured chemically). In the anaerobic environment, the enhanced biological phosphorus process was investigated. In this case, the measured sensor signals (HPR and CTR) resulting from acetate uptake were used to determine the ratio of the rates of carbon dioxide production by competing groups of microorganisms, which consequently is a measure of the activity of these organisms. The sensor involves the use of expensive equipment such as a mass spectrometer and requires special gases to operate, thus incurring significant capital and operational costs. This makes the sensor more an advanced laboratory tool than an on-line sensor.

Bacteria, Aerobic↗

Theory of pH-stat titration.

Innovative techniques are being studied to assess the activity of bioreactors and to improve the performance and operational stability of biological processes. Among these techniques, the pH-stat titration is applicable to any bioreaction involving pH variations. Up to now, the main application of the pH-stat titration has been for nitrification monitoring. In this article, we present a theoretical model of pH-stat titration, which predicts the response to any reaction involving the production or consumption of protons, hydroxyl ions, or inorganic carbon chemical species (CO(2), HCO(3)(-), CO(3)(=)). This model is a useful tool to understand pH-stat titrations, to define their applicability and limits, and to select the best experimental conditions for specific applications. Tests have been performed to compare experimental pH-stat titration rates in the presence of carbon dioxide and HCO(3)(-) producing reactions to the values predicted by the model and a very satisfying correspondence was found.

Bicarbonates↗

Analysis of size distribution and areal cell density of ammonia-oxidizing bacterial microcolonies in relation to substrate microprofiles in biofilms.

A fine-scale in situ spatial organization of ammonia-oxidizing bacteria (AOB) in biofilms was investigated by combining molecular techniques (i.e., fluorescence in situ hybridization (FISH) and 16S rDNA-cloning analysis) and microelectrode measurements. Important parameters of AOB microcolonies such as size distribution and areal cell density of the microcolonies were determined and correlated with substrate microprofiles in the biofilms. In situ hybridization with a nested 16S rRNA-targeted oligonucleotide probe set revealed two different populations of AOB, Nitrosomonas europaea-lineage and Nitrosospira multiformis-lineage, coexisting in an autotrophic nitrifying biofilm. Nitrosospira formed looser microcolonies, with an areal cell density of 0.51 cells microm(-2), which was half of the cell density of Nitrosomonas (1.12 cells microm(-2)). It is speculated that the formation of looser microcolonies facilitates substrate diffusion into the microcolonies, which might be a survival strategy to low O(2) and NH(4) (+) conditions in the biofilm. A long-term experiment (4-week cultivation at different substrate C/N ratios) revealed that the size distribution of AOB microcolonies was strongly affected by better substrate supply due to shorter distance from the surface and the presence of organic carbon. The microcolony size was relatively constant throughout the autotrophic nitrifying biofilm, while the size increased by approximately 80% toward the depth of the biofilm cultured at the substrate C/N = 1. A short-term ( approximately 3 h) organic carbon addition experiment showed that the addition of organic carbon created interspecies competition for O(2) between AOB and heterotrophic bacteria, which dramatically decreased the in situ NH(4) (+)-uptake activity of AOB in the surface of the biofilms. This result might explain the spatial distribution of AOB microcolony size in the biofilms cultured at the substrate C/N = 1. These experimental results suggest O(2) and organic carbon were the main factors controlling the spatial organization and activity of AOB in biofilms. These findings are significantly important to further improve mathematical models used to describe how the slow-growing AOB develop their niches in biofilms and how that configuration affects nitrification performance in the biofilm.

Ammonia↗

N-removal in a granular sludge sequencing batch airlift reactor.

The removal of N-compounds in the sequencing batch airlift reactor (SBAR) containing granular sludge was studied under conditions of decreased dissolved oxygen (DO). A simulation model was developed to describe and evaluate the effects of several process conditions in the SBAR on N-removal performance. The model described the experimental data reasonable well. It has been shown that nitrification, denitrification, and removal of chemical oxygen demand (COD) can occur simultaneously in a granular sludge SBR. It has also been shown that the exact location of the autotrophic biomass influences the net N-removal. The distribution of the autotrophic biomass is influenced by the DO in the reactor. The optimal DO value is expected to be around 40% air saturation. It was shown that storage and subsequent degradation of poly-beta-hydroxybutyrate (PHB) benefit the denitrification. In particular, PHB is stored in bacteria situated in deeper layers of the granules below where the autotrophic activity occurs, serves as a C-source for denitrification.

Aerobiosis↗

Modeling and simulation of oxygen-limited partial nitritation in a membrane-assisted bioreactor (MBR).

Combination of a partial nitritation process and an anaerobic ammonium oxidation process for the treatment of sludge reject water has some general cost-efficient advantages compared to nitrification-denitrification. The integrated process features two-stage autotrophic conversion of ammonium via nitrite to dinitrogen gas with lower demand for oxygen and no external carbon requirement. A nitrifying membrane-assisted bioreactor (MBR) for the treatment of sludge reject water was operated under continuous aeration at low dissolved oxygen (DO) concentrations with the purpose of generating nitrite accumulation. Microfiltration was applied to allow a high sludge retention time (SRT), resulting in a stable partial nitritation process. During start-up of the MBR, oxygen-limited conditions were induced by increasing the ammonium loading rate and decreasing the oxygen transfer. At a loading rate of 0.9 kg N m(-3) d(-1) and an oxygen concentration below 0.1 mg DO L(-1), conversion to nitrite was close to 50% of the incoming ammonium, thereby yielding an optimal effluent within the stoichiometric requirements for subsequent anaerobic ammonium oxidation. A mathematical model for ammonium oxidation to nitrite and nitrite oxidation to nitrate was developed to describe the oxygen-limited partial nitritation process within the MBR. The model was calibrated with in situ determinations of kinetic parameters for microbial growth, reflecting the intrinsic characteristics of the ammonium oxidizing growth system at limited oxygen availability and high sludge age. The oxygen transfer coefficient (K(L)a) and the ammonium-loading rate were shown to be the appropriate operational variables to describe the experimental data accurately. The validated model was used for further steady state simulation under different operational conditions of hydraulic retention time (HRT), K(L)a, temperature and SRT, with the intention to support optimized process design. Simulation results indicated that stable nitrite production from sludge reject water was feasible with this process even at a relatively low temperature of 20 degrees C with HRT down to 0.25 days.

Bacteria, Anaerobic↗