Integrating food waste management into urban wastewater treatment: Economic and environmental impacts.

Food waste is the main component of municipal solid waste (MSW) and its sustainable management is a global challenge. Co-treatment of food waste and urban wastewater in wastewater treatment plants (WWTPs) could be a plausible management strategy to reduce the MSW amount that is disposed in landfills, while converting its organic fraction into biogas in the WWTP. However, the increased organic load in the wastewater influent would impact the capital and operating costs of the WWTP, mainly due to the increase in sludge production. In this work, different scenarios for co-treatment of food waste and wastewater were studied from both economic and environmental perspectives. These scenarios were designed based on different sludge disposal and management options. The results showed that the co-treatment of food waste and wastewater would be more environmentally friendly than their separate treatment, but its economic feasibility strongly depends on the ratio between the management costs of MSW and sewage sludge.

Microalgae-bacterial biomass outperforms PN-anammox biomass for oxygen saving in continuous-flow granular reactors facing extremely low-strength freshwater aquaculture streams.

The dissolved oxygen (DO) concentration in water streams is one of the most important and critical quality parameters in aquaculture farms. The main objective of this study was to evaluate the potential of two Continuous Flow Granular Reactors, one based on Partial Nitrification-Anammox biomass (Aquammox CFGR) and the other on Microalgae-Bacteria biomass (AquaMab CFGR), for improving dissolved oxygen availability in the recirculation aquaculture systems (RAS). Both reactors treated the extremely low-strength effluents from a freshwater trout farm (1.39 mg NH4+-N/L and 7.7 mg TOC/L). The Aquammox CFGR, removed up to 68% and 100% of ammonium and nitrite, respectively, but the DO concentration in the effluent was below 1 mg O2/L while the anammox activity was not maintained. In the AquaMab CFGR, bioaugmentation of aerobic granules with microalgae was attained, producing an effluent with DO concentrations up to 9 mg O2/L and removed up to 77% and 80% of ammonium and nitrite, respectively, which is expected to reduce the aeration costs in fish farms.

Membrane Fouling Mitigation in MBR via the Feast-Famine Strategy to Enhance PHA Production by Activated Sludge.

Fouling is considered one of the main drawbacks of membrane bioreactor (MBR) technology. Among the main fouling agents, extracellular polymeric substances (EPS) are considered one of the most impactful since they cause the decrease of sludge filterability and decline of membrane flux in the long term. The present study investigated a biological strategy to reduce the membrane-fouling tendency in MBR systems. This consisted of seeding the reactor with activated sludge enriched in microorganisms with polyhydroxyalkanoate (PHA) storage ability and by imposing proper operating conditions to drive the carbon toward intracellular (PHA) rather than extracellular (EPS) accumulation. For that purpose, an MBR lab-scale plant was operated for 175 days, divided into four periods (1-4) according to different food to microorganisms' ratios (F/M) (0.80 kg COD kg TSS-1 d-1 (Period 1), 0.13 kg COD kg TSS-1 d-1 (Period 2), 0.28 kg COD kg TSS-1 d-1 (Period 3), and 0.38 kg COD kg TSS-1 d-1 (Period 4)). The application of the feast/famine strategy favored the accumulation of intracellular polymers by bacteria. The increase of the PHA accumulation inside the cells corresponded to the decrease of EPS and an F/M of 0.40-0.50 kg COD kg TSS-1 d-1 was found as optimum to maximize the PHA production, while minimizing EPS. The lowest EPS content in the sludge (18% of total suspended solids) that corresponded to the maximum content of PHA (9.3%) was found in Period 4 and determined significant mitigation of the fouling rate, whose value was close to 0.10 × 1011 m-1 h-1. Thus, by imposing proper operating conditions, it was possible to drive the organic matter toward PHA accumulation. Moreover, a lower EPS content corresponded to a decrease in the irreversible fouling mechanism, which would imply a lower frequency of the extraordinary cleaning operations. This study highlighted the possibility of obtaining a double benefit by applying an MBR system in the frame of wastewater valorization: minimizing the fouling tendency of the membrane and recovery precursors of bioplastics from wastewater in line with the circular economy model.

Simplified engineering design towards a competitive lipid-rich effluents valorization.

Medium- and long-chain fatty acids and glycerol contained in the oily fraction of many food-industry effluents are excellent candidates to produce biobased high-value triacylglycerides (TAGs) and polyhydroxyalkanoates (PHAs). The typical process configuration for TAGs recovery from lipid-rich streams always includes two steps (culture enrichment plus storage compounds accumulation) whereas, for PHAs production, an additional pretreatment of the substrate for the obtainment of soluble volatile fatty acids (VFAs) is required. To simplify the process, substrate hydrolysis, culture enrichment, and accumulation (TAG and PHA storage) were coupled here in a single sequencing batch reactor (SBR) operated under the double growth limitation strategy (DGL) and fed in pulses with industrial waste fish oil during the whole feast phase. When the SBR was operated in 12 h cycles, it was reached up to 51 wt % biopolymers after only 6 h of feast (TAG:PHA ratio of 50:51; 0.423 CmmolBIOP/CmmolS). Daily storage compound production was observed to be over 25% higher than the reached when enrichment and accumulation stages were carried in separate operational units. Increasing the feast phase length from 6 to 12 h (18 h cycle) negatively affected the DGL strategy performance and hence system storage capacity, which was recovered after also extending the famine phase in the same proportion (24 h cycle). Besides, the carbon influx during the feast phase was identified as a key operational parameter controlling storage compounds production and, together with the C/N ratio, culture selection. The different cycle configurations tested clearly modulated the total fungal abundances without no significant differences in the size of the bacterial populations. Several PHA and TAG producers were found in the mixed culture although the PHA and TAG productions were poorly associated with the increased relative abundances (RAs) of specific operational taxonomic units (OTUs).

Dynamics of PHA-Accumulating Bacterial Communities Fed with Lipid-Rich Liquid Effluents from Fish-Canning Industries.

The biosynthesis of polyhydroxyalkanoates (PHAs) from industrial wastes by mixed microbial cultures (MMCs) enriched in PHA-accumulating bacteria is a promising technology to replace petroleum-based plastics. However, the populations' dynamics in the PHA-accumulating MMCs are not well known. Therefore, the main objective of this study was to address the shifts in the size and structure of the bacterial communities in two lab-scale sequencing batch reactors (SBRs) fed with fish-canning effluents and operated under non-saline (SBR-N, 0.5 g NaCl/L) or saline (SBR-S, 10 g NaCl/L) conditions, by using a combination of quantitative PCR and Illumina sequencing of bacterial 16S rRNA genes. A double growth limitation (DGL) strategy, in which nitrogen availability was limited and uncoupled to carbon addition, strongly modulated the relative abundances of the PHA-accumulating bacteria, leading to an increase in the accumulation of PHAs, independently of the saline conditions (average 9.04 wt% and 11.69 wt%, maximum yields 22.03 wt% and 26.33% SBR-N and SBR-S, respectively). On the other hand, no correlations were found among the PHAs accumulation yields and the absolute abundances of total Bacteria, which decreased through time in the SBR-N and did not present statistical differences in the SBR-S. Acinetobacter, Calothrix, Dyella, Flavobacterium, Novosphingobium, Qipengyuania, and Tsukamurella were key PHA-accumulating genera in both SBRs under the DGL strategy, which was revealed as a successful tool to obtain a PHA-enriched MMC using fish-canning effluents.

Valorization of lipid-rich wastewaters: A theoretical analysis to tackle the competition between polyhydroxyalkanoate and triacylglyceride-storing populations.

The lipid fraction of the effluents generated in several food-processing activities can be transformed into polyhydroxyalkanoates (PHAs) and triacylglycerides (TAGs), through open culture biotechnologies. Although competition between storing and non-storing populations in mixed microbial cultures (MMCs) has been widely studied, the right selective environment allowing for the robust enrichment of a community when different types of accumulators coexist is still not clear. In this research, comprehensive metabolic analyses of PHA and TAG synthesis and degradation, and concomitant respiration of external carbon, were used to understand and explain the changes observed in a laboratory-scale bioreactor fed with the lipid-rich fraction (mainly oleic acid) of a wastewater stream produced in the fish-canning industry. It was concluded that the mode of oxygen, carbon, and nitrogen supply determines the enrichment of the culture in specific populations, and hence the type of intracellular compounds preferentially accumulated. Coupled carbon and nitrogen feeding regime mainly selects for TAG producers whereas uncoupled feeding leads to PHA or TAG production function of the rate of carbon supply under specific aeration rates and feast and famine phases lengths.

Monitoring morphological changes from activated sludge to aerobic granular sludge under distinct organic loading rates and increasing minimal imposed sludge settling velocities through quantitative image analysis.

Quantitative image analysis (QIA) was used for monitoring the morphology of activated sludge (AS) during a granulation process and, thus, to define and quantify, unequivocally, structural changes in microbial aggregates correlated with the sludge properties and granulation rates. Two sequencing batch reactors fed with acetate at organic loading rates of 1.1 ± 0.6 kgCOD m-3 d-1 (R1) and 2.0 ± 0.2 kgCOD m-3 d-1 (R2) and three minimal imposed sludge settling velocities (0.27 m h-1, 0.53 m h-1, and 5.3 m h-1) induced distinct granulation processes and rates. QIA results evidenced the turning point from flocculation to granulation processes by revealing the differences in the aggregates' stratification patterns and quantifying the morphology of aggregates with equivalent diameter (Deq) of 200 µm ≤ Deq ≤ 650 µm. Multivariate statistical analysis of the QIA data allowed to distinguish the granulation status in both systems, by clustering the observations according to the sludge aggregation and granules maturation status, and successfully predicting the sludge volume index measured at 5 min (SVI5) and 30 min (SVI30). These results evidence the possibility of defining unequivocally the granulation rate and anticipating the sludge settling properties at early stages of the process using QIA data. Hence, QIA could be used to predict episodes of granules disruption and hindered settling ability in aerobic granulation sludge processes.

Wastewater Valorization: Practice around the World at Pilot- and Full-Scale.

Over the last few years, wastewater treatment plants (WWTPs) have been rebranded as water resource recovery facilities (WRRFs), which recognize the resource recovery potential that exists in wastewater streams. WRRFs contribute to a circular economy by not only producing clean water but by recovering valuable resources such as nutrients, energy, and other bio-based materials. To this aim, huge efforts in technological progress have been made to valorize sewage and sewage sludge, transforming them into valuable resources. This review summarizes some of the widely used and effective strategies applied at pilot- and full-scale settings in order to valorize the wastewater treatment process. An overview of the different technologies applied in the water and sludge line is presented, covering a broad range of resources, i.e., water, biomass, energy, nutrients, volatile fatty acids (VFA), polyhydroxyalkanoates (PHA), and exopolymeric substances (EPS). Moreover, guidelines and regulations around the world related to water reuse and resource valorization are reviewed.

Sequencing versus continuous granular sludge reactor for the treatment of freshwater aquaculture effluents.

Ammonium and nitrite levels in water are crucial for fish health preservation and growth maintenance in freshwater aquaculture farms, limiting water recirculation. The aim of the present work was the evaluation and comparison of two granular sludge reactors which were operated to treat freshwater aquaculture streams at laboratory-scale: an Aerobic Granular Sludge - Sequencing Batch Reactor (AGS-SBR) and a Continuous Flow Granular Reactor (CFGR). Both units were fed with a synthetic medium mimicking an aquaculture recycling water (1.9-2.9 mg N/L), with low carbon content, and operational temperature varied between 17 and 25 °C. The AGS-SBR, inoculated with mature granules from a full-scale wastewater treatment plant, achieved high carbon and ammonium removal during the 157 operational days. Even at low hydraulic retention time (HRT), varying from 474 to 237 min, ammonium removal efficiencies of approximately 87-100% were observed, with an ammonium removal rate of approximately 14.5 mg NH4+-N/(L⋅d). Partial biomass washout occurred due to the extremely low carbon and nitrogen concentrations in the feeding, which could only support the growth of a small portion of bacteria, but no major changes on the reactor removal performance were observed. The CFGR was inoculated with activated sludge and operated for 98 days. Biomass granulation occurred in 7 days, improving the settling properties due to a high up-flow velocity of 11 m/h and an applied HRT of 5 min. The reactor presented mature granules after 32 days, achieving an average diameter of 1.9 mm at day 63. The CFGR ammonium removal efficiencies were of approximately 10-20%, with ammonium removal rates of 90.0 mg NH4+-N/(L⋅d). The main biological processes taking place in the AGS-SBR were nitrification and heterotrophic growth, while in the CFGR the ammonium removal occurred only by heterotrophic assimilation, with the reactor also presenting complete and partial denitrification, which caused nitrite production. Comparing both systems, the CFGR achieved 6 times higher ammonium removal rates than the AGS-SBR, being suitable for treating extremely high flows. On the other hand, the AGS-SBR removed almost 100% of ammonium content in the wastewater, discharging a better quality effluent, less toxic for the fish but treated lower flows.

Strategies for the valorisation of a protein-rich saline waste stream into polyhydroxyalkanoates (PHA).

Saline Mussels Cooking Wastewater was valorised to produce PHA with Mixed Microbial Cultures (MMC). Due to the high protein content (1.8-5.7 g CODPROT/L), PHA accumulating capacity was below 10%, so several strategies were tested. In the acidification unit, Na(HCO3) was added, increasing protein conversion into Volatile Fatty Acids (VFA) from 10.3% to 69.2% and subsequent PHA accumulation from 6.9 to 14.7%. In the enrichment unit, the incorporation of a settling stage after the feast phase provoked a shift in the proteins' oxidation from the feast to the famine phase, where the nitrogen released in the famine is used by the MMC for growth. This increased the biomass concentration and the tolerated COD (from 1.6 to 4.2 g VSS/L and from 2.2 to 4.38 g COD/L). Finally, varying the proteins/VFA ratio for MMC acclimation to proteins allowed increasing PHA accumulation from 8.8 to 41.5%.

Assessment of an aerobic granular sludge system in the presence of pharmaceutically active compounds by quantitative image analysis and chemometric techniques.

In this study, a sequencing batch reactor (SBR) with aerobic granular sludge (AGS) was operated with synthetic wastewater containing environmental relevant concentrations of 17ß-estradiol (E2), 17α-ethinylestradiol (EE2) and sulfamethoxazole (SMX). Despite the presence of the studied PhAC, the granular fraction clearly predominated (TSSgran/TSS ranging from 0.82 to 0.98) throughout the monitoring period, presenting aggregates with high organic fraction (VSS/TSS above 0.83) and good settling characteristics (SVI5 ranging from 15 to 39 mL/gTSS). A principal component analysis (PCA) with quantitative image analysis (QIA) based data allowed to distinguish the different operational periods, namely with mature granules (CONT), and the E2, EE2, and SMX feeding periods. It further revealed a positive relationship between the biomass density, sludge settling ability, overall and granular biomass contents, granulation properties, granular biomass fraction and large granules fraction and size. Moreover, a discriminant analysis (DA) allowed to successfully discriminate not only the different operational periods, mainly by using the floccular apparent density, granular stratification and contents data, but also the PhAC presence in samples. The filamentous bacteria contents, sludge settling properties, settling properties stability and granular stratification, structure and contents parameters were found to be crucial for that purpose.

Is the ammonia stripping pre-treatment suitable for the nitrogen removal via partial nitritation-anammox of OFMSW digestate?

Treating the organic fraction of municipal solid waste (OFMSW) can be performed by coupling the anaerobic digestion (AD) and partial nitritation-anammox (PN-AMX) processes for organic matter and nitrogen removal, respectively. Besides, an ammonia stripping (AS) step before the AD benefit the removal of organic matter. In the present study, the operation of two PN-AMX sequencing batch reactors with and without AS pre-treated OFMSW digestate (AS-SBR and nAS-SBR, respectively) was assessed. The specific anammox activity decreased by 90 % for increasing proportions of fed OFMSW in both cases, indicating no differences over the anammox activity whether the AS pre-treatment is implemented or not. For 100 % OFMSW proportion, the AS-SBR achieved better effluent quality than the nAS-SBR (127 ± 88 vs. 1050 ± 23 mg N/L) but with lower nitrogen removal rates (58 ± 8 vs. 687 ± 32 g N/(L·d)). Still, the latter required successive re-inoculations to obtain higher removal rates. Changes in the microbial communities were mainly correlated to sCOD/N ratios in the OFMSW, being Candidatus Brocadia the dominant anamnmox species. The results proved the AS to be a suitable pre-treatment, despite the higher sCOD/N ratios in the OFMSW digestate, achieving good synergy between the PN-AMX and heterotrophic denitrification processes.

A novel strategy for triacylglycerides and polyhydroxyalkanoates production using waste lipids.

Lipids are one of the main components of the organic matter present in the effluents of the food-processing industry. These waste streams can be biotransformed into valuable triacylglycerides (TAGs) and polyhydroxyalkanoates (PHAs), precursors of biofuels and biomaterials alternative to petroleum-based products. These compounds are yielded by mixed microbial cultures, and considering that both TAG and PHA accumulators may coexist within the community, it seems crucial to define those operational strategies that might control the selection of the dominant metabolic pathways (TAG or PHA accumulation). In this work, residual fish-canning oil was used as a carbon source in a two-stage process (culture selection and intracellular compounds accumulation) in which the substrate was simultaneously hydrolyzed in these two stages without the need for a previous fermentation unit. It was pretended to maximize preferential TAG or PHA storage in the accumulation reactor by the imposition of certain selective pressures in the enrichment one. Uncoupling C and N feedings and limiting nitrogen availability in the medium, allowed to maximize PHA production (82.3 wt% of PHAs, 0.80 CmmolPHA/CmmolS). Besides, when low pH in the famine phase was considered as additional selective pressure, it was possible to shift the ratio TAG:PHA from 4:96 obtaining 43.0 wt% of TAGs (0.67 CmmolTAG/CmmolS). Therefore, this novel and simplified process demonstrated versatility and efficiency in the storage of TAGs and PHAs from a unique residual feedstock and using an open culture proving that product selection can be harnessed if choosing the right operational conditions in the enrichment stage.

Volatile fatty acid production from saline cooked mussel processing wastewater at low pH.

The production of VFA using as substrate the wastewater produced in a cooked mussel processing factory, containing large COD (13.7 ±â€¯3.2 g COD/L), salt concentrations (21.8 ±â€¯2.8 g NaCl/L) and characterized by low pH (4.6 ±â€¯0.6) was evaluated. This wastewater was fed to a 5-L completely stirred tank reactor operated in continuous mode. The conversion efficiency of its COD content into volatile fatty acids (VFA) was evaluated. The maximum acidification of 43% (total VFA on soluble COD basis) was obtained when an organic loading rate of 2.5 ±â€¯0.4 g COD/(L·d) was applied to the reactor and corresponded to a VFA volumetric productivity of 0.72 ±â€¯0.07 g CODVFA/(L·d). Under steady-state conditions, the obtained mixture of VFA was composed by 80:18:2 as acetic:propionic:butyric acids (percentage of VFA on soluble COD basis). Carbohydrates were degraded up to 96% while protein fermentation did not take place, probably due to the low pH value, limiting the maximum acidification of the wastewater. Batch experiments showed that the increase of the pH from 4.2 to 4.9 by the addition of NaHCO3 resulted in the improvement of the acidification and changed the VFA mixture composition. Thus, this study demonstrates the opportunity of using complex substrates, as cooked mussel processing wastewater, to produce rich-VFA streams under unfavourable operational conditions, such as high salinity and low pH.

Performance of partial nitritation-anammox processes at mainstream conditions in an IFAS system.

The partial nitritation-anammox processes implementation in the main line of wastewater treatment plants would lead them closer to the energy autarky. With this purpose, an integrated fixed film activated sludge (IFAS) reactor was operated at pilot scale. Efficient nitrogen removal (72 ±â€¯11%) was achieved for anaerobically pre-treated municipal wastewater at low temperature (21 - 15 °C), with a nitrogen removal rate of 37 ±â€¯3 g N/(m3·d) at 15 °C. The ammonium oxidizing bacteria were more abundant in the activated sludge, while anammox bacteria were primarily located in biofilm attached onto the carriers surface. Nitrite oxidizing bacteria (NOB) activity was similar between both fractions and its specific activity decreased more than that of other populations when the operating temperature was reduced. Furthermore, the IFAS operational strategy (aerobic/anoxic periods) allowed an efficient NOB activity suppression inside the reactor, which accounted only for the 10 - 20% of the maximum potential activity.

Predicting Accumulation of Intermediate Compounds in Nitrification and Autotrophic Denitrification Processes: A Chemical Approach.

Nitrification and sulfur-based autotrophic denitrification processes can be used to remove ammonia from wastewater in an economical way. However, under certain operational conditions, these processes accumulate intermediate compounds, such as elemental sulphur, nitrite, and nitrous oxide, that are noxious for the environment. In order to predict the generation of these compounds, an analysis based on the Gibbs free energy of the possible reactions and on the oxidative capacity of the bulk liquid was done on case study systems. Results indicate that the Gibbs free energy is not a useful parameter to predict the generation of intermediate products in nitrification and autotrophic denitrification processes. Nevertheless, we show that the specific productions of nitrous oxide during nitrification, and of elemental sulphur and nitrite during autotrophic denitrification, are well related to the oxidative capacity of the bulk liquid.

Influence of biomass acclimation on the performance of a partial nitritation-anammox reactor treating industrial saline effluents.

The performance of the partial nitritation/anammox processes was evaluated for the treatment of fish canning effluents. A sequencing batch reactor (SBR) was fed with industrial wastewater, with variable salt and total ammonium nitrogen (TAN) concentrations in the range of 1.75-18.00 g-NaCl L-1 and 112 - 267 mg-TAN L-1. The SBR operation was divided into two experiments: (A) progressive increase of salt concentrations from 1.75 to 18.33 g-NaCl L-1; (B) direct application of high salt concentration (18 g-NaCl L-1). The progressive increase of NaCl concentration provoked the inhibition of the anammox biomass by up to 94% when 18 g-NaCl L-1 were added. The stable operation of the processes was achieved after 154 days when the nitrogen removal rate was 0.021 ± 0.007 g N/L·d (corresponding to 30% of removal efficiency). To avoid the development of NOB activity at low salt concentrations and to stabilize the performance of the processes dissolved oxygen was supplied by intermittent aeration. A greater removal rate of 0.029 ± 0.017 g-N L-1 d-1 was obtained with direct exposure of the inoculum to 18 g-NaCl L-1 in less than 40 days. Also, higher specific activities than those from the inoculum were achieved for salt concentrations of 15 and 20 g-NaCl L-1 after 39 days of operation. This first study of the performance of the partial nitritation/anammox processes, to treat saline wastewaters, indicates that the acclimation period can be avoided to shorten the start-up period for industrial application purposes. Nevertheless, further experiments are needed in order to improve the efficiency of the processes.

Performance and microbial features of the partial nitritation-anammox process treating fish canning wastewater with variable salt concentrations.

The partial nitritation-anammox (PN-AMX) process applied to wastewaters with high NaCl concentration was studied until now using simulated media, without considering the effect of organic matter concentration and the shift in microbial populations. This research work presents results on the application of this process to the treatment of saline industrial wastewater. Obtained results indicated that the PN-AMX process has the capability to recover its initial activity after a sudden/acute salt inhibition event (up to 16 g NaCl/L). With a progressive salt concentration increase for 150 days, the PN-AMX process was able to remove the 80% of the nitrogen at 7-9 g NaCl/L. The microbiological data indicated that NaCl and ammonia concentrations and temperature are important factors shaping PN-AMX communities. Thus, the NOB abundance (Nitrospira) decreases with the increase of the salt concentration, while heterotrophic denitrifiers are able to outcompete anammox after a peak of organic matter in the feeding.

Integration of the Anammox process to the rejection water and main stream lines of WWTPs.

Nowadays the application of Anammox based processes in the wastewater treatment plants has given a step forward. The new goal consists of removing the nitrogen present in the main stream of the WWTPs to improve their energetic efficiencies. This new approach aims to remove not only the nitrogen but also to provide a better use of the energy contained in the organic matter. The organic matter will be removed either by an anaerobic psychrophilic membrane reactor or an aerobic stage operated at low solids retention time followed by an anaerobic digestion of the generated sludge. Then ammonia coming from these units will be removed in an Anammox based process in a single unit system. The second strategy provides the best results in terms of operational costs and would allow reductions of about 28%. Recent research works performed on Anammox based processes and operated at relatively low temperatures and/or low ammonia concentrations were carried out in single-stage systems using biofilms, granules or a mixture of flocculent nitrifying and granular Anammox biomasses. These systems allowed the appropriated retention of Anammox and ammonia oxidizing bacteria but also the proliferation of nitrite oxidizing bacteria which seems to be the main drawback to achieve the required effluent quality for disposal. Therefore, prior to the implementation of the Anammox based processes at full scale to the water line, a reliable strategy to avoid nitrite oxidation should be defined in order to maintain the process stability and to obtain the desired effluent quality. If not, the application of a post-denitrification step should be necessary.