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Campos, J. L., Crutchik, D., Franchi, O., Pavissich, J. P., Belmonte, M., Pedrouso, A., et al. (2019). Nitrogen and Phosphorus Recovery From Anaerobically Pretreated Agro-Food Wastes: A Review. Front. Sustain. Food Syst., 2, 11 pp.
Abstract: Anaerobic digestion (AD) is commonly used for the stabilization of agro-food wastes and recovery of energy as methane. Since AD removes organic C but not nutrients (N and P), additional processes to remove them are usually applied to meet the stringent effluent criteria. However, in the past years, there was a shift from the removal to the recovery of nutrients as a result of increasing concerns regarding limited natural resources and the importance given to the sustainable treatment technologies. Recovering N and P from anaerobically pretreated agro-food wastes as easily transportable and marketable products has gained increasing importance to meet both regulatory requirements and increase revenue. For this reason, this review paper gives a critical comparison of the available and emerging technologies for N and P recovery from AD residues.
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Campos, J. L., del Rio, A. V., Pedrouso, A., Raux, P., Giustinianovich, E. A., & Mosquera-Corral, A. (2017). Granular biomass floatation: A simple kinetic/stoichiometric explanation. Chem. Eng. J., 311, 63–71.
Abstract: Floatation events are commonly observed in anammox, denitrifying and anaerobic granular systems mostly subjected to overloading conditions. Although several operational strategies have been proposed to avoid floatation of granular biomass, until now, there is no consensus about the conditions responsible for this phenomenon. In the present study, a simple explanation based on kinetic and stoichiometric principles defining the aforementioned processes is provided. The operational zones corresponding to evaluated parameters where risk of floatation exists are defined as a function of substrate concentration in the bulk liquid and the radius of the granule. Moreover, the possible control of biomass floatation by changing the operating temperature was analyzed. Defined operational zones and profiles fit data reported in literature for granular biomass floatation events. From the study the most influencing parameter on floatation occurrence has been identified as the substrate concentration in the bulk media. (C) 2016 Elsevier B.V. All rights reserved.
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Campos, J. L., Dumais, J., Pavissich, J. P., Franchi, O., Crutchik, D., Belmonte, M., et al. (2019). Predicting Accumulation of Intermediate Compounds in Nitrification and Autotrophic Denitrification Processes: A Chemical Approach. Biomed Res. Int., 2019, 9 pp.
Abstract: 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.
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Campos, J. L., Mosquera-Corral, A., del Rio, A. V., & Pedrouso, A. (2022). Sustainable Wastewater Management and Treatment. In Sustainability (Vol. 14, 9137).
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Campos, J. L., Valenzuela-Heredia, D., Pedrouso, A., del Rio, A. V., Belmonte, M., & Mosquera-Corral, A. (2016). Greenhouse Gases Emissions from Wastewater Treatment Plants: Minimization, Treatment, and Prevention. J. Chem., 3796352, 12 pp.
Abstract: The operation of wastewater treatment plants results in direct emissions, from the biological processes, of greenhouse gases (GHG) such as carbon dioxide (CO2), methane (CH4), and nitrous oxide (N-2 O), as well as indirect emissions resulting from energy generation. In this study, three possible ways to reduce these emissions are discussed and analyzed: (1) minimization through the change of operational conditions, (2) treatment of the gaseous streams, and (3) prevention by applying new configurations and processes to remove both organic matter and pollutants. In current WWTPs, to modify the operational conditions of existing units reveals itself as possibly the most economical way to decrease N-2 O and CO2 emissions without deterioration of effluent quality. Nowadays the treatment of the gaseous streams containing the GHG seems to be a not suitable option due to the high capital costs of systems involved to capture and clean them. The change of WWTP configuration by using microalgae or partial nitritation-Anammox processes to remove ammonia from wastewater, instead of conventional nitrification-denitrification processes, can significantly reduce the GHG emissions and the energy consumed. However, the area required in the case of microalgae systems and the current lack of information about stability of partial nitritation-Anammox processes operating in the main stream of the WWTP are factors to be considered.
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Cofre, C., Campos, J. L., Valenzuela-Heredia, D., Pavissich, J. P., Camus, N., Belmonte, M., et al. (2018). Novel system configuration with activated sludge like-geometry to develop aerobic granular biomass under continuous flow. Bioresour. Technol., 267, 778–781.
Abstract: A novel continuous flow system with “flat geometry” composed by two completely mixed aerobic tanks in series and a settler was used to promote the formation of aerobic granular sludge. Making similarities of this system with a typical sequencing batch reactor (SBR), for aerobic granules cultivation, the value of the tank 1/tank 2 vol ratio and the biomass recirculation rate would correspond with the feast/famine length ratio and the length of the operational cycle, respectively, while the settler upflow liquid velocity imposed would be related to the settling time. From the three experiments performed the best results were obtained when the tank 1/tank 2 vol ratio was of 0.28, the sludge recycling ratio of 0.25 and the settler upflow velocity of 2.5 m/h. At these conditions the aggregates had settling velocities between 29 and 113 m/h, sludge volume index at 10 min (SVI10) of 70 mL/g TSS and diameters between 1.0 and 5.0 mm.
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Crutchik, D., Barboza, J., Vazquez-Padin, J. R., Pedrouso, A., del Rio, A. V., Mosquera-Corral, A., et al. (2023). Integrating food waste management into urban wastewater treatment: Economic and environmental impacts. J. Environ. Manage., 345, 118517.
Abstract: 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.
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Crutchik, D., Franchi, O., Caminos, L., Jeison, D., Belmonte, M., Pedrouso, A., et al. (2020). Polyhydroxyalkanoates (PHAs) Production: A Feasible Economic Option for the Treatment of Sewage Sludge in Municipal Wastewater Treatment Plants? Water, 12(4), 12 pp.
Abstract: Sludge is a by-product of municipal wastewater treatment plants (WWTPs) and its management contributes significantly to the operating costs. Large WWTPs usually have anaerobic sludge digesters to valorize sludge as methane and to reduce its mass. However, the low methane market price opens the possibility for generating other high value-added products from the organic matter in sludge, such as polyhydroxyalkanoates (PHAs). In this work, the economic feasibility of retrofitting two types of WWTPs to convert them into biofactories of crude PHAs was studied. Two cases were analyzed: (a) a large WWTP with anaerobic sludge digestion; and (b) a small WWTP where sludge is only dewatered. In a two-stage PHA-production system (biomass enrichment plus PHAs accumulation), the minimum PHAs cost would be 1.26 and 2.26 US$/kg PHA-crude for the large and small WWTPs, respectively. In a single-stage process, where a fraction of the secondary sludge (25%) is directly used to accumulate PHAs, the production costs would decrease by around 15.9% (small WWTPs) and 19.0% (large WWTPs), since capital costs associated with bioreactors decrease. Sensitivity analysis showed that the PHA/COD (Chemical Oxygen Demand) yield is the most crucial parameter affecting the production costs. The energy, methane, and sludge management prices also have an essential effect on the production costs, and their effect depends on the WWTP's size.
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Crutchik, D., Franchi, O., Jeison, D., Vidal, G., Pinto, A., Pedrouso, A., et al. (2022). Techno-Economic Evaluation of Ozone Application to Reduce Sludge Production in Small Urban WWTPs. Sustainability, 14(5), 2480.
Abstract: In Chile, small wastewater treatment plants (WWTPs) (treatment capacity of less than 4,800 m(3)/d) are normally not designed with consideration for the potential valorization of generated sludge. For this reason, they are generally operated at high solids residence times (SRT) (15 d) to promote the decay of biomass, promoting less sludge production and reducing the costs associated with biomass management. Operation at high SRT implies the need for a larger activated sludge system, increasing capital costs. The implementation of a sludge-disintegration unit by ozonation in future WWTPs could enable operation at an SRT of 3 d, with low sludge generation. In this work, we evaluate how the implementation of a sludge-ozonation system in small WWTPs (200-4000 m(3)/d) would affect treatment costs. Four scenarios were studied: (1) a current WWTP operated at an SRT of 15 d, without a sludge ozonation system; (2) a WWTP operated at an SRT of 15 d, with a sludge-ozonation system that would achieve zero sludge production; (3) a WWTP operated at an SRT of 3 d, with a sludge-ozonation system that would provide the same sludge production as scenario 1; (4) a WWTP operated at an SRT of 15 d, with a sludge-ozonation system that would achieve zero sludge production. Economic analysis shows that the treatment costs for scenarios 1 and 2 are similar, while a reduction in cost of up to 47% is obtained for scenarios 3 and 4.
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del Rio, A. V., Campos, J. L., Da Silva, C., Pedrouso, A., & Mosquera-Corral, A. (2019). Determination of the intrinsic kinetic parameters of ammonia-oxidizing and nitrite-oxidizing bacteria in granular and flocculent sludge. Sep. Purif. Technol., 213, 571–577.
Abstract: The different oxygen affinities of ammonia-oxidizing (AOB) and nitrite-oxidizing bacteria (NOB) are often used to define the operational strategy to achieve partial nitritation (PN) required before the anammox (AMX) process. For this purpose, apparent kinetic parameters are mainly used in the case of granular sludge, which can lead to errors when defining the operational conditions to obtain only nitritation (avoiding nitratation). In the present study, a mathematical methodology is proposed to determine the intrinsic kinetic parameters of AOB and NOB in granular sludge based on data obtained by respirometric assays. Additionally, the oxygen affinity constant (K-O2) and maximum specific rate (r(max)) of flocculent and granular sludge sample, produced under mainstream and sidestream conditions were determined at various temperatures (15, 20 and 30 degrees C). The results show that for granules, the intrinsic K-O2 and r(max) values were lower and higher, respectively, than the apparent values. Furthermore, the K-O2 values for flocs and granules at all of the tested temperatures were lower for NOB than for AOB. The values obtained for the kinetic parameters indicated that it is impossible to maintain partial nitritation by only controlling the dissolved oxygen concentration.
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del Rio, A. V., Pichel, A., Fernandez-Gonzalez, N., Pedrouso, A., Fra-Vazquez, A., Morales, N., et al. (2018). Performance and microbial features of the partial nitritation-anammox process treating fish canning wastewater with variable salt concentrations. J. Environ. Manage., 208, 112–121.
Abstract: 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 aftet a peak of organic matter in the feeding. (C) 2017 Elsevier Ltd. All rights reserved.
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Diaz, C., Belmonte, M., Campos, J. L., Franchi, O., Faundez, M., Vidal, G., et al. (2020). Limits of the anammox process in granular systems to remove nitrogen at low temperature and nitrogen concentration. Process Saf. Environ. Protect., 138, 349–355.
Abstract: When partial nitritation-anammox (PN-AMX) processes are applied to treat the mainstream in wastewater treatment plants (WWTPs), it is difficult to fulfil the total nitrogen (TN) quality requirements established by the European Union (<10g TN/m(3)). The operation of the anammox process was evaluated here in a continuous stirred tank reactor operated at 15 degrees C and fed with concentrations of 50 g TN/m(3) (1.30 +/- 0.23 g NO2- -N/g NH4+-N). Two different aspects were identified as crucial, limiting nitrogen removal efficiency. On the one hand, the oxygen transferred from the air in contact with the mixed liquor surface favoured the nitrite oxidation to nitrate (up to 75 %) and this nitrate, in addition to the amount produced from the anammox reaction itself, worsened the effluent quality. On the other hand, the mass transfer of ammonium and nitrite to be converted inside the anammox granules involves relatively large values of apparent affinity constants (k(NH4+app) : 0.50 g NH4+-N/m(3) ; k(NO2-app) 0.17 g NO2--N/m(3)) that favour the presence of these nitrogen compounds in the produced effluent. The careful isolation of the reactor from air seeping and the fixation of right hydraulic and solids retention times are expected to help the maintenance of stability and effluent quality. (C) 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
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Franchi, O., Alvarez, M. I., Pavissich, J. P., Belmonte, M., Pedrouso, A., del Rio, A. V., et al. (2024). Operational variables and microbial community dynamics affect granulation stability in continuous flow aerobic granular sludge reactors. J. Water Process Eng., 59, 104951.
Abstract: Retrofitting wastewater treatment plants with continuous aerobic granular sludge reactors is a promising alternative to enhance treatment capacities and reduce footprint. This study investigates the main variables influencing granulation and microbial dynamics in two reactor configurations (25 L): stirred tanks in series (R1) and a plug-flow-like system (R2). Granule formation was achieved by increasing the organic loading rate (OLR) from 0.7 to 4.1 kg COD/(m3 & sdot;d) and the up-flow velocity in the biomass selector from 1.4 to 6.9 m/h. However, irreversible granule destabilization occurred at day 68 for R1 and day 108 for R2. Principal component analysis and examination of food-to-microorganisms (F/M) ratio medians identified the F/M ratio as the primary variable associated with instability. Microbial analysis revealed that a high F/M ratio induced significant increases in the abundance of specific genera such as Arcobacter, Cloacibacterium, Rikenella, Aquaspirillum and Sphaerotillus, whose overgrowth may negatively impact granule stability. Based on these findings, maximum F/M ratio thresholds were obtained to establish operational conditions allowing the maintenance of stable aerobic granules on continuous flow reactor configurations.
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Pavissich, J. P., Camus, N., Campos, J. L., Franchi, O., Pedrouso, A., Carrera, P., et al. (2021). Monitoring the stability of aerobic granular sludge using fractal dimension analysis. Environ. Sci-Wat Res., 7(4), 706–713.
Abstract: Cyclic episodes of granules formation and disintegration took place in two lab-scale aerobic granular sludge sequencing batch reactors, one fed with synthetic wastewater (COD: 0.6 g L-1 and NH4+-N: 0.06 g L-1) and operated at a constant organic loading rate (2.5 g COD per L d), and the other fed with real wastewater (soluble COD: 0.27-1.37 and NH4+-N: 0.02-0.16 g L-1) and with a variable loading rate (between 1.1 and 5.5 g CODsoluble per L d). The sludge volume index, density and diameter (mean value and relative standard deviation) of the granular biomass showed great fluctuations, without any clear tendency during the operational period. However, changes in granules fractal dimension values (both mean and relative standard deviation) matched with the formation and disintegration dynamics of the granular biomass. Statistical data analysis showed that the relative standard deviation of the granules fractal dimension could be a useful parameter for monitoring the granules status. Indeed, an increase of its value during the maturation or steady-state granulation stages is an early warning of disintegration episodes. A control strategy to maintain granules integrity based on this parameter is proposed.
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Pedrouso, A., Aiartza, I., Morales, N., Vazquez-Padin, J. R., Rogalla, F., Campos, J. L., et al. (2018). Pilot-scale ELAN (R) process applied to treat primary settled urban wastewater at low temperature via partial nitritation-anammox processes. Sep. Purif. Technol., 200, 94–101.
Abstract: A single stage partial nitritation and anammox granular pilot scale reactor (600 L) was operated to treat primary settled sewage in an urban wastewater treatment plant. The fed wastewater contained low total nitrogen concentrations of 6-25 mg TN/L and the system operated without temperature control ranging from 18 to 12 degrees C. A control strategy, based on the pH value, was applied to stop the aeration supply. The pH set-point was fixed at 6.0 and allowed obtaining a total nitrogen removal efficiency approximately of 50% treating a load of 67 mg TN/(L.d) without the addition of any chemicals. Although nitrite oxidizing bacteria were present in the inoculated sludge, when the pH-based control was implemented (day 30) the ammonium oxidation was favored compared to the nitrite oxidation activity. Then, the system operated stable the rest of the operational period (days 30-94) despite the presence of organic matter in the wastewater and the high variability of nitrogen load and temperature during the operation. Nitrogen was autotrophically removed accomplishing the stringent discharge limits (10 mg TN/L) and nitrate concentrations in the effluent lower than 3 mg NO3--N/L. Both biomass concentration and granules size increased during the operational period indicating the growth of the biomass inside the reactor and therefore the potential treatment capacity.
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Pedrouso, A., Correa-Galeote, D., Maza-Marquez, P., Juarez-Jimenez, B., Gonzalez-Lopez, J., Rodelas, B., et al. (2021). Understanding the microbial trends in a nitritation reactor fed with primary settled municipal wastewater. Sep. Purif. Technol., 256, 117828.
Abstract: Partial nitritation was pointed out as the key step to implement the autotrophic nitrogen removal processes at low temperature. This study investigated the initiation and maintenance of a nitritation process with simultaneous COD removal in a sequencing batch reactor (SBR) run at 15 degrees C and fed with primary settled urban wastewater characterized by 42 +/- 10 mg TOC/L and 45 +/- 4 mg NH4+-N/L. A nitrite accumulation ratio of nearly 100% was observed and the long-term (354 days) process stability was successfully maintained despite the municipal wastewater composition fluctuations. The absence of nitrite oxidizing bacteria (NOB) activity was attributed to the free nitrous acid (FNA) in-situ accumulated at high levels (0.02-0.20 mg HNO2-N/L). Despite nitrate production was not observed, the quantification of bacterial groups indicated that NOB were present in the SBR sludge throughout the entire operational period. Ammonium oxidizing bacteria (AOB) abundance and community structure were significantly influenced by the organic matter present in the feeding. Average organic matter removal efficiencies of 80% were obtained without observing any detrimental effect over the nitritation process performance, due to the functional redundancy within both the chemoheterotrophic and AOB communities.
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Pedrouso, A., del Rio, A. V., Campos, J. L., Mendez, R., & Mosquera-Corral, A. (2017). Biomass aggregation influences NaN3 short-term effects on anammox bacteria activity. Water Sci. Technol., 75(5), 1007–1013.
Abstract: The main bottleneck to maintain the long-term stability of the partial nitritation-anammox processes, especially those operated at low temperatures and nitrogen concentrations, is the undesirable development of nitrite oxidizing bacteria (NOB). When this occurs, the punctual addition of compounds with the capacity to specifically inhibit NOB without affecting the process efficiency might be of interest. Sodium azide (NaN3) is an already known NOB inhibitor which at low concentrations does not significantly affect the ammonia oxidizing bacteria (AOB) activity. However, studies about its influence on anammox bacteria are unavailable. For this reason, the objective of the present study was to evaluate the effect of NaN3 on the anammox activity. Three different types of anammox biomass were used: granular biomass comprising AOB and anammox bacteria (G1), anammox enriched granules (G2) and previous anammox granules disaggregated (F1). No inhibitory effect of NaN3 was measured on G1 sludge. However, the anammox activity decreased in the case of G2 and F1. Granular biomass activity was less affected (IC50 90 mg/L, G2) than flocculent one (IC50 5 mg/L, F1). Summing up, not only does the granular structure protect the anammox bacteria from the NaN3 inhibitory effect, but also the AOB act as a barrier decreasing the inhibition.
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Pedrouso, A., del Rio, A. V., Morales, N., Vazquez-Padin, J. R., Campos, J. L., Mendez, R., et al. (2017). Nitrite oxidizing bacteria suppression based on in-situ free nitrous acid production at mainstream conditions. Sep. Purif. Technol., 186, 55–62.
Abstract: The application of autotrophic nitrogen removal processes in the main line of wastewater treatment plants will contribute to achieve its self-energy-sufficiency. However, the effective suppression of nitrite oxidizing bacteria (NOB) activity at the conditions of low temperature and low ammonium concentration (mainstream conditions) was identified as one of the main bottlenecks. In this study, stable partial nitritation at 16 degrees C and 50 mg NH4+-N/L was achieved maintaining inside the reactor free nitrous acid (FNA) concentrations inhibitory for NOB (>0.02 mg HNO2-N/L), without dissolved oxygen concentration control, The FNA inhibitory concentration was generated by the partial nitritation process, and its stimulation was studied with two different inhibitors: sodium azide and nitrite. The microbiological analysis revealed that, throughout the operational period with inhibitory FNA levels, the NOB populations (dominated by Nitrospira) were effectively washed out from the reactor. This is an advantage that allowed maintaining a good stability of the process, even when the FNA concentration was not enough to inhibit the NOB, taking about 40 days to develop significant activity. The observed delay on the NOB development is expected to enable the establishment of corrective actions to avoid the partial nitritation destabilization. The use of the FNA to achieve a stable partial nitritation process is recommended to profit from the natural pH decrease associated to the nitritation process and from its favoured accumulation at low temperatures as those from the mainstream. In this research study an analysis about the influence of ammonium and alkalinity concentrations was also performed to know in which scenarios the FNA inhibitory concentration can be achieved. (C) 2017 Elsevier B.V. All rights reserved.
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Pedrouso, A., del Rio, A. V., Morales, N., Vazquez-Padin, J. R., Campos, J. L., & Mosquera-Corral, A. (2021). Mainstream anammox reactor performance treating municipal wastewater and batch study of temperature, pH and organic matter concentration cross-effects. Process Saf. Environ. Protect., 145, 195–202.
Abstract: The anammox process is an energy efficient promising alternative to biologically remove the nitrogen. Thus, a 5-L anammox granular reactor was inoculated with sludge coming from a sidestream partial nitritation and anammox reactor (>200 mg TN/L and 30 degrees C) and it was directly subjected to 15 +/- 1 degrees C treating mimicked municipal wastewater (50 mg TN/L). Results indicated that an acclimation period (commonly used) to progressive reach the mainstream conditions is not needed, shortening the start-up periods. The long-term anammox process stability was proved to treat synthetic wastewater with decreasing alkalinities and nitritified primary settled municipal wastewater. The low pH values (6.2 +/- 0.1) of the municipal wastewater fed did not affect the process stability. Residual organic matter concentrations augmented the nitrogen removal efficiency from 80 % (with the synthetic medium) to 92 % achieving effluent concentrations below 10 mg TN/L. Finally, the effect of pH (6-8), temperature (15-30 degrees C) and organic matter concentration (0-75 mg TOC/L) over the specific anammox activity (SA(Amx)) was evaluated at short-term. pH and temperature and their interactions exerted significant influence on the SAAmx value while the TOC concentrations itself did not significantly change the SA(AMX). (C) 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
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Pedrouso, A., Morales, N., Rodelas, B., Correa-Galeote, D., del Rio, A. V., Campos, J. L., et al. (2023). Rapid start-up and stable maintenance of the mainstream nitritation process based on the accumulation of free nitrous acid in a pilot-scale two-stage nitritation-anammox system. Sep. Purif. Technol., 317, 123851.
Abstract: Two-stage partial nitritation (PN) and anammox (AMX) systems showed promising results for applying auto-trophic nitrogen removal under mainstream conditions. In this study, a pilot-scale (600 L per reactor) two-stage PN/AMX system was installed in a municipal wastewater treatment plant (WWTP) provided with a high-rate activated sludge (HRAS) system for organic carbon removal. The PN/AMX system was operated without tem-perature control (ranging from 11 to 28 degrees C) and was subjected to the same variations in wastewater charac-teristics as the WWTP (22 to 63 mg NH4+- N/L). The developed strategy is simple, does not require the addition of chemicals and is characterised by short start-up periods. The PN process was established by applying a high hydraulic load and maintained by in situ accumulated free nitrous acid (FNA) of 0.015-0.2 mg HNO2-N/L. Based on pH value, a controlled aeration strategy was applied to achieve the target nitrite to ammonium ratio in the effluent (1.1 g NO2--N/g NH4+-N) to feed the AMX reactor. Although NOB were not fully washed out from the system, nitrite accumulation remained (>99 %) stable with no evidence of NOB activity. In the AMX reactor, an overall nitrogen removal efficiency of 80 % was achieved. Regarding effluent quality, 12 +/- 3 mg TN/L was obtained, but 5 mg NO3--N/L was already in the HRAS effluent. The relative abundance of NOB showed a strong negative correlation with the FNA concentration, providing a good strategy for establishing PN under main-stream conditions.
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