Nitrite Removal Rate Research Articles

Simultaneous aerobic nitrogen and phosphorus removal by novel halotolerant fungus Mucor circinelloides SNDM1: Function and metabolism pathway

Fungi capable of simultaneous nitrogen and phosphorus removal from wastewater is rarely found. Here, a novel fungal strain (SNDM1) performing heterotrophic nitrification, aerobic denitrification, and phosphate removal was isolated and identified as Mucor circinelloides. The favorable nutrient removal conditions by the strain using glucose were C/N ratios of 25–30, salinities of 0 %–3 %, and pH of 7.5. Strain SNDM1 achieved ammonium, nitrite, nitrate, and phosphate removal rates of 5.23, 10.08, 4.88, and 0.97 mg/L/h. Nitrogen balance indicated that gaseous (18.60 %–24.55 %) and intracellular nitrogen (43.76 %–70.63 %) were primary fate of initial nitrogen. Enzyme activity revealed that ammonium removal occurred through heterotrophic nitrification and aerobic denitrification. Removed phosphorus was mainly transformed into cell membranes (56 %–64 %) and extracellular polymeric substances (20 %–26 %). Orthophosphate was the major intracellular phosphorus species, while polyphosphate and pyrophosphate existed extracellularly. These findings highlight the potential of this fungal strain for bioremediating polluted wastewater.

Nitrite-driven anaerobic ethane oxidation

Ethane, the second most abundant gaseous hydrocarbon in vast anoxic environments, is an overlooked greenhouse gas. Microbial anaerobic oxidation of ethane can be driven by available electron acceptors such as sulfate and nitrate. However, despite nitrite being a more thermodynamically feasible electron acceptor than sulfate or nitrate, little is known about nitrite-driven anaerobic ethane oxidation. In this study, a microbial culture capable of nitrite-driven anaerobic ethane oxidation was enriched through the long-term operation of a nitrite-and-ethane-fed bioreactor. During continuous operation, the nitrite removal rate and the theoretical ethane oxidation rate remained stable at approximately 25.0 mg NO2–N L−1 d−1 and 11.48 mg C2H6 L−1 d−1, respectively. Batch tests demonstrated that ethane is essential for nitrite removal in this microbial culture. Metabolic function analysis revealed that a species affiliated with a novel genus within the family Rhodocyclaceae, designated as 'Candidatus Alkanivoras nitrosoreducens', may perform the nitrite-driven anaerobic ethane oxidation. In the proposed metabolic model, despite the absence of known genes for ethane conversion to ethyl-succinate and succinate-CoA ligase, 'Ca. A. nitrosoreducens' encodes a prospective fumarate addition pathway for anaerobic ethane oxidation and a complete denitrification pathway for nitrite reduction to nitrogen. These findings advance our understanding of nitrite-driven anaerobic ethane oxidation, highlighting the previously overlooked impact of anaerobic ethane oxidation in natural ecosystems.

Optimization of degradation conditions and analysis of degradation mechanism for nitrite by Bacillus aryabhattai 47

Excessive nitrite levels cause significant damage to aquaculture, making it crucial to explore green and reliable nitrite removal technologies. In this study, A Bacillus aryabhattai (designated as the strain 47) isolated from aquaculture wastewater was used as the experimental strain. The nitrite degradation conditions of the strain 47 were optimized, and the optimal conditions are: glucose was 12.74 g/L, fermented special soybean meal was 21.27 g/L, MgCl2 369 mg/L, pH 7.0, incubated at 30 °C with the inoculum size of 2 % and the rotation speed of 170 rpm. Under the optimal conditions, the nitrite concentration of the culture solution was 200 mg/L, and the nitrite removal rate reached 91.4 %. Meanwhile, the mechanism by which Mg2+ enhanced the nitrite degradation ability of the strain 47 was investigated by transcriptomics. An operon structure directed cellular trafficking of Mg2+, and then, the Mg2+-mediated catalytic reaction of multiple enzymes enhanced and improved cellular metabolic processes (e.g. the transport and metabolism of nitrite, central carbohydrate metabolism oxidative phosphorylation). At the same time, with the progress of cell metabolism, cells secreted a series of enzymes related to nitrite transport and metabolism to promote the metabolism of nitrite. And the process of the assimilated nitrate reduction pathway of nitrite degradation in the strain 47 was elaborated at the transcriptome level. This study provided a new insight into nitrite treatment mediated by microbial organisms.

Enhanced UV/H2O2 System for the Oxidation of Organic Contaminants and Ammonia Transformation from Tannery Effluents

In this work, a UV/H2O2 system in real tannery wastewater was evaluated by an experimental design with optimal stage 2-level I-optimal reaction surface using Design Expert software to analyze the effects of temperature, pH, UV lamp power (W), and H2O2 concentration on COD removal and nitrification. It was found that pH and temperature were the variables that affected the process the most. It was found that an acidic pH of 4.5–5.5 and temperatures between 50 and 70 °C favored improved COD and ammonium oxidation. The process conditions—temperature 54.6 °C, pH 4, pW-UV 60 W and hydrogen peroxide 0.5—were confirmed in the next phase of the study using a one-way statistical analysis ANOVA. Under these conditions, the nitrite removal rate was 98.4%, ammonium 94.53%, chromium 92.3%, chlorides 62.4%, BOD 67.4%, COD 44.5%, and color 48%.

The existence of ferric hydroxide links the carbon and nitrogen cycles by promoting nitrite-coupled methane anaerobic oxidation.

Microorganism-mediated anaerobic oxidation of methane can efficiently mitigate methane atmospheric emissions and is a key process linking the biogeochemical cycles of carbon, nitrogen, and iron. The results showed that methane oxidation and nitrite removal rates in the CF were 1.12 and 1.28 times higher than those in CK, respectively, suggesting that ferric hydroxide can enhance nitrite-driven AOM. The biochemical process was mediated by the enrichment of methanogens, methanotrophs, and denitrifiers. Methanobacterium and Methanosarcina were positively correlated with Fe3+ and Fe2+, whereas Methylocystis and Methylocaldum were positively correlated with methane, and denitrifiers were positively correlated with nitrite. Metagenomic analysis revealed that the genes related to methane oxidation, nitrogen reduction, and heme c-type cytochrome were upregulated in CF, indicating that a synergistic action of bacteria and methanogens drove AOM via diverse metabolic pathways, within which ferric hydroxide played a crucial role. This study provides novel insights into the synergistic mechanism of ferric iron and nitrite-driven AOM.

Potential of biological sulphur recovery from thiosulphate by haloalkaliphilic Thioalkalivibrio denitrificans

ABSTRACT The aim of this study was to investigate the potential for elemental sulphur recovery from sulphurous solutions under aerobic and anoxic conditions by haloalkalophilic Thioalkalivibrio denitrificans at 0.8–19.6 g S2O3 2−-S L−1 and 0.2–0.58 g NO2 L−1, respectively. The experiments were conducted as batch assays with haloalkaline (pH 10 and ≥ 14 g Na+ L−1) thiosulphate solution. Aerobically, the highest biotransformation rate of thiosulphate obtained was 0.03 h−1 at 8.5 g L S2O3 2−-S. Based on Monod model, the maximum substrate utilisation rate (qm) was 0.024 h−1 with half saturation constant (Ks) 0.42 g S2O3 2−-S L−1 at initial [S2O3 2--S] of 14 g L−1. S0 accumulated at [S2O3 2−-S] ≥ 1.5 g L−1 (10% yield at initial 9.5 g S2O3 2−-S L−1) and the highest S0 yield estimated with the model was 61% with initial [S2O3 2--S] of 16.5 g L−1. Anoxically, the maximum nitrite removal rate based on Monod modelling was 0.011 h−1 with Ks = 0.84 g NO2 − L−1. Aerobically and anoxically the maximum specific growth rates (µm) were 0.046 and 0.022 h−1, respectively. In summary, high-rate aerobic biotransformation kinetics of thiosulphate were demonstrated, whereas the rates were slower and no S0 accumulated under anoxic conditions. Thus, future developments of biotechnical applications for the recovery of S0 from haloalkaline streams from the process industry should focus on aerobic treatment. Highlights Haloalkaline S2O3 2− biotransformations kinetics by Thioalkalivibrio denitrificans Aerobic thiosulphate-S bioconversion up to 0.024 h−1 with Ks = 0.42 g S2O3 2−-S L−1 10% S0 yield with initial 9.5 g S2O3 2--S L−1 in aerobic condition Anoxic NO2 removal up to 0.01 h−1 with Ks = 0.84 g NO2 − L−1

Enhancement of denitrifying anaerobic methane oxidation via applied electric potential.

In this study, single-chamber three-electrode electrochemical sequencing batch reactor (ESBR) was set up to investigate the impact of applying potential on denitrifying anaerobic methane oxidation (DAMO) process. When the applied potential was +0.8V, the conversion rate of nitrite to nitrogen was superior to those of other potentials. With the optimal potential of +0.8V for 60 days, the nitrite removal rate of ESBR could reach 3.34±0.28mgN/L/d, which was 4.5 times more than that of the non-current control (0.74±0.16mgN/L/d). The DAMO functional bacteria Candidatus Methylomirabilis exhibited noticeable enrichment under applying potential, and its functional gene of pmoA was significantly expressed. Through untargeted LC-MS metabolomics analysis, applied potential was shown to affect the regulation of prior metabolites including spermidine, spermine and glycerophosphocholine that were related to the metabolic pathways of glycerophospholipid metabolism and arginine and proline metabolism, which had positive effects on DAMO process. These results show that applying electric potential could be a useful strategy in DAMO process used for methane and nitrogen removal.

Effects of formaldehyde ( Parasite‐S ®) on biofilter nitrification from a cold and a warm freshwater RAS

The effect of Parasite-S® (an aqueous formaldehyde solution) on the nitrification processes of biofilters was evaluated in two recirculating aquaculture systems (RASs). Rearing tanks in the warmwater RAS contained yellow perch (Perca flavescens) and grass carp (Ctenopharyngodon idella) with an initial weight of 166.8 kg and a mean density of 39.5 kg/m3. Rearing tanks in the coldwater RAS contained rainbow trout (Oncorhynchus mykiss) and lake trout (Salvelinus namaycush) with an initial weight of 1377.8 kg at a system density of 41.9 kg/m3. Parasite-S® was administered to the entire system on four consecutive days in both trials to achieve a nominal concentration of 14.8 mg/L formaldehyde (40 mg/L formalin) at the biofilter. Removal efficiencies for total ammonia nitrogen (TAN) and nitrite nitrogen were measured as indicators of biofilter nitrification processes. The active ingredient in Parasite-S®, formaldehyde, was measured until it was below the method detection limit of 0.8 mg/L. TAN volumetric removal rate was significantly decreased in both systems after formaldehyde addition and remained below pre-exposure efficiency in the coldwater RAS. Nitrite nitrogen volumetric removal rate was not significantly different, but the slope and intercepts were less after formaldehyde addition indicating an effect on the nitrifying bacteria. Although removal rates were decreased, no mortality occurred after four consecutive formaldehyde indefinite bath exposures in either system.

Deciphering the Inhibition of Ethane on Anaerobic Ammonium Oxidation.

Anaerobic ammonium oxidation (anammox) and nitrification, two common biological ammonium oxidation pathways, are critical for the microbial nitrogen cycle. Short chain alkanes (C2-C8) have been well-known as inhibitors for nitrification through interaction with the ammonia monooxygenase, while whether these alkanes affect anammox is an open question. Here, this work demonstrated significant inhibition of ethane on anammox and revealed the inhibitory mechanism. The acute inhibition of ethane on anammox was concentration-dependent and reversible; 0.86 mM dissolved ethane caused 50% inhibition (IC50), and 1.72 mM ethane almost completely inhibited anammox. After long-term exposure to 0.09 mM ethane for 30 days, the ammonium (nitrite) removal rate dropped from 202 (267) mg N L-1 d-1 to 1 (1) mg N L-1 d-1, and the abundance of anammox bacteria decreased from 61.9% to 9.5%. The intercellular ammonium concentration of anammox bacteria decreased after ethane exposure, while metatranscriptome analysis showed significant upregulation of genes for ammonium transport of anammox bacteria. Thus, ethane could suppress ammonium uptake resulting in the inhibition of anammox activities. As ethane is the second most prevalent alkane after methane in various anoxic environments, ethane may have an important effect on the nitrogen cycle driven by anammox that should be investigated in future research.

Operating strategies to optimize a membrane bioreactor enriched in nitrite-dependent anaerobic methane-oxidizing bacteria

The use of N-damo bacteria, which can simultaneously remove nitrite and dissolved methane, could represent a cost-effective and sustainable alternative to minimize the environmental impact of effluents from methanogenic reactors treating domestic sewage. The operation of N-damo bacteria reactors is often not stable, and their activity decrease with no proven reason over time. This study aims to optimize the nitrite removal capacity of a lab-scale MBR by using different operating strategies for 878 days. The reactor was operated in continuous mode and with biomass highly enriched in these microbes. It was found that adjustments in the mineral medium concentration (reduction of Ca, P, and K) and composition (EDTA addition, increasing the Cu and Fe bioavailability), contributed to achieving remarkable and prolonged nitrite removal rates of up to 285.7 mg NO2–-N/L/d. N-damo bacteria dominated the culture, up to 57 %. To the best of our knowledge, the rates attained are the highest reported in an N-damo bacteria enrichment.

Evaluation of Non-Biodegradable Organic Matter and Microbial Community’s Effects on Achievement of Partial Nitrification Coupled with ANAMMOX for Treating Low-Carbon Livestock Wastewater

After the anaerobic digestion of livestock manure, high concentrations of nutrients still remain. Treatment of livestock wastewater through partial nitrification coupled with anaerobic ammonium oxidation (ANAMMOX) could be a useful technology depending on the investigation of microorganism enrichment and partial nitrification coupled with achievement of the ANAMMOX process. The results show 78.4% and 64.7% nitrite accumulation efficiency was successfully obtained in an intermittent aeration sequencing batch reactor and a continuous aeration sequencing batch reactor, respectively, at a loading rate of 0.93 kg ammonium/(m3·d). The main reason for the high nitrite accumulation efficiency was the intermittent aeration strategy which generated a 20–30 min lag reaction for nitrite oxidation and promoted the growth of the dominant ammonium oxidation bacteria (Nitrosomonas). Non-biodegradable organic matter in the effluents of partial nitrification did not have obvious influence on ANAMMOX activity at low loading rates (118 ± 13 mg COD/L and 168 ± 9 mg COD/L), and up to 87.4% average nitrite removal rate was observed. However, with the influent COD concentration increasing to 242 ± 17 mg/L, the potential inhibition of ANAMMOX activity was exerted by non-biodegradable organic matter.

Ammonium and hydroxylamine can be preferentially removed during simultaneous nitrification and denitrification by Pseudomonas taiwanensis EN-F2

To overcome a large amount of nitrite accumulation and poor removal rate for hydroxylamine, a simultaneous nitrification and denitrification (SND) bacterium was isolated and identified as Pseudomonas taiwanensis EN-F2 by DNA sequencing. Strain EN-F2 could remove 100% of ammonium (52.90 mg/L), 100% of hydroxylamine (23.32 mg/L), 86.99% of nitrite (56.32 mg/L) and 89.21% of nitrate (56.18 mg/L) with a maximum removal rate of 8.72, 2.12, 4.55 and 5.80 mg/L/h, respectively. Ammonium and hydroxylamine could be preferentially removed during the SND process. The nitrite removal rate and cell growth were substantially enhanced by 2.10 mg/L/h and 0.45 after supplementation of hydroxylamine. The specific activities of ammonia monooxygenase (AMO), hydroxylamine oxidoreductase (HAO), nitrate reductase (NR), nitrite reductase (NIR) were successfully detected as 0.95, 0.31, 0.42 and 0.03 U/mg protein, respectively. All results demonstrated that strain EN-F2 could perform SND to remove multiple nitrogen sources from wastewater.

Isolation and Identification of an Efficient Aerobic Denitrifying Pseudomonas stutzeri Strain and Characterization of Its Nitrite Degradation

Nitrogen pollution in water bodies is becoming increasingly serious, and how to remove nitrogen from water bodies economically and effectively has become a research hotspot. Especially in recent years, with the gradual expansion of aquaculture in China, the content of nitrite and other nitrogen-containing substances in water bodies has been increasing, which inhibits the growth of farm animals and is one of the causes of eutrophication in water bodies. In this study, a strain of bacteria was isolated from the sludge of an aquaculture fishpond and identified as Pseudomonas stutzeri, which can efficiently degrade nitrite. After continuous domestication in nitrite mixed solution, the nitrite nitrogen reduction capacity of P. stutzeri was significantly improved. Univariate experiments aiming to optimize the degradation conditions indicate that the optimal culture conditions for strain F2 are: medium with a carbon source of sodium succinate; C/N of 18; pH of 8; culture temperature of 28 °C; and shaking speed of 210 rpm in the shaker. Under the optimal culture conditions, the NO2−-N concentration of the culture solution was 300 mg/L, and the nitrite removal rate reached 98.67%. Meanwhile, the results of the nitrogen balance test showed that the strain converted 6.1% of the initial nitrogen into cellular organic nitrogen and 62.3% into gaseous nitrogen.

Unraveling the nitrogen removal properties and microbial characterization of “Candidatus Scalindua”-dominated consortia treating seawater-based wastewater

“Candidatus Scalindua”, as known as marine anammox bacteria (MAB), was engineered to remove nitrogen from seawater-based wastewater (SWW). In this study, “Candidatus Scalindua” was successfully enriched within 106 days with marine sediments as inoculated sludge. The operating temperature was 20 ± 2 °C, and influent pH was 7.5 ± 0.1. Ammonia (NH4+-N) removal rate (ARR) was 0.53 kg/(m3·d) with the NH4+-N loading rate of 0.68 kg/(m3·d), and nitrite (NO2−-N) removal rate (NRR) was 0.57 kg/(m3·d) at 0.89 kg/(m3·d) NO2−-N loading rate. Nitrogen removal was negatively affected at an influent NO2− above 224 mg/L, which decreased the ARR and NRR to 0.36 and 0.31 kg/(m3·d), respectively. The genus “Ca. Scalindua” dominated the reactor, and it synergistically coexisted with Marinicella to achieve efficient nitrogen removal. This work would help to better understand the nitrogen removal properties and microbial characterization of MAB in SWW wastewater treatment under low temperature.

Effect of water velocity on ammonium and nitrite removal in pilot scale fixed bed biofilters

The effect of water velocity on nitrification rates in fixed bed biofilters was investigated in three freshwater pilot scale RAS with rainbow trout. Removal of total ammonia nitrogen (TAN) and nitrite-nitrogen were assessed by NH4Cl spikes and tested at four different water velocities in the biofilters (1.4, 5.4, 10.8 and 16.2 m h−1) under identical conditions. Water velocities below 10.8 m h−1 significantly reduced TAN- and nitrite removal rates. The surface specific TAN removal rates correlated with the TAN concentrations at the water velocities 10.8 and 16.2 m h−1, and the first order surface removal rate constant was estimated at 0.45 m h−1. However, no correlations between TAN removal and TAN concentrations were found at the lowest velocities. Up to five-fold elevated nitrite levels were found in the RAS when biofilters were operated at 1.4 m h−1 compared to the trials at other water velocities, substantiating the significant effect of water velocity on both nitrification processes. The importance of biofilter hydraulics documented in this pilot scale RAS probably have implications for design and operation in larger scale RAS.

Deciphering nitrogen removal mechanism through marine anammox bacteria treating nitrogen-laden saline wastewater under various phosphate doses: Microbial community shift and phosphate crystal

The effect of phosphate on marine anammox bacteria (MAB)-dominated anammox process in nitrogen-laden saline wastewater was first investigated. The activity of MAB was enhanced by dosing low concentrations of phosphate (5–30 mg/L PO43−-P), and the time of complete ammonium removal was shortened by 0.5 h. When PO43−-P exceeded 160 mg/L, the calcium magnesium phosphate precipitation was formed in the reactor. The contact between substrates and biomass was hindered by the sediments, and the nitrogen removal performance of MAB was also worsened. At 400 mg/L PO43−-P, the ammonium removal rate and nitrite removal rate decreased to 0.45 and 0.43 kg/(m3⋅d), respectively. During the 158-day operation, MAB was still the dominant strain, but its relative abundance decreased by 15.4% at 400 mg/L PO43−-P. Besides, the presence of sediments stimulated the production of extracellular polymeric substances and the maximum yield reached 11.25 mg/g⋅wet weight at 200 mg/L PO43−-P.

Application of xylitol on nitrogen removal from saline wastewater through "Candidatus Brocadia sinica"-dominated anammox process under low temperature.

Xylitol was first applied to enhance nitrogen removal from saline wastewater through "Candidatus Brocadia sinica"-dominated anammox process under low temperature. The reactor was maintained at 15°C, and the salinity of wastewater was 35g/L. Ammonium removal rate (ARR) and nitrite removal rate (NRR) were stable at around 0.27kg/(m3 d) without xylitol addition. As an osmotic pressure regulator and cryoprotective agent, optimal ARR and NRR were 0.51kg/(m3 d) and 0.63kg/(m3 d) at 0.3mM xylitol. At the addition of 1mM high-dosage xylitol, there existed dissimilatory reduction in nitrate to ammonium nitrogen and heterotrophic denitrification in the reactor. Remodified logistic model was suitable to simulate removal process with xylitol addition. As a result, xylitol dose should be controlled within 0.3mM, which greatly promoted the nitrogen removal from saline wastewater under low temperature. PRACTITIONER POINTS: Xylitol could be used as osmotic pressure regulator and cryoprotective agent to enhance nitrogen removal. The optimal dose was achieved at 0.3mM xylitol for "Candidatus Brocadia sinica" in low-temperature saline wastewater. High-dosage xylitol could interfere with nitrogen removal efficiency due to the presence of DNAR and HB. Remodified logistic model was suitable for the analysis and prediction of nitrogen removal process with xylitol addition.

Combined genome-centric metagenomics and stable isotope probing unveils the microbial pathways of aerobic methane oxidation coupled to denitrification process under hypoxic conditions

Obligate aerobic methanotrophs have been proven to oxidize methane and participate in denitrification under hypoxic conditions. However, this phenomenon and its metabolic mechanism have not been investigated in detail in aerobic methane oxidation coupled to denitrification (AME-D) process. In this study, a type of hypoxic AME-D consortium was enriched and operated for a long time in a CH4-cycling bioreactor with strict anaerobic control and the nitrite removal rate reached approximately 50 mg N/L/d. Metagenomics combined with DNA stable-isotope probing demonstrated that the genus Methylomonas, which constitutes type I aerobic methanotrophs, was the dominant member and contributed to methane oxidation and partial denitrification. Metagenomic binning recovered a near-complete (98%) draft genome affiliated with the family Methylococcaceae containing essential genes that encode nitrite reductase (nirK), nitric oxide reductase (norBC) and hydroxylamine dehydrogenase (hao). Metabolic reconstruction of the selected Methylococcaceae genomes also revealed a potential link between methanotrophy and partial denitrification.

Biostimulation of a marine anammox bacteria-dominated bioprocess by Co(II) to treat nitrogen-rich, saline wastewater

The biostimulation of a marine anammox bacteria (MAB)-dominated bioprocess with Co(II) was studied in a sequencing batch reactor (SBR) treating nitrogen-rich saline wastewater at 15 °C. The low Co(II) load of 0.0015 kgCo2+added/(m3.d) had little effect on the removal of nitrogen. The nitrite removal rate (NRR), ammonia removal rate (ARR), and specific anammox activity (SAA) reached 0.73 kg/(m3·d), 0.59 kg/(m3·d), and 0.23 kg/(kg·d), respectively, under the Co(II) load of 0.009 kgCo2+added/(m3.d). However, the loadings of Co(II) at 0.024–0.03 kgCo2+added/(m3.d) negatively affected the activity of MAB. Besides, the values of ΔNO2−-N/ΔNH4+-N (1.15–1.29) were lower than the theoretical ratio values (around 1.32) likely because of the marine commamox process. The removal of nitrogen from nitrogen-rich saline wastewater was achieved by the synergy between Candidatus Scalindua (27.11%) and Candidatus Kuenenia (9.55%). The nitrogen removal with Co(II) addition could be well described by a modified Logistic model.

Cometabolic removal of organic micropollutants by enriched nitrite-dependent anaerobic methane oxidizing cultures

The innovative and recently discovered n-damo process, based on anaerobic methane oxidation with nitrite, was developed in a membrane-based bioreactor and evaluated in terms of organic micropollutants (OMPs) removal. The main singularity of this study consisted in the evaluation of organic micropollutants (OMPs) removal in the biological reactor. A strategy consisting on progressively increasing the nitrogen loading rate in order to increase the specific denitrification activity was followed to check if the selected OMPs were co-metabolically biotransformed. Significant nitrite removal rate (24.1 mg N L−1 d−1) was achieved after only 30 days of operation. A maximum specific removal of 186.3 mg N gVSS−1 d−1 was obtained at the end of the operation, which is one of the highest previously reported. A successfully n-damo bacteria enrichment was achieved, being Candidatus Methylomirabilis the predominant bacteria during the whole operation attaining a maximum relative abundance of about 40 %. The natural hormones (E1 and E2) were completely removed in the bioreactor. The specific removal rates of erythromycin (ERY), fluoxetine (FLX), roxithromycin (ROX) and sulfamethoxazole (SMX) were successfully correlated with the specific nitrite removal rates, suggesting a co-metabolic biotransformation.