Short-term batch studies on biological removal of chromium from synthetic wastewater using activated sludge biomass

Biological treatment of Cr(VI)-containing wastewater has drawn much attention recently as a method to treat environmental Cr(VI) contamination. The activated sludge method, due to its convenient operation and easy-to-scale-up, has been widely applied to treat municipal wastewater and some industrial wastewaters. In order to develop a suitable technique using activated sludge as the biomass to continuously remove Cr(VI) from wastewater, this paper investigated in short-term batch experiments the environmental elements affecting chromium biological removal from synthetic wastewater. The dissolved oxygen (DO), Cr(VI) initial concentration, biomass density, temperature, glucose content in the influent and contact time were observed to strongly influence chromium removal. It was found that the chromium removal efficiency decreased with the increase of DO and Cr(VI) initial concentration as well as glucose content in the feed, but increases in temperature and contact time improved the chromium removal efficiency. Although raising biomass concentration resulted in an increased chromium removal efficiency under both anaerobic and aerobic conditions, its influence on specific chromium removal was not significant.     

Biosorption of Heavy Metals from Wastewater by Biosolids

In a study where the removal of heavy metals from wastewater is the primary aim, the biosorption of heavy metals onto biosolids prepared as Pseudomonas aeruginosa immobilized onto granular activated carbon was investigated in batch and column systems. In the batch system, adsorption equilibriums of heavy metals were reached between 20 and 50 min, and the optimal dosage of biosolids was 0.3 g/L. The biosorption efficiencies were 84, 80, 79, 59 and 42 % for Cr(VI), Ni(II), Cu(II), Zn(II) and Cd(II) ions, respectively. The rate constants of biosorption and pore diffusion of heavy metals were 0.013–0.089 min^–1 and 0.026–0.690 min^–0.5. In the column systems, the biosorption efficiencies for all heavy metals increased up to 81–100 %. The affinity of biosorption for various metal ions towards biosolids was decreased in the order: Cr = Ni > Cu > Zn > Cd.     

Hexavalent chromium removal by viable, granular anaerobic biomass

Hexavalent chromium in industrial wastewater is a major concern due to its extreme toxicity. This study investigates the removal of Cr(VI) using viable anaerobic granular biomass as a biosorbent. The effect of Cr(VI) concentration on biogas content and COD removal using batch studies indicated that the phase II (methanogenic-rich) culture was more sensitive than the phase I (acidogenic-rich) culture. Toxicity indices for both cultures using COD removal were developed based on linear-log interpolation. The median inhibition Cr(VI) concentration (IC(50)), for phase II cultures was found to be 263mg/L, while that for phase I cultures was 309mg/L. A sorption study was conducted on viable and non-viable (dried) phase I-rich biomass: both followed the Langmuir model. In addition, the biosorption capacity for metabolically inhibited biomass was 25% less indicating some level of cellular uptake associated with Cr(VI) removal. This study demonstrated the potential for a two-phase anaerobic treatment system for a Cr(VI)-contaminated effluent.     

Estimation of Kinetic Parameters for Bioremediation of Cr(VI) from Wastewater Using Pseudomonas taiwanensis,an Isolated Strain from Enriched Mixed Culture

An aerobic mixed culture collected in the form of activated sludge was enriched for Cr(VI) reduction. An indigenous microorganism was isolated from the enriched aerobic mixed culture and identified as Pseudomonas taiwanensis. Bioremediation studies were carried out for treating Cr(VI)-contaminated wastewater using the indigenous microorganism. The kinetic studies were carried out for initial Cr(VI) concentrations ranging from 20 to 200 mg L^?1. The maximum consumption of Cr(VI) obtained was 108.3 mg L^?1 for an initial Cr(VI) concentration of 150 mg L^?1 at a solution pH of 7.0. The effect of nutrient dosage and pH were studied to get their optimum values. The same isolated bacterial strain was also used to treat Cr(VI)-contaminated industrial wastewater collected from a local plating industry. Various growth kinetic models, such as Monod, Powell, Haldane, Luong, and Edward models, were fitted with the obtained experimental data. The obtained results for different growth kinetic models indicate that the growth kinetics of Pseudomonas taiwanensis for bioremediation of Cr(VI) can be better understood by the Luong model (R² = .913). The rate kinetic analysis was performed using zero-order and three-half-order kinetic models. The three-half-order kinetic model was found to be suitable for the present bioremediation study.     

Aerobic granules: a novel zinc biosorbent

AIMS: Aerobic granules are aggregates with a compact and porous microbial structure. In view of the potential use of aerobic granules as biosorbents for Zn(II) removal from industrial wastewater, this study investigated the effects of initial Zn(II) and aerobic granule concentrations on the kinetics of Zn(II) biosorption on the aerobic granule surface. METHODS AND RESULTS: Acetate-fed aerobic granules with a mean diameter of 1.0 mm were used as biosorbents. Results showed that the kinetics of Zn(II) biosorption on the aerobic granule surface were related to both initial Zn(II) and granule concentrations. It was found that the real driving force for Zn(II) biosorption on the aerobic granule surface could be described by the ratio of initial Zn(II) concentration (Co) to initial granule concentration (Xo), rather than individual Co or Xo. The Co/Xo ratio provides a unified basis for interpretation of the biosorption data obtained under different initial conditions. The maximum biosorption capacity of Zn(II) by aerobic granules was 270 mg g(-1). CONCLUSIONS: It appears that the aerobic granule can be used as an effective biosorbent for efficient removal of Zn(II) or other types of heavy metals from industrial wastewater. SIGNIFICANCE AND IMPACT OF THE STUDY: This study could lead to the development of a novel granular biosorbent for the removal of heavy metals from wastewater. A simple and compact aerobic granule-based biosorber could be expected.     

Aerobic selectors in slaughterhouse activated sludge systems: a preliminary investigation

Filamentous bulking at a slaughterhouse activated sludge treatment plant significantly reduced mixed liquor settling properties, which caused many operational problems and worsening in effluent quality. The main cause of this condition was attributed to significant levels of influent readily biodegradable COD, which was present primarily in the form of organic acids. An aerobic selector was chosen to eradicate the usual bulking incidents of slaughterhouse wastewater treatment plants. Other plant enhancements included increased aeration batch reactor volume, and provision of step feed capability. Comparison of data before and after aerobic selector installation showed a significant improvement in mixed liquor settleability, which eradicated the need for chemicals that had been used to control filaments and to control effluent solids loss. The additional volume of the aeration and chemicals eliminations from the activated sludge system also served to eliminate aquatic toxicity in the treated effluent.     

Start-up procedures and analysis of heavy metals inhibition on methanogenic activity in EGSB reactor

The effectiveness of operating an industrial UASB reactor, treating wastewater from the beer industry, with flows containing heavy metals was evaluated. A pilot-scale UASB reactor, already used to simulate the industrial reactor, was unsuccessfully employed. An easy start-up was obtained arranging it as an EGSB reactor. Considerations about this modification are reported. The effects of Cu(II), Ni(II) and Cr(III) ions on the anaerobic activity were analyzed by measurements of methane production rate and COD removal. The employed biomass was the sludge of the industrial UASB reactor, while a solution of ethanol and sodium acetate with COD of 3000 mg/L and a heavy metal concentration of 50 mg/L were continuously fed. Experimental results proved higher biomass sensitivity for copper and much slighter for nickel and chromium. Moreover, copper inhibition has been demonstrated to be less significant if a metal-free feed was provided to the system before copper addition.     

Cr(VI) removal from aqueous solution by thermophilic denitrifying bacterium <Emphasis Type="Italic">Chelatococcus daeguensis</Emphasis> TAD1 in the presence of single and multiple heavy metals

Cr(VI) pollution is increasing continuously as a result of ongoing industrialization. In this study, we investigated the thermophilic denitrifying bacterium Chelatococcus daeguensis TAD1, isolated from the biofilm of a biotrickling filter used in nitrogen oxides (NO_X) removal, with respect to its ability to remove Cr(VI) from an aqueous solution. TAD1 was capable of reducing Cr(VI) from an initial concentration of 10 mg/L to non-detectable levels over a pH range of 7–9 and at a temperature range of 30–50°C. TAD1 simultaneously removed both Cr(VI) and NO₃^?-N at 50°C, when the pH was 7 and the initial Cr(VI) concentration was 15 mg/L. The reduction of Cr(VI) to Cr(III) correlated with the growth metabolic activity of TAD1. The presence of other heavy metals (Cu, Zn, and Ni) inhibited the ability of TAD1 to remove Cr(VI). The metals each individually inhibited Cr(VI) removal, and the extent of inhibition increased in a cooperative manner in the presence of a combination of the metals. The addition of biodegradable cellulose acetate microspheres (an adsorption material) weakened the toxicity of the heavy metals; in their presence, the Cr(VI) removal efficiency returned to a high level. The feasibility and applicability of simultaneous nitrate removal and Cr(VI) reduction by strain TAD1 is promising, and may be an effective biological method for the clean-up of wastewater.     

Reducing effect of aerobic selector on the toxicity of synthetic organic compounds in activated sludge process

The effects of phenol, 2-chlorophenol (2-CP), 2,4-dichlorophenol (2,4-DCP) and 1,2,4-trichlorobenzene (1,2,4-TCB) on the biodegradation kinetics of the conventional activated sludge system (CASS) and the selector activated sludge system (SASS) were investigated. Experiments were carried out using a respirometric method on unacclimated biomass from two lab-scale systems that were operated with the sludge age of 8 days. Toxicity of the test compounds for both reactors were arranged according to EC₅₀ (effective concentration) values in order as: 1,2,4-TCB > 2,4-DCP > 2-CP > phenol. All selected test compounds induced higher inhibition effect in the CASS. The SASS appeared to reduce inhibition effect in comparison to the CASS, by 21.36%, 66.95%, 64.37% and 33.33% for phenol, 2-CP, 2,4-DCP and 1,2,4-TCB, respectively. Consequently, the SASS may be recommended as a promising configuration alternative for the waste streams containing toxic compounds.     

Effect of Zn(II) on the reduction and accumulation of Cr(VI) by <Emphasis Type="Italic">Arthrobacter</Emphasis> species

Natural habitats are often characterized by the coexistence of Zn and Cr. This study assessed the potential of two Gram-positive, Cr(VI)-reducing, aerobic bacterial strains belonging to Arthrobacter genera, which were isolated from basalt samples taken from the most polluted region of the Republic of Georgia, to remediate Cr(VI) in environments in the presence of Zn(II). Our batch experiments revealed that the addition of Zn(II) to the tested bacterial cells significantly enhanced the accumulation of Cr. According to electron spin resonance (ESR) measurements, the presence of Zn(II) ions did not change the nature of Cr(V) and Cr(III) complexes generated during the microbial reduction of Cr(VI). The efficiency of Cr(VI) reduction also remained unchanged after the addition of 50 mg/l of Zn(II) to the bacterial cells. However, at high concentrations of Zn(II) (higher than 200 mg/l), the transformation of Cr(VI) to Cr(V) and Cr(III) complexes decreases significantly. In addition, it was shown that the accumulation pattern of Zn in the tested bacterial species in the presence of 100 mg/l of Cr(VI) fits the Langmuir–Freundlich model well. The two tested bacterial strains exhibited different characteristics of Zn accumulation.     

Influence of nickel (II) and chromium (VI) on the laboratory scale rotating biological contactor

High concentration of heavy metals is toxic for most microorganisms and cause strict damage in wastewater treatment operations and often a physico-chemical pretreatment prior to biological treatment is considered necessary. However, in this study it has been shown that biological systems can adapt to Ni (II) and Cr (VI) when their concentration is below 10 and 20 mg/L, respectively. The aim of this study was to evaluate the effect of Ni (II) and Cr (VI) on the lab-scale rotating biological contactor process. It was found that, addition of Ni (II) up to 10 mg/L did not reduce the chemical oxygen demand removal efficiency and on the contrary concentrations below 10 mg/L improved the performance. The influent Ni (II) concentration of 1 mg/L was the concentration where the treatment efficiency produced a maximum COD removal of 86.5%. Moreover, Ni (II) concentration above 10 mg/L was relatively toxic to the system and produced lower treatment efficiencies than the baseline study without Ni (II). Turbidity and suspended solids removals were not stimulated to a great extent with nickel. Addition of Ni (II) did not seem to affect the pH of the system during treatment. The dissolved oxygen concentration did not drop below 4 mg/L at all concentrations of Ni (II) indicating aerobic conditions prevailed in the system. Experiments conducted with Cr (VI) revealed that addition of Cr (VI) up to 20 mg/L did not reduce the COD removal efficiency and on the contrary concentrations below 20 mg/L improved the performance. The influent Cr (VI) concentration of 1 mg/L was the concentration where the treatment efficiency produced a maximum COD removal of 88%. Turbidity and SS removals were more efficient at 5 mg/L Cr (VI) concentration, rather than 1 mg/L, which lead to the conclusion that 5 mg/L Cr (VI) concentration is the optimum concentration, in terms of COD, turbidity and SS removals. Similar with Ni (II) experiments, addition of Cr (VI) did not significantly affect the pH value of the effluent. The DO concentration remained above 5 mg/L.     

Optimization and/or acclimatization of activated sludge process under heavy metals stress

The present study aimed to overcome the toxicity of the heavy metals load, discharged with the industrial effluents into Alexandria sewerage network, on the activated sludge treatment system through effective acclimation for organic matter and heavy metals removal. Optimization and/or acclimatization of the activated sludge process in the presence of Cu, Cd, Co and Cr contaminating mixed domestic-industrial wastewater was investigated. Acclimatization process was performed through abrupt and stepwise addition of tested metals using sequencing batch reactors treatment approach and evaluated as microbial oxygen uptake rate (OUR), dehydrogenase activity (DHA), organic matter (COD) and heavy metals removal. Abrupt addition of metals adversely affected sludge bioactivity leading to decline in the removal efficiency of the targeted contaminants and loss of floc structure. Metals IC₅₀ confirmed that copper possessed the highest toxicity towards the OUR, DHA activity and COD removal with orders Cu > Cd > Cr > Co; Cu > Cd > Co = Cr and Cu > Cd > Cr > Co, respectively. The highest metal removal was recorded for Cd followed by Co, Cu and finally Cr, most of which was retained in the dissolved influent. However, controlled stepwise application of the tested metals exhibited high sensitivity of DHA and OUR activities only at the highest metal concentrations although enhanced at the lowest concentrations while COD removal was not significantly affected. In conclusion, this approach resulted in adaptation of the system where sludge microbes acquired and developed natural resistance to such metals leading to remarkable enhancement of both organic matter and heavy metals removal.     

Effects of combining biological treatment and activated carbon on hexavalent chromium reduction

The objectives of the present work were: (a) to analyze the Cr(VI) removal by combining activated sludge (AS) with powdered activated carbon (PAC), (b) to analyze the effect of PAC and Cr(VI) on the growth kinetics of activated sludge, and (c) to determine if the combined method (AS-PAC) for Cr(VI) removal can be considered additive or synergistic with respect to the individual processes. Chromate removal was improved by increasing PAC concentrations in both PAC and AS-PAC systems. Cr(VI) removal using the AS-PAC system was higher than using AS or PAC. The increase of Cr(VI) caused longer lag phase and lower observed specific growth rate (μ_obs), biomass yield (Y_X/S), and specific growth substrate consumption rate (q_S) of activated sludge; additionally, PAC did not enhance the growth kinetic parameters (μ_obs, Y_X/S, q_S). Cr(VI) reduction in AS-PAC system was the result of the additive effect of each individual Cr(VI) removal process.     

Preliminary studies on continuous chromium(VI) biological removal from wastewater by anaerobic-aerobic activated sludge process

The long-term continuous chromium(VI) removal from synthetic wastewater affected by influent hexavalent chromium (Cr(VI)) and glucose concentrations were studied with an anaerobic-aerobic activated sludge process. It was observed that before activated sludge was acclimated, the chromium in the effluent increased immediately as the influent chromium increased. However, both Cr(VI) and total chromium (TCr) in the effluent significantly decreased after acclimation. In the acclimated activated sludge, the chromium removal efficiency was 100% Cr(VI) and 98.56% TCr at influent Cr(VI) levels of 20 mg/day, 100% Cr(VI) and 98.92% TCr at influent Cr(VI) levels of 40 mg/day, and 98.64% Cr(VI) and 97.16% TCr at influent Cr(VI) levels of 60 mg/day. The corresponding effluent Cr(VI) and TCr concentrations were 0 and 0.012 mg/l, 0 and 0.018 mg/l, and 0.034 mg/l and 0.071 mg/l, respectively. When the influent glucose increased from 1125 to 1500 mg/l at influent Cr(VI) dosage of 60 mg/day, the Cr(VI) and TCr removal efficiency with the acclimated activated sludge improved from 98.64% and 97.16% to 100% and 98.48%, respectively, and the chromium concentration in the effluent decreased from 0.034 mg/l of Cr(VI) and 0.071 mg/l of TCr to 0 (Cr(VI)) and 0.038 mg/l (TCr). The effluent COD and turbidity was around 40 mg/l and 0, respectively, after the activated sludge was acclimated. Further studies showed that after the activated sludge was acclimated, its specific dehydrogenases activity (SDA) and protein contents increased. The SDA and protein increased respectively 15% and 10% when influent Cr(VI) increased from 20 to 60 mg/day.     

Evaluation of methods to solubilize and analyze cell-associated ectoenzymes

A protocol for production, storage, and use of Shock 1 (Shk1) bioreporter cells for toxicity monitoring in wastewater treatment facilities was developed. Shk1 is a bioluminescent toxicity bioreporter for activated sludge previously constructed by the incorporation of lux genes into an activated sludge microorganism.

A number of factors affecting Shk1 growth and bioluminescence were examined including the growth medium, tetracycline concentration, storage conditions, and test media. Based on the results of these experiments, a toxicity testing protocol was developed that involved growth of cultures in nutrient broth with tetracycline, storage of cultures at 4 °C, cell activation by reinoculation into nutrient broth, and toxicity testing by cell injection into the test media. Effective use of this approach required standardized time intervals for cell growth, storage, activation and exposure in the test media.

Bioluminescence from Shk1 cells was measured in nutrient broth and influent wastewater and activated sludge mixed liquor from a municipal wastewater treatment plant. Using the Shk1 toxicity testing protocol, Zn EC₅₀ values for bioluminescence in nutrient broth, influent wastewater, and activated sludge mixed liquor were approximately 42, 7, and 32 mg/l, respectively. Zn concentrations as low as 1 mg/l could be detected in influent wastewater. The detection limit in influent wastewater is below the Zn concentrations typically reported to affect the activated sludge process.     

Ochrobactrum tritici strain 5bvl1 - characterization of a Cr(VI)-resistant and Cr(VI)-reducing strain

Bacterial strain 5bvl1, isolated from a chromium-contaminated wastewater treatment plant and identified as Ochrobactrum tritici, was resistant to a broad range of antibiotics, to Cr(VI), Ni(II), Co(II), Cd(II), and Zn(II), and was able to grow in the presence of 5% NaCl and within the pH range 4-10. Characterization showed that strain 5bvl1 could be considered a halotolerant and alkalitolerant microorganism resistant to high concentrations of Cr(VI). This strain was able to grow aerobically in up to 10 mmolxL(-1) Cr(VI). Cr(VI) resistance was independent of sulphate concentration. Under aerobic conditions strain 5bvl1 was also able to reduce high Cr(VI) concentrations (up to 1.7 mmolxL(-1)). Increasing concentrations of Cr(VI) in the medium lowered the growth rate of strain 5bv11 but the reduction in growth rate could not be directly correlated with the amount of Cr(VI) reduced. Unlike the type strain, which was only able to reduce Cr(VI), strain 5bvl1 was resistant to Cr(VI) and able to reduce it. Moreover, in strain 5bvl1, the rate and extent of Cr(VI)-reduction were higher than in the other strains of the genus Ochrobactrum. Ochrobactrum strain 5bvl1 resists high Cr(VI) concentrations and has a high Cr(VI)-reducing ability, making it a valuable tool in bioremediation.     

Anaerobic digestion of cattail with rumen culture in the presence of heavy metals

The anaerobic digestion of cattail by rumen cultures in the presence of Cu(II), Cd(II) or Cr(VI) was investigated in this study. Three cases were respectively observed for the different metal dosages: promoted cattail degradation and methane production at a low heavy metal concentration, e.g., Cu(II) 2.4 mg/l, Cd(II) 1.6 mg/l, Cr(VI) 4.0 mg/l; reduced cattail degradation efficiency and methane production at a middle metal level; a severe inhibition to the cattail degradation at a high heavy metal dosage. The inhibition kinetics of Cu(II) on the digestion of cattail by rumen cultures was also analyzed and a simplified Andrews equation was used to describe such an inhibition. The inhibition constants for Cu(II) on the degradation of cattail, production of volatile fatty acids and formation of methane were estimated as 7.4, 9.5 and 6.4 mg/l, respectively. Comparative experimental results suggest that the order of toxicity degree of the tested metals on the rumen cultures was: Cd(II) > Cu(II) > Cr(VI).     

Cost-effective production of Bacillus thuringiensis biopesticides by solid-state fermentation using wastewater sludge: effects of heavy metals

This study demonstrated the feasibility to produce Bacillus thuringiensis subsp. kurstaki (Btk) based biopesticides using wastewater sludge as raw materials under solid-state fermentation (SSF). More than 10¹⁰ CFU/g viable cells of Btk were obtained using sludge or its mixture with agricultural wastes. This study well considered the effect of heavy metals on Btk growth and their changes of chemical speciation caused by SSF. The IC₅₀ (concentration causing 50% inhibition in total cell biomass) for Pb(II), Cu(II), Cd(II) and Cr(III) on Btk were determined to be 227, 82, 15 and 263 mg/L, respectively. Exposure to 150 mg/L of Cu(II) severely reduced the amount and size of toxin crystals, which decreased the endotoxin synthesis and entomotoxicity potency of Btk cells. Using Tessier’s sequential extraction procedure, the exchangeable heavy metals in sludge were shown to be transformed into residual fractions after SSF, and thus significantly reduced their bioavailability and potential environmental risks.     

Characteristics of a novel Acinetobacter sp. and its kinetics in hexavalent chromium bioreduction

Cr-B2, a Gram-negative hexavalent chromium [Cr(VI)] reducing bacteria, was isolated from the aerator water of an activated sludge process in the wastewater treatment facility of a dye and pigment based chemical industry. Cr- B2 exhibited a resistance for 1,100 mg/l Cr(VI) and, similarly, resistance against other heavy metal ions such as Ni2+ (800 mg/l), Cu2+ (600 mg/l), Pb2+ (1,100 mg/l), Cd2+ (350 mg/l), Zn2+ (700 mg/l), and Fe3+ (1,000 mg/l), and against selected antibiotics. Cr-B2 was observed to efficiently reduce 200 mg/l Cr(VI) completely in both nutrient and LB media, and could convert Cr(VI) to Cr(III) aerobically. Cr(VI) reduction kinetics followed allosteric enzyme kinetics. The Km values were found to be 43.11 mg/l for nutrient media and 38.05 mg/l for LB media. Vmax values of 13.17 mg/l/h and 12.53 mg/l/h were obtained for nutrient media and LB media, respectively, and the cooperativity coefficients (n) were found to be 8.47 and 3.49, respectively, indicating positive cooperativity in both cases. SEM analysis showed the formation of wrinkles and depressions in the cells when exposed to an 800 mg/l Cr(VI) concentration. The organism was seen to exhibit pleomorphic behavior. Cr-B2 was identified on the basis of morphological, biochemical, and partial 16S rRNA gene sequencing chracterizations and found to be Acinetobacter sp.     

Experimental investigation of the external nitrification biological nutrient removal activated sludge (ENBNRAS) system

A systematic lab-scale experimental investigation is reported for the external nitrification (EN) biological nutrient removal (BNR) activated sludge (ENBNRAS) system, which is a combined fixed and suspended medium system. The ENBNRAS system was proposed to intensify the treatment capacity of BNR-activated sludge (BNRAS) systems by addressing two difficulties often encountered in practice: (a) the long sludge age for nitrification requirement; and (b) sludge bulking. In the ENBNRAS system, nitrification is transferred from the aerobic reactor in the suspended medium activated sludge system to a fixed medium nitrification system. Thus, the sludge age of the suspended medium activated sludge system can be reduced from 20 to 25 days to 8 to 10 days, resulting in a decrease in reactor volume per ML wastewater treated of about 30%. Furthermore, the aerobic mass fraction can also be reduced from 50% to 60% to <30% and concommitantly the anoxic mass fraction can be increased from 25% to 35% to >55% (if the anaerobic mass fraction is 15%), and thus complete denitrification in the anoxic reactors becomes possible. Research indicates that both the short sludge age and complete denitrification could ameliorate anoxic aerobic (AA) or low food/microorganism (F/M) ratio filamentous bulking, and hence reduce the surface area of secondary settling tanks or increase the treatment capacity of existing systems. The lab-scale experimental investigations indicate that the ENBNRAS system can obtain: (i) very good chemical oxygen demand (COD) removal, even with an aerobic mass fraction as low as 20%; (ii) high nitrogen removal, even for a wastewater with a high total kjeldahl nitrogen (TKN)/COD ratio, up to 0.14; (iii) adequate settling sludge (diluted sludge volume index [DSVI] <100 mL/g); and (iv) a significant reduction in oxygen demand.