Biological treatment of real textile wastewater containing sulphate, salinity, and surfactant through an anaerobic-aerobic system.

Real textile wastewater containing high salinity (up to 12.6 g·kg-1) and surfactant (up to 5.9 mg·L-1 of linear alkylbenzene sulfonate - LAS) was submitted to biological treatment for colour (up to 652 mg Pt-Co·L-1) and sulphate (up to 1,568.6 mg SO4-2·L-1) removal. The influence of ethanol and molasses supplementation was firstly evaluated in anaerobic batch reactors for the removal of dyes and sulphate. Subsequently, aiming to remove aromatic amines (dye degradation by-products), an anaerobic-aerobic continuous system supplemented with molasses was applied. Supplementation had no influence on colour removal (maximum efficiencies around 70%), while it improved sulphate reduction (23% without supplementation against 87% with supplementation), and conferred robustness to the reactors, which recovered quickly after higher salinity impact. The aerobic reactor removed aromatic amines when the level of surfactants was lower than 1.0 mg LAS·L-1, but the performance of the system was hindered when the concentration was increased to 5.9 mg LAS·L-1. Findings suggest that the supplementation of an easily biodegradable organic matter might be a strategy to overcome wastewater fluctuation in composition.

Hacking biofilm developed in a structured-bed reactor (SBRRIA) with integrated processes of nitrogen and organic matter removal.

Biomass samples from a structured-bed reactor subjected to recirculation and intermittent aeration (SBRRIA) were analyzed to investigate the bacterial community shift along with the changes in the C/N ratio. The C/N ratios tested were 7.6 ± 1.0 (LNC) and 2.9 ± 0.4 (HNC). The massive sequencing analyses revealed that the microbial community adjusted itself to different organic and nitrogenous applied loads, with no harm to reactor performance regarding COD and Total-N removal. Under LNC, conventional nitrification and heterotrophic denitrification steered the process, as indicated by the detection of microorganisms affiliated with Nitrosomonadaceae, Nitrospiraceae, and Rhodocyclaceae families. However, under HNC, the C/N ratio strongly affected the microbial community, resulting in the prevalence of members of Saprospiraceae, Chitinophagaceae, Xanthomonadaceae, Comamonadaceae, Bacillaceae, and Planctomycetaceae. These families include bacteria capable of using organic matter derived from cell lysis, ammonia-oxidizers under low DO, heterotrophic nitrifiers-aerobic denitrifiers, and non-isolated strains of Anammox. The DO profile confirmed that the stratification in aerobic, anoxic, and anaerobic zones enabled the establishment of different nitrogen degradation pathways, including the Anammox.

Two-stage partial nitrification-Anammox process for nitrogen removal from slaughterhouse wastewater: Evaluation of the nitrogen loading rate and microbial community analysis.

The production of inputs for animal feed using slaughterhouse byproducts is a predominant waste valorization route of the meat industry. This practice generates complex effluents containing high concentrations of organic matter and nutrients. The partial nitrification process followed by the Anammox process (PN/A) has been shown to be a viable technology for nitrogen removal from wastewaters with high concentrations of ammonia and low COD/N ratios, as found in Upflow Anaerobic Sludge Blanket (UASB) effluent from animal feed inputs industries. However, its application has not been assessed for slaughterhouse byproducts processing wastewaters. This work aimed at evaluating the influence of the nitrogen loading rate (NLR) on the removal of total nitrogen (TN) of a PN/A process treating real animal feed industry wastewater. The NLR in the Anammox reactor varied from 1.3 to 6.3 g N L-1.d-1, with a constant COD/N ratio of 0.5 ± 0.1 mg COD.mg N-1. An average removal efficiency of TN of 84.2 ± 9.8% was observed throughout 440 days of operation. Microbiological analyses of the granular Anammox sludge performed before and after the operation revealed an increase in the population of heterotrophic denitrifying bacteria, while the relative abundance of Anammox species decreased. It was demonstrated that although both microbial groups can coexist synergistically, the presence of organic matter contributed to the growth of heterotrophic denitrifying species and impaired the growth of Anammox bacteria, without affecting system performance.

Sulfide-driven denitrification: detecting active microorganisms in fed-batch enrichment cultures by DNA stable isotope probing.

A microbial community was enriched in the anoxic compartment of a pilot-scale bioreactor that was operated for 180 days, fed with sewage and designed for organic matter, nitrogen and sulfide removal by coupling anaerobic digestion, nitrification and mixotrophic denitrification. Denitrification occurred with endogenous electron donors, mainly sulfide and residual organic matter, coming from the anaerobic compartment. The microorganisms involved in denitrification with sulfide as electron donor were identified by DNA-stable isotope probing with [U-13C]-labelled CO2 and NaHCO3. Complete denitrification occurred every two days, and the applied NO3-/S2- ratio was 1.6. Bacteria belonging to the Sulfurimonas denitrificans was identified as a chemoautotrophic denitrifier, and those related to Georgfuchisa toluolica, Geothrix fermentans and Ferritrophicum radicicola were most probably associated with heterotrophic denitrification using endogenous cells and/or intermediate metabolites. This study showed that DNA-SIP was a suitable technique to identify the active microbiota involved in sulfide-driven denitrification in a complex environment, which may contribute to improve design and operation of bioreactors aiming for carbon-nitrogen-sulfur removal.

Proof of concept and improvement of a triple chamber biosystem coupling anaerobic digestion, nitrification and mixotrophic endogenous denitrification for organic matter, nitrogen and sulfide removal from domestic sewage.

In this study, two versions of a triple chamber biosystem, coupling anaerobic digestion, nitrification and mixotrophic endogenous denitrification (ADNMED), were evaluated and compared. They were designed to maximize the use of endogenous electron donors produced by anaerobic digestion (residual organic matter and sulfide) to abate a portion of the influent nitrogen contained in domestic sewage while removing the inconvenience of effluent sulfide. The first version was able to abate 40% of the influent nitrogen but presented operational and hydrodynamic problems, which resulted in sulfide emissions. A modified second version was proposed, improving the first approach and achieving a nitrogen abatement of more than 60% and a sulfide-free effluent, complying with local emission standards. The results demonstrated that endogenous electron donors produced by anaerobic digestion should not be neglected, and a significant cost reduction in nitrogen removal from domestic sewage could be achieved by exploiting their potential with novel reactor configurations.

Effects of intermittent aeration periods on a structured-bed reactor continuously fed on the post-treatment of sewage anaerobic effluent.

This study assessed the simultaneous nitrification and denitrification processes and remaining organic matter removal from anaerobic reactor effluent treating wastewater in a single reactor. A structured-bed reactor, with polyurethane foam as support media, was subjected to intermittent aeration and effluent recirculation. Aerated/non-aerated periods varied in the range of 2/1-1/3 h. The chemical oxygen demand (COD) in the effluent remained between 26 and 42 mg L-1 throughout all the aeration conditions. Aeration periods of 1/2 h removed 80 and 26% of Total Kjeldahl Nitrogen and Total Nitrogen, respectively. A low solid production was observed during the 300 days of operation, resulting in a solid retention time of 139 days. The results indicate that the non-aerated periods generated alkalinity that favored nitrification, maintaining low COD concentrations in the effluent. The structured bed reactor presented a low solid production and effluent loss below 20 mgSSV L-1, similar to concentrations obtained in secondary decanters.

Carbon-nitrogen removal in a structured-bed reactor (SBRRIA) treating sewage: Operating conditions and metabolic perspectives.

The present study evaluated the efficiency of a structured-bed reactor subjected to recirculation and intermittent aeration (SBRRIA) to promote nitrogen and carbon removal from domestic sewage. The intermittent aeration and the recycling rate of 3 keeps the desired mixing degree inside the SBRRIA. Four different operational conditions were tested by varying the hydraulic retention time (HRT) from 12 to 8 h and aerated and non-aerated periods (A/NA) from 2 h/1 h and 3 h/1 h. At the THD of 8 h and A/NA of 2 h/1 h there was a decrease in the nitrification process (77.5%) due to the increase of organic matter availability, affecting the total-N removal performance. However, by increasing the aerated period from 2 h to 3 h, the nitrification efficiency rose to 91.1%, reaching a total-N removal efficiency of 79%. The system reached a maximum total-N loading removed of 0.117 kgN.m-3.d-1 by applying an HRT of 8 h and an intermittent aeration cycle of 3 h, aerated and 1 h non-aerated. The simultaneous nitrification and denitrification (SND) process was related to a complex interplay among microorganisms affiliated mostly to Acidovorax sp., Comamonas sp., Dechloromonas sp., Hydrogenophaga sp., Mycobacterium sp., Rhodobacter sp., and Steroidobacter sp.

Down-flow fixed-structured bed reactor: An innovative reactor configuration applied to acid mine drainage treatment and metal recovery.

A down-flow fixed-structured bed reactor (DFSBR) was operated for 277 days treating a mixture of synthetic substrates simulating an iron-rich acid mine drainage (AMD) and the soluble fraction of a sugarcane vinasse. The synthetic sugarcane vinasse was used as electron donor for biological sulfate-reduction, resulting in influent chemical oxygen demand (COD) close to 4000 mg L-1 and volumetric organic loading rate of 4.8 g L-1d-1. The influent sulfate concentration was kept close to 2000 mg L-1 (volumetric sulfate loading rate of 2.5 g L-1d-1) while a gradual increase of iron concentration (2-400 mg L-1) was applied. COD removal efficiencies were higher than 93% and the sulfate removal efficiencies were close to 100%. With the highest iron concentration (400 mg L-1) applied, the DFSBR achieved 95% of iron removal efficiency. The precipitate collected at the reactor bottom showed increasing concentrations of fixed suspended solids (FSS), as well as an increasing proportion of iron, indicating the possibility of metal recovery from the system. The association between sulfidogenic and methanogenic processes also enables energy recovery from the methane-rich biogas produced.

Microbial community in a pilot-scale bioreactor promoting anaerobic digestion and sulfur-driven denitrification for domestic sewage treatment.

A pilot-scale reactor treating domestic sewage was operated to promote anaerobic digestion and denitrification using endogenous electron donors. While 55 % of organic matter was removed, nitrogen and sulfur showed a different dynamics during the operation. Pyrosequencing analysis clarified this behavior revealing that specific microbial communities inhabited the anaerobic (47.05 % of OTUs) and anoxic (31.39 % of OTUs) chambers. Analysis of 16S rRNA gene partial sequences obtained through pyrosequencing revealed a total of 1727 OTUs clustered at a 3 % distance cutoff. In the anaerobic chamber, microbial community was comprised of fermentative, syntrophic and sulfate-reducing bacteria. The majority of sequences were related to Aminobacterium and Syntrophorhabdus. In the anoxic chamber, the majority of sequences were related to mixotrophic and strictly autotrophic denitrifiers Arcobacter and Sulfuricurvum, respectively, both involved in sulfur-driven denitrification. These results show that pyrosequencing was a powerful tool to investigate the microbial panorama of a complex system, providing new insights to the improvement of the system.

Influence of COD/N ratio and carbon source on nitrogen removal in a structured-bed reactor subjected to recirculation and intermittent aeration (SBRRIA).

This study aimed to evaluate the influence of COD/N ratio and carbon source on simultaneous nitrogen and carbon removal processes. A continuous up-flow structured-bed reactor subjected to recirculation and intermittent aeration (SBRRIA) was operated with hydraulic retention time (HRT) of 11.2 ± 0.6 h. The carbon sources were meat peptone and sucrose. The COD/N ratio varied by maintaining the organic loading rate fixed at 1.07 kg COD m(-3) d(-1) and changing the total-N concentration. The COD/N ratios tested were 9.7 ± 1 (sucrose); 7.6 ± 1 (meat peptone); 2.9 ± 1 (meat peptone) and 2.9 ± 0.4 (sucrose). COD removal efficiencies remained above 90% in all experimental phases. At lower COD/N ratios, NH4(+)-N oxidation efficiencies were higher than 90%. An autotrophic metabolism by anammox process was observed in Phases III and IV, which was responsible for 35% and 27% of total-N loading removal rates, respectively. Therefore, the system achieved total nitrogen removal efficiencies of 84.6 ± 10.1 and 81.5 ± 5.3%, under low availability of organic electron donors.

Removal of COD and nitrogen from animal food plant wastewater in an intermittently-aerated structured-bed reactor.

This study evaluated the performance of a continuous flow structured-bed reactor in the simultaneous removal of total nitrogen (TN) and chemical oxygen demand (COD) in the effluent from an animal food plant. The reactor had an intermittent aeration system; hydraulic retention time (HRT) of one day; temperature of 30 °C; and recirculation ratio of five times the flow. An experimental central composite rotational delineation (CCRD) type design was used to define the aeration conditions and nitrogen load (factors) to be studied. Response surface methodology was used to analyse the influence of the factors above the results, the removal of TN and COD. It was observed that the aeration factor showed the greatest significance for the results and that the affluent TKN concentration did not have a significant effect, at a 95% level of confidence, on COD removal. Throughout the experiment, the COD/N ratio remained between 3.2 and 3.8. The best results for COD and TN removal, 80% and 88%, respectively, were obtained with 158 min of aeration on a cycle of 180 min and 255 mg L(-1) of Total Kjeldahl Nitrogen (TKN) in the substrate.

Removing organic matter from sulfate-rich wastewater via sulfidogenic and methanogenic pathways.

The simultaneous organic matter removal and sulfate reduction in synthetic sulfate-rich wastewater was evaluated for various chemical oxygen demand (COD)/sulfate ratios applied in a horizontal-flow anaerobic immobilized sludge (HAIS) reactor. At higher COD/sulfate ratios (12.5 and 7.5), the removal of organic matter was stable, likely due to methanogenesis. A combination of sulfate reduction and methanogenesis was clearly established at COD/sulfate ratios of 3.0 and 1.9. At a COD/sulfate ratio of 1.0, the organic matter removal was likely influenced by methanogenesis inhibition. The quantity of sulfate removed at a COD/sulfate ratio of 1.0 was identical to that obtained at a ratio of 1.9, indicating a lack of available electron donors for sulfidogenesis. The sulfate reduction and organic matter removal were not maximized at the same COD/sulfate ratio; therefore, competitive inhibition must be the predominant mechanism in establishing an electron flow.

BTEX removal in a horizontal-flow anaerobic immobilized biomass reactor under denitrifying conditions.

Because benzene, toluene, ethylbenzene, and xylenes (BTEX) and ethanol are important contaminants present in Brazilian gasoline, it is essential to develop technology that can be used in the bioremediation of gasoline-contaminated aquifers. This paper evaluates the performance of a horizontal-flow anaerobic immobilized biomass (HAIB) reactor fed with water containing gasoline constituents under denitrifying conditions. Two HAIB reactors filled with polyurethane foam matrices (5 mm cubes, 23 kg/m(3) density and 95 % porosity) for biomass attachment were assayed. The reactor fed with synthetic substrate containing protein, carbohydrates, sodium bicarbonate and BTEX solution in ethanol, at an Hydraulic retention time (HRT) of 13.5 h, presented hydrocarbon removal efficiencies of 99 % at the following initial concentrations: benzene 6.7 mg/L, toluene 4.9 mg/L, m-xylene and p-xylene 7.2 mg/L, ethylbenzene 3.7 mg/L, and nitrate 60 mg N/L. The HAIB reactor fed with gasoline-contaminated water at an HRT of 20 h showed hydrocarbon removal efficiencies of 96 % at the following initial concentrations: benzene, 4.9 mg/L; toluene, 7.2 mg/L; m-xylene, 3.7 mg/L; and nitrate 400 mg N/L. Microbiological observations along the length of the HAIB reactor fed with gasoline-contaminated water confirmed that in the first segment of the reactor, denitrifying metabolism predominated, whereas from the first sampling port on, the metabolism observed was predominantly methanogenic.

A most-probable number technique for methanotrophic bacteria determination in biological reactors using methane as an electron donor for denitrification.

The most-probable number (MPN) technique along with methane uptake determinations were used to estimate the density of methanotrophic organisms in the biological reactors used for wastewater treatment. The experimental technique was conducted using serum bottles seeded with an inoculum taken from an aerobic sequencing batch reactor that used methane as the sole carbon source. To verify the presence ofmethanotrophic organisms in the support media, biomass samples were subjected to molecular cloning and sequencing techniques. When compared with the sequences published in databanks, the nucleotide sequences obtained showed a phylogenetic similarity of 98% to Methylomonas sp. (access number AF150792) and a phylogenetic similarity of 96% to Chryseobacterium sp. (access number AB264124), which are type I methanotrophs and denitrifiers, respectively.

Unique biofilm structure and mass transfer mechanisms in the foam aerated biofilm reactor (FABR).

The foam-aerated biofilm reactor (FABR) is a novel biofilm process that can simultaneously remove carbon and nitrogen from wastewater. A porous polyurethane foam sheet forms an interface between wastewater and aerated water, making it a counter-diffusional biofilm process similar to the membrane-aerated biofilm reactor (MABR). However, it is not clear how biofilm develops the foam interior, and how this impacts mass transfer and performance. This research explored biofilm development within the foam sheet and determined whether advective transport within the sheet played a significant role. Foam sheets with 2-, 4.5- and 9-mm thicknesses were explored. Oxygen, nitrate, nitrite and ammonia profiles in the sheet were measured using microsensors, and biofilm imaging studies were carried out using optical coherence tomography (OCT). On the foam's aerated side, a dense nitrifying biofilm formed. Beyond the aerobic zone, much less biomass was observed, with a high porosity foam-biofilm layer. The higher effective diffusivity within the foam for the 4- and 9-mm sheets suggested advective transport within the foam channel structures. Using an effective diffusivity factor in conventional 1-D biofilm models reproduced the measured substrate concentration profiles within the foam. Four different practical conditions were modelled. The maximum TN removal efficiency was about 70% and a nitrogen removal flux of 1.25 gN.m-2.d-1. We conclude that mass transfer resistance occurred primarily in the dense, nitrifying layer near the aerated side. The rest of the foam sheet was porous, allowing the advective mass transfer.

Enhanced biological nitrogen and phosphorus removal from sewage driven by fermented glycerol: comparative assessment between sequencing batch- and continuously fed-structured fixed bed reactor.

The nutrient biological removal from sewage, especially from anaerobic reactor effluents, still represents a major challenge in conventional sewage treatment plants. In this work, the nitrogen and phosphorus removal from anaerobic pre-treated domestic sewage in an up-flow anaerobic sludge blanket (UASB) reactor was assessed in a structured fixed bed reactor (SFBR) operated in a continuous and in a batch mode using polyurethane foam as material support for biomass and fermented glycerol as the exogenous carbon source. The SFBR was operated as a sequencing batch reactor with cycles of 90, 120, and 150 min under anaerobic, oxic, and anoxic conditions, respectively, reaching average efficiencies for total nitrogen and phosphorus removal of 88% and 56%, respectively. Fermented glycerol was added during the non-aerated periods. Under continuous feeding, the SFBR was operated with aeration/non-aeration periods of 2/1 (h) and 3/1 (h), hydraulic retention time of 12 h, and a recirculation ratio of 3. Without fermented glycerol addition, the maximum removal of total nitrogen (TN) reached 42%, while adding glycerol in the non-aerated period improved TN removal to 64.9% (2/1 h) and 69.5% (3/1 h). During continuous operation, no phosphorus removal was observed, which was released during the non-aerated period, remaining in the effluent. Optical microscopy analyses confirmed the presence of polyphosphate granules and of the phosphorus accumulating organisms in the reactor biofilm. It was concluded that the batch feeding method was determinant for phosphorus removal. The structured fixed bed reactor with polyurethane foam proved to be feasible in the removal of organic matter and nutrients remaining in the UASB reactor effluent.

Nitrogen and carbon removal from synthetic wastewater in a vertical structured-bed reactor under intermittent aeration.

The removal of nitrogen and organic matter using a single reactor has been a common focus of investigation, and reactors operated in batch mode and under intermittent aeration have attracted special attention. This study aimed to evaluate the application of a new reactor configuration consisting of a fixed-bed reactor that was operated under conditions of continuous feeding and intermittent aeration. The reactor was built using acrylic, with a working volume of 6.1L. The fixed bed used for biomass support was composed of polyurethane foam cylinders vertically oriented inside the reaction zone. The reactor was operated under intermittent aeration (2h aerated and 1h non-aerated) and a recirculation ratio Q(r)/Q=5. Three different operating conditions (Phase I, Phase II, and Phase III) corresponding to hydraulic retention times (HRT) of 12h, 8h, and 10h, respectively, were tested. In Phase I, the system achieved total nitrogen (TN) and chemical oxygen demand (COD) removal efficiencies of 82% and 89%, respectively. At HRTs of 8 h and 10 h, the reactor was unstable with respect to TN removal, and the average resultant removal efficiencies were 49% and 45%, respectively. However, COD removal efficiencies remained high with mean values of 85% and 88% for Phases II and III, respectively. Based on these results, it can be concluded that this new reactor configuration constitutes an alternative method for effective removal of organic matter and nitrogen from wastewater.

Foam aerated biofilm reactor: a novel counter-diffusional process for COD and nitrogen removal from low COD/N effluents.

Counter-diffusional biofilms are efficient in the removal of nitrogen from low strength wastewaters. Although counter-diffusion is usually established using expensive gas-permeable membranes, a polyurethane sheet is used to separate the aerobic and anoxic environments in the novel foam aerated biofilm reactor (FABR). Foam sheets with thicknesses of 10, 5 and 2 mm and synthetic wastewater with COD/N ratios of 5 and 2.5 were evaluated. The 2 mm thick foam reactor did not show good biomass adherence and, therefore, did not show N removal efficiency. The 5 and 10 mm reactors, in both COD/N ratios, showed similar total nitrogen and COD removal performance, up to 60% and 80%, respectively. The denitrification efficiency was close to 100% throughout the experimental period. Nitrification efficiency decreased with microbial growth, which was recovered after removal of excessive biomass. Lower values of polyurethane foam thickness and COD/N ratio did not provide a higher nitrification rate, as expected. The increase in resistance to mass transfer was associated with the growth of biomass attached to the foam rather than to its thickness and resulted in specialization of the microbial communities as revealed by 16S amplicon sequencing. FABR reveals as a promising alternative for simultaneous removal of nitrogen and COD from low COD/N ratio wastewaters.

Temperature and feed strategy effects on sulfate and organic matter removal in an AnSBB.

The objective of this work was to analyze the interaction effects between temperature, feed strategy and COD/[SO(4)(2-)] levels, maintaining the same ratio, on sulfate and organic matter removal efficiency from a synthetic wastewater. This work is thus a continuation of Archilha et al. (2010) who studied the effect of feed strategy at 30 °C using different COD/[SO(4)(2-)] ratios and levels. A 3.7-L anaerobic sequencing batch reactor with recirculation of the liquid phase and which contained immobilized biomass on polyurethane foam (AnSBBR) was used to treat 2.0 L synthetic wastewater in 8 h cycles. The temperatures of 15, 22.5 and 30 °C with two feed strategies were assessed: (a) batch and (b) batch followed by fed-batch. In strategy (a) the reactor was fed in 10 min with 2 L wastewater containing sulfate and carbon sources. In strategy (b) 1.2 L wastewater (containing only the sulfate source) was fed during the first 10 min of the cycle and the remaining 0.8 L (containing only the carbon source) in 240 min. Based on COD/[SO(4)(2-)] = 1 and on the organic matter (0.5 and 1.5 gCOD/L) and sulfate (0.5 and 1.5 gSO(4)(2-)/L) concentrations, the sulfate and organic matter loading rates applied were 1.5 and 4.5 g/L.d, i.e., same COD/[SO(4)(2-)] ratio (=1) but different levels (1.5/1.5 and 4.5/4.5 gCOD/gSO(4)(2-)). When reactor feed was 1.5 gCOD/L.d and 1.5 gSO(4)(2-)/L.d, gradual feeding (strategy b) showed to favor sulfate and organic matter removal in the investigated temperature range, indicating improved utilization of the electron donor for sulfate reduction. Sulfate removal efficiencies were 87.9; 86.3 and 84.4%, and organic matter removal efficiencies 95.2; 86.5 and 80.8% at operation temperatures of 30; 22.5 and 15 °C, respectively. On the other hand, when feeding was 4.5 gCOD/L.d and 4.5 gSO(4)(2-)/L.d, gradual feeding did not favor sulfate removal, indicating that gradual feeding of the electron donor did not improve sulfate reduction.

Interaction effects of organic load and cycle time in an AsBr applied to a personal care industry wastewater treatment.

A mechanically stirred anaerobic sequencing batch reactor (ASBR) containing granular biomass was applied to the treatment of a wastewater simulating the effluent from a personal care industry. The ASBR was operated with cycle lengths (t(C)) of 8, 12 and 24 h and applied volumetric organic loads (AVOL) of 0.75, 0.50 and 0.25 gCOD/L.d, treating 2.0 L liquid medium per cycle. Stirring frequency was 150 rpm and the reactor was kept in an isothermal chamber at 30 °C. Increase in t(C) resulted in efficiency increase at constant AVOL, reaching 77% at t(C) of 24 h versus 69% at t(C) of 8 h. However, efficiency decreased when AVOL decreased as a function of increasing t(C), due to the lack of substrate in the reaction medium. Moreover, replacing part of the wastewater by a chemically balanced synthetic one did not yield the expected effect and system efficiency dropped.