Aerobic granulation in sequencing batch reactors with different reactor height/diameter ratios

Effects of reactor height/diameter ratios ranged from 24 to 4 corresponding to reactor settling velocities from 12 to 2 m h^?1 on aerobic granulation were investigated. It was found that granules appeared after 1-week operation and granule volume percentages exceeded 50% after 2–3 weeks in four reactors. In addition, similar granule fraction of 94–96% was found at steady state in all four reactors. Sludge volume index (SVI), average sludge size, biomass density and granule settling velocity at steady state were around 50 ml g^?1, 1800 μm, 53 g l^?1 and 40 m h^?1, respectively, in four reactors. Extracellular polymeric substances (EPS) and specific oxygen uptake rate (SOUR) were around 38 mg g^?1 VSS and 40 mg O₂ g^?1 VSS h^?1, respectively. Denaturing gradient gel electrophoresis (DGGE) fingerprint of sludge in four reactors showed the same microbial population shift during the start-up period and same microbial community structure during steady-state period. These results recommended strongly that reactor height/diameter ratio or reactor setting velocity in the used range in this study did not affect granule formation, physical characteristics, microbial community structure of granules and stable operation of granular sludge reactor. Reactor height/diameter ratio thus can be very flexible in the practice, which is important for the application of aerobic granule technology.     

Investigation on the formation and kinetics of glucose-fed aerobic granular sludge

Aerobic granular sludge was cultivated in a glass sequencing batch reactor (SBR) with glucose synthetic wastewater. The spherical shaped granules were observed on 4th day with the mean diameter of 0.1 mm. With the increase of chemical oxygen demand (COD) concentration of the influent, aerobic granules grew matured, the size of which ranged from 1.2 to 1.9 mm. The aerobic granular sludge could sustain high organic loading rate (about 4.0 g COD L⁻¹ d⁻¹), with good settling ability (settling velocity 36 m/h) and high biomass concentration (MLSS 6.7 ±0.2 g/L). Experimental data indicated that the substrate utilization and biomass growth kinetics followed Monod's kinetics model approximately. The corresponding kinetic coefficients of maximum specific substrate utilization rate (k), half velocity coefficient (K_s), growth yield coefficient (Y) and decay coefficient (K_d) were 13.2 d⁻¹, 275.8 mg/L, 0.183–0.250 mg MLSS/mg COD and 0.023–0.075 d⁻¹, respectively, which made aerobic granules have short setup period, high rate of substrate utilization and little surplus sludge.     

A key cultivation technology for denitrifying granular sludge

Well-formed denitrifying granular sludge with a biomass concentration of 24.8 gVSS L^?1 and a specific nitrate removal rate of 0.19 gNO₃-N gVSS^?1 d^?1 was obtained in an upflow sludge blanket (USB) reactor by cultivating seeded aerobic flocculent sludge for 6–8 weeks. Regularity phenomena exist in the granulation including flotation of flocculent sludge, formation of fine granules, occurrence of channelling, and formation of mature granular sludge. The granulation is similar to crystal growth, that the non-denitrifying bacteria evolve into the carriers (fine granules), on the surface of which denitrifying bacteria proliferate and develop into mature granular sludge.There are several key parameters that must be considered when developing a good denitrifying granular sludge. First, the proper seed sludge must be chosen (VSS/SS at 0.65–0.75, SRT over 25 days) to accelerate the granulation process. Secondly, any floating sludge should be stirred, and the sludge loading rate should be within the range of 0.05–0.15 gNO₃-N gVSS^?1 d^?1 until fine granules emerge. Additionally, spontaneous gas agitation or interval air-blowing should be used to effectively eliminate channelling; Finally, the sludge loading rate should be less than 0.25 gNO₃-N gVSS^?1 d^?1 until dense, mature granular sludge appears. This study could support and promote the full-scale application of denitrifying granular sludge.     

Characterization of aerobic granular sludge at various organic loading rates

Aerobic granular sludge was cultivated from activated sludge with two types of supports, namely bivalve shell carrier (BSC) and anaerobic granules (ANG). Granules were characterized at different organic loading rates (OLRs) ranging from 2.5 to 15 kg COD/m³ d and these granules were observed to withstand high OLRs. The physico-chemical characteristics of the aerobic granules were better than those of seed sludge. The granule formation with ANG support was found to be similar to that of non-support cultivation, i.e. formation from activated sludge only. By contrast, BSC support showed better performance in terms of faster settleability, compactness and especially resistance against organic shock loading. It also enabled self-cleaning effect by removing biofilm attached on the reactor wall during the start-up phase resulting rapid granulation process.     

Distribution of anammox bacteria in domestic WWTPs and their enrichments evaluated by real-time quantitative PCR

Enrichment of anaerobic ammonium oxidation (anammox) bacteria using five activated sludges in three domestic wastewater treatment plants (WWTPs) were processed in a short term of 70 days and evaluated by real-time quantitative PCR (RTQ-PCR). Before the enrichment, building phylogenetic trees of Planctomycetes phylum in four reactors of sequencing batch reactor (SBR), anoxic and oxic reactors of anaerobic–anoxic–oxic (A2O) process, and rotating biological contactor (RBC) revealed six groups of distantly relative genera of Planctomyces, Pirellula, Gemmata, Isophaera, Candidatus and putative anammox bacteria. All clones of Candidatus sp. were affiliated with anammox bacteria and the majority of anammox clones were related to Planctomycete KSU-1 (AB057453). The discovery of anammox bacteria in raw activated sludges provided a partial rationale for the utilization of activated sludge as a seeding source of the anammox process. To verify the activity of anammox bacteria in the activated sludges, enrichment cultivations were conducted using SBRs. The enrichment of anammox bacteria resulted in the significant anammox activity of three samples. Quantification of 16S rRNA gene of anammox bacteria using RTQ-PCR showed the highest concentration of anammox bacteria of 2.48 ± 0.22 × 10⁹ copies of 16S rRNA gene/mg-volatile suspended solids (VSS), which was the same order of magnitude as that of the referential granular anammox sludge, 6.23 ± 0.59 × 10⁹ copies of 16S rRNA gene/mg-VSS, taken from an anammox upflow anaerobic sludge blanket (UASB) reactor. The doubling time of anammox bacteria enriched in this study was 1.18 days. The growth yield of anammox bacteria enriched in this study was 4.75 ± 0.57 × 10⁶ copies of 16S rRNA gene/mg of ammonium- and nitrite-nitrogen, which was similar to 4.50 ± 0.61 × 10⁶ copies of 16S rRNA gene/mg of ammonium- and nitrite-nitrogen for the referential anammox sludge. Substrate uptake rates of three successful enrichments at the end of the enrichment were comparable to those of granular and suspended anammox sludges. Rapid enrichment of anammox bacteria using activated sludge could offer an alternative method for obtaining a large volume of seeding anammox sludge.     

Phenol and sulfide oxidation in a denitrifying biofilm reactor and its microbial community analysis

A microbial consortium attached onto a polyethylene support was used to evaluate the simultaneous oxidation of sulfide and phenol by denitrification. The phenol, sulfide and nitrate loading rates applied to an inverse fluidized bed reactor were up to 168 mg phenol–C/(l d), 37 mg S^2?/(l d) and 168 mg NO₃^?–N/(l d), respectively. Under steady state operation the consumption efficiencies of phenol, sulfide and nitrate were 100%. The N₂ yield (g N₂/g NO₃^?–N) was 0.89. The phenol was mineralized resulting in a yield of 0.82 g bicarbonate–C/g phenol–C and sulfide was completely oxidized to sulfate with a yield of 0.99 g SO₄^2?–S/g S^2?. 16S rRNA gene-based microbial community analysis of the denitrifying biofilm showed the presence of Thauera aromatica, Thiobacillus denitrificans, Thiobacillus sajanensis and Thiobacillus sp. This is the first work reporting the simultaneous oxidation of sulfide and phenol in a denitrifying biofilm reactor.     

A half-submerged integrated two-phase anaerobic reactor for agricultural solid waste codigestion

Anaerobic digestion is widely used in bioenergy recovery from waste. In this study, a half-submerged, integrated, two-phase anaerobic reactor consisting of a top roller acting as an acidogenic unit and a recycling bottom reactor acting as a methanogenic unit was developed for the codigestion of wheat straw (WS) and fruit/vegetable waste (FVW). The reactor was operated for 21 batches (nearly 300 d). Anaerobic granular sludge was inoculated into the methanogenic unit. The residence time for the mixed waste was maintained as 10 d when the operation stabilized, and the temperature was kept at 35 °C. The highest organic loading rate was 1.37 kg VS/(m³ d), and the maximum daily biogas production was 328 L/d. Volatile solid removal efficiencies exceeded 85%. WS digestion could be confirmed, and efficiency was affected by both the ratio of WS to FVW and the loading rate. The dominant bacteria were Bacteroides-like species, which are involved in glycan and cellulose decomposition. Methanogenic community structures, pH levels, and volatile fatty acid concentrations in the acidogenic and methanogenic units differed, indicating successful phase separation. This novel reactor can improve the mass transfer and microbial cooperation between acidogenic and methanogenic units and can efficiently and steady codigest solid waste.     

Urine nitrification with a synthetic microbial community

During long-term extra-terrestrial missions, food is limited and waste is generated. By recycling valuable nutrients from this waste via regenerative life support systems, food can be produced in space. Astronauts’ urine can, for instance, be nitrified by micro-organisms into a liquid nitrate fertilizer for plant growth in space. Due to stringent conditions in space, microbial communities need to be be defined (gnotobiotic); therefore, synthetic rather than mixed microbial communities are preferred. For urine nitrification, synthetic communities face challenges, such as from salinity, ureolysis, and organics.In this study, a synthetic microbial community containing an AOB (Nitrosomonas europaea), NOB (Nitrobacter winogradskyi), and three ureolytic heterotrophs (Pseudomonas fluorescens, Acidovorax delafieldii, and Delftia acidovorans) was compiled and evaluated for these challenges. In reactor 1, salt adaptation of the ammonium-fed AOB and NOB co-culture was possible up to 45 mS cm⁻¹, which resembled undiluted nitrified urine, while maintaining a 44 ± 10 mg NH₄⁺–N L⁻¹ d⁻¹ removal rate. In reactor 2, the nitrifiers and ureolytic heterotrophs were fed with urine and achieved a 15 ± 6 mg NO₃⁻–N L⁻¹ d⁻¹ production rate for 1% and 10% synthetic and fresh real urine, respectively. Batch activity tests with this community using fresh real urine even reached 29 ± 3 mg N L⁻¹ d⁻¹. Organics removal in the reactor (69 ± 15%) should be optimized to generate a nitrate fertilizer for future space applications.     

Investigation on the properties and kinetics of glucose-fed aerobic granular sludge

Aerobic granulation is a process in which suspended biomass aggregate and form discrete well-defined granules in aerobic systems. To investigate the properties and kinetics of aerobic granular sludge, aerobic granules were cultivated with glucose synthetic wastewater in a series of sequencing batch reactors (SBR). The spherical shaped granules were observed on 8th day with the mean diameter of 0.1 mm. With the organic loading rate (OLR) being increased to 4.0 g COD L⁻¹ d⁻¹, aerobic granules grew matured with spherical shape. The size of granules ranged from 1.2 to 1.8 mm, and the corresponding settling velocity of individual granule was 24.2–36.4 m h⁻¹. The oxygen utilization rate (OUR) of mature granules was 41.90 g O₂ kg MLSS⁻¹ h⁻¹, which was two times higher than that of activated sludge (18.32 g O₂ kg MLSS⁻¹ h⁻¹). The experimental data indicated that the substrate utilization and biomass growth kinetics generally followed Monod's kinetics model. The corresponding kinetic coefficients of k (maximum specific substrate utilization rate), K_s (half velocity coefficient), Y (growth yield coefficient) and K_d (decay coefficient) were determined as follows, k_c = 23.65 d⁻¹, K_c = 3367.05 mg L⁻¹, K_N = 0.038 d⁻¹, K_N = 29.65 mg L⁻¹, Y = 0.1927–0.2022 mg MMLS (mg COD)⁻¹ and K_d = 0.00845–0.0135 d⁻¹, respectively. Those properties of aerobic granules made aerobic granules system had a short setup period, high substrate utilization rate and low sludge production.     

Research on soybean protein wastewater treatment by the integrated two-phase anaerobic reactor

The start-up tests of treating soybean protein wastewater by the integrated two-phase anaerobic reactor were studied. The results showed that the soybean protein wastewater could be successfully processed around 30 days when running under the situation of dosing seed sludge with the influent of approximately 2000 mg/L and an HRT of 40 h. When the start-up was finished, the removal rate of COD by the reactor was about 80%. In the zone I, biogas mainly revealed carbon dioxide (CO₂) and hydrogen (H₂). Methane was the main component in the zone 2 which ranged from 53% to 59% with an average of 55%. The methane content in biogas increased from the zone I to II. It indicated that the methane-producing capacity of the anaerobic sludge increased. It was found that the uniquely designed two-phase integrated anaerobic reactor played a key role in treating soybean protein wastewater. The acidogenic fermentation bacteria dominated in the zone I, while methanogen became dominant in the zone II. It realized the relatively effective separation of hydrolysis acidification and methanogenesis process in the reactor, which was benefit to promote a more reasonable space distribution of the microbial communities in the reactor. There were some differences between the activities of the sludge in the two reaction zones of the integrated two-phase anaerobic reactor. The activity of protease was higher in the reaction zone I. And the coenzyme F₄₂₀ in the reaction zone II was twice than that in the reaction zone I, which indicated that the activity of the methanogens was stronger in the reaction zone II.     

Anaerobic granule development for removal of pentachlorophenol in an upflow anaerobic sludge blanket (UASB) reactor

The granulation process using synthetic wastewater containing pentachlorophenol (PCP) in four 1.1 l laboratory scale upflow anaerobic sludge blanket (UASB) reactors was studied, and the anaerobic biotransformation of PCP during the granulation process investigated. After 110 days granular sludge was developed and up to 160 and 180 mg/l of PCP was added into the reactors R1 and R2, respectively, when they were inoculated with acclimated anaerobic sludge from an anaerobic digester of a citric acid plant. The inoculum was predominately composed of bacilli and filamentous bacteria. Granulation did not occur in reactors R3 and R4 which were inoculated with acclimated anaerobic sludge from aerobic sludge of the municipal sewage treatment plant which consisted mainly of cocci. Despite similar bacilli in the granule, the filamentous bacteria from reactor R1 were thicker than those of reactor R2. The granular sludge had a maximum diameter of 2.5 and 2.2 mm, and SMA of 1.44 and 1.32 gCOD/gTVS per day for reactors R1 and R2, respectively. Over 98% chemical oxygen demand (COD) removal rate and 99% of PCP removal rate were achieved when reactors R1 and R2 were operated at PCP and COD loading rates of 150 and 7.5 g/l per day, respectively. H₂-producing acetogens were the dominant anaerobes in the granular sludge.     

Thermophilic treatment of bulk drug pharmaceutical industrial wastewaters by using hybrid up flow anaerobic sludge blanket reactor

The hybrid up flow anaerobic sludge blanket reactor was evaluated for efficacy in reduction of chemical oxygen demand (COD) and biochemical oxygen demand (BOD) of bulk drug pharmaceutical wastewater under different operational conditions. The start-up of the reactor feed came entirely with glucose, applied at an organic loading rate (OLR) 1 kg COD/m³ d. Then the reactor was studied at different OLRs ranging from 2 to 11 kg COD/m³ d with pharmaceutical wastewater. The optimum OLR was found to be 9 kg COD/m³ d, where we found 65–75% COD and 80–94% of BOD reduction with biogas production containing 60–70% of methane and specific methanogenic activity was 320 ml CH₄/g-VSS d. By the characterization studies of effluent using GC–MS, the hazardous compounds like phenol, l,2-methoxy phenol, 2,4,6-trichloro phenol, dibutyl phthalate, 1-bromo naphthalene, carbamazepine and antipyrine were present. After the treatment, these compounds degraded almost completely except carbamazepine. Thermophilic methanothrix and methanosaetae like bacteria are present in the granular sludge.     

Identification of microorganisms in the granules generated during methane fermentation of the syrup wastewater produced while canning fruit

The wastewater produced in the process of canning fruit contains a syrup that consists mainly of sucrose. This syrup wastewater was treated by methane fermentation in an upflow anaerobic sludge blanket reactor. The organic loading rate of syrup wastewater was increased gradually as fermentation progressed. The higher the organic loading rate, the more methane gas evolved until the organic loading rate reached 30.3 kg COD m^?3 d^?1, at which point methane generation abruptly diminished because the loading rate was too high to stably operate the reactor. The changes in the microbial community, that of both bacteria and archaea in the granules, were analyzed simultaneously using PCR-DGGE during the fermentation process. Methanosaeta spp., which are methanogenic archaea that produce extracellular polymers indispensable for the formation of granules, were dominant when the methane gas vigorously evolved, and the iron-reducing bacterium belonging to genus Geobacter, which outcompetes methanogens, grew proportionally with the deterioration of methane fermentation.     

High-rate denitrifying sulfide removal process in expanded granular sludge bed reactor

An expanded granular sludge bed (EGSB) reactor was adopted to incubate bio-granules that could simultaneously convert 4.8 kg-S m^?3 d^?1 of sulfide in 97% efficiency; 2.6 kg-N m^?3 d^?1 of nitrate in 92% efficiency; and 2.7 kg-C m^?3 d^?1 acetate in 95% efficiency. Mass balance calculation of sulfur, nitrogen, and carbon over the EGSB reactor confirmed the performance results. This noted reactor performance is much higher than those reported in literature. Stoichiometric relation suggests that the nitrate was reduced to nitrite via autotrophic denitrification pathway, then the formed nitrite was converted via heterotrophic denitrification pathway to N₂.     

Removal of malachite green (MG) from aqueous solutions by native and heat-treated anaerobic granular sludge

The performance of native and heat-treated anaerobic granular sludge in removing of malachite green (MG) from aqueous solution was investigated with different conditions, such as pH, ionic strength, initial concentration and temperature. The maximum biosorption was both observed at pH 5.0 on the native and heat-treated anaerobic granular sludge. The ionic strength had negative effect on MG removal. Kinetic studies showed that the biosorption process followed pseudo-second-order and q_e for native and heat-treated anaerobic granular sludge is 61.73 and 59.17 mg/g at initial concentration 150 mg/L, respectively. Intraparticle diffusion model could well illuminate adsorption process and faster adsorption rate of native anaerobic granular sludge than heat-treated anaerobic granular sludge. The equilibrium data were analyzed using Langmuir and Freundlich model, and well fitted Langmuir model. The negative values of ΔG° and ΔH° suggested that the interaction of MG adsorbed by native and heat-treated anaerobic granular sludge was spontaneous and exothermic. Desorption studies revealed that MG could be well removed from anaerobic granular sludge by 1% (v/v) of HCl–alcohol solution.     

Anaerobic treatment of saline wastewater by Halanaerobium lacusrosei

An upflow anaerobic packed bed reactor was operated continuously with synthetic saline wastewater at different initial COD concentrations (COD₀ = 1900–6300 mg/L), salt concentrations (0–5%, w/v) and hydraulic retention times (θ_H = 11–30 h) to investigate the effect of those operating parameters on COD removal from saline synthetic wastewater. Anaerobic salt tolerant bacteria, Halanaerobium lacusrosei, were used as dominant microbial culture in the process. The percent COD removal reached up to 94% at COD₀ = 1900 mg/L, 19 h hydraulic retention time and 3% salt concentration. No substrate inhibition effect was observed at high feed CODs. Increasing hydraulic retention time from 11 h to 30 h resulted in a substantial improvement in the COD removal from 60% to 84% at around COD₀ = 3400 mg/L and 3% salt concentration. Salt inhibition effect on COD utilization was observed at above 3% salt concentration. Modified Stover–Kincannon model was applied to the experimental data to determine the biokinetic coefficients. Saturation value constant, and maximum utilization rate constant of Stover–Kincannon model for COD were determined as K_B = 5.3 g/L day, U_max = 7.05 g/L day, respectively.     

Two ways to achieve an anammox influent from real reject water treatment at lab-scale: Partial SBR nitrification and SHARON process

A comparative study to produce the correct influent for Anammox process from anaerobic sludge reject water (700–800 mg NH₄⁺-N L⁻¹) was considered here. The influent for the Anammox process must be composed of NH₄⁺-N and NO₂⁻-N in a ratio 1:1 and therefore only a partial nitrification of ammonium to nitrite is required. The modifications of parameters (temperature, ammonium concentration, pH and solid retention time) allows to achieve this partial nitrification with a final effluent only composed by NH₄⁺-N and NO₂⁻-N at the right stoichiometric ratio. The equal ratio of HCO₃⁻/NH₄⁺ in reject water results in a natural pH decrease when approximately 50% of NH₄⁺ is oxidised. A Sequencing batch reactor (SBR) and a chemostat type of reactor (single-reactor high activity ammonia removal over nitrite (SHARON) process) were studied to obtain the required Anammox influent. At steady state conditions, both systems had a specific conversion rate around 40 mg NH₄⁺-N g⁻¹ volatile suspended solids (VSS) h⁻¹, but in terms of absolute nitrogen removal the SBR conversion was 1.1 kg N day⁻¹ m⁻³, whereas in the SHARON chemostat was 0.35 kg N day⁻¹ m⁻³ due to the different hydraulic retention time (HRT) used. Both systems are compared from operational (including starvation experiments) and kinetic point of view and their advantages/disadvantages are discussed.     

Morphology and bloom dynamics of Cochlodinium geminatum (Schütt) Schütt in the Pearl River Estuary,South China Sea

An unarmored dinoflagellate bloom of Cochlodinium geminatum (Schütt) Schütt has been identified in the Pearl River Estuary, South China Sea during the severe dry season, from late October to early November, 2009, when temperature and salinity ranged between 20.0–27.2 °C and 10.6–33.4, respectively. Light and scanning electron microscopy were used to identify the characteristics of C. geminatum and provided the clear morphological structure for this species. The organism was primarily found in chains of two cells or single cell, and no longer chains were observed. Cells were irregularly spherical or slightly dorso-ventrally, with size ranged between 28 and 36 μm and longer than wide. A large nucleus in the center with numerous golden chloroplasts was present, and the cingulum made 1.5 turns around the cell. The concentration of C. geminatum ranged from 10² to greater than 10⁷ cells l⁻¹ during the bloom period. Nutrient concentration ranges during the bloom were 1.29–81.00 μM NO₃, 0.14–12.14 μM NO₂, 0.21–6.29 μM NH₄, 0.23–6.26 μM PO₄ and 3.29–171.43 μM SiO₃, respectively. Total biomass expressed in terms of chlorophyll a ranged from 2.44 to 135.45 μg l⁻¹, with an average 19.9 μg l⁻¹ in surface water throughout the PRE. Two main clusters corresponding to the water sectors were defined with multivariate analysis (cluster and nMDS). Based on the composition and abundance of phytoplankton, spatial variations were observed at a significant level (ANOSIM, R = 0.44, P < 0.01). Although the pairwise correlation analysis detected no significant effect of any single environmental variable on the abundance of C. geminatum, the multivariate analysis (BIO-ENV) between biotic and abiotic variables resulted in the best variables combination with all measured factors involved (temperature, salinity, turbidity, NO₃, NO₂, NH₄, PO₄ and SiO₃) which showed a combined effect during the bloom of C. geminatum in the Pearl River Estuary (ρ_w = 0.477).     

Production of gluconic acid from glucose by Aspergillus niger: growth and non-growth conditions

Batch fermentation of glucose to gluconic acid was conducted using Aspergillus niger under growth and non-growth conditions using pure oxygen and air as a source of oxygen for the fermentation in 2 and 5 l stirred tank reactors (batch reactor). Production of gluconic acid under growth conditions was conducted in a 5 l batch reactor. Production and growth rates were higher during the period of supplying pure oxygen than that during supplying air, and the substrate consumption rate was almost constant. For the production of gluconic acid under non-growth conditions, conducted in the 2 l batch reactor, the effect of the pure oxygen flow rate and the biomass concentration on the gluconic acid production was investigated and an empirical equation suggested to show the dependence of the production rate r_p on the biomass concentration C_x and oxygen flow rate Q, at constant operating conditions (30 °C, 300 rpm and pH 5.5). Biomass concentration had a positive effect on the production rate r_p, and the effect of Q on r_p was positive at high biomass concentrations.     

Anaerobic submerged membrane bioreactor (AnSMBR) treating low-strength wastewater under psychrophilic temperature conditions

An anaerobic submerged membrane bioreactor (AnSMBR) treating low-strength wastewater was operated for 90 days under psychrophilic temperature conditions (20 °C). Besides biogas sparging, additional shear was created by circulating sludge to control membrane fouling. The critical flux concept was used to evaluate the effectiveness of this configuration. Biogas sparging with a gas velocity (U_G) of 62 m/h together with sludge circulation (94 m/h) led to a critical flux of 7 L/(m² h). Nevertheless, a further increase in the U_G only minimally enhanced the critical flux. A low fouling rate was observed under critical flux conditions. The cake layer represented the main fouling resistance after 85 days of operation. Distinctly different volatile fatty acid (VFA) concentrations in the reactor and in the permeate were always observed. This fact suggests that a biologically active part of the cake layer contributes to degrade a part of the daily organic load. Hence, chemical oxygen demand (COD) removal efficiencies of up to 94% were observed. Nevertheless, the biogas balance indicates that even considering the dissolved methane, the methane yield were always lower than the theoretical value, which indicates that the organic compounds were not completely degraded but physically retained by the membrane in the reactor.