Analysis of fouling mechanisms in anaerobic membrane bioreactors

In this paper, we investigate the fouling mechanisms responsible for MF and UF membrane flux decline in Anaerobic Membrane Bioreactors (AnMBR). We have used the fouling mechanism models proposed by Hermia (1982), namely pore constriction, cake formation, complete blocking and intermediate blocking. Based on an optimization approach and using experimental data extracted from the literature, we propose a systematic procedure for identifying the most likely fouling mechanism in play. Short-term as well as long-term experiments are considered and discussed. It was found that short-term experiments are usually characterized by two fouling phases during which the same fouling mechanism or two different mechanisms affect the process. In contrast, in long-term experiments involving cleaning cycles, membrane fouling appears to be better ascribed to one phase only.The impact of abiotic parameters on membrane fouling mechanisms is reviewed and discussed in the light of these results. Finally, it is shown that the mechanism most responsible for membrane fouling in an AnMBR is cake formation. This main result will be useful for the future development of simple integrated models for optimization and control.     

Evaluation of molecular methods used for establishing the interactions and functions of microorganisms in anaerobic bioreactors

Molecular techniques have unveiled the complexity of the microbial consortium in anaerobic bioreactors and revealed the presence of several uncultivated species. This paper presents a review of the panoply of classical and recent molecular approaches and multivariate analyses that have been, or might be used to establish the interactions and functions of these anaerobic microorganisms. Most of the molecular approaches used so far are based on the analysis of small subunit ribosomal RNA but recent studies also use quantification of functional gene expressions. There are now several studies that have developed quantitative real-time PCR assays to investigate methanogens. With a view to improving the stability and performance of bioreactors, monitoring with molecular methods is also discussed. Advances in metagenomics and proteomics will lead to the development of promising lab-on chip technologies for cost-effective monitoring.     

Enrichment of acetogenic bacteria in high rate anaerobic reactors under mesophilic and thermophilic conditions

The objective of the current study was to expand the knowledge of the role of acetogenic Bacteria in high rate anaerobic digesters. To this end, acetogens were enriched by supplying a variety of acetogenic growth supportive substrates to two laboratory scale high rate upflow anaerobic sludge bed (UASB) reactors operated at 37 °C (R1) and 55 °C (R2). The reactors were initially fed a glucose/acetate influent. Having achieved high operational performance and granular sludge development and activity, both reactors were changed to homoacetogenic bacterial substrates on day 373 of the trial. The reactors were initially fed with sodium vanillate as a sole substrate. Although % COD removal indicated that the 55 °C reactor out performed the 37 °C reactor, effluent acetate levels from R2 were generally higher than from R1, reaching values as high as 5023 mg l⁻¹. Homoacetogenic activity in both reactors was confirmed on day 419 by specific acetogenic activity (SAA) measurement, with higher values obtained for R2 than R1.Sodium formate was introduced as sole substrate to both reactors on day 464. It was found that formate supported acetogenic activity at both temperatures. By the end of the trial, no specific methanogenic activity (SMA) was observed against acetate and propionate indicating that the methane produced was solely by hydrogenotrophic Archaea. Higher SMA and SAA values against H₂/CO₂ suggested development of a formate utilising acetogenic population growing in syntrophy with hydrogenotrophic methanogens. Throughout the formate trial, the mesophilic reactor performed better overall than the thermophilic reactor.     

Functional bacterial and archaeal community structures of major trophic groups in a full-scale anaerobic sludge digester

Functional Bacteria and Archaea community structures of a full-scale anaerobic sludge digester were investigated by using a full-cycle 16S rRNA approach followed by microautoradiography (MAR)-fluorescent in situ hybridization (FISH) technique and micromanipulation. FISH analysis with a comprehensive set of 16S and 23S rRNA-targeted oligonucleotide probes based on 16S rRNA clone libraries revealed that the Gram-positive bacteria represented by probe HGC69A-hybridized Actinobacteria (8.5+/-1.4% of total 4', 6-diamidino-2-phenylindole (DAPI)-stained cells) and probe LGC354-hybridized Firmicutes (3.8+/-0.8%) were the major phylogenetic bacterial phyla, followed by Bacteroidetes (4.0+/-1.2%) and Chloroflexi (3.7+/-0.8%). The probe MX825-hybridized Methanosaeta (7.6+/-0.8%) was the most abundant archaeal group, followed by Methanomicrobiales (2.8+/-0.6%) and Methanobacteriaceae (2.7+/-0.4%). The functional community structures (diversity and relative abundance) of major trophic groups were quantitatively analyzed by MAR-FISH. The results revealed that glucose-degrading microbial community had higher abundance (ca. 10.6+/-4.9% of total DAPI-stained cells) and diversity (at least seven phylogenetic groups) as compared with fatty acid-utilizing microbial communities, which were more specialized to a few phylogenetic groups. Despite the dominance of Betaproteobacteria, members of Chloroflexi, Smithella, Syntrophomonas and Methanosaeta groups dominated the [(14)C]glucose-, [(14)C]propionate-, [(14)C]butyrate- and [(14)C]acetate-utilizing microorganism community, and accounted for 27.7+/-4.3%, 29.6+/-7.0%, 34.5+/-7.6% and 18.2+/-9.5%, respectively. In spite of low abundance (ca. 1%), the hitherto unknown metabolic functions of Spirochaeta and candidate phylum of TM7 as well as Synergistes were found to be glucose and acetate utilization, respectively.     

Continuous treatment of the organic fraction of municipal solid waste in an anaerobic two-stage membrane process with liquid recycle

The stability and performance of a two-stage anaerobic membrane process was investigated at different organic loading rates (OLRs) and Hydraulic Retention Times (HRTs) over 200 days. The Hydrolytic Reactor (HR) was fed with the Organic Fraction of Municipal Solid Waste (OFMSW), while the leachate from the HR was fed continuously to two Submerged Anaerobic Membrane Bioreactors (SAMBR1 and 2). The Total COD (TCOD) of the leachate varied over a wide range, typically between 4000 and 26,000 mg/L while the Soluble COD (SCOD) in the permeate was in the range 400-600 mg/L, achieving a COD removal greater than 90% at a HRT of 1.6-2.3 days in SAMBR1. The operation was not sustainable below this HRT due to a membrane flux limitation at 0.5-0.8 L/m² h (LMH), which was linked to the increasing MLTSS. SCOD in the recycled permeate did not build up indicating a slow degradation of recalcitrants over time. SAMBR2 was run in parallel with SAMBR1 but its permeate was treated aerobically in an Aerobic Membrane Bioreactor (AMBR). The AMBR acted as a COD-polishing and ammonia removal step. About 26% of the recalcitrant SCOD from SAMBR2 could be aerobically degraded in the AMBR. In addition, 97.7 % of the ammonia-nitrogen was converted to nitrate in the AMBR at a maximum nitrogen-loading rate of 0.18 kg NH₄⁺-N/m³ day. GC-MS analysis was performed on the reactor effluents to determine their composition and what compounds were recalcitrant.     

Anaerobic biodegradability and treatment of grey water in upflow anaerobic sludge blanket (UASB) reactor

Feasibility of grey water treatment in an upflow anaerobic sludge blanket (UASB) reactor operated at different hydraulic retention time (HRT) of 16, 10 and 6h and controlled temperature of 30 degrees C was investigated. Moreover, the maximum anaerobic biodegradability without inoculum addition and maximum removal of chemical oxygen demand (COD) fractions in grey water were determined in batch experiments. High values of maximum anaerobic biodegradability (76%) and maximum COD removal in the UASB reactor (84%) were achieved. The results showed that the colloidal COD had the highest maximum anaerobic biodegradability (86%) and the suspended and dissolved COD had similar maximum anaerobic biodegradability of 70%. Furthermore, the results of the UASB reactor demonstrated that a total COD removal of 52-64% was obtained at HRT between 6 and 16 h. The UASB reactor removed 22-30% and 15-21% of total nitrogen and total phosphorous in the grey water, respectively, mainly due to the removal of particulate nutrients. The characteristics of the sludge in the UASB reactor confirmed that the reactor had a stable performance. The minimum sludge residence time and the maximum specific methanogenic activity of the sludge ranged between 27 and 93 days and 0.18 and 0.28 kg COD/(kg VS d).     

Electricity assisted anaerobic treatment of salinity wastewater and its effects on microbial communities

High salinity wastewater is often difficult to treat using common anaerobic technologies. Considering that high conductivity of salinity wastewater may enhance electrodes reaction to accelerate the decomposition of volatile fatty acids produced in anaerobic digestion, a pair of electrodes was packed into an anaerobic reactor (R1) with the aim to enhance the treatment of salinity wastewater. With increasing the salt concentration (NaCl) gradually from 0 to 50 g/L in 137 days' operation, COD removal in this reactor under the voltage for the electrodes of 1.2 V was well maintained at 93%, while the COD removal in a reference anaerobic reactor without electrodes (R2) decreased to 53%. When the voltage for R1 was cut off, about 10% COD removal was declined, which was still 30 percentage points higher than that in R2. The electrodes enhanced the biodegradation of volatile fatty acids, especially propionate. Fluorescence in situ hybridization analysis confirmed that the relative abundance of propionate-utilizing bacteria in R1 was significantly higher than that in R2. PCR-DGGE analysis of bacteria and archaea domains indicated that the electric field stimulation effectively enriched salt-adapted microorganisms during the treatment.     

Sludge properties and their effects on membrane fouling in submerged anaerobic membrane bioreactors (SAnMBRs)

Two submerged anaerobic membrane bioreactors (SAnMBRs) (thermophilic vs. mesophilic) were operated for a period of 3.5 months with kraft evaporator condensate at a feed chemical oxygen demand of 10,000 mg/L. The results show that the filtration behavior of the two systems was significantly different. The filtration resistance in the thermophilic SAnMBR was about 5–10 times higher than that of the mesophilic system when operated under similar hydrodynamic conditions. Comparison of sludge properties and cake layer structure from the two systems was made to elucidate major factors governing the different filtration characteristics. There were more soluble microbial products (SMP) and biopolymer clusters (BPC) produced and a larger portion of fine flocs (<15 μm) in the thermophilic SAnMBR. Analysis of bound extracellular polymeric substances (EPS) showed that the thermophilic sludge had a higher protein/polysaccharide ratio in EPS, as compared to that in the mesophilic sludge. A series of analyses, including Fourier transform infrared (FTIR) spectroscopy, energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), atomic force microscopy (AFM) and particle size analyzer showed that the cake layer formed in the thermophilic SAnMBR contained higher levels of both organic and inorganic foulants, smaller particle sizes, and especially, a denser and more compact sludge cake structure. These results indicate that floc size, SMP, BPC, bound EPS as well as cake layer structure are the major factors governing membrane fouling in SAnMBR systems.     

A comprehensive microbial insight into two-stage anaerobic digestion of food waste-recycling wastewater

Microbial community structures were assessed in a two-stage anaerobic digestion system treating food waste-recycling wastewater. The reactors were operated for 390 d at 10 different hydraulic retention times (HRTs) ranging from 25 to 4 d. Stable operation was achieved with the overall chemical oxygen demand (COD) removal efficiency of 73.0-85.9% at organic loading rate of up to 35.6 g COD/L·d. Performance of the acidogenic reactors, however, changed significantly during operation. This change coincided with transition of the bacterial community from one dominated by Aeriscardovia- and Lactobacillus amylovorus-related species to one dominated by Lactobacillus acetotolerans- and Lactobacillus kefiri-like organisms. In methanogenic reactors, the microbial community structures also changed at this stage along with the shift from Methanoculleus- to Methanosarcina-like organisms. This trend was confirmed by the non-metric multidimensional scaling joint plot of microbial shifts along with performance parameters. These results indicated that the overall process performance was relatively stable compared to the dynamic changes in the microbial structures and the acidogenic performance.     

Nylon fibers as supporting media in anaerobic hybrid reactors: it's effects on system's performance and microbial distribution

The performances of three anaerobic hybrid reactors with various nylon fiber densities per packed bed volume (33, 22, and 11 kg/m(3) in R1, R2, and R3, respectively) as supporting media were evaluated through their ability to remove organic compounds in cassava starch wastewater. In addition, the distributions of non-methanogenic and methanogenic population in the reactors were investigated. During a 6-month operation, the organic loading rate was increased in stepwise from 0.5 to 4.0 kg COD/m3/day and the hydraulic retention time (HRT) shortened to 5.4 days. The COD removal efficiency was more favorable in R1 (87%) and R2 (84%) than in R3 (70%). The total biomass in the reactors with greater nylon fiber densities was also higher and increased from 20.4 to 67.3 g VSS and to 57.5 g VSS in R1 and R2, respectively. When the HRT was further shortened to 3 days, however, the efficiency of both reactors demonstrated a declining trend and reached 74% in R1 and 61% in R2. The distribution of microbial populations involved in the reactors was determined using the Most Probable Number technique. The result showed the lowest number of methanogens in R3 which correlated well to its relatively low efficiency. The number of non-methanogens in all reactors was, nonetheless, comparable. By shortening the HRT to 3 days, the methanogenic population in R2 diminished in both attached and suspended biomass whereas a slight reduction was detected only in the attached biomass of R1.     

Integrated application of upflow anaerobic sludge blanket reactor for the treatment of wastewaters

The UASB process among other treatment methods has been recognized as a core method of an advanced technology for environmental protection. This paper highlights the treatment of seven types of wastewaters i.e. palm oil mill effluent (POME), distillery wastewater, slaughterhouse wastewater, piggery wastewater, dairy wastewater, fishery wastewater and municipal wastewater (black and gray) by UASB process. The purpose of this study is to explore the pollution load of these wastewaters and their treatment potential use in upflow anaerobic sludge blanket process. The general characterization of wastewater, treatment in UASB reactor with operational parameters and reactor performance in terms of COD removal and biogas production are thoroughly discussed in the paper. The concrete data illustrates the reactor configuration, thus giving maximum awareness about upflow anaerobic sludge blanket reactor for further research. The future aspects for research needs are also outlined.     

Characterization of dairy waste streams, current treatment practices, and potential for biological nutrient removal

Fifteen milk processing plants in the Upper Midwest of the United States participated in a study to obtain information on general process operation, waste generation and treatment practices, chemical usage, and wastewater characteristics. Long term data on wastewater characteristics were obtained for 8 of the 15 dairy plants, and a 24-h composite wastewater sample was characterized in detail for each plant. Wastewater flow rates and characteristics varied greatly among and within plants and were not easily predictable even when detailed information on processing operations was available. In addition, the contribution of milk and milk products to the waste streams was underestimated by plant operators. The use of caustic soda, phosphoric acid, and nitric acid for cleaning had a significant impact on wastewater characteristics, despite the implementation of changes in chemical usage practices during recent years. In particular, the use of phosphoric acid based cleaning products has been reduced to eliminate or decrease discharge fines. It was determined that most of the on site treatment facilities require renovations and/or operational changes to comply with current and future discharge regulations, especially with respect to nutrient (nitrogen and phosphorus) levels in their waste streams. It was concluded that biological nutrient removal of dairy wastewaters should be feasible given the relatively high concentrations of easily degradable organics, the generally favorable organic matter to total phosphorus ratio, and the very favorable organic matter to nitrogen ratio.     

Effect of polyelectrolytes and quaternary ammonium compounds on the anaerobic biological treatment of poultry processing wastewater

Quaternary ammonium compounds (QACs) and polyelectrolytes are extensively used in poultry processing facilities as sanitizing agents and flocculants, respectively. These chemicals may affect the performance of biological treatment systems resulting in low effluent quality. The impact of these chemicals on the anaerobic treatment of poultry processing wastewater (PPWW) samples, collected before and after a solids separation process, was tested in batch assays using a mixed, mesophilic (35 degrees C) methanogenic culture. The results of this study showed that Vigilquat (VQ), a commercial mixture of four QACs, has a high affinity for the organic solids in the PPWW. Cationic and anionic polyelectrolytes, alone or in combination, did not have any adverse effect on the anaerobic biodegradation of PPWW at concentrations typically used in poultry processing facilities (20 and 5 mg/L, respectively). In spite of the high affinity of VQ for the PPWW solids, VQ at a concentration of 50mg/L and above adversely affected the anaerobic degradation of the PPWW, which resulted in a significantly reduced methane production and accumulation of volatile fatty acids. In the absence of any inhibition, the methane yield varied from 0.76 to 0.98 L methane at STP per g volatile solids added. VQ was not biodegraded under the batch, methanogenic conditions used in this study.     

Quantitative and qualitative analyses of methanogenic community development in high-rate anaerobic bioreactors

Methanogenic community structure and population dynamics were investigated in two anaerobic reactors treating a dairy wastewater, an Inverted Fluidized Bed (IFB) and Expanded Granular Sludge Bed (EGSB). A combination of real-time PCR, denaturing gradient gel electrophoresis and statistical techniques was employed. Distinct methanogenic communities developed in the IFB and EGSB reactors reflecting step-wise reductions in the applied hydraulic retention time from 72 to 12 h during the 200-day trial. The aceticlastic family Methanosarcinaceae was only detected in the IFB and the order Methanomicrobiales was also much more abundant in this reactor, while the aceticlastic family Methanosaetaceae was more abundant in the EGSB. The hydrogenotrophic order, Methanobacteriales, predominated in both reactors under all applied operational conditions. Non-metric multidimensional scaling (NMS) and moving-window analyses, based on absolute and relative abundance quantification data, demonstrated that the methanogenic communities developed in a different manner in the IFB, compared to the EGSB reactor. In our study, relative abundance-based quantification by NMS and moving-window analysis appeared to be a valuable molecular approach that was more applicable to reflect the changes in the anaerobic digestion process than approaches based either on qualitative analysis, or solely on absolute quantification of the various methanogenic groups. The overall results and findings provided a comparative, quantitative and qualitative insight into anaerobic digestion processes, which could be helpful for better future reactor design and process control.     

Identification of biofoulant of membrane bioreactors in soluble microbial products

To reveal primary biofoulant in soluble microbial products (SMP) and/or soluble extracellular polymeric substances (EPS), after removal of sludge particles, activated sludge samples were subjected to microfiltration tests in a submerged MBR. Filtration resistance directly correlates with the saccharide concentration. Saccharides in wastewater from several sources contained uronic acids, which increased the filtration resistance. When the microfiltration test liquids contained saccharides over 80 mg l⁻¹, a gelatinous mass remained on the membrane surface after filtration and contained concentrations of saccharides and uronic acids 50 times higher than the original test liquid while only trace amounts of these substances were contained in the filtrate. The gelatinous mass contained high molecular weight substances of 10⁶-10⁸ Da, suggesting the presence of polysaccharides. However, molecules of this size were calculated to be much smaller than the pore size of the membrane. Ethylenediaminetetraacetic acid decreased filtration resistance, suggesting that polysaccharides containing uronic acid units could undergo intermolecular or intramolecular ionic cross-linking by polyvalent cations and form the gel, thus clogging the membrane pores as an actual biofoulant.     

The performance enhancements of upflow anaerobic sludge blanket (UASB) reactors for domestic sludge treatment--a state-of-the-art review

Nowadays, carbon emission and therefore carbon footprint of water utilities is an important issue. In this respect, we should consider the opportunities to reduce carbon footprint for small and large wastewater treatment plants. The use of anaerobic rather than aerobic treatment processes would achieve this aim because no aeration is required and the generation of methane can be used within the plant. High-rate anaerobic digesters receive great interests due to their high loading capacity and low sludge production. Among them, the upflow anaerobic sludge blanket (UASB) reactors have been most widely used. However, there are still unresolved issues inhibiting the widespread of this technology in developing countries or countries with climate temperature fluctuations (such as subtropical regions). A large number of studies have been carried out in order to enhance the performance of UASB reactors but there is a lack of updated documentation. In face of the existing limitations and the increasing importance of this technology, the authors present an up-to-date review on the performance enhancements of UASB reactors over the last decade. The important aspects of this article are: (i) enhancing the start-up and granulation in UASB reactors, (ii) coupling with post-treatment unit to overcome the temperature constraint, and (iii) improving the removal efficiencies of the organic matter, nutrients and pathogens in the final effluent. Finally the authors have highlighted future research direction based on their critical analysis.     

Submerged anaerobic membrane bioreactor for low-strength wastewater treatment: Effect of HRT and SRT on treatment performance and membrane fouling

Three 6-L submerged anaerobic membrane bioreactors (SAnMBRs) with solids retention times (SRTs) of 30, 60 and infinite days were setup for treating synthetic low-strength wastewater at hydraulic retention times (HRTs) of 12, 10 and 8 h. Total COD removal efficiencies higher than 97% were achieved at all operating conditions. Maximum biogas production rate was 0.056 L CH₄/g MLVSS d at an infinite SRT. A shorter HRT or longer SRT increased biogas production due to increased organic loading rate or enhanced dominancy of methanogenics. A decrease in HRT enhanced growth of biomass and accumulation of soluble microbial products (SMP), which accelerated membrane fouling. A drop in carbohydrate to protein ratio also inversely affected fouling. At 12-h HRT, the effect of SRT on biomass concentration in SAnMBRs was negligible and membrane fouling was controlled by variant surface modification due to different SMP compositions, i.e., higher carbohydrate and protein concentrations in SMP at longer SRT resulted in higher membrane fouling rate. At 8 and 10-h HRTs, infinite SRT in SAnMBR caused highest MLSS and SMP concentrations, which sped up particle deposition and biocake/biofilm development. At longer SRT, lower extracellular polymeric substances reduced flocculation of particulates and particle sizes, further aggravated membrane fouling.     

Long-term effect of ZnO nanoparticles on waste activated sludge anaerobic digestion

The increasing use of zinc oxide nanoparticles (ZnO NPs) raises concerns about their environmental impacts, but the potential effect of ZnO NPs on sludge anaerobic digestion remains unknown. In this paper, long-term exposure experiments were carried out to investigate the influence of ZnO NPs on methane production during waste activated sludge (WAS) anaerobic digestion. The presence of 1 mg/g-TSS of ZnO NPs did not affect methane production, but 30 and 150 mg/g-TSS of ZnO NPs induced 18.3% and 75.1% of inhibition respectively, which showed that the impact of ZnO NPs on methane production was dosage dependant. Then, the mechanisms of ZnO NPs affecting sludge anaerobic digestion were investigated. It was found that the toxic effect of ZnO NPs on methane production was mainly due to the release of Zn²⁺ from ZnO NPs, which may cause the inhibitory effects on the hydrolysis and methanation steps of sludge anaerobic digestion. Further investigations with enzyme and fluorescence in situ hybridization (FISH) assays indicated that higher concentration of ZnO NPs decreased the activities of protease and coenzyme F₄₂₀, and the abundance of methanogenesis Archaea.     

Quantification and molecular characterization of enteric viruses detected in effluents from two hospital wastewater treatment plants

Hospital wastewater has been described as an important source of spreading pathogenic microorganisms in the environment. However, there are few studies reporting the presence and concentrations of gastroenteric viruses and hepatitis A viruses in these environmental matrices. The aim of this study was to assess the contamination by viruses responsible for acute gastroenteritis and hepatitis derived from hospital wastewater treatment plants (WWTPs). Rotavirus A (RV-A), human adenoviruses (HAdV), norovirus genogroup I and II (NoV GI/GII) and hepatitis A viruses (HAV) were detected and quantified in sewage samples from two WWTPs located in Rio de Janeiro (Brazil) that operates different sewage treatments. WWTP-1 uses an Upflow Anaerobic Sludge Blanket (UASB reactor) and three serial anaerobic filters while WWTP-2 uses aerobic processes, activated sludge with extended aeration and final chlorination of the effluents. Viruses’ detection was investigated by using conventional PCR/RT-PCR, quantitative real-time PCR (qPCR) and partial sequencing of the genome of the viruses detected. Rate of viruses detection ranged from 7% (NoV GI in WWTP-1) to 95% (RV-A in WWTP-2) and genome from all viruses were detected. The most prevalent genotypes were RV-A SG I, HAdV species D and F, NoV GII/4 and HAV subgenotype IA. Mean values of viral loads (genome copies (GC)/ml) obtained in filtered effluents from anaerobic process was 1.9 × 10³ (RV-A), 2.8 × 10³ (HAdV) and 2.4 × 10³ (NoV GII). For chlorinated effluents from activated sludge process, the mean values of viral loads (GC/ml) was 1.2 × 10⁵ (RV-A), 1.4 × 10³ (HAdV), 8.1 × 10² (NoV GII) and 2.8 × 10⁴ (HAV). Data on viral detection in treated effluents of hospital WWTPs confirmed the potential for environmental contamination by viruses and could be useful to establish standards for policies on wastewater management.     

Sequential treatment of PTA wastewater in a two-stage UASB process: focusing on p-toluate degradation and microbial distribution

Two-stage upflow anaerobic sludge blanket (UASB) process was investigated as an efficient process configuration option for the treatment of purified terephthalic acid (PTA) wastewater. To study its feasibility in a defined condition, synthetic wastewater containing only the major pollutants (i.e., acetate, benzoate, terephthalate and p-toluate) was used. By focusing the role of the second stage on the p-toluate degradation, improved overall COD and p-toluate removal capacities of 4.18 and 1.35 g-thCOD/L·day could be achieved together with a complete COD removal efficiency. In this situation, all the pollutants except p-toluate were completely degraded in the first stage while 38 and 62% of p-toluate originally present in the wastewater were consecutively degraded in the individual stages. The concomitant methane production rate in each stage was 0.91 and 0.35 L/L·day respectively, and the methane yield on p-toluate was determined to be 0.12 L/g-thCOD. Batch tests using the granules obtained from each stage revealed that the acidogenic microorganisms enriched in both stages had a universal ability to degrade all aromatic pollutants present in the PTA wastewater. Moreover, image analysis using scanning electron microscope and confocal laser scanning microscopy combined with fluorescence in situ hybridization technique elucidated that the distribution of acidogens and methanogens within the granule was varied in each stage, which influenced the mass transfer regime resulting in the different pollutant degradation rates during the batch tests.