Anaerobic biological treatment of phenol at 9.5-15 degrees C in an expanded granular sludge bed (EGSB)-based bioreactor

The aims of this study were to demonstrate the (1) feasibility of psychrophilic, or low-temperature, anaerobic digestion (PAD) of phenolic wastewaters at 10–15 °C; (2) economic attractiveness of PAD for the treatment of phenol as measured by daily biogas yields and (3) impact on bioreactor performance of phenol loading rates (PLRs) in excess of those previously documented (1.2 kg phenol m⁻³ d⁻¹). Two expanded granular sludge bed (EGSB)-based bioreactors, R1 and R2, were employed to mineralise a volatile fatty acid-based wastewater. R2 influent wastewater was supplemented with phenol at an initial concentration of 500 mg l⁻¹ (PLR, 1 kg m⁻³ d⁻¹). Reactor performance was measured by chemical oxygen demand (COD) removal efficiency, CH₄ composition of biogas and phenol removal (R2 only). Specific methanogenic activity, biodegradability and toxicity assays were employed to monitor the physiological capacity of reactor biomass samples. The applied PLR was increased to 2 kg m⁻³ d⁻¹ on day 147 and phenol removal by day 415 was 99% efficient, with 4 mg l⁻¹ present in R2 effluent. The operational temperature of R1 (control) and R2 was reduced by stepwise decrements from 15 °C through to a final operating temperature of 9.5 °C. COD removal efficiencies of c. 90% were recorded in both bioreactors at the conclusion of the trial (day 673), when the phenol concentration in R2 effluent was below 30 mg l⁻¹. Daily biogas yields were determined during the final (9.5 °C) operating period, when typical daily R2 CH₄ yields of c. 3.3 l CH₄ g⁻¹ COD_removed d⁻¹ were recorded. The rate of phenol depletion and methanation by R2 biomass by day 673 were 68 mg phenol g VSS⁻¹ d⁻¹ and 12–20 ml CH₄ g VSS⁻¹ d⁻¹, respectively.     

Long-term (1243 days), low-temperature (4-15 °C), anaerobic biotreatment of acidified wastewaters: Bioprocess performance and physiological characteristics

The feasibility of long-term (>3 years), low-temperature (4-15 °C) and anaerobic bioreactor operation, for the treatment of acidified wastewater, was investigated. A hybrid, expanded granular sludge bed-anaerobic filter bioreactor was seeded with a mesophilic inoculum and employed for the mineralization of moderate-strength (3.75-10 kg chemical oxygen demand (COD) m⁻³) volatile fatty acid-based wastewaters at 4-15 °C. Bioprocess performance was assessed in terms of COD removal efficiency (CODRE), methane biogas concentration, and yield, and biomass retention. Batch specific methanogenic activity assays were performed to physiologically characterise reactor biomass.Despite transient disimprovements, CODRE and methane biogas concentrations exceeded 80% and 65%, respectively, at an applied organic loading rate (OLR) of 10 kg COD m⁻³ d⁻¹ between 9.5 and 15 °C (sludge loading rate (SLR), 0.6 kg COD kg[VSS]⁻¹ d⁻¹). Over 50% of the granular sludge bed was lost to disintegration during operation at 9.5 °C, warranting a reduction in the applied OLR to 3.75-5 kg COD m⁻³ d⁻¹ (SLR, c. 0.4-0.5 kg COD kg[VSS]⁻¹ d⁻¹). From that point forward, remarkably stable and efficient performance was observed during operation at 4-10 °C, with respect to CODRE (≥82%), methane biogas concentration (>70%) and methane yields (>4 l_Methane d⁻¹), suggesting the adaptation of our mesophilic inoculum to psychrophilic operating conditions.Physiological activity assays indicated the development of psychroactive syntrophic and methanogenic populations, including the emergence of putatively psychrophilic propionate-oxidising and hydrogenotrophic methanogenic activity. The data suggest that mesophilic inocula can physiologically adapt to sub-optimal operational temperatures: treatment efficiencies and sludge loading rates at 4 °C (day, 1243) were comparable to those achieved at 15 °C (day 0). Furthermore, long-term, low-temperature bioreactor operation may act as a selective enrichment for psychrophilic methanogenic activity from mesophilic inocula. The observed efficient and stable bioprocess performance highlights the potential for long-term, low-temperature bioreactor operation.     

Low-temperature anaerobic biological treatment of toluene-containing wastewater

Two expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactors, R1 and R2, were operated at 15 degrees C for the treatment of toluene-contaminated volatile fatty acid-based wastewater. The seed inoculum and the R1 reactor were unexposed to toluene, prior to and during the trial, respectively. Both reactors were operated at a hydraulic retention time of 24h at applied organic loading rates of 0.71-1.43kg chemical oxygen demand (COD)m(-3)d(-1). Toluene was supplemented to the R2 influent at concentrations of 5-104 mg toluenel(-1) (solubilised in ethanol). Bioreactor performance was evaluated by COD and toluene removal efficiency, and the methane content of biogas (%). Specific methanogenic activity and toxicity assays were employed to investigate the activity and toluene toxicity thresholds of key trophic groups, respectively, within the seed and reactor biomass samples. COD and toluene removal efficiencies of 70-90% and 55-99%, respectively, were achieved during the 630-d trial. Metabolic assays suggested that a psychrotolerant H(2)/CO(2)-utilizing methanogenic community developed in the toluene-degrading biomass. The results indicate the viability of low-temperature anaerobic digestion for the treatment of wastewater containing toluene.     

Low-temperature anaerobic biological treatment of solvent-containing pharmaceutical wastewater

Low-temperature or psychrophilic (<20 degrees C) anaerobic digestion (PAD) has recently been demonstrated as a cost-effective option for the treatment of a range of wastewater categories. The aim of this work was 2-fold: (1) to screen three anaerobic sludges, obtained from full-scale reactors, with respect to suitability for PAD of pharmaceutical-like, solvent-contaminated wastewater; (2) to assess the feasibility of PAD of this wastewater category. Toxicity thresholds of key trophic groups within three candidate biomass samples were assessed against solvents prevalent in pharmaceutical wastewaters (propanol, methanol and acetone). Specific methanogenic activity (SMA) assays indicated that the metabolic optimum of each candidate biomass was within the mesophilic range. One biomass sample exhibited higher SMA assays than the other candidate samples and was also the sample least methanogenically inhibited by the addition of solvents to batch cultures. This sludge was selected as the biomass of choice for laboratory-scale trials. Two identical expanded granular sludge bed (EGSB)-based anaerobic reactors were used for the treatment of solvent-contaminated wastewater at 15 degrees C, and at applied organic loading rates (OLRs) of 5-20 kg chemical oxygen demand (COD) m(-3)d(-1). COD removal efficiencies of 60-70% were achieved during the 450 day trial. In addition, SMA assays carried out at the conclusion of the trial indicated the development of a putatively psychrophilic hydrogenotrophic methanogenic community.     

Psychrophilic and mesophilic anaerobic digestion of brewery effluent: a comparative study

Two expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactors (3.38 l active volume) were used to directly compare psychrophilic (15 degrees C), anaerobic digestion (PAD) to mesophilic (37 degrees C) anaerobic digestion (MAD) for the treatment of a brewery wastewater (chemical oxygen demand (COD) concentration of 3,136+/-891 mg l(-1)). Bioreactor performance was evaluated by COD removal efficiency and biogas yields at a range of hydraulic and organic loading rates. Specific methanogenic activity (SMA) assays were also employed to investigate the activity of the biomass in the bioreactors. No significant difference in the COD removal efficiencies (which ranged from 85-93%) were recorded between PAD and MAD during the 194-d trial at maximum organic and hydraulic loading rates of 4.47 kg m(-3) day(-1) and 1.33 m(3) m(-3) day(-1), respectively. In addition, the methane content (%) of the biogas was very similar. The volumetric biogas yield from the PAD bioreactor was approximately 50% of that from the MAD bioreactor at an organic loading rate of 4.47 kg COD m(-3) day(-3) and an applied liquid up-flow velocity (V(up)) of 2.5 m h(-1). Increasing the V(up) in the PAD bioreactor to 5 m h(-1) resulted in a volumetric biogas production rate of approximately 4.1 l d(-1) and a methane yield of 0.28 l CH(4) g(-1) COD d(-1), which were very similar to the MAD bioreactor. Significant and negligible biomass washout was observed in the mesophilic and psychrophilic systems, respectively, thus increasing the sludge loading rate applied to the former and underlining the robustness of the latter, which appeared underloaded. A psychrotolerant mesophilic, but not truly psychrophilic, biomass developed in the PAD bioreactor biomass, with comparable maximum SMA values to the MAD bioreactor biomass. PAD, therefore, was shown to be favourably comparable to MAD for brewery wastewater treatment and biogas generation.     

Methanogenic community development in anaerobic granular bioreactors treating trichloroethylene (TCE)-contaminated wastewater at 37 °C and 15 °C

Four expanded granular sludge bed (EGSB) bioreactors were seeded with a mesophilically-grown granular sludge and operated in duplicate for mesophilic (37 °C; R1 & R2) and low- (15°; R3 & R4) temperature treatment of a synthetic volatile fatty acid (VFA) based wastewater (3 kg COD m⁻³ d⁻¹) with one of each pair (R1 & R3) supplemented with increasing concentrations of trichloroethylene (TCE; 10, 20, 40, 60 mg l⁻¹) and one acting as a control. Bioreactor performance was evaluated by % COD removal efficiency and % biogas methane (CH₄) content. Quantitative Polymerase Chain Reaction (qPCR) was used to investigate the methanogenic community composition and dynamics in the bioreactors during the trial, while specific methanogenic activity (SMA) and toxicity assays were utilized to investigate the activity and TCE/dichloroethylene (DCE) toxicity thresholds of key trophic groups, respectively. At both 37 °C and 15 °C, TCE levels of 60 mg l⁻¹ resulted in the decline of % COD removal efficiencies to 29% (Day 235) and 37% (Day 238), respectively, and in % biogas CH₄ to 54% (Day 235) and 5% (Day 238), respectively. Despite the inhibitory effect of TCE on the anaerobic digestion process, the main drivers influencing methanogenic community development, as determined by qPCR and Non-metric multidimensional scaling analysis, were (i) wastewater composition and (ii) operating temperature. At the apical TCE concentration both SMA and qPCR of methanogenic archaea suggested that acetoclastic methanogens were somewhat inhibited by the presence of TCE and/or its degradation derivatives, while competition by dechlorinating organisms may have limited the availability of H₂ for hydrogenotrophic methanogenesis. In addition, there appeared to be an inverse correlation between SMA levels and TCE tolerance, a finding that was supported by the analysis of the inhibitory effect of TCE on two additional biomass sources. The results indicate that low-temperature anaerobic digestion is a feasible approach for the treatment of TCE-containing wastewater.     

Low-temperature (7 °C) anaerobic treatment of a trichloroethylene-contaminated wastewater: microbial community development

The feasibility of low-temperature (7 °C) anaerobic digestion for the treatment of a trichloroethylene (TCE) contaminated wastewater was investigated. Two expanded granular sludge bed (EGSB) bioreactors (R1 and R2) were employed for the mineralisation of a synthetic volatile fatty acid based wastewater at an initial organic loading rate (OLR) of 3 kg COD m⁻³ d⁻¹, and an operating temperature of 15 °C. Successive reductions in OLR to 0.75 kg COD m⁻³ d⁻¹, and operational temperature to 7 °C, resulted in stable bioreactor operation by day 417, with COD removal efficiency and biogas CH₄ content ≥74%, for both bioreactors. Subsequently, the influent to R1 was supplemented with increasing concentrations (10, 20, 30 mg l⁻¹) of TCE, while R2 acted as a control. At an influent TCE concentration of 30 mg l⁻¹, although phase average TCE removal rates of 79% were recorded, a sustained decrease in R1 performance was observed, with COD removal of 6%, and % biogas CH₄ of 3% recorded on days 595 and 607, respectively. Specific methanogenic activity (SMA) assays identified a general shift from acetate- to hydrogen-mediated methanogenesis in both R1 and R2 biomass, while toxicity assays confirmed an increased sensitivity of the acetoclastic community in R1 to TCE and dichloroethylene (DCE), which contributed to acetate accumulation. Quantitative Polymerase Chain Reaction (qPCR) analysis of the methanogenic community confirmed the dominance of hydrogenotrophic methanogens in both R1 and R2, representing 71-89% of the total methanogenic population, however acetoclastic Methanosaeta were the dominant organisms, based on 16S rRNA gene clone library analysis of reactor biomass. The greatest change in the bacterial community, as demonstrated by UPGMA analysis of DGGE banding profiles, was observed in R1 biomass between days 417 and 609, although 88% similarity was retained between these sampling points.     

Development of microbial community structure and actvity in a high-rate anaerobic bioreactor at 18 degrees C

Anaerobic digestion in the psychrophilic (< 20 degrees C) or sub-mesophilic temperature range has recently been proven as an effective treatment option for the mineralization of a wide variety of problematic wastewaters. In this study, an expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactor was seeded with a full-scale, mesophilic sludge and employed to evaluate the long-term operational potential, and underlying microbial ecology, of this approach for the treatment of a medium-strength (5 g chemical oxygen demand [COD] l(-1)), synthetic, volatile fatty acid-based wastewater. Throughout the trial period of 625 days, extended intervals of consistently stable and efficient wastewater treatment were sustained. These results were highlighted by a short start-up period (21 d), low hydraulic retention times (4.88h), high organic (up to 24.64kg CODm(-3)d(-1)), and volumetric loading rates (up to 4.92 m3 m(-3) d(-1)). A stable, well-settling granular sludge bed was maintained in the bioreactor for the majority of the trial; however, reduced treatment efficiency and biomass washout were observed at an imposed OLR of 36.96 kg COD m(-3) d(-1). The microbial biomass in the bioreactor was investigated using maximum specific methanogenic activity assays and polymerase chain reaction-denaturing gradient gel electrophoresis. A temporal succession of both the bacterial and archaeal populations was noted during the trial, compared to the seed sludge, in response to bioreactor operation at lower temperatures, loading rate increases and to VFA accumulation in the bioreactor. During the trial, an increased contribution of hydrogenotrophic methanogenesis as a pathway of methane production was observed, along with the overall emergence of a highly active psychrotolerent-though still mesophilic biomass.     

中温两相厌氧消化工艺处理混合污泥的效能

采用中温(35℃)两相厌氧消化工艺处理初沉污泥与剩余污泥的混合样(1∶1),以实现污泥的稳定化。结果表明,在水力停留时间约为10 d、有机负荷为2.75 kgVS/(m3.d)时,对TCOD的去除率可达46%,对VS的去除率为41%;产甲烷相反应器的最大比产甲烷活性为0.19LCH4/(gVS.d),并保持相对稳定;两相反应器内污泥的比脱氢酶活性都出现了增长,并维持在25~32μgINTF/(mgVS.h)之间;经消化后污泥的沉降性和脱水性变差。     

Anaerobic treatment of municipal wastewater at ambient temperature: Analysis of archaeal community structure and recovery of dissolved methane

Anaerobic treatment is an attractive option for the biological treatment of municipal wastewater. In this study, municipal wastewater was anaerobically treated with a bench-scale upflow anaerobic sludge blanket (UASB) reactor at temperatures from 6 to 31 °C for 18 months to investigate total chemical oxygen demand (COD) removal efficiency, archaeal community structure, and dissolved methane (D-CH₄) recovery efficiency. The COD removal efficiency was more than 50% in summer and below 40% in winter with no evolution of biogas. Analysis of the archaeal community structures of the granular sludge from the UASB using 16S rRNA gene-cloning indicated that after microorganisms had adapted to low temperatures, the archaeal community had a lower diversity and the relative abundance of acetoclastic methanogens decreased together with an increase in hydrogenotrophic methanogens. D-CH₄, which was detected in the UASB effluent throughout the operation, could be collected with a degassing membrane. The ratio of the collection to recovery rates was 60% in summer and 100% in winter. For anaerobic treatment of municipal wastewater at lower temperatures, hydrogenotrophic methanogens play an important role in COD removal and D-CH₄ can be collected to reduce greenhouse gas emissions and avoid wastage of energy resources.     

Anaerobic treatment of phenolic coal conversion wastewater in semicontinuous cultures

High strength wastewater (7600 mg 1⁻¹ phenolics) from the H-coal liquefaction process was diluted and fed to anaerobic, methane-producing cultures. Total phenolic concentrations of 150 and 300 mg 1⁻¹ were added to 50 ml semicontinuous cultures with hydraulic retention times of 12.5, 16.7 and 25 days. The rates of methane production and effluent concentrations of three fermentable phenolics (phenol, p-cresol and m-cresol) were monitored over a 188-day period. After acclimation to the wastewater, stable periods followed during which each of the six cultures removed essentially all of these fermentable phenolics. The duration of the stable periods decreased with increasing phenolic mass loading rates. m-Cresol was the first phenolic to appear in the effluent and its presence served as the first indicator of reduced phenolic removal capability. The effluent m-cresol concentrations from cultures receiving 300 mg 1⁻¹ total phenolics followed simple washout curves suggesting that its degradation stopped abruptly. Later, p-cresol and ultimately phenol appeared in the effluents from the cultures which received the highest phenolic mass loadings.     

聚季胺物对厌氧污泥特性的影响研究

通过对聚季胺物的生物化学甲烷势(BMP)和厌氧微生物在不同聚季胺物浓度下的毒性测定(ATA)，评价了聚季胺物的厌氧生物可降解程度，对比了不同聚季胺物浓度对厌氧微生物的比产甲烷活性(SMA)和沉降性能(SVI)的影响。试验结果表明：聚季胺物适宜作为厌氧微生物自身固定化的促进剂，建议采用间隔9～10d的多次投加方式，厌氧反应器中聚季胺物浓度为10～20mg／L，聚季胺物与厌氧接种污泥的质量比≤3．2。     

Anaerobic digestion of wine distillery wastewater in down-flow fluidized bed

In down-flow fluidization, particles with a specific density smaller than the liquid are fluidized downward by a concurrent flow of liquid. This paper describes the application of the down-flow (or inverse) fluidization technology for the anaerobic digestion of red wine distillery wastewater. The carrier employed was ground perlite, an expanded volcanic rock. Before starting-up the reactor, physical and fluidization properties of the carrier material were determined. 0.968 mm perlite particles were found to have a specific density of 280 kg m⁻³ and a minimum fluidization velocity of 2.3 m h⁻¹. Once the down-flow anaerobic fluidized bed system reached the steady-state, organic load was increased stepwise by reducing HRT, from 3.3–1.3 days, while maintaining constant the feed TOC concentration. The system achieved 85% TOC removal, at an organic loading rate of 4.5 kg TOC m³ d⁻¹. It was found that the main advantages of this system are: low energy requirement, because of the low fluidization velocities required; there is no need of a settling device, because solids accumulate at the bottom of the reactor so they can be easily drawn out, and particles with high-biomass content, whose specific density have become larger than 1000 kg m⁻³ can be easily recovered.     

Anoxic treatment of phenolic wastewater in sequencing batch reactor

Studies were conducted on the anoxic phenol removal using granular denitrifying sludge in sequencing batch reactor at different cycle lengths and influent phenol concentrations. Results showed that removal exceeded 80% up to an influent phenol concentration of 1050 mg/l at 6 h cycle length, which corresponded to 6.4 kg COD/m3/d. Beyond this, there was a steep decrease in phenol and COD removal efficiencies. This was accompanied by an increase in nitrite concentration in the effluent. On an average, 1 g nitrate-N was consumed per 3.4 g phenol COD removal. Fraction of COD available for sludge growth was calculated to be 11%.     

Anaerobic treatment of phenol in wastewater under thermophilic condition

Over 99% of phenol was effectively degraded in an upflow anaerobic sludge blanket (UASB) reactor at 55 degrees C with 40 h of hydraulic retention time (HRT) for a wastewater containing 630 mg/L of phenol, corresponding to 1500 mg/L of chemical oxygen demand (COD) and a loading rate of 0.9 g-COD/L/d. The maximum specific methanogenic activity (SMA) of the phenol-degrading sludge was 0.09 g-CH4-COD/g-volatile suspended solids (VSS)/d. Based on 16S rDNA analysis, a total of 21 operational taxonomy units (OTUs) were found in the sludge, of which eight (42.6% of the total population) were related to the sequences in the GenBank with similarity of over 97%, and 13 (79.6%) were affiliated with the known thermophilic species. Additional SMA data and phylogenetic analysis suggest that the degradation pathway of phenol for thermophilic sludge was likely via caproate, instead of benzoate as for the mesophilic sludge.     

Continuous electrochemical treatment of phenolic wastewater in a tubular reactor

The electrochemical treatment of phenolic wastewater in a continuous tubular reactor, constructed from a stainless steel tube with a cylindrical carbon anode at the centre, was investigated in this study, being first in literature. The effects of residence time on phenol removal was studied at 25 degrees C, 120 g l(-1) electrolyte concentration for 450 and 3100 mg l(-1) phenol feed concentrations with 61.4 and 54.7 mA cm(-2) current densities, respectively. The change in phenol concentration and pH of the reaction medium was monitored in every run and GC/MS analyses were performed to determine the fate of intermediate products formed during the electrochemical reaction in a specified batch run. During the electrolysis mono, di- and tri-substituted chlorinated phenol products were initially formed and consumed along with phenol thereafter mainly by polymerization mechanism. For 10 and 20 min of residence time phenol removal was 56% and 78%, respectively, with 450 mg l(-1) phenol feed concentration and above 40 min of residence time all phenol was consumed within the column. For 1, 1.5, 2 and 3h of residence time, phenol removal achieved was 42%, 71%, 81% and 98%, respectively, at 3100 mg l(-1) phenol feed concentration. It is noteworthy that more than 95% of the initial phenol was converted into a non-passivating polymer without hazardous end products in a comparatively fast and energy-efficient process, being a safe treatment.     

Treatment of phenol and cresols in upflow anaerobic sludge blanket (UASB) process: a review

Upflow anaerobic sludge blanket (UASB) process has been successfully applied in the treatment of municipal and industrial wastewaters. Several researchers have investigated the suitability of the process for the treatment of phenols and phenolic wastewaters. The anaerobic treatment of phenols is still at an investigative stage. With increasing recognition of the UASB process, feasibility studies on the treatment of wastewater containing phenol and cresols (o-, m- and p- isomers) in UASB have been reviewed. It is reported that phenol concentration up to a range of 500-750 mg/L is generally not inhibitory to the UASB process. Phenol concentrations greater than 500 mg/L can be effectively treated with acclimatization of inocula, recirculation of the treated effluent and/or supplementing with co-substrates such as glucose, VFA and dilute molasses. The degradability of phenol is more than p-cresol, which in turn is more than m- and o-cresol.     

Fuzzy control of disturbances in a wastewater treatment process

This paper describes a real-time process control scheme to cope with the problem of input disturbances in wastewater treatment processes, based on a fuzzy inferential control system. This can detect the presence of a dangerous input conditions whenever either organic overload or inhibitory/toxic compounds are present in the raw wastewater. Based on this diagnosis, a set of fuzzy rules are implemented to divert the process flow and bring the system back into a safe state. The control system was designed and tested using a pilot plant, to which a toxic disturbance was applied. Its behaviour with and without fuzzy control are compared, and the process reliability improvement in the controlled case is demonstrated.     

Methanogen community structure-activity relationship and bioaugmentation of overloaded anaerobic digesters

Accumulation of acids in anaerobic digesters after organic overload can inhibit or stop CH₄ production. Therefore, methods to reduce acid concentrations would be helpful. One potential method to improve recovery involves bioaugmentation, addition of specific microorganisms to improve performance. In this study, transiently overloaded digesters were bioaugmented with a propionate-degrading enrichment culture in an effort to decrease recovery time. Biomass samples from 14 different, full-scale anaerobic digesters were screened for specific methanogenic activity (SMA) against propionate; the microbial communities were also compared. SMA values spanned two orders of magnitude. Principal component analysis of denaturing gradient gel electrophoresis (DGGE) banding patterns for a functional gene (mcrA) suggested an underlying community structure-activity relationship; the presence of hydrogenotrophic methanogens closely related to Methanospirillum hungatei and Methanobacterium beijingense was associated with high propionate SMA values. The biomass sample demonstrating the highest SMA was enriched for propionate degrading activity and then used to bioaugment overloaded digesters. Bioaugmented digesters recovered more rapidly following the organic overload, requiring approximately 25 days (2.5 solids retention times (SRTs)) less to recover compared to non-bioaugmented digesters. Benefits of bioaugmentation continued for more than 12 SRTs after organic overload. Bioaugmentation is a promising approach to decrease recovery time after organic overload.     

Quantitative image analysis as a diagnostic tool for identifying structural changes during a revival process of anaerobic granular sludge

Due to unspecified operational problems, the specific acetoclastic activity (SAA) of the anaerobic granular sludge present in an industrial UASB reactor was considerably damaged (from 250 to less than 10mL CH(4)@STP/gVSS.d), significantly reducing the biogas production of that industrial unit. The hydrogenotrophic methanogenic activity exhibited a value of 600mL CH4@STP/gVSS.d, the settling velocity was 31.4+/-9.8m/h, the average equivalent diameter was 0.92+/-0.43mm, and about 70% of the VSS were structured in aggregates larger than 1mm. In order to study the recovery of the SAA, this sludge was collected and inoculated in a lab-scale expanded granular sludge blanket (EGSB) reactor. Ethanol was fed as the sole carbon source during a trial period of 106 days. Process monitoring included COD removal efficiency, methane production, and periodic determination of the specific methanogenic activity in the presence of acetate, propionate, butyrate, ethanol and H(2)/CO(2). Quantitative image analysis allowed for information to be obtained on granular fragmentation/erosion and filaments release. During the first operational period, biogas production was mainly due to the hydrogenotrophic activity. However, after 40 days, the SAA steadily increased achieving a maximum value of 183+/-13mL CH4@STP/gVSS.d. The onset of SAA recovery, granules breakdown and filaments release to the bulk occurred simultaneously. Further increase in SAA was accompanied by granular growth. In the last 25 days of operation, the size distribution was stable with more than 80% of projected area of aggregates corresponding to granules larger than 1mm (equivalent diameter). Confocal images from FISH hybridized sections of the granules showed that after SAA recovery, the granules developed an organized structure where an acidogenic/acetogenic external layer was apparent. Granular fragmentation and increase of filaments in the bulk, simultaneously with the increase in the acetoclastic activity are described for the first time and might represent a structural response of granular sludge to promote the optimal substrate uptake at minimal diffusion limitations.