Stimulation of methanol degradation in UASB reactors: in situ versus pre-loading cobalt on anaerobic granular sludge

The effect of pre-loading and in situ loading of cobalt onto a cobalt-limited granular sludge on the performance of methanol fed bioreactors was investigated. One upflow anaerobic sludge bed (UASB) reactor was inoculated with cobalt pre-loaded sludge (24h; 30 degrees C; 1 mM CoCl2) and a second UASB with unloaded sludge. The UASB reactors (30 degrees C; pH 7) were operated for 77 days at 8 h hydraulic retention time and organic loading rates ranging from 5 to 20 g COD.L reactor(-1).d(-1). Cobalt pre-loading clearly stimulated the methanogenic activity of the sludge with methanol as the substrate, e.g., after 30 days of reactor operation this activity was 5.8 times higher than that of the cobalt unloaded sludge. During the experiment, part of the cobalt leached from the pre-loaded sludge, i.e., 54% of the cobalt content was lost during the 77 days of reactor operation. Sequential metal extraction showed that losses mainly occurred from the exchangeable and carbonate fraction and in the sludge remaining cobalt was mainly present in the organic/sulfide fraction of the sludge. In situ loading of cobalt in the unloaded UASB reactor on day 57 by adding 31 microM cobalt to the influent for a 24-h period (16% of the cobalt present in the loaded sludge at day 11) resulted in a 4 time increase of the methanogenic activity of the sludge with methanol as the substrate at the end of the reactor experiment, while the accumulated amount of cobalt in the sludge only amounted to 6% of the cobalt accumulated in the loaded sludge (on day 11). This study showed that both pre-loading sludge and in situ loading are adequate for achieving an increased reactor performance of methanol fed UASB reactors operating under cobalt limitation. However, the in situ dosing procedure needs substantially lower amounts of cobalt, while it also gives significantly smaller losses of cobalt with the effluent.     

Anaerobic dechlorination of pentachlorophenol in fixed-film and upflow anaerobic sludge blanket reactors using different inocula

Longterm performance and stability of two upflow anaerobic sludge blanket (UASB) reactors inoculated with granular sludge and treating a synthetic waste water containing pentachlorophenol (PCP) and phenol were studied. A similar system consisting of two fixed-film reactors inoculated with anaerobic digested sewage sludge were further studied. One reactor in each series received glucose in addition to the phenols. Dechlorination of PCP proceeded via two different dominating pathways in the respective reactor systems, suggesting that two distinct microbial populations were present, probably originating from the different inocula. Dechlorinating activity was maintained for more than 18 months in the UASB reactors and was generally higher than in the fixed-film reactors. In the fixed-film reactors, dechlorination of PCP suddenly decreased after 15.5 months of operation compared to earlier performance. Since no operational parameters had been changed, this indicated that the enriched culture was unstable on a longterm basis. Addition of yeast extract to the medium restored activity. General process stability in both reactor systems was clearly enhanced by the addition of glucose and was superior in the UASB/granular sludge system. The better performance and the higher stability in the UASB/granular sludge reactor highlights the importance of thorough screening of inocular prior to start-up of processes treating waste waters containing xenobiotic compounds.Abbreviations PCP pentachlorophenol - TeCP tetrachlorophenol - TCP trichlorophenol - DCP dichlorophenol - UASB upflow anaerobic sludge blanket - HRT hydraulic retention time     

Performance of upflow anaerobic sludge blanket (UASB) reactor treating wastewaters containing carbon tetrachloride

The anaerobic biodegradation of carbon tetrachloride (CT) was investigated during the granulation process by reducing the hydraulic retention time, increasing the chemical oxygen demand (COD) and CT loadings in a 2l laboratory-scale upflow anaerobic sludge blanket (UASB) reactor. Anaerobic unacclimated sludge and glucose were used as seed and primary substrate, respectively. Granules were developed 4 weeks after start-up, which grew at an accelerated rate for 8 months, and then became fully grown. The effect of operational parameters such as influent CT concentrations, COD, CT loading, food to biomass ratio and specific methanogenic activity (SMA) were also considered during granulation. The granular sludge cultivated had a maximum diameter of 2.1 mm and SMA of 1.6 g COD/g total suspended solid (TSS) day. COD and CT removal efficiencies of 92 and 88% were achieved when the reactor was firstly operating at CT and COD loading rates of 17.5 mg/l day and 12.5 g/l day, respectively. This corresponds to hydraulic retention time of 0.28 day and food to biomass ratio of 0.5 g COD/g TSS day. Kinetic coefficients of maximum specific substrate utilization rate, half velocity coefficient, growth yield coefficient and decay coefficient were determined to be 2.4 × 10^–3 mg CT/TSS day^–1, 1.37 mg CT/l, 0.69 mg TSS/mg CT and 0.046 day^–1, respectively for CT biotransformation during granulation.     

Assessment of the addition of Thiobacillus denitrificans and Thiomicrospira denitrificans to chemolithoautotrophic denitrifying bioreactors

The aim of the present study was to assess the impact of adding cultures of Thiobacillus denitrificans and Thiomicrospira denitrificans to two upflow anaerobic sludge bed (UASB) reactors: one inoculated with granular sludge and the other filled only with activated carbon (AC). The performances of the bioreactors and the changes in biomass were compared with a non-bioaugmented control UASB reactor inoculated with granular sludge. The reactors inoculated with granular sludge achieved efficiencies close to 90% in nitrate and thiosulfate removal for loading rates as high as 107 mmol-NO3 -/l per day and 68 mmol-S2O3 2-/l per day. Bioaugmentation with Tb. denitrificans and Tm. denitrificans did not enhance the efficiency compared to that achieved with non-bioaugmented granular sludge. The loading rates and efficiencies were 30-40% lower in the AC reactor. In all the reactors tested, Tb. denitrificans became the predominant species. The results strongly suggest that this bacterium was responsible for denitrification and sulfoxidation within the reactors. We additionally observed that granules partially lost their integrity during operation under chemolithoautotrophic conditions, suggesting limitations for long-term operation if bioaugmentation is applied in practice.     

Role of nickel in high rate methanol degradation in anaerobic granular sludge bioreactors

The effect of nickel deprivation from the influent of a mesophilic (30 degrees C) methanol fed upflow anaerobic sludge bed (UASB) reactor was investigated by coupling the reactor performance to the evolution of the Methanosarcina population of the bioreactor sludge. The reactor was operated at pH 7.0 and an organic loading rate (OLR) of 5-15 g COD l(-1) day(-1) for 191 days. A clear limitation of the specific methanogenic activity (SMA) on methanol due to the absence of nickel was observed after 129 days of bioreactor operation: the SMA of the sludge in medium with the complete trace metal solution except nickel amounted to 1.164 (+/-0.167) g CH(4)-COD g VSS(-1) day(-1) compared to 2.027 (+/-0.111) g CH(4)-COD g VSS(-1) day(-1) in a medium with the complete (including nickel) trace metal solution. The methanol removal efficiency during these 129 days was 99%, no volatile fatty acid (VFA) accumulation was observed and the size of the Methanosarcina population increased compared to the seed sludge. Continuation of the UASB reactor operation with the nickel limited sludge lead to incomplete methanol removal, and thus methanol accumulation in the reactor effluent from day 142 onwards. This methanol accumulation subsequently induced an increase of the acetogenic activity in the UASB reactor on day 160. On day 165, 77% of the methanol fed to the system was converted to acetate and the Methanosarcina population size had substantially decreased. Inclusion of 0.5 muM Ni (dosed as NiCl(2)) to the influent from day 165 onwards lead to the recovery of the methanol removal efficiency to 99% without VFA accumulation within 2 days of bioreactor operation.     

Production of PHA from starchy wastewater via organic acids

Polyhydroxyalkanoate (PHA) was produced from a starchy wastewater in a two-step process of microbial acidogenesis and acid polymerization. The starchy organic waste was first digested in a thermophilic upflow anaerobic sludge blanket (UASB) reactor to form acetic (60-80%), propionic (10-30%) and butyric (5-40%) acids. The total volatile fatty acids reached 4000 mg l(-1) at a chemical oxygen demand (COD) loading rate of 25-35 g l(-1) day(-1). A carbon balance indicates that up to 43% of the organic carbon in the starchy waste went to the organic acids and the rest to biogas, volatile suspended solids and residual sludge accumulated in the reactor. The acid composition profile was affected by COD loading rate: a medium rate around 9 g l(-1) day(-1) gave a high propionic acid content (29% wt) and a high rate around 26 g l(-1) day(-1) led to a high butyric acid content (34% wt). The acids in the effluent solution after microfiltration were utilized and polymerized into PHA by bacterium Alcaligenes eutrophus in a second reactor. Fifty grams of PHA was produced from 100 g total organic carbon (TOC) utilized, a yield of 28% based on TOC, which is comparable with 55 g PHA per 100 g TOC of pure butyric and propionic acids used. PHA formation from individual acids was further investigated in a semi-batch reactor with three acid feeding rates. With a limited nitrogen source (80-100 mg NH(3) per liter), the active biomass of A. eutrophus, not including the accumulated PHA in cells, was maintained at a constant level (8-9 g l(-1)) while PHA content in the cell mass increased continuously in 45 h; 48% PHA with butyric acid and 53% PHA with propionic acid, respectively. Polyhydroxybutyrate was formed from butyric acid and poly(hydroxybutyrate-hydroxyvalerate) formed from propionic acid with 38% hydroxyvalerate.     

A further study of the anaerobic biotreatment of malt whisky distillery pot ale using an UASB system

Pot ale from a pilot-scale malt whisky distillery was treated using a mesophilic upflow anaerobic sludge blanket (UASB) digester. Stable operation was observed at organic loading rates (OLRs) of 5.46 kg COD/m3 day or less when the pot ale was diluted with tap water. Digester failure occurred when undiluted pot ale was used, even though OLR was less than 5 kg COD/m3 day. Overall performance was worse than that observed previously when UASB digesters were used to treat pot ale from a different source supplemented with trace elements. A substantial proportion of effluent chemical oxygen demand (COD) was present as volatile fatty acids (VFA), particularly during periods of reactor stress, indicating that overall performance was limited by the rate of VFA conversion. Wastewater alkalinity rose during digestion. The sludge which developed in the reactor was flocculent but did not form compact granules.     

Effect of long-term cobalt deprivation on methanol degradation in a methanogenic granular sludge bioreactor

The effect of the trace metal cobalt on the conversion of methanol in an upflow anaerobic sludge bed (UASB) reactor was investigated by studying the effect of cobalt deprivation from the influent on the reactor efficiency and the sludge characteristics. A UASB reactor (30 degrees C; pH 7) was operated for 261 days at a 12-h hydraulic retention time (HRT). The loading rate was increased stepwise from 2.6 g chemical oxygen demand (COD) x L reactor(-1) x d(-1) to 7.8 g COD x L reactor(-1) x d(-1). Cobalt deprivation had a strong impact on the methanogenic activity of the sludge. In batch tests, the methanogenic activity of the sludge with methanol as the substrate increased 5.3 (day 28) and 2.1 (day 257) times by addition of 840 nM of cobalt. The sludge had an apparent K(m) for cobalt of 948 nM after 28 days of operation and 442 nM at the end of the run. Cobalt deprivation during 54 days of operation led to a methanol conversion efficiency of only 55%. Continuous addition of cobalt (330 nM) for 33 days improved the methanol removal efficiency to 100%. In this period of cobalt dosing, the cobalt concentration in the sludge increased 2.7 times up to 32 microg x g TSS(-1). Upon omission of the cobalt addition, cobalt washed-out at a stable rate of 0.1 microg x g VSS(-1) x d(-1). At the end of the run, the cobalt concentration of the sludge was similar to that of the seed sludge.     

Reactivation of effluent granular sludge from a high-rate Anammox reactor after storage

In this study, effluent sludge from a high-rate Anammox reactor was used to re-start new Anammox reactors for the reactivation of Anammox granular sludge. Different start-up strategies were evaluated in six upflow anaerobic sludge blanket (UASB) reactors (R₁–R₆) for their effect on nitrogen removal performance. Maximal nitrogen removal rates (NRRs) greater than 20 kg N/m³/day were obtained in reactors R₃–R₅, which were seeded with mixed Anammox sludge previously stored for approximately 6 months and 1 month. A modified Boltzmann model describing the evolution of the NRR fit the experimental data well. An amount of sludge added to the UASB reactor or decreasing the loading rate proved effective in relieving the substrate inhibition and increasing the NRR. The modified Stover–Kincannon model fit the nitrogen removal data in the Anammox reactors well, and the simulation results showed that the Anammox process has great nitrogen removal potential. The observed inhibition in the Anammox reactors may have been caused by high levels of free ammonia. The sludge used to seed the reactors did not settle well; sludge flotation was observed even after the reactors were operated for a long time at a floating upward velocity (F_s) of greater than 100 m/h. The settling sludge, however, exhibited good settling properties. Scanning electron microscopy showed that the Anammox granules consisted mainly of spherical and elliptical bacteria with abundant filaments on their surface. Hollows in the granules were also present, which may have contributed to sludge floatation.     

Flocculants effect in biomass retention in a UASB reactor treating dairy manure

The performance and biomass retention of an upflow anaerobic sludge bed (UASB) reactor treating liquid fraction of dairy manure has been investigated at several organic loading rates. Two identical UASB reactors were employed. The biomass of one UASB reactor (FBR) had previously been treated with a cationic polyacrylamide, the other reactor was operated as a control reactor (CR). At 3 and 2 days of HRT both reactors functioned similarly, but at 1.5 days HRT some differences were observed between both effluents. Mean COD(T) removal percentages were 83.4% and 76.5%; COD(VFA) values in effluents were 977 and 2682 mg l(-1) for the FBR and the CR respectively. The VSS initial value in both reactors was 25.66 g VSS, whereas after the experiment the quantities were 31.83 g VSS in the FBR and 23.18 g VSS in the CR reactors. Polymer addition resulted in a higher degree of biomass retention and better performance in the FBR reactor.     

A study of the effects of methanol in start-up of UASB reactors

Upflow anaerobic sludge blanket reactors were started up, using a stepped loading regime, on a dairy waste prepared from raw ice-cream production waste from an ice-cream manufacturing plant. One UASB was supplemented with methanol. Granules were detected in the methanol-fed UASB at 3 weeks after start-up. The maximum load of 6·7 kgCOD m⁻³ day⁻¹ with an HRT of 0·75 days could not be sustained during the 73 days of operation. Complete granulation had, however, occurred by the 4th week. Methanol addition at start-up aided rapid biomass granulation, and enhanced the settling velocity and specific activity of the sludge. However, wash-out was severe and residual biomass was sensitive to methanol withdrawal and produced no net growth.     

Development of anaerobic sludge bed (ASB) reactor technologies for domestic wastewater treatment: motives and perspectives

During the treatment of raw domestic wastewater in the upflow anaerobic sludge blanket (UASB) reactor, the suspended solids (SS) present in the wastewater tend to influence negatively the methanogenic activity and the chemical oxygen demand (COD) conversion efficiency. These problems led to the emergence of various anaerobic sludge bed systems such as the expanded granular sludge bed (EGSB), the upflow anaerobic sludge blanket (UASB)-septic tank, the hydrolysis upflow sludge bed (HUSB), the two-stage reactor and the anaerobic hybrid (AH) reactor. However, these systems have, like the UASB reactor, limited performance with regard to complete treatment (e.g., removal of pathogens). In this respect, a new integrated approach for the anaerobic treatment of domestic wastewater is suggested. This approach combines a UASB reactor and a conventional completely stirred tank reactor (CSTR) for the treatment of the wastewater low in SS and sedimented primary sludge, respectively. The principal advantages of the proposed system are energy recovery from organic waste in an environmentally friendly way; lowering the negative effect of suspended solids in the UASB reactor; production of a high quality effluent for irrigation; and prevention of odour problems.     

Zinc deprivation of methanol fed anaerobic granular sludge bioreactors

The effect of omitting zinc from the influent of mesophilic (30 degrees C) methanol fed upflow anaerobic sludge bed (UASB) reactors, and latter zinc supplementation to the influent to counteract the deprivation, was investigated by coupling the UASB reactor performance to the microbial ecology of the bioreactor sludge. Limitation of the specific methanogenic activity (SMA) on methanol due to the absence of zinc from the influent developed after 137 days of operation. At that day, the SMA in medium with a complete trace metal solution except Zn was 3.4 g CH4-COD g VSS(-1) day(-1), compared to 4.2 g CH4-COD g VSS(-1) day(-1) in a medium with a complete (including zinc) trace metal solution. The methanol removal capacity during these 137 days was 99% and no volatile fatty acids accumulated. Two UASB reactors, inoculated with the zinc-deprived sludge, were operated to study restoration of the zinc limitation by zinc supplementation to the bioreactor influent. In a first reactor, no changes to the operational conditions were made. This resulted in methanol accumulation in the reactor effluent after 12 days of operation, which subsequently induced acetogenic activity 5 days after the methanol accumulation started. Methanogenesis could not be recovered by the continuous addition of 0.5 microM ZnCl2 to the reactor for 13 days. In the second reactor, 0.5 microM ZnCl2 was added from its start-up. Although the reactor stayed 10 days longer methanogenically than the reactor operated without zinc, methanol accumulation was observed in this reactor (up to 1.1 g COD-MeOH L(-1)) as well. This study shows that zinc limitation can induce failure of methanol fed UASB reactors due to acidification, which cannot be restored by resuming the continuous supply of the deprived metal.     

Feasibility studies on the treatment of dairy wastewaters with upflow anaerobic sludge blanket reactors

The feasibility of using upflow anaerobic sludge blanket (UASB) reactors for the treatment of dairy wastewaters was explored. Two types of UASBs were used--one operating on anaerobic sludge granules developed by us from digested cowdung slurry (DCDS) and the other on the granules obtained from the reactors of M/s EID Parry treating sugar industry wastewaters. The reactors were operated at HRT of 3 and 12 h and on COD loading rates ranging from 2.4 kg per m3 of digester volume, per day to 13.5 kg m(-3) d(-1). At the 3 h HRT, the maximum COD reduction in the DCDS-seeded and the industrial sludge-seeded reactors was 95.6% and 96.3%, respectively, better than at 12 h HRT (90% and 92%, respectively). In both the reactors, the maximum, the second best, and the third best COD reduction occurred at the loading rates of 10.8, 8.6 and 7.2 kg m3 d(-1), respectively. At loading rates higher than 10.8 kg, the reactor performance dropped precipitously. Whereas in the first few months the reactors operating on sludge from EID Parry achieved better biodegradation of the waste, compared to the reactors operated on DCDS, the performance of the latter gradually improved and matched with the performance of the former.     

Effects of cationic polymer on performance of UASB reactors treating low strength wastewater

The effect of cationic polymer additives on biomass granulation and COD removal efficiency had been examined in lab-scale upflow anaerobic sludge blanket (UASB) reactors, treating low strength synthetic wastewater (COD 300-630 mg/l). Under identical conditions, two reactors were operated with and without polymer additives in inoculum under four different organic loading rates (OLRs). The optimum polymer dose was adopted based upon the results of jar test and settling test carried out with inoculum seed sludge. With the use of thick inoculum, SS greater than 110 g/l and VSS/SS ratio less than 0.3, granulation was observed in UASB reactor treating synthetic wastewater as well as actual sewage, when OLR was greater than 1.0 kg COD/m(3) d. Polymer additive with such thick inoculum was observed to deteriorate percentage granules and COD removal efficiency compared to inoculum without polymer additives. At OLR less than 1.0 kg COD/m(3) d, proper granulation could not be achieved in both the reactors inoculated with and without polymer additive. Also, under this low loading, drastic reduction in COD removal efficiency was observed with polymer additives in inoculum. Hence, it is rational to conclude that biomass granulation for treatment of low strength biodegradable wastewater depends on the applied loading rate and selection of thick inoculum sludge.     

Effect of sulfur source on the performance and metal retention of methanol-fed UASB reactors

The effect of a sulfur source on the performance and metal retention of methanol-fed upflow anaerobic sludge bed (UASB) reactors was investigated. For this purpose, two UASB reactors were operated with cobalt preloaded granular sludge (1 mM CoCl2; 30 degrees C; 24 h) at an organic loading rate (OLR) of 5 g COD.L reactor(-1).d(-1). One UASB reactor (R1) was operated without a sulfur source in the influent during the first 37 days. In this period the methanol conversion to methane remained very poor, apparently due to the absence of a sulfur source, because once cysteine, a sulfur-containing amino acid, was added to the influent of R1 (day 37) a full conversion of methanol to methane occurred within 6 days. The second reactor (R2) was operated with sulfate (0.41 mM) in the influent during the first 86 days of operation, during which no limitation in the methanol conversion to methane manifested. Cobalt washed out from the sludge at similar rates in both reactors. The leaching of cobalt occurred at two distinct rates, first at a high rate of 22 microg.g TSS(-1).d(-1), which proceeded mainly from the exchangeable and carbonate fraction and later at a relatively slow rate of 9 mug.g TSS(-1).d(-1) from the organic/sulfide fraction. This study showed that the supply of the sulfur source L-cysteine has a pronounced positive effect on the methanogenic activity and the retention of metals such as iron, zinc and molybdenum.     

Anaerobic ammonium oxidation process in an upflow anaerobic sludge blanket reactor with granular sludge selected from an anaerobic digestor

The purpose of this work was to evaluate the development of the anammox process by the use of granular sludge selected from a digestion reactor as a potential seed source in a lab-scale UASB (upflow anaerobic sludge blanket) reactor system. The reactor was operated for approximately 11 months and was fed by synthetic wastewater. After 200 days of feeding with NH₄ ⁺ and NO₂ ⁻ as the main substrates, the biomass showed steady signs of ammonium consumption, resulting in over 60% of ammonium nitrogen removal. This report aims to present the results and to more closely examine what occurs after the onset of anammox activity, while the previous work described the start-up experiment and the presence of anammox bacteria in the enriched community using the fluorescencein situ hybridization (FISH) technique. By the last month of operation, the consumed NO₂ ⁻N/NH₄ ⁺-N ratio in the UASB reactor was close to 1.32, the stoichiometric ratio of the anammox reaction. The obtained results from the influentshutdown test suggested that nitrite concentration would be one key parameter that promotes the anammox reaction during the start-up enrichment of anammox bacteria from granular sludge. During the study period, the sludge color gradually changed from black to red-brownish.     

Design,Construction, and Starting-up of an Anaerobic Reactor for the Stabilisation,Handling, and Disposal of Excess Biological Sludge Generated in a Wastewater Treatment Plant

Design, construction, and starting-up of an upflow anaerobic sludge blanket reactor was carried out. This system was proposed for excess sludge stabilisation, particularly that generated at an activated sludge wastewater treatment facility installed in a sugarcane mill. The upflow anaerobic sludge blanket (UASB) reactor built, had a working volume of 22.3 m³and a hydraulic residence time of 22 days. Methane production was at a maximum of 79% volume with an average of 60% for this treatment. For starting up the anaerobic reactor, a suitable inoculum from a neighboring plant was used. As the waste characteristics in both plants were different, an acclimation procedure was followed to achieve granulation. Control and stability of anaerobic reactions were monitored with alkalinity data, using the so-called ‘alfa alkalinity’ to try to keep its value at around 0.4. Once pseudosteady-state conditions were reached (chemical oxygen demand reduction and methane-rich biogas production within ±10 percent), the organic load was steadily increased up to feeding 100% excess sludge. The UASB reactor used to stabilise the excess biomass generated a sludge with a much lower volume than that originally fed. Its design ensured adequate hydraulic flow and biogas production with a high methane content. The bacteria were attached constituting spheres and very minor maintenance operations were required.     

The inhibitory effects and removal of dieldrin in continuous upflow anaerobic sludge blanket reactors

The inhibitory effects and removal efficiency of dieldrin (DLD) in anaerobic reactors were investigated. Anaerobic toxicity assay (ATA) experiments conducted in batch reactors revealed that 30 mg/l DLD had inhibitory effects on the unacclimated mixed anaerobic cultures. Continuous reactor experiments performed in a lab-scale two-stage upflow anaerobic sludge blanket (UASB) reactor system which was fed with ethanol as the sole carbon source, indicated that anaerobic granular cultures could be successfully acclimated to DLD. Chemical oxygen demand (COD) removal efficiencies were 88-92% for the two-stage system. The influent DLD concentration of 10 mg/l was removed by 44-86% and 86-94% in the second stage and overall UASB system, respectively. Biosorption of DLD on granular anaerobic biomass was found to be a significant mechanism for DLD removal in the UASB system. The maximum DLD loading rate and minimum HRT achievable for the first stage UASB reactor were 0.5 mg/lday (76 microg DLD/g VSS.day) and 10 h, respectively, which resulted in the overall COD removal efficiency of 85%.     

Rapid granulation and sludge retention for tetrachloroethylene removal in an upflow anaerobic sludge blanket reactor

A laboratory scale upflow anaerobic sludge blanket (UASB) reactor was operated at 35 °C for over 200 days to investigate the granulation mechanism during tetrachloroethylene (TCE) biodegradation. Anaerobic, unacclimated sludge and glucose were used as seed and primary substrate, respectively. TCE-degrading granules developed after 1.5 months of start-up. They grew at an accelerated pace for 7 months. The TCE-degrading granules had a maximum diameter of 2.5 mm and specific methanogenic activity of 1.32 g chemical oxygen demand (COD) g^–1 total suspended solid (TSS) day^–1. 94% COD and 90% TCE removal efficiencies were achieved when the reactor was operating at loading rates as high as 160 mg TCE l^–1 day^–1 and 14 g COD l^–1 day^–1, after 230 days of continuous operation.