Search

This study was designed to evaluate a treatment system for high strength wastewater (vinasse) from a sugarcane molasses-based bio-ethanol plant in Thailand. A laboratory-scale two-phase treatment system composed of a sulfate reducing (SR) tank and multi-staged up-flow anaerobic sludge blanket (MS-UASB) reactor was used as the pre-treatment unit. Conventional UASB and down-flow hanging sponge (DHS) reactors were used as the post-treatment unit. The treatment system was operated for 300 days under ambient temperature conditions (24.6-29.6 °C). The hydraulic retention time (HRT) in each unit was kept at 25 h for the two-phase system and 23 h for the UASB&DHS. The influent concentration was allowed to reach up to 15,000 mg chemical oxygen demand (COD)/L. COD removal efficiency (based on influent COD) of the two-phase MS-UASB and the UASB&DHS was 54.9 and 18.7%, respectively. Due to the effective removal of sulfide in the SR tank, the MS-UASB achieved a high methane conversion ratio of up to 97%. In DHS, nitrification occurred at the outside portion of the sponge media while denitrification occurred at the inside. Consequently, 27% of the total nitrogen (TN) was removed. An amount of 32% of residual nitrogen (28 mgN/L) was in the form of nitrate, a better nitrogen state for fertilizer. [ABSTRACT FROM AUTHOR]

A combination of an acidification reactor and an up-flow staged sludge bed (USSB) reactor was applied to treatment of molasses wastewater. The USSB, equipped with three gas solid separators, was selected because of superior organic removal by means of good retention of granular sludge. The combined system was continuously operated at mesophilic (35°C) conditions for 600 days. The USSB achieved a higa organic removal rate of 37 kgCOD/m³ day (organic loading rate of 43 kgCOD/m³ day with 86.2% COD -removal). The USSB retained high sludge concentration of approximately 58 gVSS/L based on the reactor volume. The retained granular sludge had good settle-ability with a sludge volume index (SVI) cf 4 to 20 ml/gSS. The retained sludge possessed a sufficient level of methanogenic activities for acetate (1.2 gCOD/gVSS day) and for hydrogen (1.7 gCOD/gVSS day). Analysis of microbial community revealed that genus Methanobacterium as hydrogen utilizing methanogen and order Methanosarcinales as acetate utilizing methanogen were detected in the retained sludge of the USSB reactor. The superior performance of the USSB was attributed to good retention of a large amount of granular sludge with high methanogenic activity. [ABSTRACT FROM AUTHOR]

The biodegradation characteristics of palm oil mill effluent (POME) and the related microbial community were studied in both actual sequential anaerobic ponds in Malaysia and enrichment cultures. The significant degradation of the POME was observed in the second pond, in which the temperature was 35-37 WC. In this pond, biodegradation of major long chain fatty acids (LCFA), such as palmitic acid (C16:0) and oleic acid (C18:1), was also confirmed. The enrichment culture experiment was conducted with different feeding substrates, i.e. POME, C16:0 and C18:1, at 35 WC. Good recovery of methane indicated biodegradation of feeds in the POME and C16:0 enrichments. The methane production rate of the C18:1 enrichment was slower than other substrates and inhibition of methanogenesis was frequently observed. Denaturing gradient gel electrophoresis (DGGE) analyses indicated the existence of LCFA-degrading bacteria, such as the genus Syntrophus and Syntorophomonas, in all enrichment cultures operated at 35 WC. Anaerobic degradation of the POME under mesophilic conditions was stably processed as compared with thermophilic conditions. [ABSTRACT FROM AUTHOR]

In this study, continuous operation of a pilot-scale upflow anaerobic sludge blanket (UASB) reactor for sewage treatment was conducted for 630 days to investigate the physical and microbial characteristics of the retained sludge. The UASB reactor with a working volume of 20.2 m³ was operated at ambient temperature (16-29 °C) and seeded with digested sludge. After 180 days of operation, when the sewage temperature had dropped to 20 °C or lower, the removal efficiency of both total suspended solids (TSS) and total biochemical oxygen demand (BOD) deteriorated due to washout of retained sludge. At low temperature, the cellulose concentration of the UASB sludge increased owing to the rate limitation of the hydrolytic reaction of suspended solids in the sewage. However, after an improvement in sludge retention (settleability and concentration) in the UASB reactor, the process performance stabilized and gave sufficient results (68% of TSS removal, 75% of total BOD removal) at an hydraulic retention time (HRT) of 9.7 h. The methanogenic activity of the retained sludge significantly increased after day 246 due to the accumulation of Methanosaeta and Methanobacterium following the improvement in sludge retention in the UASB reactor. Acid-forming bacteria from phylum Bacteroidetes were detected at high frequency; thus, these bacteria may have an important role in suspended solids degradation. [ABSTRACT FROM AUTHOR]

A lab-scale expanded granular sludge bed (EGSB) reactor was operated at 20°C with low strength wastewater (0.6-0.8 g COD/L) for over 200 days. Reactor was inoculated with mesophilic granular sludge. The up-flow velocity was set to 5 m/h by effluent recirculation. The COD loading was increased up to 12 kg COD/m³/day until the day 76, resulting in hydraulic retention time of 1.5 hours. Physical properties (settleability and diameter) of retained sludge tended to deteriorate during the first 2-3 months, however sludge settleability kept sufficiently in the later part of experiment due to the reconstruction of granular sludge. The growth yield (Yg) of retained sludge (0.13 g VSS/g COD) was about two times higher than mesophilic and thermophilic granular sludge processes while the endogenous decay constant (Kd) is very low (0.0001/day) as compared with those processes. The sludge retention time of retained sludge reduced from 100 days to 40 days by the reduction of hydraulic retention time from 4 hours to 1.5 hours. Maintenance of 40 days of sludge retention time caused the stable retainment of biomass and the significant increase of methanogenic activity of the retained sludge. [ABSTRACT FROM AUTHOR]

In order to establish the appropriate methane fermentation technology for low strength wastewater, a 2.0 L EGSB reactor was operated at 20°C with 0.3 - 0.4 g COD/L of sucrose-based synthetic wastewater for 500 days. At the start up period, the reactor was operated in EGSB mode with a 5 m/h up flow velocity by continuous effluent recirculation. However, the EGSB reactor exhibited insufficient COD removal (50-60%) at COD loading of 7.2 - 9.6 kg COD/m3day due to the low COD concentration in the sludge bed. Therefore, we proposed the new operation mode by switching to UASB mode (without recirculation, 0.7 m/h up flow velocity) for 30 minutes and EGSB mode for 10 minutes in 40 minutes cycle. Moreover we added sodium sulfide to make the low ORP condition. In this operation, COD removal increased dramatically, from 65% to 91%. Additionally, physical properties of the retained sludge were well maintained in this operation mode. Furthermore, the retained sludge possessed appropriate levels of methanogenic activity (0.2-0.4 g COD/g VSS/ day) at 20°C. [ABSTRACT FROM AUTHOR]

A 2.0 L volume of EGSB reactor was operated at 208C for more than 500 days with 0.3-0.4 g COD/L of sucrose base wastewater to investigate the influence of effluent-recirculation on the process performance. At the start up period, the reactor was operated in EGSB mode with 5 m/h upflow velocity by continuous effluent recirculation. The COD loading was set to 7.2-9.6 kg COD/m3 day with HRT of 1 hour. However, in this mode, EGSB reactor exhibited insufficient COD removal efficiency, i.e., 50-60%. Therefore, UASB mode (without recirculation, 0.7 m/h upflow velocity) was used for 30 minutes in every 40 minutes cycle to increase the COD concentration in the sludge bed. As a result, an excellent process performance was shown. The COD removal efficiency increased from 65% to 91% and the reactor could maintain a good physical property of retained sludge (sludge concentration: 33.4 g VSS/L and SVI: 25 mL/g VSS). Furthermore, retained sludge possessed sufficient level of methanogenic activity at 208C. [ABSTRACT FROM AUTHOR]

In this study, a lab scale EGSB reactor was operated for 400 days to investigate the influence of temperature-decrease on the microbial characteristic of retained sludge. The EGSB reactor was started-up at 15°C seeding with 20°C-grown granular sludge. The influent COD of synthetic wastewater was set at 0.6-0.8 gCOD/L. The process-temperature was stepwise reduced from 15°C to 5°C during 400 days operation. Decrease of temperature of the reactor from 15°C to 10°C caused the decline of COD removal efficiency. However, continuous operation of the EGSB reactor led the efficient treatment of wastewater (70% of COD removal, 50% of methane recovery) at 10°C. We confirmed that the both acetate-fed and hydrogen-fed methanogenic activities of retained sludge clearly increased under 15 to 20°C. Changes of microbial profiles of methanogenic bacteria were analyzed by 16S rDNA-targeted DGGE analysis and cloning. It shows that genus Methanospirillum as hydrogen-utilizing methanogen proliferated due to low temperature operation of the reactor. On the other hand, genus Methanosaeta presented in abundance as acetoclastic-methanogen throughout the experiment. [ABSTRACT FROM AUTHOR]

Water quality parameters influence the abundance of pathogenic bacteria. The genera Aeromonas, Arcobacter, Klebsiella, and Mycobacterium are among the representative pathogenic bacteria identified in wastewater. However, information on the correlations between water quality and the abundance of these bacteria, as well as their reduction rate in existing wastewater treatment facilities (WTFs), is lacking. Hence, this study aimed to determine the abundance and reduction rates of these bacterial groups in WTFs. Sixty-eight samples (34 influent and 34 non-disinfected, treated, effluent samples) were collected from nine WTFs in Japan and Thailand. 16S rRNA gene amplicon sequencing analysis revealed the presence of Aeromonas, Arcobacter, and Mycobacterium in all influent wastewater and treated effluent samples. Quantitative real-time polymerase chain reaction (qPCR) was used to quantify the abundance of Aeromonas, Arcobacter, Klebsiella pneumoniae species complex (KpSC), and Mycobacterium. The geometric mean abundances of Aeromonas, Arcobacter, KpSC, and Mycobacterium in the influent wastewater were 1.2 × 104-2.4 × 105, 1.0 × 105-4.5 × 106, 3.6 × 102-4.3 × 104, and 6.9 × 103-5.5 × 104 cells mL-1, respectively, and their average log reduction values were 0.77-2.57, 1.00-3.06, 1.35-3.11, and -0.67-1.57, respectively. Spearman's rank correlation coefficients indicated significant positive or negative correlations between the abundances of the potentially pathogenic bacterial groups and Escherichia coli as well as water quality parameters, namely, chemical/biochemical oxygen demand, total nitrogen, nitrate-nitrogen, nitrite-nitrogen, ammonium-nitrogen, suspended solids, volatile suspended solids, and oxidation-reduction potential. This study provides valuable information on the development and appropriate management of WTFs to produce safe, hygienic water., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Aoki et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The management of pathogenic bacteria in waterways is a public health issue. Here, we investigated the concentrations of potentially pathogenic bacteria, Arcobacter spp. and Campylobacter spp., and Escherichia coli, by quantifying species-specific genes in surface water samples from canals and the Chao Phraya River from June 2017 to June 2018 in Bangkok, Thailand. We assessed the relationship between the specific bacterial concentrations, water quality, and seasonal changes. Arcobacter spp. were detected at high density in all samples and showed seasonal fluctuations according to analyses based on 16S rDNA and the invasion gene ciaB. High levels of 16S rDNA and dut gene of E. coli were detected in the polluted drainage canals. A high correlation was observed between E. coli and chemical and biochemical oxygen demand (COD and BOD), suggesting that untreated domestic wastewater was the source of the E. coli. In contrast, Arcobacter spp. were detected with high density even in water samples with relatively low COD, suggesting that Arcobacter spp. are more likely than E. coli to survive in the water environment. The analysis of 16S rDNA and ciaB gene sequence analyses indicated that the Arcobacter spp. isolated from the drainage canals were A. butzleri and A. cryaerophilus.

In this study, a pilot-scale down-flow hanging sponge (DHS) reactor was operated in the community plant of Bangkok for the treatment of domestic wastewater (COD 285 mg/L, BOD 105 mg/L) collected by separate sewer to evaluate the reactor's feasibility as a decentralized treatment system. The DHS reactor was operated for 600 days at ambient temperatures of 25-30 °C, both with constant flow conditions and with fluctuating flow conditions that simulated wastewater discharge patterns of the community. The results indicate that under constant flow at an HRT of 5 h, the volumetric loading rates of 0.36 kgBOD/m 3 -sponge/day and 0.16 kgN/m 3 -sponge/day were the optimum operational conditions of the DHS reactor in order to satisfy the effluent discharge standards. The DHS achieved removal rates of 89, 95, 91 and 90% for COD, BOD, TSS and NH 4 -N. Under the fluctuating flow condition, improvement of denitrification was confirmed at volumetric loading rates of 0.50 kgBOD/m 3 -sponge/day and 0.18 kgN/m 3 -sponge/day. The fluctuating flow of wastewater positively affects retained sludge activities in terms of homogenizing sludge concentration and stimulating oxygen uptake rates. These results suggest that the DHS reactor can be applied as a decentralized treatment system for domestic wastewater with fluctuating flow rates in tropical regions.

This study assesses the performance of an aerobic trickling filter, down-flow hanging sponge (DHS) reactor, as a decentralized domestic wastewater treatment technology. Also, the characteristic eukaryotic community structure in DHS reactor was investigated. Long-term operation of a DHS reactor for direct treatment of domestic wastewater (COD = 150-170 mg/L and BOD = 60-90 mg/L) was performed under the average ambient temperature ranged from 28°C to 31°C in Bangkok, Thailand. Throughout the evaluation period of 550 days, the DHS reactor at a hydraulic retention time of 3 h showed better performance than the existing oxidation ditch process in the removal of organic carbon (COD removal rate = 80-83% and BOD removal rate = 91%), nitrogen compounds (total nitrogen removal rate = 45-51% and NH 4 + -N removal rate = 95-98%), and low excess sludge production (0.04 gTS/gCOD removed). The clone library based on the 18S ribosomal ribonucleic acid gene sequence revealed that phylogenetic diversity of 18S rRNA gene in the DHS reactor was higher than that of the present oxidation ditch process. Furthermore, the DHS reactor also demonstrated sufficient COD and NH 4 + -N removal efficiency under flow rate fluctuation conditions that simulates a small-scale treatment facility. The results show that a DHS reactor could be applied as a decentralized domestic wastewater treatment technology in tropical regions such as Bangkok, Thailand.

This study investigated down-flow hanging sponge (DHS) technology as a promising trickling filter (TF) using sponge media as a biomass carrier with an emphasis on protection of the biomass against macrofauna overgrazing. A pilot-scale DHS reactor fed with low-strength municipal sewage was operated under ambient temperature conditions for 1 year at a sewage treatment plant in Bangkok, Thailand. The results showed that snails (macrofauna) were present on the surface of the sponge media, but could not enter into it, because the sponge media with smaller pores physically protected the biomass from the snails. As a result, the sponge media maintained a dense biomass, with an average value of 22.3 gVSS/L sponge (58.1 gTSS/L sponge) on day 370. The snails could graze biomass on the surface of the sponge media. The DHS reactor process performance was also successful. The DHS reactor requires neither chemical treatments nor specific operations such as flooding for snail control. Overall, the results of this study indicate that the DHS reactor is able to protect biomass from snail overgrazing.

A pilot-scale experiment of a down-flow hanging sponge (DHS) reactor for treatment of low-strength municipal wastewater was conducted over 1 year in Bangkok, Thailand, to establish an appropriate method for treatment under tropical climate conditions. Municipal wastewater with an average BOD of 19 mg/L was fed directly into the DHS reactor. Superior effluent quality (5.1 ± 3.4 mg/L TSS, 21.1 ± 9.0 mg/L COD, 2.8 ± 1.4 mg/L BOD, and 4.1 ± 1.0 mg/L TN) was achieved at a hydraulic retention time (HRT) of 1 h under an average temperature of 30 °C. The DHS reactor reached an actual HRT of 19.0 min, indicating good contact efficiency between wastewater and retained sludge. The DHS reactor retained dense sludge at 15.3-26.4 g VSS/L based on the sponge media volume. The sludge activity in terms of specific oxygen uptake rate was good. Excess sludge was produced as 0.051 g TSS/g COD removed (0.11 g TSS/g BOD removed), and a good SVI of 28 mL/g was observed. The sufficient performance was attributed to dense sludge with high activity, regardless of the low-strength wastewater. Overall, the DHS was advantageous owing to its simple operation, lack of operational problems, and low power consumption.

The process performance of a down-flow hanging sponge (DHS) reactor for treating low strength sewage (biochemical oxygen demand (BOD) 20-50 mg/L) was investigated in Bangkok, Thailand. The hydraulic retention time (HRT) was set at 4 h during the start-up period and was reduced to 1.5 h in a stepwise manner. Throughout the 300-day operational period, the DHS reactor shows high performance with respect to the removal of total suspended solid (>90% total suspended solid removal efficiency). No clogging of sponge media was observed in response to the self-digestion phenomena of the biofilm. At a HRT of 1.5 h, the BOD removal efficiency was sufficiently high (about 85%). The pathogen Escherichia coli and other coliform bacteria were removed almost completely as well (removal was 99.4% and 98.1%, respectively). Regarding the retained sludge activity measurement, the nitrite oxidation rate was higher than the ammonium oxidation rate (0.031 and 0.022 gram of nitrogen per gram of volatile suspended solids per day, respectively). In the 300 days of operation, the amount of excess sludge production was negligible. Thus, no sludge treatment system is required. Introduction of the DHS system in developing countries is recommended because this system requires a relatively small area, and has low electricity consumption and operation costs.

This study evaluated the process performance of a novel treatment system consisting of an acidification reactor, an upflow staged sludge bed (USSB) reactor, an upflow anaerobic sludge blanket reactor, and an aerobic trickling filter for the treatment of a high-strength molasses wastewater with a chemical oxygen demand (COD) of up to 120,000mg/L. The USSB operating at 35°C was capable of achieving an organic loading rate of 11kgCOD/m(3) day with a methane recovery of 62.4% at an influent COD of 120,000mg/L. The final effluent COD was 4520mg/L. The system was effective with regard to nitrification and sulfur removal. Fifty percent inhibition of the bacterial activity of the retained sludge by the cations was determined at 8gK/L for sucrose degradation, 16gK/L for sulfate reduction, and 12gK/L or 9gNa/L for acetoclastic methane production. Cation inhibition of anaerobic degradation reduced the process performance of the USSB., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In this study, the performance of a proposed treatment system consisting of an anaerobic process (acidification, methane fermentation) and an aerobic process (trickling filter) was evaluated for treating high concentrations of molasses-based wastewater (43-120 gCOD/L) by a continuous flow experiment. An anaerobic up-flow staged sludge bed (USSB) reactor, equipped with multiple gas solid separators, was used as the main treatment/methane recovery process. The USSB showed good efficiency of both COD removal (80-87%) and methane recovery (70-80%) at an organic loading rate of 11-43 kgCOD/m(3) day. As the influent COD concentration was increased, the organic loading rate for stable operation of the USSB was reduced due to cation inhibition. However, the COD removal efficiency of the whole treatment system (including the aerobic post-treatment process) was 96% even at an influent COD concentration of 120 gCOD/L. Use of the treated wastewater as a fertilizer and/or irrigation-water for sugarcane was evaluated by a field cultivation test. Both growth of sugarcane and emission of greenhouse gases from the field soil were measured. A relatively high methane flux (352 μgCH4/m(2) h) was observed when the treated wastewater from day 0 was used. By day 3, however, this value was reduced to the same level as the control. In addition, growth of sugarcane was satisfactory when the treated wastewater was used. The treated wastewater was found to be useful for cultivation of sugarcane in terms of both a low risk of greenhouse gas emission from the field soil and effectiveness for growth of sugarcane.

A combination of an acidification reactor and an up-flow staged sludge bed (USSB) reactor was applied for treatment of molasses wastewater containing a large amount of organic compounds and sulfate. The USSB reactor had three gas-solid separators (GSS) along the height of the reactor. The combined system was continuously operated at mesophilic temperature over 400 days. In the acidification reactor, acid formation and sulfate reduction were effectively carried out. The sugars contained in the influent wastewater were mostly acidified into acetate, propionate, and n-butyrate. In addition, 10-30% of influent sulfur was removed from the acidification reactor by means of sulfate reduction followed by stripping of hydrogen sulfide. The USSB achieved a high organic loading rate (OLR) of 30 kgCOD m(-3) day(-1) with 82% COD removal. Vigorous biogas production was observed at a rate of 15 Nm(3) biogas m(-3) reactor day(-1). The produced biogas, including hydrogen sulfide, was removed from the wastewater mostly via the GSS. The GSS provided a moderate superficial biogas flux and low sulfide concentration in the sludge bed, resulting in the prevention of sludge washout and sulfide inhibition of methanogens. By advantages of this feature, the USSB may have been responsible for achieving sufficient retention (approximately 60 gVSS L(-1)) of the granular sludge with high methanogenic activity (0.88 gCOD gVSS(-1) day(-1) for acetate and as high as 2.6 gCOD gVSS(-1) day(-1) for H(2)/CO(2)). Analysis of the microbial community revealed that sugar-degrading acid-forming bacteria proliferated in the sludge of the USSB as well as the acidification reactor at high OLR conditions.

The anaerobic biological treatment of sucrose-based, low-strength wastewater was investigated in expanded granular sludge bed (EGSB) reactors at low temperatures over a 300-day trial period. During the trial, the operating temperature was lowered in a stepwise manner from 20 degrees C to 5 degrees C. As a result, the reactors exhibited sufficient performances until 10 degrees C operation. The COD removal rate was 3.1-3.8 kgCOD m(-3) day(-1) at 10 degrees C. In particular, the COD removal rate increased gradually through the low-temperature operation; indeed, the later stages of the 10 degrees C operation attained a rate similar to those achieved at 20 degrees C and 15 degrees C. This finding is especially practical for applications of psychrophilic methane fermentation. Additionally, the structure of the microbial community in the granular sludge was analyzed by clone analysis based on 16S rRNA genes and fluorescence in situ hybridization (FISH). As a result, the percentage of the phylum Firmicutes, which were assumed to be Anaerobivrio sp. and Lactococcus sp., greatly increased from 0.7% to 8.0% of the total cells, especially in the surface layer of the granular sludge. These bacteria would contribute to the degradation of the sucrose substrate anaerobically at ambient temperatures. Moreover, the results suggest that a Methanospirillum species, which is a H2-utilizing methanogen, increased from 0.5% to 6.7% during the low-temperature incubation, with a significant increase of methanogenic activity from H2/CO2 at 20 degrees C. Thus, the Methanospirillum species detected in this study may have a key role as hydrogen scavenger during hydrogen-metabolism in low-temperature conditions.

The influence of the sugar content of wastewater on changes in the characteristics of the retained sludge was investigated by using two lab-scale granular sludge bed reactors at 20 degrees C. Both reactors were inoculated with granular sludge grown at 20 degrees C and were fed with synthetic wastewater containing sucrose and volatile fatty acids (VFAs). On day 70, the sucrose content of the wastewater was changed to 90% (based on wastewater COD value) for the first reactor and 0% (VFA 90%) for the second. After this change in feed composition, the COD removal efficiency became about 91% for the sucrose-fed reactor and 95% for the VFA-fed reactor. The growth yield (Yg) of the sucrose-fed sludge increased more than that of the VFA-fed sludge. Consequently, deterioration of the settleability of the sucrose-fed sludge was observed. The sucrose-degrading activity of the retained sludge obtained from the sucrose-fed reactor increased significantly from 3.7 g COD g VSS(-1) day(-1) on day 62 to 36.8 g COD g VSS(-1) day(-1) on day 230, in accordance with the predominant growth of sugar-degrading bacteria--namely, Lactococcus, Clostridium and Chloroflexi--in the retained sludge. The excessive growth of these sugar-degrading bacteria in the retained sludge caused unstable process performance in the sucrose-fed reactor at 20 degrees C.

A lab-scale expanded granular sludge bed (EGSB) reactor was seeded with granular sludge and operated to investigate the influence of temperature decrease on both process performance and the microbial community structure of the granular sludge. Synthetic wastewater containing sucrose and volatile fatty acids was used as feed. The EGSB reactor was brought online at a starting temperature of 15 degrees C and was reduced stepwise to a final temperature of 5 degrees C. The reactor exhibited sufficient COD removal efficiency between 10 degrees C and 15 degrees C. However at 5 degrees C serious deterioration of process performance was observed. The methane-producing activity of the retained sludge increased when it was compared to the activity of the seed sludge (day 0) during 10 degrees C to 15 degrees C operation. When hydrogen fed, sludge showed much higher methanogenic activity as compared with seed sludge activity at test temperatures of 15 degrees C and 20 degrees C on day 196 of reactor operation. At this time, proliferation of the genus Methanospirillum in the retained sludge was observed and a decrease in Methanobacterium species was also measured. Throughout the experiment, the genus Methanosaeta was detected in abundance and the community structure of the Domain Bacteria was stably maintained. The sugar-degrading acid-forming bacteria, Lactococcus and Anaerovibrio were detected in the retained sludge throughout the experiment as well and the propionate-degrading acetogen Syntrophobacter fumaroxidans was also detected, although its population size decreased at 5 degrees C.