Interactions between methanogenic,sulfate-reducing and syntrophic acetogenic bacteria in the anaerobic degradation of benzoate

Effect of sulfate on the anaerobic degradation of benzoate was investigated by using the chemostat-type reactors at 35°C. The benzoate concentrations were equivalent to 1250–10000 mg.l⁻¹ in COD (chemical oxygen demand) and the sulfate concentrations were equivalent to 167–1670 mg.l⁻¹ in sulfur (S). Interactions between the methane-producing bacteria (MPB) and sulfate-reducing bacteria (SRB) were dependent strongly on the ratio of COD/S in wastewater. The MPB consumed 99% of the available electron donors at COD/S ratio of 60, but consumed only 69% at ratio of 1.5, and 13% at 0.75. The biochemical reactions and the bacterial composition in the biomass were also governed by the COD/S ratio. At high COD/S ratios (3.0 or higher), benzoate was degraded mainly to methane via acetate and hydrogen/formate. The degradation of benzoate required the syntrophic association between the hydrogen-producing acetogens such as Syntrophus buswellii and hydrogen-consuming MPB, plus Methanothrix-like MPB. On the other hand, at low COD/S ratio (1.5 or lower), benzoate was consumed mainly by SRB, converting sulfate into sulfide and suppressing the methane production. The anaerobic degradation of benzoate was partially inhibited when sulfide concentration was high.     

Long term competition between sulfate-reducing and methane-producing bacteria for acetate in anaerobic biofilm

A long term competition between sulfate-reducing bacteria (SRB) and methane-producing bacteria (MPB) for acetate was investigated using a laboratory scale anaerobic fluidized bed. When the synthetic wastewater composed of acetate and sulfate was fed at a low organic loading rate, averages of the remaining acetate and sulfate concentrations were 1.7 mg Cl⁻¹ and 78.5 mg l⁻¹, respectively. During several months of this acetate limited operation the methane production rate as well as the microbial mass of MPB declined gradually, whereas the amount of reduced sulfate along with the microbial mass of SRB increased, which apparently indicates that SRB out-compete MPB in the biofilm at lower acetate concentrations. On the other hand, MPB were able to form a biofilm faster than SRB at higher acetate concentrations presumably due to MPB's higher ability to adhere carrier surfaces compared with SRB. Kinetic constants for both species in the biofilm were determined and compared with those reported for pure MPB and SRB cultures. Based on the kinetic mechanism of this competition, operational conditions which would support methanogenesis by suppressing sulfate reduction were identified.     

Effect of specific gas loading rate on thermophilic (55 degrees C) acidifying (pH 6) and sulfate reducing granular sludge reactors

The effect of the specific gas loading rate on the acidifying, sulfate reducing and sulfur removal capacity of thermophilic (55 degrees C; pH 6.0) granular sludge bed reactors treating partly acidified wastewater was investigated. A comparison was made between a regular UASB reactor and a UASB reactor continuously sparged with N(2) at a specific gas loading rate of 30 m(3)m(-2)d(-1). Both UASB reactors (upflow velocity 1.0 mh(-1), hydraulic retention time about 5h) were fed a synthetic wastewater containing starch, sucrose, lactate, propionate and acetate and a low sulfate concentration (COD/SO(4)(2-) ratio of 10) at volumetric organic loading rates (OLR) ranging from 4.0 to 49.8 gCODl(-1) reactord(-1). Immediately after imposing an OLR of 25 gCODl(-1) reactord(-1), the acidification and sulfate reduction efficiency dropped to 80% and 30%, respectively, in the UASB reactor. Both efficiencies recovered slowly to 100% during the course of the experiment. In the N(2) sparged reactor, both the acidification and sulfate reduction efficiency remained 100% following the OLR increase to 25 gCODl(-1) reactord(-1). However, the sulfate reduction efficiency gradually decreased to about 20% at the end of the experiment. The biogas (CO(2) and CH(4)) production rate in the UASB was very low, i.e. <3l biogasl(-1) reactorday(-1), resulting in negligible amounts (<20%) of H(2)S stripped from the reactor liquid. The total H(2)S concentration of the N(2) sparged UASB reactor effluent was always below 25 mgl(-1), but incomplete sulfate reduction kept the maximal H(2)S stripping efficiency below 70%.     

Adaptation of denitrifying bacteria to acetate and methanol in activated sludge

Adaptation of denitrifying bacteria to the external carbon sources acetate and methanol was studied in bench-scale sequencing batch reactors. The reactors were seeded with sludge from a full-scale pre-denitrifying activated sludge plant and operated in cycles consisting of a 23.5 h anoxic period followed by 23 h aeration. A control reactor received no additional carbon. Potential denitrification rate, measured with the acetylene inhibition technique, and the most probable number of bacteria denitrifying with the specific carbon source were followed for 52 days. Actual rates in the reactors were estimated from cumulative gas production during anoxic operation. A period of adaptation was necessary when methanol or acetate was the supplemental carbon source. Adaptation to acetate was also observed in the control reactor. The acetate sludge was, however, probably better adapted to acetate as evidenced by the greater increase in activity per bacterium. The increase continued even after the maximum denitrification rate was reached. During adaptation to methanol the methanol denitrifying capacity per bacterium was fairly constant while the potential rate increased after an initial lag-phase. Sewage water without external carbon did not cause an increased methanol activity that could be interpreted as adaptation. We suggest that the bacteria denitrifying with acetate could be the same as those using the electron donors in the sewage. The bacteria in the methanol reactor seemed to consist of one population denitrifying with methanol and another with compounds in the sewage.     

Conversion of acetate, propionate and butyrate to methane under thermophilic conditions in batch reactors

These experiments were performed to determine the degradation of VFA (acetate, propionate and butyrate) and the maximum methanogenic activity of granular sludge from the thermophilic anaerobic digestion of pure molasses. The compositions of acetate, propionate and butyrate used as substrate were 25:35:40. The tests were performed at constant temperature (55°C) and pH 7 on two duplicate batch reactors (I and II) running in parallel and were repeated to show the effect of acclimatization. During the first feeding, there was a significant lag phase and after about 23 h incubation the volumes of CH₄ gases produced from two reactors were only about 20 and 490 ml, respectively. In this experiment, propionate was converted to acetate only after the initial concentrations of butyrate and acetate had completely degraded. Acetate formed from propionate was immediately converted to methane and carbon dioxide. The maximum methanogenic activities of the first feeding were not high because the natural populations of the propionate-degrading bacteria were low and the sludge adapted itself to the VFAs' substrate very slowly. In a second experiment with the same sludge, the maximum methanogenic activities of the second feedings were about 1.60 times higher than those of the first feedings because of the adaptation of the sludge and increase of populations of the propionate-degrading bacteria.     

Sulfidogenic fluidized-bed treatment of metal-containing wastewater at 8 and 65 degrees C temperatures is limited by acetate oxidation

Acetate utilization in sulfidogenic fluidized-bed reactors (FBRs) was investigated for the treatment of iron containing wastewater at low (8 degrees C) and high (65 degrees C) temperatures. The FBRs operated at low and high temperatures were inoculated with cultures of sulfate-reducing bacteria (SRB) originally enriched from arctic and hot mining environments, respectively. Acetate was not utilized as a carbon and electron source for SRB at 8 degrees C. With ethanol, hydrogen sulfide was produced from ethanol to acetate oxidation, which precipitated the iron. Then, several attempts were made to obtain acetate oxidation at 8 degrees C. Inoculation of two different low temperature enrichments and operating the FBR for a long period of time (321 days) did not result in enrichment of acetate oxidizing SRB. Due to the absence of acetate oxidation at 8 degrees C, external alkalinity addition was required to keep the pH neutral. At 65 degrees C, average acetate and sulfate removals were 52+/-12% and 24+/-8% at 670 mg/Ld acetate and 1500 mg/Ld sulfate loadings, respectively. The produced alkalinity from acetate oxidation increased the pH from 6.4 to around 7.5 and electron flow to sulfate reduction averaged 65%. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed quite stable SRB community at 8 degrees C, whereas, at 65 degrees C SRB community was dynamic. In the FBRs, Desulfomicrobium apsheronum and Desulfosporosinus sp. at 8 degrees C and Desulfotomaculum sp. at 65 degrees C were detected.     

厌、好氧周期循环下的污泥颗粒化过程

为研究厌氧／好氧周期循环条件下厌氧快速吸收工艺中的污泥颗粒化过程、成因及影响因素，分别采用葡萄糖、乙酸钠人工配水及实际城市污水进行了试验。结果发现，形成的好氧颗粒污泥呈球形或椭球形，致密且边界清晰，其中葡萄糖配水的污泥粒径为0．5～0．8mm，最大可达1．0mm，SVI值为25～30mL／g；乙酸钠配水的污泥粒径为0．2～0．4mm，SVI值为40mL／g左右；实际城市污水的污泥经过短期运行即开始出现小颗粒，SVI值为60mL／g左右。3种污泥均具有良好的厌氧COD吸收活性。     

Tannery effluent as a carbon source for biological sulphate reduction

Tannery effluent was assessed as a carbon source for biological sulphate reduction in a pilot-scale upflow anaerobic sludge blanket (UASB), stirred tank reactor (STR) and trench reactor (TR). Sulphate removals of between 60-80% were obtained in all three reactors at total sulphate feed levels of up to 1800 mg l(-1). Sulphate removal in the TR (400-500 mg SO4 l(-1) day(-1)) and UASB (up to 600 mg SO4 l(-1) day(-1)) were higher than those obtained in the STR (250 mg SO4 l(1) day(-1)). A change in operation mode from a UASB to a STR had a large impact on chemical oxygen demand (COD) removal efficiencies. COD removal rates decreased by 25% from 600-700 mg COD l(-1) day(-1) to 200-600 mg COD l(-1) day(-1). The TR had an average COD removal rate of 500 mg COD l(-1) day(-1). Large quantities of sulphide were produced in the reactors (up to 1500 mg l(-1)). However due to the elevated pH in the reactor, only a small amount was in the form of H2S and thus the odour problem normally associated with biological sulphate reduction was not present.     

Optimization of the hydrolytic-acidogenic anaerobic digestion stage (55 degrees C) of sewage sludge: influence of pH and solid content

In conventional single-stage anaerobic digestion processes, hydrolysis is regarded as the rate-limiting step in the degradation of complex organic compounds, such as sewage sludge. Two-stage systems have been proposed to enhance this process. However, so far it is not clear which are the best conditions for a two-stage anaerobic digestion process of sewage sludge, in terms of temperature and hydraulic retention time of each stage. The aim of this work was to determine the optimal conditions for the hydrolytic-acidogenic stage treating real sludge with a high concentration of total solids (40-50gL(-1)) and volatile solids (25-30gL(-1)), named high concentration sludge. The variables considered for this first stage were: hydraulic retention time (1-4 days) and temperature (55 and 65 degrees C). Maximum volatile fatty acids generation was obtained at 4 days and 3 days hydraulic retention time for 55 degrees C and 65 degrees C, respectively. Consequently, 4 days hydraulic retention time and temperature of 55 degrees C were set as the working conditions for the hydrolytic-acidogenic stage treating high concentration sludge. The results obtained when operating with high concentration sludge were compared with a low concentration sludge consisting of 17-28gL(-1) total solids and 13-21gL(-1) volatile solids. The effect of decreasing the influent sludge pH, when working at the optimal conditions established, was also evaluated.     

Mixed carbon sources for nitrate reduction in activated sludge-identification of bacteria and process activity studies

Mixtures of methanol and acetate as carbon source were investigated in order to determine their capacity to enhance denitrification and for analysis of the microbial composition and carbon degradation activity in activated sludge from wastewater treatment plants. Laboratory batch reactors at 20 degrees C were used for nitrate uptake rate (NUR) measurements in order to investigate the anoxic activity, while single and mixed carbon substrates were added to activated sludge. Microautoradiography (MAR) in combination with fluorescence in situ hybridisation (FISH) were applied for microbial analysis during exposure to different carbon sources. The NUR increased with additions of a mixture of acetate and methanol compared with additions of a single carbon source. MAR-FISH measurements demonstrated that the probe-defined group of Azoarcus was the main group of bacteria utilising acetate and the only active group utilising methanol under anoxic conditions. The present study indicated an improved denitrification potential by additions of a mixed carbon source compared with commonly used single-carbon additions. It is also established that Azoarcus bacteria are involved in the degradation of both acetate and methanol in the anoxic activated sludge.     

Silage supports sulfate reduction in the treatment of metals- and sulfate-containing waste waters

Silage was used as source of carbon and electrons for enrichment of silage-degrading and sulfate reducing bacteria (SRB) from boreal, acidic, metals-containing peat-bog samples and to support their use in batch and semi-batch systems in treatment of synthetic waste water. Sulfidogenic silage utilization resulted in a rapid decrease in lactate concentrations; concentrations of acetate, butyrate and propionate increased concomitantly. Synthetic waste water consisting of Mn, Mg and Fe (II) ions inhibited sulfate reduction at concentrations of 6 g/l, 8 g/l and 1 g/l respectively. During treatment, Mn and Mg ions remained in solution while Fe ions partially precipitated. Up to 87 mg sulfate was reduced per gram of silage. Sulfate reduction rates of 34, 22 and 6 mg/l/day were obtained at temperatures of 30, 20 and 9 °C respectively. In semi-batch reactors operated at low pH, the iron precipitation capacity was controlled by sulfate reduction rates and by partial loss of hydrogen sulfide to the gas phase. Passive reactor systems should, therefore, be operated at neutral pH. Metals tolerant, silage-fermenting (predominantly species belonging to genus Clostridium) and sulfate reducing bacteria (including a species similar to the psychrotolerant Desulfovibrio arcticus) were obtained from the peat bog samples. This work demonstrates that silage supports sulfate reduction and can be used as a low cost carbon and electron source for SRB in treatment of metals-containing waste water.     

Comparative study of laboratory-scale thermophilic and mesophilic activated sludge processes

Laboratory-scale mesophilic (20-35 degrees C) and thermophilic (55 degrees C) activated sludge processes (ASPs) treating diluted molasses wastewater were compared in effluent quality, removal of different COD fractions, sludge yield, floc size, and sludge settleability. The effect of polyaluminium chloride (PAC) with high cationic charge on sludge settleability and effluent quality was also studied. In the ASPs, the hydraulic retention time was 12h in both processes, corresponding to a volumetric loading rate of 3.2+/-1.0 kg COD(filt) m(-3)d(-1). The mesophilic ASP gave 79+/-18% and 90+/-2% and the thermophilic ASP gave 50+/-6% and 67+/-11% total COD (COD(tot)) and GF/A-filtered COD (COD(filt)) removals, respectively. The higher COD values in the thermophilic effluent were due to dispersed particles, such as free bacteria, measured as colloidal COD (COD(col)=difference between soluble COD (COD(sol)) and COD(filt)). Both ASPs gave high (90%) COD(sol) removals. Aeration (24-h) of the thermophilic ASP effluent at 35 degrees C reduced the density of the free bacteria by half, which was also measured as 90% COD(col) removal. Post-aeration of thermophilic effluent at 55 degrees C had no influence either on the density of free bacteria or on the COD(col) values. The use of PAC did not increase COD(tot) or COD(filt) removals in either process, but reduced the mesophilic and increased the thermophilic sludge volume index values.     

Effect of EDTA and Fe-EDTA complex concentration on TCF Kraft mill effluent degradability. Batch and continuous treatments

The effect of ethylenediaminetetracetic acid (EDTA) and Fe-EDTA complex on synthetic totally chlorine-free (TCF) effluent degradability in batch and continuously operating reactors was evaluated. Under batch treatment, the addition of EDTA and Fe-EDTA complex was studied in the range of 80 to 320 mg l(-1). Under continuously operated reactors, the Fe-EDTA complex concentration varied from 20 to 80 mg l(-1), and the hydraulic retention time (HRT) varied from 48 to 24 h. Sludge oxygen uptake rate (OUR) and chemical oxygen demand (COD) removal decreased when EDTA concentration increased in the influent under batch treatment; however, this inhibitory effect was reduced by the addition of Fe-EDTA complex. Without the addition of EDTA, COD removal decreased from 71% to 8%. The most efficient EDTA removal treatment (almost 10%) was the treatment of 80 mg l(-1) Fe-EDTA. Under continuously operated reactors, COD removal was greater than 57% in the synthetic TCF effluent with a Fe-EDTA concentration that varied from 20 to 80 mg l(-1); however, EDTA removal was lower than 25% in all cases. Synthetic TCF effluent with a Fe -EDTA concentration higher than 80 mg l(-1) could not be treated by the activated sludge treatment due to EDTA's inhibitory effect on the sludge.     

Effects of carbon source on denitrification efficiency and microbial community structure in a saline wastewater treatment process

Two different denitrifying reactors were monitored in order to evaluate the effects of carbon source on denitrification efficiency and microbial community structure under various saline conditions. Nitrogen removal performances were determined when salinity concentrations increase gradually in acetate- or methanol-fed denitrifying reactor. As a result, acetate-fed process attained high nitrate removal at 0-10% NaCl, while methanol was proven beneficial electron donors at 0-3% NaCl. A parallel analysis of T-RFLP and cloning in the acetate-fed sludge showed that a specialized microbial population (i.e. the genera Halomonas and Marinobacter) adapted to a high saline environment. Meanwhile, there were no major changes of bacterial populations in the methanol-fed reactor at 4% NaCl, although the relative abundances of the genera Azoarcus and Methylophaga increased when salinity concentration was at 1-3% NaCl, indicating that methanol-utilizing populations in activated sludge was unable to adapt to a high saline environments (>4% NaCl).     

Effects of substrate concentrations on the growth of heterotrophic bacteria and algae in secondary facultative ponds

This paper presents the effect of substrate concentration on the growth of a mixed culture of algae and heterotrophic bacteria in secondary facultative ponds (SFPs) utilizing settled domestic sewage as a sole source of organic carbon. The growth of the mixed culture was studied at the concentrations ranging between 200 and 800 mg COD/l in a series of batch chemostat reactors. From the laboratory data, the specific growth rate (micro) was determined using the modified Gompertz model. The maximum specific growth rate ( micro(max)) and half saturation coefficients (K(s)) were calculated using the Monod kinetic equation. The maximum observed growth rate ( micro(max)) for heterotrophic bacteria was 3.8 day(-1) with K(s) of 200 mg COD/l. The micro(max) for algal biomass based on suspended volatile solids was 2.7 day(-1) with K(s) of 110 mg COD/l. The micro(max) of algae based on the chlorophyll-a was 3.5 day(-1) at K(s) of 50mg COD/l. The observed specific substrate removal by heterotrophic bacteria varied between the concentrations of substrate used and the average value was 0.82 (mg COD/mg biomass). The specific substrate utilization rate in the bioreactors was direct proportional to the specific growth rate. Hence, the determined Monod kinetic parameters are useful for the definition of the operation of SFPs.     

Integration of anammox into the aerobic granular sludge process for main stream wastewater treatment at ambient temperatures

Anaerobic ammonium oxidation, nitrification and removal of COD was studied at ambient temperature (18 °C ± 3) in an anoxic/aerobic granular sludge reactor during 390 days. The reactor was operated in a sequencing fed batch mode and was fed with acetate and ammonium containing medium with a COD/N ratio of 0.5 [g COD/gN]. During influent addition, the medium was mixed with recycled effluent which contained nitrate in order to allow acetate oxidation and nitrate reduction by anammox bacteria. In the remainder of the operational cycle the reactor was aerated and controlled at a dissolved oxygen concentration of 1.5 mg O₂/l in order to establish simultaneous nitritation and Anammox. Fluorescent in-situ hybridization (FISH) revealed that the dominant Anammox bacterial population shifted toward Candidatus “Brocadia fulgida” which is known to be capable of organotrophic nitrate reduction. The reactor achieved stable volumetric removal rates of 900 [g N₂-N/m³/day] and 600 [g COD/m³/day]. During the total experimental period Anammox bacteria remained dominant and the sludge production was 5 fold lower than what was expected by heterotrophic growth suggesting that consumed acetate was not used by heterotrophs. These observations show that Anammox bacteria can effectively compete for COD at ambient temperatures and can remove effectively nitrate with a limited amount of acetate. This study indicates a potential successful route toward application of Anammox in granular sludge reactors on municipal wastewater with a limited amount of COD.     

Involvement of protozoa in anaerobic wastewater treatment process

It is only very rarely recognised in literature that anaerobic reactors may contain protozoa in addition to various bacterial and archeal groups. The role of protozoa in anaerobic degradation was studied in anaerobic continuous stirred tank reactors (CSTR) and batch tests. Anaerobic protozoa, especially the ciliated protozoa, have direct influence on the performance of CSTR at all organic loading rates (1–2 g COD l⁻¹ d⁻¹) and retention times (5–10 days). The studies revealed that chemical oxygen demand (COD) removal is strongly correlated to ciliate density in CSTR fed with oleate (suspended COD) and acetate (soluble COD). There was no significant difference in COD removal between reactors fed suspended COD and those fed soluble COD. However, the diversity and number of ciliates is greater in CSTR fed with particulate feed. The mixed liquor suspended solids (MLSS) representing biomass was significantly lower (16–34%) in CSTR with protozoa. In batch tests, increased COD removal and methane production was observed in sludge having ciliates as compared with sludge without protozoa. Methane production increased linearly with number of ciliates (R²=0.96) in batch tests with protozoa. Direct utilization of COD by flagellates and ciliates was observed in bacteria-suppressed cultures. The technological importance of these results is that reactors with protozoa-rich sludge can enhance the rate of mineralization of complex wastewater, especially wastewater containing particulate COD.     

Anaerobic stabilisation and conversion of biopolymers in primary sludge--effect of temperature and sludge retention time

The effect of sludge retention time (SRT) and process temperature on the hydrolysis, acidification and methanogenesis of primary sludge was investigated in completely stirred tank reactors (CSTRs). The CSTRs were operated to maintain SRTs of 10, 15, 20 and 30 days at process temperatures of 25 degrees C and 35 degrees C. The rates of hydrolysis and the biodegradability of primary sludge were assessed in batch reactors incubated at 15 degrees C, 25 degrees C and 35 degrees C. The results revealed that the major amount of sludge stabilisation occurred between 0 and 10 days at 35 degrees C and 10 and 15 days at 25 degrees C. Hydrolysis was found to be the rate limiting-step of the overall digestion process, for the reactors operated at 35 degrees C and 25 degrees C, except for the reactor operated at 10 days and 25 degrees C. At the latter conditions, methanogenesis was the rate-limiting step of the overall digestion process. Proteins hydrolysis was limited to a maximum value of 39% at 30 days and 35 degrees C due to proteins availability in the form of biomass. The biodegradability of primary sludge was around 60%, and showed no temperature dependency. The hydrolysis of the main biopolymers and overall particulate COD of the primary sludge digested in CSTRs were well described by first-order kinetics, in case hydrolysis was the rate-limiting step. Similarly, the hydrolysis of the overall particulate COD of the primary sludge digested in batch reactors were described by first-order kinetics and revealed strong temperature dependency, which follows Arrhenius equation.     

Study of anaerobic lactate metabolism under biosulfidogenic conditions

Biological sulfate reduction (BSR) has been reported to have potential for the treatment of acid mine drainage (AMD). The provision of a suitable carbon source and electron donor for this process remains a challenge. Lactate offers potential advantages as carbon source and electron donor in the biological sulfate reduction process. As this substrate is utilized by both fermentative bacteria and oxidative sulfate-reducing bacteria (SRB), the effect of feed sulfate concentration on the lactate pathways utilized under biosulfidogenic conditions was investigated. Studies were carried out in chemostat bioreactors across a range of residence times, using an enriched culture of SRB. The stoichiometry of biological sulfate reduction was affected by feed sulfate concentration and dilution rate. Incomplete oxidation of lactate was dominant at low feed sulfate concentration (1.0 g/L), while the yield of propionate from lactate metabolism increased at feed sulfate concentrations of 2.5-10.0 g/L, indicating the occurrence of lactate fermentation. Furthermore, at each sulfate feed concentration, in the range 2.5-10.0 g/L, the ratio in which lactate was metabolized by the oxidative and fermentative pathways varied with varying dilution rates. Lactate oxidation was higher at a feed sulfate concentration of 10.0 g/L relative to 2.5 and 5.0 g/L. The volumetric lactate utilization rate was enhanced by increasing the feed sulfate concentration. However, the proportion of total lactate consumed that was channelled into providing electrons for other activities apart from sulfate reduction also increased over the range of increasing sulfate concentrations studied and appeared to be a function of residual lactate and sulfide concentrations.     

Mesophilic and thermophilic activated sludge post-treatment of paper mill process water

Increasing system closure in paper mills and higher process water temperatures make the applicability of thermophilic treatment systems increasingly important. The use of activated sludge as a suitable thermophilic post-treatment system for anaerobically pre-treated paper process water from a paper mill using recycled wastepaper was studied. Two lab-scale plug flow activated sludge reactors were run in parallel for 6 months; a thermophilic reactor at 55 degrees C and a reference reactor at 30 degrees C. Both reactors were operated simultaneously at 20, 15 and 10 days SRT. The effects of temperature and SRT on sludge settleability and chemical oxygen demand (COD) removal efficiencies of different fractions were studied. Total COD removal percentages over the whole experimental period were 58+/-5% at 30 degrees C and 48 +/- 10% at 55 degrees C. The effect of the SRT on the total COD removal was negligible. Differences in total COD removal between both systems were due to a lesser removal of soluble and colloidal COD at 55 degrees C compared to the reference system. At 30 degrees C, colloidal COD removal percentages were 65+/-25%, 75+/-17% and 86+/-22% at 20, 15 and 10 days SRT, respectively. At 55 degrees C, these percentages were 48+/-34%, 40+/-28% and 70+/-25%, respectively. The effluent concentrations of colloidal COD in both systems were related to the influent concentration of colloidal material. The thermophilic sludge was not able to retain influent colloidal material as well as the mesophilic sludge causing a higher thermophilic effluent turbidity. Sludge settling properties were excellent in both reactor systems. These were neither temperature nor SRT dependent but were rather caused by extensive calcium precipitation in the aeration tanks creating a very dense sludge. For application in the board industry, a thermophilic in line treatment system seems feasible. The higher effluent turbidity is most likely offset by the energy gains of treatment under thermophilic conditions.