Formation and growth of heterotrophic aerobic biofilms on small suspended particles in airlift reactors

In this article, the conditions for aerobic biofilm formation on suspended particles, the dynamics of biofilm formation, and the biomass production during the start-up of a Biofilm Airlift Suspension reactor (BAS reactor) have been studied. The dynamics of biofilm formation during start up in the biofilm airlift suspension reactor follows three consecutive stages: bare carrier, microcolonies or patchy biofilms on the carrier, and biofilms completely covering the carrier. The effect of hydraulic retention time and of substrate loading rate on the formation of biofilms were investigated. To obtain in a BAS reactor a high biomass concentration and predominantly continuous biofilms, which completely surround the carrier, the hydraulic retention time must be shorter than the inverse of the maximum growth rate of the suspended bacteria. At longer hydraulic retention times, a low amount of attached biomass can be present on the carrier material as patchy biofilms. During the start-up at short hydraulic retention times the bare carrier concentration decreases, the amount of biomass per biofilm particle remains constant, and biomass increase in the reactor is due to increasing numbers of biofilm particles. The substrate surface loading rate has effect only on the amount of biomass on the biofilm particle. A higher surface load leads to a thicker biofilm.A strong nonlinear increase of the concentration of attached biomass in time was observed. This can be explained by a decreased abrasion of the biofilm particles due to the decreasing concentration of bare carriers. The detachment rate per biofilm area during the start-up is independent of the substrate loading rate, but depends strongly upon the bare carrier concentration.The Pirt-maintenance concept is applicable to BAS reactors. Surplus biomass production is diminished at high biomass concentrations. The average maximal yield of biomass on substrate during the experiments presented in this article was 0.44 +/- 0.08 C-mol/C-mol, the maintenance value 0.019 +/- 0.012 C-mol/(C-mol h). The lowest actual biomass yield measured in this study was 0.15 C-mol/C-mol. (c) 1994 John Wiley & Sons, Inc.     

Influence of detachment,substrate loading and reactor scale on the formation of biofilms in airlift reactors

 For a stable and reliable operation of the biofilm airlift suspension reactor (BAS reactor) means to control biomass concentration, biofilm thickness and biofilm morphology are required. For this reason, the influence of applied detachment forces and surface substrate loading on the formation of heterotrophic biofilms in laboratory-scale BAS reactors was studied. Detachment forces were altered by variation of the initial bare carrier concentration or the superficial air velocity. In addition, the dynamics of biofilm formation during start-up of a full scale BAS reactor (300 m³) was monitored and compared with the laboratory-scale start-up (3 l). This study shows that the biofilm morphology and strength were influenced to a large extent by the surface substrate loading and applied detachment forces. A moderate surface substrate loading and a high detachment force yielded smooth and strong biofilms. The combination of a high surface substrate loading and low detachment forces did lead to rough biofilms, but did not lead to the expected high amount of biomass on the carrier, apparently because of the formation of weaker biofilms. The strength of the bio-films appeared to be related to the detachment forces applied during biofilm formation, in combination with the surface substrate loading. The biofilm morphology and biomass on carrier in the BAS reactor can be controlled using the carrier concentration, substrate loading rate and the superficial air velocity as parameters. The dynamics of biofilm formation during the start-up of a full-scale BAS reactor proved to be similar to heterotrophic biofilm formation in laboratory-scale reactors. This indicates that a model system on the laboratory scale can successfully be applied to predict dynamic phenomena in the full-scale reactor. Received: 31 March 1995/Received revision: 11 August 1995/Accepted: 22 August 1995     

Potential of biogas recirculation to enhance biomass accumulation on supporting media

Two lab-scale anaerobic hybrid reactors (AHR) were operated to investigate the effect of recirculated biogas on the development of biomass on supporting media during the start-up. The reactor comprised of two distinct zones; sludge bed on the bottom and packed bed using nylon fiber as the media on the upper half of the reactor. Both reactors were continuously fed with cassava starch wastewater. The organic loading rate (OLR) was increased from 0.3 to 5.5 g COD/L/day by gradually decreasing the hydraulic retention time (HRT) from 37 to 3.5 days in two months. The biogas at 2.6 L/L/day was recirculated merely in the first month of the operation in order to allow the attached biomass to grow according to the organic matters present in the reactor at the final stage of the start up. Chemical oxygen demand (COD) removal efficiency of over 80% was achieved throughout the study. The result demonstrated a better COD removal efficiency for the reactor with biogas recirculation, especially at low HRTs. The amounts of biomass accumulated on the media in both reactors were slightly different with 11.9 gVSS found on the one with biogas recirculation compared to 9.8 gVSS on the other. In addition, 16.3% increase of the sludge bed was achieved with biogas recirculation as opposed to 9% in the control one. The attached biomass activity test indicated a greater amount and more favorable ratio of the methanogenic bacterial group on the media with the recirculation correlating well to a relatively higher methane content in biogas. As a result, the recirculation of biogas has a potential of improving the characteristics of the AHR especially in terms of biomass accumulation.     

固定载体卧式厌氧反应器处理糖蜜废水的快速启动

为高效处理高浓度有机废水而设计了固定载体卧式厌氧反应器R1和R2, 它是厌氧折流板反应器(ABR)的改进, 以活性炭纤维作为生物膜载体固定并充当反应器的折流板, 在实验室规模上对R1和R2处理糖蜜废水进行快速启动运行。HRT和ORL是影响R1和R2稳定高效运行及启动的2个重要工艺参数。实验证明: HRT为2 d时, 反应器运行最佳。在第30天时, R1的COD去除率达到84.88%, R2达到81.72%。随着进水ORL由1.25 kg/(m3·d)提升到10 kg/(m3·d), 沼气容积产气率由0.35 L/(L·d)逐渐增加到4.98 L/(L·d)。进水pH值为3.9?4.5之间, 整个启动运行过程中, 未调节pH值, R1和R2的出水pH值均在6.7?7.6之间, 2个反应器均有较强的抗酸能力, R1的pH波动更为平缓。在整个实验过程中, 污泥流失量小, 没有发生堵塞现象, 在处理酸性高浓度有机废水时, 2个反应器均表现出较强的抗负荷冲击能力。     

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.     

Organic Matter Degradation Kinetics in an Anaerobic Thermophilic Fluidised Bed Bioreactor

This paper describes the thermophilic anaerobic biodegradation of wine distillery wastewater (vinasses) in a laboratory fluidised bed reactor (AFB) with a porous support medium. The experimental protocol was defined to examine the effect of increasing organic loading rate on the efficiency of AFB and to report on its steady-state performance. Moreover, in order to evaluate treatment efficiency and to investigate fermentation kinetics in an AFB reactor, experimental data were used to estimate the ‘active biomass’ concentration using an autocatalytic kinetic model proposed in this paper, since viable biomass in AFB reactors is very difficult to measure experimentally. The AFB reactor was subjected to a program of steady-state operation over a range of hydraulic retention time (HRTs) of 2.5–0.37 days and organic loading rate (OLRs) up to 5.88 kgCOD/m³/day in order to evaluate its treatment capacity. The AFB reactor was initially operated with organic loading rate of 5.88 kgCOD/m³/day and HRT of 2.5 days. The chemical oxygen demand (COD) removal efficiency was found to be 96.5% in the reactor while the methane content of biogas produced in the digester reached 1.08 m³/m³digester/day. Over 94 days operating period, an OLR of 32 kgCOD/m³/day at a food-to-micro-organisms (F:M) ratio of 0.55 kgCOD/kgVS_att/day was achieved with 81.5% COD removal efficiency in the experimental AFB reactor. At this moment, the methane content of biogas produced in the digester reached 9.0 m³/m³digester/day. The proposed kinetic model is able to estimate kinetic constants of the biodegradation process: non-biodegradable substrate (S_nb) and active adhered biomass concentration (X_a). The parameters of the model were obtained by the curve-fitting method to the proposed kinetic model using the COD as substrate of the anaerobic process and assuming a maximum specific μ_max: 0.72 per day. The comparison of the measured concentration of volatile attached solids (VS_att) with the estimated ‘active’ biomass concentration indicated that extremely high ‘active biomass’ concentrations can be maintained in the system because biofilm thickness is limited by the liquid flow rate applied. This is due to the fact that the anaerobic fluidised bed system retains the growth support medium in suspension by drag forces exerted by upflowing wastewater, and the distribution of biomass holdup (in the form of a biofilm) is thus relatively uniform.     

Performance of inverse anaerobic fluidized bed reactor for treating high strength organic wastewater during start-up phase

The aim of this work is to report on the physical characteristics of carrier material (perlite), biomass growth on the carrier material and the biogas production during an apparent steady state period in an inverse anaerobic fluidized bed reactor (IAFBR) for treating high strength organic wastewater. Before starting up the reactor, physical properties of the carrier material were determined. One millimeter diameter perlite particle is found to have a wet specific density of 295 kg/m(3) with specific surface area of 7.010 m(2)/g. This material has provided a good surface for biomass attachment and development. The biofilm concentration (in terms of attached volatile solids (AVS)) attached to carrier material was found to be 0.66 g(AVS)/g(solid). Most particles have been covered with a thin biofilm of uniform thickness. Once the inverse anaerobic fluidized bed system reached the steady state, the organic load was increased step wise by reducing hydraulic retention time (HRT) from 2 days to 0.16 day, while maintaining the constant feed of chemical oxygen demand (COD) concentration. This system has achieved 84% COD removal and reached the biogas production of 13.22 l/l/d at an organic loading rate (OLR) of 35 kgCOD/m(3)/d.     

A modeling study of anaerobic biofilm systems: II. Reactor modeling

A rigorous steady-state model of anaerobic biofilm reactors taking into account acid-base and gas-phase equilibria in the reactor in conjunction with detailed chemical equilibria and mass transfer in acetate-utilizing methanogenic biofilms is presented. The performances of ideal completely stirred tank reactors (CSTRs) and plug-flow reactors, as well as reactors with nonideal hydraulic conditions, are simulated. Decreasing the surface loading rate increases the acetate removal efficiency, while decreasing the influent pH and increasing the buffering capacity improves the removal efficiency only if the bulk pH of the reactor shifts toward more optimal values between 6.8 to 7.0. The reactor can have negative or positive removal efficiencies depending on the start-up conditions. The respiration coefficient plays a critical role in determining the minimum influent pH required for reactor recovery after failure. Having multiple CSTRs-in-series generally increases the overall removal efficiency for the influent conditions investigated. Monitoring of the influent feed quality is critical for plug-flow reactors, becasue failure of the initial sections of the reactor may cause a cascading effect that may lead to a rapid reactor failure. (c) 1995 John Wiley & Sons, Inc.     

Evaluation of inverse anaerobic fluidized bed reactor for treating high strength organic wastewater

The aim of this work is to evaluate the feasibility of an inverse fluidized bed reactor for the anaerobic digestion of distillery effluent, with a carrier material that allows low energy requirements for fluidization, providing also a good surface for biomass attachment and development. Inverse fluidization particles having specific gravity less than one are carried out in the reactor. The carrier particles chosen for this study was perlite having specific surface area of 7.010 m2/g and low energy requirements for fluidization. Before starting up the reactor, physical properties of the carrier material were determined. One millimeter diameter perlite particle is found to have a wet specific density of 295 kg/m3. It was used for the treatment of distillery waste and performance studies were carried out for 65 days. Once the down flow anaerobic fluidized bed system reached the steady state, the organic load was increased step wise by reducing hydraulic retention time (HRT) from 2 days to 0.19 day, while maintaining the constant feed of chemical oxygen demand (COD) concentration. Most particles have been covered with a thin biofilm of uniform thickness. This system achieved 84% COD removal at an organic loading rate (OLR) of 35 kg COD/m3/d.     

Start-up of an anaerobic hybrid (UASB/filter) reactor treating wastewater from a coffee processing plant

The ability of an anaerobic hybrid reactor, treating coffee wastewater, to achieve a quick start-up was tested at pilot scale. The unacclimatized seed sludge used showed a low specific methanogenic activity of 26.47 g CH4 as chemical oxygen demand (COD)/kg volatile suspended solids (VSS) x day. This strongly limited the reactor performance. After a few days of operation, a COD removal of 77.2% was obtained at an organic loading rate (OLR) of 1.89 kg COD/m3 x day and a hydraulic retention time (HRT) of 22 h. However, suddenly increasing OLR above 2.4 kg COD/m3 x day resulted in a deterioration in treatment efficiency. The reactor recovered from shock loads after shutdowns of 1 week. The hybrid design of the anaerobic reactor prevented the biomass from washing-out but gas clogging in the packing material was also observed. Wide variations in wastewater strength and flow rates prevented stable reactor operation in the short period of the study.     

Granule development in a split-feed anaerobic baffled reactor

Operating anaerobic reactors at high organic loading rates during start-up can lead to instability, accumulation of volatile fatty acids and low pH, such problems being exacerbated in reactors that exhibit plug-flow characteristics. Moreover, plug-flow conditions increase the exposure of biomass to any toxic components in the feed. To overcome these limitations, an anaerobic baffled reactor (ABR), a reactor exhibiting partial plug-flow characteristics, was modified by splitting the feed between the individual compartments to produce the split-feed ABR (SFABR). Consequently, more favourable conditions were created in the initial compartments, such as lower, longer hydraulic retention time and longer cell retention time; conditions in the final compartments were also improved by the increased food availability for microorganisms. Other benefits included better gas mixing characteristics as a result of the more balanced gas production across the reactor. Granule development was compared in SFABR and normally fed ABR by analysing sludge samples, taken during start-up and continuous operation, using scanning electron microscopy. Photomicrographs allowed tentative conclusions to be made concerning the effect of split-feeding on the distribution of bacterial populations within the granule architecture and the role of extracellular polymers on granule formation.     

Biomass retention and performance of anaerobic fixed-film reactors treating acetic acid wastewater

An acetic-acid-based synthetic wastewater of different organic concentrations was successfully treated at 35 degrees C in anaerobic downflow fixed-film reactors operated at high organic loading rates and short hydraulic retention times (HRTs). Substrate removal and methane production rates close to theoretical values of complete volumetric chemical oxygen demand (COD) removal and maximum methane conversion were obtained. A high concentration of biofilm biomass was retained in the reactor. Steady-state biofilm concentration increased with increased organic loading rate and decreased HRTs, reaching a maximum of 8.3 kg VFS/m(3) at a loading rate of 17 kg COD/m(3) day. Biofilm substrate utilization rates of up to 1.6 kg COD/kg VFS day were achieved. Soluble COD utilization rates at various COD concentrations can be described by half-order reaction kinetics.     

生物膜法强化净化氨氮污染水体及其微生物群落解析

【目的】比较不同营养条件及挂膜方式下生物膜法对氨氮污染水体的净化效果及其功能微生物群落结构。【方法】设置空白(Blank)、自然成膜(Raw)、预附脱氮菌强化挂膜(PCC)3组生物膜反应器,利用末端限制性片段长度多态性(T-RFLP)技术和非度量多维标度(NMDS)分析方法对生物膜反应器转化氨氮过程中微生物群落结构及其演替过程进行动态解析。【结果】在C/N=1:1时,除PCC在起始阶段短暂具有较高的氨氮脱除效率外,Blank、Raw和PCC最终均表现出较低的氨氮转化效率(10%-20%)。改变C/N=2:1后,Raw和PCC对人工合成污水中NH4+-N的转化率均提高至95%以上,而且Raw与PCC的群落结构在C/N=2:1时具有较高的相似性,优势菌群主要为γ-变形菌纲(Gammaproteobacteria)、放线菌纲(Actinobacteria)和硝化螺菌纲(Nitrospira)。【结论】C/N是影响生物膜反应器氨氮去除效果及驱动生物膜反应器中细菌群落结构发生改变的重要因子。     

Anammox enrichment from reject water on blank biofilm carriers and carriers containing nitrifying biomass: operation of two moving bed biofilm reactors (MBBR)

The anammox bacteria were enriched from reject water of anaerobic digestion of municipal wastewater sludge onto moving bed biofilm reactor (MBBR) system carriers-the ones initially containing no biomass (MBBR1) as well as the ones containing nitrifying biomass (MBBR2). Duration of start-up periods of the both reactors was similar (about 100?days), but stable total nitrogen (TN) removal efficiency occurred earlier in the system containing nitrifying biomass. Anammox TN removal efficiency of 70% was achieved by 180?days in both 20?l volume reactors at moderate temperature of 26.0°C. During the steady state phase of operation of MBBRs the average TN removal efficiencies and maximum TN removal rates in MBBR1 were 80% (1,000?g-N/m(3)/day, achieved by 308?days) and in MBBR2 85% (1,100?g-N/m(3)/day, achieved by 266?days). In both reactors mixed bacterial cultures were detected. Uncultured Planctomycetales bacterium clone P4, Candidatus Nitrospira defluvii and uncultured Nitrospira sp. clone 53 were identified by PCR-DGGE from the system initially containing blank biofilm carriers as well as from the nitrifying biofilm system; from the latter in addition to these also uncultured ammonium oxidizing bacterium clone W1 and Nitrospira sp. clone S1-62 were detected. FISH analysis revealed that anammox microorganisms were located in clusters in the biofilm. Using previously grown nitrifying biofilm matrix for anammox enrichment has some benefits over starting up the process from zero, such as less time for enrichment and protection against severe inhibitions in case of high substrate loading rates.     

Highly efficient phenol degradation in a batch moving bed biofilm reactor: benefiting from biofilm-enhancing bacteria

In this study, the efficiency improvement of three moving bed biofilm reactors (MBBRs) was investigated by inoculation of activated sludge cells (R1), mixed culture of eight strong phenol-degrading bacteria consisted of Pseudomonas spp. and Acinetobacter spp. (R2) and the combination of both (R3). Biofilm formation ability of eight bacteria was assessed initially using different methods and media. Maximum degradation of phenol, COD, biomass growth and also changes in organic loading shock were used as parameters to measure the performance of reactors. According to the results, all eight strains were determined as enhanced biofilm forming bacteria (EBFB). Under optimum operating conditions, more than 90% of initial COD load of 2795 mg L^?1 was reduced at 24 HRT in R3 while this reduction efficiency was observed in concentrations of 1290 mg L^?1 and 1935 mg L^?1, in R1 and R2, respectively. When encountering phenol loading shock—twice greater than optimum amount-R1, R2 and R3 managed to return to the steady-state condition within 32, 24 and 18 days, respectively. SEM microscopy and biomass growth measurements confirmed the contribution of more cells to biofilm formation in R3 followed by R2. Additionally, established biofilm in R3 was more resistant to phenol loading shock which can be attributed to the enhancer role of EBFB strains in this reactor. It has been demonstrated that the bacteria with both biofilm-forming and contaminant-degrading abilities are not only able to promote the immobilization of other favorable activated sludge cells in biofilm structure, but also cooperate in contaminant degradation which all consequently lead to improvement of treatment efficiency.     

Characterization of aerobic granular sludge at various organic loading rates

Aerobic granular sludge was cultivated from activated sludge with two types of supports, namely bivalve shell carrier (BSC) and anaerobic granules (ANG). Granules were characterized at different organic loading rates (OLRs) ranging from 2.5 to 15 kg COD/m³ d and these granules were observed to withstand high OLRs. The physico-chemical characteristics of the aerobic granules were better than those of seed sludge. The granule formation with ANG support was found to be similar to that of non-support cultivation, i.e. formation from activated sludge only. By contrast, BSC support showed better performance in terms of faster settleability, compactness and especially resistance against organic shock loading. It also enabled self-cleaning effect by removing biofilm attached on the reactor wall during the start-up phase resulting rapid granulation process.     

Anaerobic fluidised bed for the purification of effluents from chemical and mechanical pulping

Anaerobic treatment has seldom been used for wastewaters from the pulp and paper industry and other branches of the chemical industry. Escape of volatile pollutants to the atmosphere, which always occurs during aerobic treatment, is avoided, and much less sludge is being produced than in an aerobic process. The greatest obstacle for using anaerobic treatment in the pulp and paper industry is the large wastewater volume, which necessitates short hydraulic detention times, because the treatment is to occur in an enclosed space. We used solid carrier particles to prevent wash-out of biomass from the reactor at high hydraulic loading, and an up-flow system in order to be able to use very small carrier particles, maximizing the surface area for biomass attachment. In this paper we describe and discuss the results obtained with this type of anaerobic reactor (fluidised bed) at bench and semitechnical scale for wastewaters from pressurized ground wood pulping and paper manufacture, sulphite pulp evaporator condensate and bleach waste. Earlier work with Kraft pulp bleaching effluent and thermomechanical pulping wastewater and evaporator condensates using anaerobic reactors is also discussed. The results obtained thus far show that there are several wastewater streams from the pulping industry, where 60 to 90% of the dissolved organic pollutants (measured as COD_Cr or TOC) was biodegraded within 4 to 24 h. The high strength waste streams (COD_Cr 2000 mg O₂ 1⁻¹) allowed organic space load of 4 to 10 kg COD_Cr m⁻³ reactor volume d⁻¹. With low strength wastes the hydraulic loading was the limiting factor.     

Initial anaerobic biofilm development

Summary The initial rate of methanogen accumulation and anaerobic biofilm accumulation increased with increasing bulk liquid organic space loading and volatile suspended solids concentration. The rates were faster at a 15 day bulk liquid SRT than 5 day SRT. The results indicate that a reduction in start-up time for anaerobic fixed film reactors might be achieved by maintaining high organic space loadings and high concentrations of active microorganisms in the bulk liquid phase during the start-up period.     

Startup of anaerobic fluidized bed reactors with acetic acid as the substrate

The startup of anaerobic fluidized bed reactors, which use Manville R-633 beads as the growth support media, acetate enriched bacterial culture as the inoculum, and acetic acid as the sole substrate, is studied. Tow startup strategies are evaluated: one based on maximum and stable substrate utilization and another based on maximum substrate loading controlled by reactor pH. The startup process is characterized using a number of operational parameters.The reactors again excellent total organic carbon (TOC) removal (i.e., > 97% at a feed concentration of 5000 mg TOC/L) and stable methane production (i.e., 0.90 L CH(4)/g TOC, where TOC(r) is TOC removed) at a early stage of the startup process, regardless of the strategies applied. The loading can be increased rapidly without the danger of being overloaded. Significant losses of growth support media and biomass caused by gas effervescence at higher loadings limits the maximum loading that can be safely applied during startup process.A high reactor immobilized biomass inventory is achievable using the porous growth support media (e.g., Manville 633 beads). A rapid increase in loading creates a substrate rich environment that yields more viable reactor biomass. Both substrate utilization rate (batch and continuous) and immobilized biomass inventory stabilize concomitantly at the late stage of the startup process, indicating the attainment of steady-state conditions in reactors. Therefore, they are better parameters that TOC removal and methane production for characterizing the entire startup process of aerobic fluidized bed reactor.The strategy based on maximum substrate loading controlled by reactor pH significantly shortens the startup time. In this case, the reactor attains steady-state conditions approximately 140 days after startup. On the other hand, a startup time of 200 days is required when the strategy based maximum substrate utilization is adopted. (c) 1993 John Wiley & Sons, Inc.     

Granule development and performance in sucrose fed anaerobic baffled reactors

Two 90 L anaerobic baffled reactors were used to study the granulation of sludge and the effect of the organic loading rate and NaHCO3/COD ratios on reactor performance. Furthermore, it was determined whether an anaerobic baffled reactor would promote phase separation and if additive of bentonite or granular active carbon was capable of enhancing granule formation. In order to minimize feed variations, and have a totally biodegradable substrate, a synthetic sucrose substrate was used. Granulation was achieved in both reactors within 75 days. However, the granules from the granular active carbon amended reactor appeared earlier and were larger and more compact. The reactors were maintained at a hydraulic retention time of 20 h during performance study stage. The results showed that when organic loading rate were changed from 2.15 to 6.29 kg COD m(-3)day(-1), chemical oxygen demand (COD) removal was not decreased (91-93%), but a slight increase in effluent COD was observed. It was found that the COD removals were generally good (87-92%) and had not obviously change with the decreasing NaHCO3/COD ratios. From the bacterial distribution and the concentration of volatile fatty acids in four compartments, it was concluded that a separation of phases occurred within the anaerobic baffled reactors.