Effect of bioaugmentation of activated sludge with white-rot fungi on olive mill wastewater detoxification

AIMS: To test the potential use of Phanerochaete chrysosporium and other white-rot fungi to detoxify olive mill wastewaters (OMW) in the presence of a complex activated sludge. To combine the aerobic with anaerobic treatment to optimize the conversion of OMW in biogas. METHODS AND RESULTS: A 25-l air lift reactor was used to pretreat OMW by white-rot fungi. Detoxification of the OMW was monitored by size exclusion HPLC analysis, chemical oxygen demand (COD)/biological oxygen demand (BOD(5)) ratio evolution, and bioluminescence toxicity test. Anaerobic treatment of OMW was performed in a 12-l anaerobic filter reactor. Efficiency of the treatment was evaluated by organic matter removal, and biogas production. By comparison with the pretreatment by activated sludge only, the bioaugmentation with Phanerochaete chrysosporium or Trametes versicolor led to high removal of organic matter, decreased the COD/BOD(5) ratio and the toxicity. The subsequent anaerobic digestion of the OMW pretreated with activated sludge-white-rot fungi showed higher biomethanization yields than that pretreated with activated sludge only. Higher loading rates (7 g COD l(-1) day(-1)) were reached without any acidification or inhibition of biomethanization. CONCLUSIONS: The use of white-rot fungi, even in the presence of complex biological consortia to detoxify OMW, proved to be possible and made the anaerobic digestion of OMW for methane production feasible. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of fungi for OMW reuse and energy production could be adapted to industrial applications.     

The effect of pretreatment on anaerobic activity of olive mill wastewater using batch and continuous systems

This study was focused on several physico-chemical and biological treatment methods that may affect the reduction of the organic load in olive mill wastewater (OMW). In this study, removal of 95% of the phenolic compounds present in OMW was achieved using sand filtration and subsequent treatment with powdered activated carbon in a batch system. This pretreatment for OMW was found to enhance the anaerobic activity of the sludge in the batch system significantly. The efficiency of organic load removal achieved by the anaerobic treatment of untreated OMW in batch reactors with tap water dilution factors below 1:10, reached approximately 65% chemical oxygen demand (COD) removal. However, in the up-flow sludge anaerobic blanket (UASB) reactor, COD removal efficiency of 80–85% was reached at a hydraulic retention time (HRT) of 5 days with an influent COD concentration of 40 g l⁻¹ and organic loading rate (OLR)=8 g⁻¹ COD l⁻¹ per day.     

Performances and microbial features of an aerobic packed-bed biofilm reactor developed to post-treat an olive mill effluent from an anaerobic GAC reactor

Background  

Olive mill wastewater (OMW) is the aqueous effluent of olive oil producing processes. Given its high COD and content of phenols, it has to be decontaminated before being discharged. Anaerobic digestion is one of the most promising treatment process for such an effluent, as it combines high decontamination efficiency with methane production. The large scale anaerobic digestion of OMWs is normally conducted in dispersed-growth reactors, where however are generally achieved unsatisfactory COD removal and methane production yields. The possibility of intensifying the performance of the process using a packed bed biofilm reactor, as anaerobic treatment alternative, was demonstrated. Even in this case, however, a post-treatment step is required to further reduce the COD. In this work, a biological post-treatment, consisting of an aerobic biological "Manville" silica bead-packed bed aerobic reactor, was developed, tested for its ability to complete COD removal from the anaerobic digestion effluents, and characterized biologically through molecular tools.     

Long-term impact of dissolved O(2) on the activity of anaerobic granules

The impact of influent dissolved O(2) on the characteristics of anaerobic granular sludge was investigated at various dissolved O(2) concentrations (0.5-8.1 ppm) in 1- and 5-L laboratory-scale upflow anaerobic sludge bed (UASB)-like anaerobic/aerobic coupled reactors with a synthetic wastewater (carbon sources containing 75% sucrose and 25% acetate). The rate of dissolved O(2) supplied to the coupled reactor was as high as 0.40 g O(2)/L(rx).d, and the anaerobic/aerobic coupled reactors maintained excellent methanogenic performances at a COD loading rate of 3 g COD/L(rx).d even after the reactors had been operated with dissolved O(2) for 3 months. The activities of granular sludge on various substrates (glucose, propionate, and hydrogen) were not impaired, and acetate activity was even improved over a short term. However, after 3 months of operation, slight declines on the acetoclastic activities of granules were observed in the coupled reactor receiving the recirculated fluid containing 8.1 ppm dissolved O(2).Methane yield in the anaerobic control reactor and anaerobic/aerobic coupled reactors revealed that a significant aerobic elimination (up to 30%) of substrate occurred in the coupled reactors, as expected. The presence of dissolved O(2) in the recirculated fluid resulted in the development of fluffy biolayers on the granule surface, which imposed a negative impact on the settleability of granular sludge and caused a slightly higher sludge washout. This research shows that the anaerobic/aerobic coupled reactor can be successfully operated under O(2)-limited conditions and is an ideal engineered ecosystem integrating oxic and anaerobic niches. (c) 1996 John Wiley & Sons, Inc.     

Anaerobic granule development for removal of pentachlorophenol in an upflow anaerobic sludge blanket (UASB) reactor

The granulation process using synthetic wastewater containing pentachlorophenol (PCP) in four 1.1 l laboratory scale upflow anaerobic sludge blanket (UASB) reactors was studied, and the anaerobic biotransformation of PCP during the granulation process investigated. After 110 days granular sludge was developed and up to 160 and 180 mg/l of PCP was added into the reactors R1 and R2, respectively, when they were inoculated with acclimated anaerobic sludge from an anaerobic digester of a citric acid plant. The inoculum was predominately composed of bacilli and filamentous bacteria. Granulation did not occur in reactors R3 and R4 which were inoculated with acclimated anaerobic sludge from aerobic sludge of the municipal sewage treatment plant which consisted mainly of cocci. Despite similar bacilli in the granule, the filamentous bacteria from reactor R1 were thicker than those of reactor R2. The granular sludge had a maximum diameter of 2.5 and 2.2 mm, and SMA of 1.44 and 1.32 gCOD/gTVS per day for reactors R1 and R2, respectively. Over 98% chemical oxygen demand (COD) removal rate and 99% of PCP removal rate were achieved when reactors R1 and R2 were operated at PCP and COD loading rates of 150 and 7.5 g/l per day, respectively. H₂-producing acetogens were the dominant anaerobes in the granular sludge.     

Distribution and change of microbial activity in combined UASB and AFB reactors for wastewater treatment

A thermophilic upflow anaerobic sludge blanket (UASB) reactor was combined with a mesophilic aerobic fluidized bed (AFB) reactor for treatment of a medium strength wastewater with 2,700?mg COD?l^?1. The COD removal efficiency reached 75% with a removal rate of 0.2 g COD?l^?1 h^?1 at an overall hydraulic retention time 14 hours. The distribution of microbial activity and its change with hydraulic retention time in the two reactors were investigated by measuring ATP concentration in the reactors and specific ATP content of the biomass. In the UASB reactor, the difference in specific ATP was significant between the sludge bed and blanket solution (0.02?mg ATP g VS^?1 versus 0.85?mg ATP g VS^?1) even though the ATP concentrations in these two zones were similar. A great pH gradient up to 4 was developed along the UASB reactor. Since a high ATP or biological activity in the blanket solution could only be maintained in a narrow pH range from 6.5 to 7.5, the sludge granules showed a high pH tolerance and buffering capacity up to pH 11. The suspended biomass in AFB reactor had a higher specific ATP than the biomass fixed in polyurethane carriers (1.6?mg ATP g VS^?1 versus 1.1?mg ATP g VS^?1), which implies a starvation status of the immobilized cells due to mass transfer limitation. The aerobes had to work under starvation conditions in this polishing reactor. The anaerobic biomass brought into AFB reactor contributed to an increase in suspended solids, but not the COD removal because of its fast deactivation under aerobic conditions. A second order kinetic model was proposed for ATP decline of the anaerobes. The results on distribution of microbial activity in the two reactors as well as its change with hydraulic retention time lead to further performance improvement of the combined anaerobic/aerobic reactor system.     

Metabolic transformations and characterisation of the sludge community in an enhanced biological phosphorus removal system

Enhanced biological phosphorus removal was performed in a continuous laboratory-scale two-reactor system with sludge recirculation over a 75-day period. Influent wastewater was a synthetic medium based on acetate, and the sludge age was kept at 12 days. The adapted sludge stored poly-β-hydroxyalkanoic acids (PHA) in the anaerobic reactor with a conversion ratio of 1.45 PHA/acetic acid (based on chemical O₂ demand: COD/COD) and gave ratio of a phosphate-P release to acetic acid uptake of 0.51 P/CH₃COOH (w/w). Fractionation of anaerobic and aerobic sludges showed that the main part of phosphorus taken up, was eluted in the trichloroacetic acid fraction indicating that it was polyphosphate. A total of 60% of the phosphorus in the aerobic sludge was solubilized in the trichloroacetic acid fraction, whereas this fraction accounted for only 32% of the phosphorus in the anaerobic sludge. Only 4% of the total phosphorus in the aerobic sludge and 2% in the anaerobic sludge was found in the EDTA fraction, indicating low amounts of metal-bound phosphates. Isolation on acetate-based agar medium showed that Acinetobacter strains were present in the sludge. However, a more complete analysis of the bacterial community of the sludge was obtained by creating a clone library based on the 16S rRNA gene. A total of 51 partial clone sequences were phylogenetically evaluated. The predominating group was found in the high-(G+C) (mol%) gram-positive bacterial subphylum (31% of the sequenced clones), while the gamma proteobacteria only constituted 9.8% of the clones. Received: 12 June 1997 / Received revision: 26 September 1997 / Accepted: 28 September 1997     

Enzymatic pre-hydrolysis and anaerobic degradation of wastewaters with high fat contents

Dairy wastewaters containing elevated fat and grease levels (868 mg l^–1) were treated in an upflow anaerobic sludge blanket reactor (UASB) and resulted in effluents of high turbidity (757 nephelometric turbidity units), volatile suspended solids up to 944 mg l^–1 and COD removal below 50%. When the same dairy wastewater was pre-treated with 0.1% (w/v) of fermented babassu cake containing Penicillium restrictum lipases, turbidity and volatile suspended solids were decreased by 75% and 90%, respectively, and COD removal was as high as 90%.     

Challenge of psychrophilic anaerobic wastewater treatment

Psychrophilic anaerobic treatment is an attractive option for wastewaters that are discharged at moderate to low temperature. The expanded granular sludge bed (EGSB) reactor has been shown to be a feasible system for anaerobic treatment of mainly soluble and pre-acidified wastewater at temperatures of 5--10 degrees C. An organic loading rate (OLR) of 10--12 kg chemical oxygen demand (COD) per cubic meter reactor per day can be achieved at 10--12 degrees C with a removal efficiency of 90%. Further improvement might be obtained by a two-module system in series. Stabile methanogenesis was observed at temperatures as low as 4--5 degrees C. The specific activity of the mesophilic granular sludge was improved under psychrophilic conditions, which indicates that there was growth and enrichment of methanogens and acetogens in the anaerobic system. Anaerobic sewage treatment is a real challenge in moderate climates because sewage belongs to the 'complex' wastewater category and contains a high fraction of particulate COD. A two-step system consisting of either an anaerobic up-flow sludge bed (UASB) reactor combined with an EGSB reactor or an anaerobic filter (AF) combined with an anaerobic hybrid reactor (AH) is successful for anaerobic treatment of sewage at 13 degrees C with a total COD removal efficiency of 50% and 70%, respectively.     

Exploitation of olive oil mill wastewater for combined biohydrogen and biopolymers production

The present study aimed to the investigation of the feasibility of the combined biohydrogen and biopolymers production from OMW (Olive oil Mill Wastewater), using a two stage system. H₂ and volatile fatty acids (VFAs) were produced via anaerobic fermentation and subsequently the acidified wastewater was used as substrate for aerobic biodegradable polymer production. Two different bioreactors, one of CSTR type and a SBR were used for the anaerobic and the aerobic process respectively. The anaerobic reactor was operated at different hydraulic retention times (HRTs) with OMW, diluted 1:4 (v/v) with tap water, as feed. The main VFAs produced were acetate, butyrate and propionate, in different ratios depending on the HRT. Valerate, isovalerate and isobutyrate were also detected in small quantities. Selective effluents of the acidogenic/hydrogen producing reactor were subsequently used as feed for the aerobic reactor. The aerobic reactor was inoculated with an enriched PHAs producing bacteria culture, and was operated in sequential cycles of nitrogen offer (growth phase) and nitrogen limitation (PHAs accumulation phase). The operational program of the SBR was determined according to the results from batch test, and its performance was evaluated for a period of 100 days. During the accumulation phase butyrate was consumed preferably, indicating that the dominant PHA produced is polyhydroxybutyrate. The higher yield of PHAs observed was 8.94% (w/w) of dry biomass weight.     

Potentials of anaerobic treatment for catalytically oxidized olive mill wastewater (OMW)

The catalytically oxidized olive mill wastewater (OMW) was subjected to continuous anaerobic treatment using two treatment schemes. The 1st step in both schemes was an up-flow anaerobic sludge blanket (UASB) reactor (2 0 l). The 2nd step was either a hybrid UASB reactor or a classical one (1 0 l, each). The 1st stage was operated at constant hydraulic retention time (HRT) of 24 h. The organic loading rate (OLR) varied from 3.4 to 4.8 kgCOD/m³ d depending on the quality of the pretreated wastewater. The results obtained indicated that, the 1st step UASB reactor achieved a COD percentage removal value of 53.9%. Corresponding total BOD₅ and TSS removal were 51.5% and 68.3%, respectively.The results obtained indicated that the hybrid UASB reactor as a 2nd step produced better quality effluent as compared to the classical one. This could be attributed to the presence of the packing curtain sponge with active biomass in the sedimentation part of hybrid UASB reactor which minimizes suspended solids washout, consequently enhancement of the efficiency of the reactor.Available data showed that a two stage system consisting of a classical and a hybrid UASB reactor operated at a total HRT of 48 h and OLR of 2.0 kgCOD/m³ d provided promising results. Removal values of COD_total, BOD_5 total, TOC, VFA, oil and grease were 83%, 84%, 81%, 93% and 81%, respectively. Based on the available data, the use of a two stage anaerobic system consisting of a classical UASB reactor followed by a hybrid UASB as a post-treatment step for catalytically oxidized OMW is recommended.     

Treatment of slaughterhouse wastewater in a UASB reactor and an anaerobic filter

A study was performed to assess the feasibility of anaerobic treatment of slaughterhouse wastewaters in a UASB (Upflow Anaerobic Sludge Blanket) reactor and in an AF (Anaerobic Filter). Among the different streams generated, the slaughter line showed the highest organic content with an average COD of 8000 mg/l, of which 70% was proteins. The suspended solids content represented between 15 and 30% of the COD. Both reactors had a working volume of 21. They were operated at 37°C. The UASB reactor was run at OLR (Organic Loading Rates) of 1–6.5 kg COD/m³/day. The COD removal was 90% for OLR up to 5 kg COD/m³/day and 60% for an OLR of 6.5 kg COD/m³/day. For similar organic loading rates, the AF showed lower removal efficiencies and lower percentages of methanization. At higher OLR sludge, flotation occurred and consequently the active biomass was washed out from the filter. The results indicated that anaerobic treatment systems are applicable to slaughterhouse wastewaters and that the UASB reactor shows a better performance, giving higher COD removal efficiencies than the AF.     

Pilot study on the upgrading configuration of UASB-MBBR with two carriers: Treatment effect,sludge reduction and functional microbial identification

Two kinds of biocarriers were adopted and a combined process of “AMC (Anaerobic microorganism carrier)-UASB and PBG (Porous bio-gel)-MBBR” was operated at the pilot scale for the treatment of real textile wastewater. The influence mechanism of the two carriers on the start-up, pollutant removal and sludge reduction were investigated within 118 days of operation. The dominant functional bacteria in anaerobic and aerobic systems were identified by high-throughput sequencing, and the possible ways and related mechanisms of nutrient removal and sludge reduction were analyzed based on the data. 37.0 ± 7.5 % and 53 ± 12.7 % of COD removal efficiencies were achieved in anaerobic system and aerobic system, respectively. Ammonia nitrogen concentration decreased from 20 to 45 to 3.49 ± 0.54 mg/L after treatment. An anaerobe was found to be closely related to color removal, which existed in both anaerobic and aerobic systems, achieving 84.0 % of color removal. With the operation of the system, the sludge yield decreased gradually. The sludge yields of anaerobic and aerobic systems were calculated individually and compared with similar studies. Aging biofilms were characterized to explore the factors associated with biofilm renewal.     

The anaerobic filtration of dairy waste: Results of a pilot trial

Results are presented from a pilot scale (4·3 m³) upflow anaerobic filter for the treatment of the wastewater from ice-cream manufacture. The reactor was completely mixed by gas production but the solids or sludge held within the reactor were shown to be affected by the liquid velocities. The reactor was subject to a number of organic and hydraulic shocks and this reduced the consistency of COD removal. Daily loading rates varied from 0 to 18 kg COD/m³/day but with an average load of 5·5 kg/m³/day the mean COD removal was 70%. This was compared with previous work and shown to be a typical performance for an anaerobic filter. Alkalinity and carboxylic acid data are also presented and were within the normal, stable, operating range. Previous research on the anaerobic treatment of industrial effluents has shown alkalinity to be the most important factor controlling reliability.     

An integrated process for the treatment of CETP wastewater using coagulation, anaerobic and aerobic process

The aim of this study was to treat the wastewater collected from equalization tank of Common Effluent Treatment Plant (CETP), which was a mixture of waste coming from 525 small-scale industries manufacturing textile and dyestuff intermediate, pigments and pharmaceuticals. Initially a pretreatment using ferric chloride and lime was carried out to increase the biodegradability (BOD(5)/COD) of the effluent, which showed color removal of 74% and COD reduction of 75% at a concentration of 10 and 4 g/L. respectively. The biological treatment system using anaerobic fixed film reactor was investigated as secondary treatment. A mixture of bacterial consortium DMAB and cowdung slurry was used for the formation of biofilm. The effect of hydraulic retention time (HRT) and organic loading rate (OLR) on the efficiency of treatment of anaerobic reactor was analysed. Subsequent aerobic treatment after anaerobic step using aerobic culture Pseudomonas aeroginosa helped in further removal of COD and color. Formation of aromatic amines during anaerobic treatment was mineralized by sequential aerobic treatment.     

Experimental investigation of the external nitrification biological nutrient removal activated sludge (ENBNRAS) system

A systematic lab-scale experimental investigation is reported for the external nitrification (EN) biological nutrient removal (BNR) activated sludge (ENBNRAS) system, which is a combined fixed and suspended medium system. The ENBNRAS system was proposed to intensify the treatment capacity of BNR-activated sludge (BNRAS) systems by addressing two difficulties often encountered in practice: (a) the long sludge age for nitrification requirement; and (b) sludge bulking. In the ENBNRAS system, nitrification is transferred from the aerobic reactor in the suspended medium activated sludge system to a fixed medium nitrification system. Thus, the sludge age of the suspended medium activated sludge system can be reduced from 20 to 25 days to 8 to 10 days, resulting in a decrease in reactor volume per ML wastewater treated of about 30%. Furthermore, the aerobic mass fraction can also be reduced from 50% to 60% to <30% and concommitantly the anoxic mass fraction can be increased from 25% to 35% to >55% (if the anaerobic mass fraction is 15%), and thus complete denitrification in the anoxic reactors becomes possible. Research indicates that both the short sludge age and complete denitrification could ameliorate anoxic aerobic (AA) or low food/microorganism (F/M) ratio filamentous bulking, and hence reduce the surface area of secondary settling tanks or increase the treatment capacity of existing systems. The lab-scale experimental investigations indicate that the ENBNRAS system can obtain: (i) very good chemical oxygen demand (COD) removal, even with an aerobic mass fraction as low as 20%; (ii) high nitrogen removal, even for a wastewater with a high total kjeldahl nitrogen (TKN)/COD ratio, up to 0.14; (iii) adequate settling sludge (diluted sludge volume index [DSVI] <100 mL/g); and (iv) a significant reduction in oxygen demand.     

Two-stage biological treatment of olive mill wastewater with whey as co-substrate

Olive mill wastewater (OMW) is a highly polluting wastewater, caused by a high organic load and phenol content. These characteristics suggest that it may be suitable for aerobic treatment and anaerobic bacterial digestion. Aerobic treatment coupled with anaerobic bacterial digestion may be economically feasible as the methane produced is a valuable energy source while simultaneously purifying the OMW. In an attempt to improve the overall performance of the process, the addition of a co-substrate such as whey to the aerobic treatment pre-treatment of OMW by the yeast Candida tropicalis was studied.The two-stage system operated satisfactorily up to an organic loading rate (OLR) of 3.0 kg COD L⁻¹ day⁻¹ with a biogas production rate of 1.25 L_biogas L_reactor⁻¹ day⁻¹ and a total COD reduction in excess of 93% (62% COD reduction in aerobic pretreatment and 83% COD reduction in anaerobic digestion). Fifty-four percent of the phenol was biodegraded during the aerobic treatment stage, and biogas with between 68% and 75% methane was produced during anaerobic digestion.     

Treatment of wastewater from abattoirs before land application--a review

Pre-treatments are screening, catch basins, flotation, equalization, and settlers for recovering proteins and fats from abattoir wastewater. With chemical addition, dissolved air flotation (DAF) units can achieve chemical oxygen demand (COD) reductions ranging from 32% to 90% and are capable of removing large amounts of nutrients. Aerobic trickling towers reduced soluble COD by additional 27% but did not reduced total COD. Chemical-DAF reduced 67% of total COD and soluble COD. About 40-60% of the solids or approximately 25-35% of the biological oxygen demand (BOD) load can be separated by pre-treatment screening and sedimentation. Anaerobic systems are lagoon, anaerobic contact (AC), up-flow anaerobic sludge blanket (UASB), anaerobic sequence batch reactor (ASBR), and anaerobic filter (AF) processes. Abattoir wastewater is well suited to anaerobic treatment because it is high in organic compounds. Typical reductions of up to 97% BOD, 95% SS and 96% COD are reported. UASB's average COD removal efficiencies are of 80-85%. UASB seems to be a suitable process for the treatment of abattoir wastewater, due to its ability to maintain a sufficient amount of viable sludge. Wastewater in abattoirs can be reduced by treatment of immersion chiller effluent by membrane filtration which can produce recyclable water. Total organic C can be reduced below 100mg/L, and bacteria can not pass through the membrane pores. The abattoir waste minimization options are also discussed.     

The influence of anaerobic pretreatment on the nitrogen removal from biosynthetic pharmaceutical wastewaters

The C:N ratio of the pharmaceutical wastewaters is usually suitable for a combination of the anaerobic pretreatment with the high COD removal and aerobic posttreatment with the efficient biological N removal. This kind of anaerobic-aerobic process was tested in semipilot scale by using a UASB reactor and an activated sludge system with a predenitrification (total volume 100 1). It was found that at a total HRT of 2.3 days an average of 97.5% of COD and 73.5% of total N was removed. The UASB reactor was operated at 30°C with a volumetric loading rate of 8.7 kg.m^-3.d^-1, the efficiency of COD removal was 92.2%. The processes, which take part in the biological removal of nitrogen, especially the nitrification, were running with lower rates than usually observed in aerobic treatment systems.Abbreviations AAO anaerobic anoxic oxic configuration - AOO anaerobic oxic oxic configuration - B _V volumetric organic loading rate (kg COD.m^-3. d^-1) - dB _x specific COD removal rate (mg COD. g^-1 VSS. d^-1) - DNR denitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - E_COD efficiency of COD removal (%) - HRT hydraulic retention time (d) - NR nitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - R recirculation ratio (%) - SBP specific biogas production (m³.kg^-1 removed COD) - SRT solids retention time; sludge age (d) - SS suspended solids (g.1^-1) - UASB upflow anaerobic sludge blanket reactor - VSS volatile suspended solids (g.1^-1)     

Comparison of the performance of one stage and two stage sequential anaerobic-aerobic biological processes for the treatment of reactive-azo-dye-containing synthetic wastewaters

In the present study the performance of anaerobic-aerobic one and two stage processes for the biological treatment of synthetic wastewaters containing Reactive Black 5 (RB5) were studied and compared with each other. In both processes the majority of colour removal by biodegradation occurred under anaerobic environment. The colour change under aerobic conditions was correlated with extent of anaerobic decolourisation in the preceding phase/stage of the process. Partial mineralisation of the anaerobic dye metabolites, roughly to the same extent, was achieved aerobically in both one stage and two stage processes. The majority of COD was removed in the anaerobic stage for two stage processes and aerobic stage in one stage processes. In one stage processes, the exposure of anaerobic sludge to alternating anaerobic-aerobic environment decreased anaerobic decolourisation efficiency and COD removal; when employing activated sludge, the same exposure enhanced anaerobic substrate utilisation whereas the effect on the anaerobic decolourisation efficiency depended on RB5 concentration. The comparative performance of one and two stage processes in terms of overall dye decolourisation depended on RB5 concentration. Both types of processes brought about similar overall COD removal. Increase in RB5 concentration, in the range studied, resulted in decrease in overall COD removal for both processes.