Biological treatment of acid dyeing wastewater using a sequential anaerobic/aerobic reactor system

The treatment of the wastewater taken from a wool dyeing processing in a wool manufacturing plant was investigated using an anaerobic/aerobic sequential system. The process units consisted of an anaerobic UASB reactor and an aerobic CSTR reactor. Glucose, alkalinity and azo dyes were added to the raw acid dyeing wastewater in order to simulate the dye industry wastewater since the raw wastewater contained low levels of carbon, NaHCO₃ and color through anaerobic/aerobic sequential treatment. The UASB reactor gave COD and color removals of 51–84% and 81–96%, respectively, at a HRT of 17 h. The COD and color removal efficiencies of the UASB/CSTR sequential reactor system were 97–83% and 87–80%, respectively, at a hydraulic retention time (HRTs) of 3.3 days. The aromatic amines (TAA) formed in the anaerobic stage were effectively removed in the aerobic stage.     

The effect of cyclic anaerobic–aerobic conditions on biodegradation of azo dyes

The effect of cyclic anaerobic–aerobic conditions on the biodegradative capability of the mixed microbial culture for the azo dye Remazol Brilliant Violet 5R (RBV-5R) was investigated in the sequencing batch reactor (SBR) fed with a synthetic textile wastewater. The SBR had a 12-h cycle time with anaerobic–aerobic periods of 3/9, 6/6 and 9/3 h. General SBR performance was assessed by measurement of catabolic enzymes (catechol 2,3-dioxygenase, azo reductase), chemical oxygen demand (COD), color and amount of aromatic amines. In this study, under steady-state conditions, the anaerobic period of the cyclic SBR was found to allow the reductive decolorization of azo dye. Longer anaerobic periods resulted in higher color removal efficiencies, approximately 71% for the 3-h, 87% for 6-h and 92% for the 9-h duration. Total COD removal efficiencies were over 84% under each of the cyclic conditions and increased as the length of the anaerobic period was increased; however, the highest color removal rate was attained for the cycle with the shortest anaerobic period of 3 h. During the decolorization of RBV-5R, two sulfonated aromatic amines (benzene based and naphthalene based) were formed. Additionally, anaerobic azo reductase enzyme was found to be positively affected with the increasing duration of the anaerobic period; however; it was vice versa for the aerobic catechol 2,3-dioxygenase (C23DO) enzyme.     

Start-up procedures and analysis of heavy metals inhibition on methanogenic activity in EGSB reactor

The effectiveness of operating an industrial UASB reactor, treating wastewater from the beer industry, with flows containing heavy metals was evaluated. A pilot-scale UASB reactor, already used to simulate the industrial reactor, was unsuccessfully employed. An easy start-up was obtained arranging it as an EGSB reactor. Considerations about this modification are reported. The effects of Cu(II), Ni(II) and Cr(III) ions on the anaerobic activity were analyzed by measurements of methane production rate and COD removal. The employed biomass was the sludge of the industrial UASB reactor, while a solution of ethanol and sodium acetate with COD of 3000 mg/L and a heavy metal concentration of 50 mg/L were continuously fed. Experimental results proved higher biomass sensitivity for copper and much slighter for nickel and chromium. Moreover, copper inhibition has been demonstrated to be less significant if a metal-free feed was provided to the system before copper addition.     

Application of redox mediators to accelerate the transformation of reactive azo dyes in anaerobic bioreactors.

Azo dyes are nonspecifically reduced under anaerobic conditions but the slow rates at which reactive azo dyes are converted presents a serious problem for the application of anaerobic technology as a first stage in the complete biodegradation of these compounds. As quinones have been found to catalyze reductive transfers by acting as redox mediators, the application of anthraquinone-2,6-disulfonic acid (AQDS) during continuous anaerobic treatment of the reactive azo dye, Reactive Red 2 (RR2), was evaluated. A mixture of volatile fatty acids was used as the electron-donating primary substrate. Batch experiments demonstrated that AQDS could increase the first-order rate constant of RR2 reductive cleavage by one order of magnitude. In the continuous experiment, treatment of RR2 containing synthetic wastewater in a lab-scale upflow anaerobic sludge blanket (UASB) reactor yielded low dye removal efficiencies (<30%). Consequently, severe toxicity problems occurred, eventually resulting in almost complete inhibition of the methanogenic activity. Addition of catalytic concentrations of AQDS (19 microM) to the reactor influent caused an immediate increase in the dye removal efficiency and recovery of biological activity. Ultimately, RR2 removal efficiency stabilized at 88%, and higher AQDS loads resulted in higher RR2 removal efficiencies (up to 98% at 155 microM AQDS). Examination of the RR2 decolorizing properties of dye-adapted reactor sludge and of nonadapted reactor seed sludge revealed that RR2 decolorization was principally a biologically driven transfer of reducing equivalents from endogenous and added substrates to the dye. Hydrogen, added in bulk, was clearly the preferred electron donor. Bacteria that couple dye decolorization to hydrogen oxidation were naturally present in seed sludge. However, enrichment was required for the utilization of electrons from volatile fatty acids for dye reduction. The stimulatory effect of AQDS on RR2 decolorization by AQDS-unadapted sludge was mainly due to assisting the electron transfer from endogenous substrates in the sludge to the dye. The stimulatory effect of AQDS on RR2 decolorization by sludge from the AQDS-exposed reactor was, in addition, strongly associated with the transfer of electrons from hydrogen and acetate to the dye, probably due to enrichment of specialized AQDS-reducing bacteria.     

Anaerobic degradation of azo dye Drimaren blue HFRL in UASB reactor in the presence of yeast extract a source of carbon and redox mediator

This paper presents results on anaerobic degradation of the azo dye blue HFRL in a bench scale Upflow anaerobic sludge blanket (UASB) reactor operated at ambient temperature. The results show that the addition of yeast extract (500 mg/L) increased color removal (P < 0.05) from 62 to 93% despite the low chemical oxygen demand (COD) removal (~35%) which happened due to volatile fatty acids (VFA) accumulation. There were no differences in color removal (~91%) when yeast extract (500 mg/L) was used in the presence or absence of glucose, suggesting that yeast extract acted as source of redox mediator (riboflavin) and carbon. The specific rate of dye removal increased along the operational phases and depended on the presence of yeast extract, suggesting progressive biomass acclimatization. Analysis of bacterial diversity by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR–DGGE) method showed there was biomass selection along the bioreactor operation and no evidence of azo dye degrading bacteria predominance. This strengthens the hypothesis that color removal happens extracellularly by the reduction of azo bond by reduced redox mediators, such as riboflavin, which is present in high amount in the yeast extract.     

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

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

Continuous anaerobic treatment of wastewaters containing formaldehyde and urea

The toxicity of formaldehyde (FA) in batch assays, using volatile fatty acids (VFA) as co-substrate, and the continuous anaerobic treatment of wastewaters containing FA in upflow anaerobic sludge blanket (UASB) reactors was investigated. In batch studies, FA exerted a 50% methanogenic toxicity on VFA at concentrations of around 100 mg/l, 2.5 times lower than values reported with sucrose. Although at FA concentrations higher than 200 mg/l methanogenesis was completely inhibited, a partial recovery of the bacterial activity was observed after 250 h when the FA had been removed from the medium. The continuous anaerobic degradation of FA at concentrations up to 2 g/l, using 1.6 g/l of glucose as co-substrate, was studied in a UASB reactor. A stable and efficient operation was observed at organic loading rates (OLR) of 6.0 g COD/l·d and with a COD/FA ratio as low as 1.4. A synthetic substrate with the same characteristics as the effluents produced during fibreboard adhesives manufacturing (based on urea-FA), i.e. 0.95 g FA/l and 0.35 g urea/l, was treated in a UASB reactor. The applied OLR and nitrogen loading rate (NLR) were 3.45 g COD/l·d and 0.58 g N/l·d, respectively. COD removal efficiencies were maintained at 90–95%, FA and urea being completely degraded.     

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.     

Decolorizing kinetics of a recombinant Escherichia coli SS125 strain harboring azoreductase gene from Bacillus latrosporus RRK1

PCR amplified product containing gene responsible for dye decolorization was cloned and expressed in Escherichia coli. The resulting recombinant strain E. coli SS125 decolorized 200mg/l azo dye (Remazol Red) at 30 degrees C at 255 mg cell/l/h, while the host E. coli (DH5 alpha) had no color removal ability. The dependence of the decolorization rate on initial dye concentration and the maximum rate occurred with the dye at 100 mg l(-1). The decolorization rate of E. coli SS125 was optimal at 37-45 degrees C. Aeration strongly-inhibited the decolorization, but decolorization occurred effectively under static and anaerobic incubation conditions. The E. coli SS125 strain also exhibited excellent stability during reported batch operation.     

Anaerobic treatment of a chemical synthesis-based pharmaceutical wastewater in a hybrid upflow anaerobic sludge blanket reactor

In this study, performance of a lab-scale hybrid up-flow anaerobic sludge blanket (UASB) reactor, treating a chemical synthesis-based pharmaceutical wastewater, was evaluated under different operating conditions. This study consisted of two experimental stages: first, acclimation to the pharmaceutical wastewater and second, determination of maximum loading capacity of the hybrid UASB reactor. Initially, the carbon source in the reactor feed came entirely from glucose, applied at an organic loading rate (OLR) 1 kg COD/m(3) d. The OLR was gradually step increased to 3 kg COD/m(3) d at which point the feed to the hybrid UASB reactor was progressively modified by introducing the pharmaceutical wastewater in blends with glucose, so that the wastewater contributed approximately 10%, 30%, 70%, and ultimately, 100% of the carbon (COD) to be treated. At the acclimation OLR of 3 kg COD/m(3) d the hydraulic retention time (HRT) was 2 days. During this period of feed modification, the COD removal efficiencies of the anaerobic reactor were 99%, 96%, 91% and 85%, and specific methanogenic activities (SMA) were measured as 240, 230, 205 and 231 ml CH(4)/g TVS d, respectively. Following the acclimation period, the hybrid UASB reactor was fed with 100% (w/v) pharmaceutical wastewater up to an OLR of 9 kg COD/m(3) d in order to determine the maximum loading capacity achievable before reactor failure. At this OLR, the COD removal efficiency was 28%, and the SMA was measured as 170 ml CH(4)/g TVS d. The hybrid UASB reactor was found to be far more effective at an OLR of 8 kg COD/m(3) d with a COD removal efficiency of 72%. At this point, SMA value was 200 ml CH(4)/g TVS d. It was concluded that the hybrid UASB reactor could be a suitable alternative for the treatment of chemical synthesis-based pharmaceutical wastewater.     

Effect of nitrate on anaerobic azo dye reduction

The aim of the study was to investigate the effect of nitrate on anaerobic color removal efficiencies. For this aim, anaerobic–aerobic sequencing batch reactor (SBR) fed with a simulated textile effluent including Remazol Brilliant Violet 5R azo dye was operated with a total cycle time of 12 h, including anaerobic (6 h) and aerobic cycles (6 h). Microorganism grown under anaerobic phase of the reactor was exposed to different amounts of competitive electron acceptor (nitrate) and performance of the system was determined by monitoring color removal efficiency, nitrate removal, nitrite formation and removal, oxidation reduction potential, color removal rate, chemical oxygen demand (COD), specific anaerobic enzyme (azo reductase) and aerobic enzyme (catechol 1,2 dioxygenase), and formation and removal of aromatic amines. Variations of population dynamics of microorganisms exposed to various amount of nitrate were identified by denaturing gradient gel electrophoresis (DGGE). It was found that nitrate has adverse effect on anaerobic color removal efficiency and color removal was achieved after denitrification process was completed. It was found that nitrate stimulates the COD removal efficiency and accelerates the COD removal in the first hour of anaerobic phase. About 90 % total COD removal efficiencies were achieved in which microorganism exposed to increasing amount of nitrate. Population dynamics of microorganisms exposed to various amount of nitrate were changed and diversity was increased.     

Effect of organic loading rate on the performance of an up-flow anaerobic sludge blanket reactor treating olive mill effluent

The primary objective of this study was to evaluate the effects of the organic loading rate on the performance of an up-flow anaerobic sludge blanket (UASB) reactor treating olive mill effluent (OME), based on the following indicators: (i) chemical oxygen demand (COD) removal efficiency; and (ii) effluent variability (phenol, suspended solids, volatile fatty acids, and pH stability). The UASB reactor was operated under different operational conditions (OLRs between 0.45 and 32 kg COD/m³·day) for 477 days. The results demonstrated that the UASB reactor could tolerate high influent COD concentrations. Removal efficiencies for the studied pollution parameters were found to be as follows: COD, 47∼92%; total phenol, 34∼75%; color, 6∼46%; suspended solids, 34∼76%. The levels of VFAs in the influent varied between 310 and 1,750 mg/L. Our measurements of the VFA levels indicated that some of the effluent COD could be attributed to VFAs (principally acetate, butyrate, iso-butyrate, and propionate) in the effluent, which occurred at levels between 345 and 2,420 mg/L. As the OLRs were increased, more VFAs were measured in the effluent. A COD removal efficiency of 90% could be achieved as long as OLR was kept at a level of less than 10 kg COD/m³·day. However, a secondary treatment unit for polishing purposes is necessary to comply with discharge standards.     

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.     

Performance evaluation of a mesophilic anaerobic fluidized-bed reactor treating wastewater derived from the production of proteins from extracted sunflower flour

A study of the anaerobic digestion of wastewater derived from the production of protein isolates from extracted sunflower flour was carried out in a laboratory-scale, mesophilic (35 degrees C) fluidized-bed reactor with saponite as bacterial support. Chemical oxygen demand (COD) removal efficiencies in the range of 98.3-80.0% were achieved in the reactor at organic loading rates (OLR) of between 0.6 and 9.3 g COD/I d, hydraulic retention times (HRT) of between 20.0 and 1.1 d and average feed COD concentration of 10.6 g/l. Eighty percent of feed COD could be removed up to OLR of 9.3 g COD/l d. The yield coefficient of methane production was 0.33 l of methane (at STP) per gram of COD removed and was virtually independent of the OLR applied. Because the buffering capacity of the experimental system was maintained at favorable levels with excess total alkalinity present at all loadings, the rate of methanogenesis was not affected by loading. The experimental data indicated that a total alkalinity in the range of 2,000-2,460 mg/l as CaCO3 was sufficient to prevent the pH from dropping to below 7.0 for OLR of up to 9.3 g COD/l d. The volatile fatty acid (VFA) levels and the VFA/alkalinity ratio were lower than the suggested limits for digester failure (0.3-0.4) for OLR and HRT up to 9.3 g COD/l d and 1.1 d, respectively. For a HRT of 0.87 d (OLR of 12.1 g COD/l d) the start of acidification was observed in the reactor.     

Anaerobic treatment of pre-hydrolysate liquor (PHL) from a rayon grade pulp mill: pilot and full-scale experience with UASB reactors

Pilot studies were carried out for the treatment of pre-hydrolysate liquor (PHL), a high strength effluent (COD: 70,000-80,000 mg/l) emanating from a rayon grade pulp mill using up-flow anaerobic sludge blanket reactor (UASB). Substrate inhibition was avoided with optimum COD feed of around 25,000 mg/l. This was achieved by diluting the PHL with a low strength effluent stream known as alkali back wash (ABW) available in the plant and also by partially recycling the reactor liquid outlet. An optimum organic loading rate (OLR) of 10 could be achieved with a COD reduction of 70-75%, a BOD reduction of 85-90% and a methane yield of 0.31-0.33 m3/kg of COD reduced. The pilot scale studies also revealed that addition of milk of lime (MOL) was essential for neutralization and buffering and DAP and urea to supplement the nutrients in the PHL. Based on the pilot studies, a full-scale high rate biomethanation plant was designed and erected for treating the PHL, which after some modification showed similar performance for COD, BOD reduction and methane yield.     

Mesophilic anaerobic digestion in a fluidised-bed reactor of wastewater from the production of protein isolates from chickpea flour

A study of the anaerobic digestion of wastewater derived from the production of protein isolates from chickpea flour was carried out in a laboratory-scale, mesophilic (35 °C) fluidised-bed reactor with saponite as bacterial support. Soluble chemical oxygen demand (SCOD) removal efficiencies in the range of 96.8–85.2% were achieved in the reactor at organic loading rates (OLR) of between 0.58 and 2.10 g chemical oxygen demand (COD)/l per day, hydraulic retention times (HRT) of between 14.9 and 4.5 days and average feed COD concentration of 9.1 g/l. Eighty-five percent of feed COD could be removed up to OLR of 2.1 g COD/l per day. The yield coefficient of methane production was 0.34 l of methane (at STP) per gram COD removed and was virtually independent of the OLR applied. Because the buffering capacity of the experimental system was maintained at favourable levels with excess total alkalinity present at all loadings, the rate of methanogenesis was not affected by loading. Experimental data indicated that a total alkalinity in the range of 1090–2130 mg/l as CaCO₃ was sufficient to prevent the pH from decreasing to below 7.2 for OLR of up to 2.7 g COD/l per day. The volatile fatty acid (VFA) levels and the VFA/alkalinity ratio were lower than the suggested limits for digester failure (0.3–0.4) for OLR and HRT up to 2.7 g COD/l per day and 3.5 days, respectively. For a HRT of 2.8 days (OLR of 3.00 g COD/l per day) the start of acidification was observed in the reactor.     

Treatment of low strength soluble wastewaters in UASB reactors

Low strength wastewaters can be those with chemical oxygen demand (COD) below 2,000 mg/l. The anaerobic treatment of such wastewaters has not been fully explored so far. The suboptimal reaction rates with low substrate concentrations, and the presence of dissolved oxygen in the influent are regarded as possible constraints. In this study, the treatment of low strength soluble wastewaters containing ethanol or whey was studied in lab-scale upflow anaerobic sludged bed (UASB) reactors at 30°C. The high treatment performance obtained demonstrates that UASB reactors are viable for treating both types of wastewaters at low COD concentrations. The treatment of the ethanol containing wastewater resulted in COD removal efficiencies exceeding 95% at organic loading rates (OLR) between 0.3 to 6.8 g COD/l-d with influent concentrations in the range of 422 to 943 mg COD/l. In the case of the more complex whey containing wastewater, COD removal efficiencies exceeded 86% at OLRs up to 3.9 g COD/l·, as long as the COD influent was above 630 mg/l. Lowering the COD influent resulted in decreased efficiency with sharper decrease at values below 200 mg/l. Acidification instead of methanogenesis was found to be the rate limiting step in the COD removal at low concentrations, which was not the case when treating ethanol. The effect of dissolved oxygen in the influent as a potential danger in anaerobic treatment was investigated in reactors fed with and without dissolved oxygen. Compared with the control reactor, the reactor receiving oxygen showed no detrimental effects in the treatment performance. Thus, the presence of dissolved oxygen in dilute wastewaters is expected to be of minor importance in practice.     

Characteristics of the bioreactor landfill system using an anaerobic-aerobic process for nitrogen removal

A sequential upflow anaerobic sludge blanket (UASB) and air-lift loop sludge blanket (ALSB) treatment was introduced into leachate recirculation to remove organic matter and ammonia from leachate in a lab-scale bioreactor landfill. The results showed that the sequential anaerobic-aerobic process might remove above 90% of COD and near to 100% of NH4+ -N from leachate under the optimum organic loading rate (OLR). The total COD removal efficiency was over 98% as the OLR increased to 6.8-7.7 g/l d, but the effluent COD concentration increased to 2.9-4.8 g/l in the UASB reactor, which inhibited the activity of nitrifying bacteria in the subsequent ALSB reactor. The NO3- -N concentration in recycled leachate reached 270 mg/l after treatment by the sequential anaerobic-aerobic process, but the landfill reactor could efficiently denitrify the nitrate. After 56 days operation, the leachate TN and NH4+ -N concentrations decreased to less than 200 mg/l in the bioreactor landfill system. The COD concentration was about 200 mg/l with less than 8 mg/l BOD in recycled leachate at the late stage. In addition, it was found that nitrate in recycled leachate had a negative effect on waste decomposition.     

Thermophilic anaerobic treatment of sulphur rich forest industry wastewater

Thermophilic anaerobic treatment of sulphur-rich paper mill wastewater (0.8-3.1 gCOD/l, 340–850 mgSO₄/l; COD:SO₄ 3.4-5.3) was studied in three laboratory-scale, upflow anaerobic sludge blanket (UASB) reactors and in bioassays. The reactors were inoculated with non-adapted thermophilic granular sludge. In the bioassays, no inhibition of the inoculum was detected and about 62% COD removal (sulphide stripped) was obtained. About 70 to 80% of the removed COD was methanised. In the reactors, up to 60–74% COD removal (effluent sulphide stripped) was obtained at loading rates up to 10–30 kgCOD/m³d and hydraulic retention times down to 6 to 2 hours. The effluent total sulphide was up to 150–250 mg/l. Sulphide inhibition could not be confirmed from the reactor performances. The results from bioassays suggested that both the inoculum and sludge from the UASB reactor used acetate mainly for methane production, while sulphide was produced from hydrogen or its precursors.     

Use of the UASB reactor for the anaerobic treatment of stillage from sugar cane molasses

The feasibility of applying the UASB concept for the anaerobic treatment of stillage of distilleries in the sugar producing area of Argentina was subject to study. Results obtained in a 100-L UASB reactor treating stillages with COD values between 35 and 100 g COD/L are presented. Loading rates of up to 24 g COD/L/day were applied with an average COD removal of 75% and a biogas production of more than 9 L/L/day, with an average methane content of 58%. The settling velocity distribution of sludge particles would indicate a good formation of biomass pellets. System interruptions of months without feed and at ambient temperature (20-24 degrees C) were well tolerated.