Characteristics of granular methanogenic sludge grown on phenol synthetic medium and methanogenic fermentation of phenolic wastewater in a UASB reactor

The growth of granules on a phenol synthetic medium and the methanogenic fermentation of industrial phenolic wastewater from a steel factory in an upflow anaerobic sludge blanket (UASB) reactor were investigated. Total granular sludge concentration retained in the UASB reactor was 6.7 g MLSS/l (6.0 g MLVSS/l) during the 10 months' operation on the phenol synthetic medium. This realized a maximum phenol removal rate of 2.2 g/l·d (phenol concentration of influent = 500 mg/l), which corresponded to 5.2 g COD/l·d at space velocity (SV) of 4.4 d⁻¹. The granules formed were of relatively small size ranging from 0.61 to 0.77 mm, and had a relatively low density of 0.013–0.023 g MLVSS/cm³ and low specific gravity (1.11) due to very low ash content (8.7–11.9%). Electron microscopic analysis showed that Methanothrix spp. appeared dominantly on the granule surface as well as within it. The specific metabolic activities of bacterial trophic groups were the highest for H₂ followed by acetate, benzoate, phenol, and propionate. In the case of industrial phenolic wastewater, although phenol efficiency was only 50% at SV of 0.4 d⁻¹, when the wastewater was diluted twofold and the treated wastewater was recycled at SV of 7.3 d⁻¹, the removal efficiencies of phenol and CODcr were restored to 90% (influent=400 mg/l) and 80% (influent=5,000 mg/l), respectively. It was suggested that recycling of the treated wastewater might be improved by partly degrading unknown toxic compounds contained in phenolic wastewater.     

Anaerobic treatment of wastewater with high salt content from a pickled-plum manufacturing process

Anaerobic treatment of wastewater with a high salt content generated during a pickled-plum manufacturing process (TOC, 14g/l; ash, 150g/l; pH 2.7, hereafter called pickled-plum effluent) was investigated for its effect on the high salt content of the wastewater. The synthetic wastewater, including NaCl up to 30g/l, was treated anaerobically by a draw and fill method (treatment temperature, 37°C; volumetric loading rate of organic matter, 2g/l·d). The TOC removal efficiency and rate of gas evolution then gradually decreased as salt content increased, although stable operation was maintained. At NaCl concentrations above 30g/l, TOC removal efficiency decreased rapidly and stable operation could not be maintained. Five-fold-diluted pickled-plum effluent was treated by the same method at a volumetric TOC loading rate of 2.9g/l·d with a TOC removal efficiency of 71%. Five-fold-diluted pickled-plum effluent was also treated in an anaerobic fluidized-bed reactor (AFBR) at a maximum volumetric TOC loading rate of 3.0g/l·d, which gave almost the same results as the draw and fill method. However, ten-fold-diluted pickled-plum effluent could be treated in the AFBR at a maximum volumetric TOC loading rate of 11.1g/l·d with a TOC removal efficiency of 84.6%. The red pigment in the pickled-plum effluent was completely decolorized by the anaerobic treatment.     

Anaerobic treatment of wastewater from a food-manufacturing plant with a low concentration of organic matter and regeneration of usable pure water

Wastewater from a food-manufacturing plant with a low concentration of organic matter below 100 mg/l TOC was first treated at 37°C in an anaerobic fluidized-bed reactor (AFBR) or in an upflow anaerobic sludge blanket (UASB). The TOC removal efficiency in both reactors decreased from 85% to 65% as the influent TOC concentration decreased from 100 to 35 mg/l at a hydraulic retention time (HRT) of 6 h. Treatment at an HRT of 4 h resulted in an effluent TOC concentration of 11 to 15 mg/l. The concentration of suspended solids in the effluent could be reduced to 20 mg/l, which corresponded to 7% of that of the influent. The effluent from both reactors was then treated anaerobically in a fixed-bed reactor system. The TOC concentration and optical density (OD) of the effluent from the aerobic treatment were reduced to 5 mg/l and 0.005, respectively, at an HRT of 2 h. When anaerobically or aerobically treated effluent was passed over an activated carbon column, the effluent TOC concentration was reduced to 2 to 3 mg/l. The conductivity of 1.3 mS/cm in raw wastewater, which was not removed through the above treatments, was reduced to 0.001 mS/cm on an ion-exchange resin column. An effluent quality corresponding to that of ultra-pure water for industrial use was finally attained by the treatment in this multi-step system.     

Efficient removal of NH4+ accumulated during anaerobic treatment of distillery wastewater from Shochu making

Propionate and NH₄⁺ were accumulated in the effluent during anaerobic treatment of five-fold diluted distillery wastewater from shochu making. Propionate could be removed efficiently during biological denitrification by the addition of NO₃⁻ (4.2 g/l) to the anaerobically treated wastewater. At a hydraulic retention time of more than 2 h, a TOC removal efficiency of 90% could be achieved. The wastewater was then treated aerobically by biological nitrification. With a hydraulic retention time of more than 14 h the efficiency of reduction of NH₄⁺ could be maintained above 97%. In order to reduce the amount of NO₃⁻ addition necessary for the removal of propionate, simultaneous removal of propionate and NH₄⁺ was studied by recirculating the effluent from a nitrification process to a denitrification process using denitrification and nitrification reactors connected in series. At a recirculation ratio of 2, the amount of NO₃⁻ that had to be added was reduced to 0.3 g/l of anaerobically treated wastewater, which corresponds to 6.9% of the theoretical value. Under the same conditions except for the addition of NO₃⁻ at 1.0 g/l, TOC and BOD in the effluent from the nitrification were 23 and 5 mg/l respectively, which are sufficiently low to allow discharge into river water. Moreover, the NO₃⁻ concentration in the effluent decreased with increases in the recirculation ratio.     

Characteristics of granular methanogenic sludge grown on glucose in a UASB reactor

The formation of granules grown on glucose in an upflow anaerobic sludge blanket (UASB) reactor was investigated. Total granular sludge concentration retained in the UASB reactor was 34.5 g MLSS/l (30.0 g MLVSS/l) during 240 d operation on glucose minimum medium with the supplementation of 1.07 g NaHCO₃ per 1 g glucose. This realized a high-rate methanogenic fermentation of glucose of 17.6 g COD/l-reactor-d at 3.4 d⁻¹ of space velocity. The granules formed were relatively small, ranging mainly from 0.4 to 0.5 mm, had a relatively low cell density of 0.0542–0.0560 g MLVSS/ml, and had low specific gravity (0.97–1.19) due to very low ash content (11–13%). Electron microscopic analysis showed that Methanothrix spp. appeared dominant over the granules. The specific metabolic activities of bacterial trophic groups were the highest for H₂ followed by glucose, acetate, and propionate.     

Treatment and phosphorus removal from high-concentration organic wastewater by the yeast Hansenula anomala J224 PAWA

A flocculent yeast, Hansenula anomala J224 PAWA, bred in this study, accumulated twice as much phosphorus as the wild type. Over a 30-d period, PAWA removed 70–80% of dissolved total phosphorus from sweet-potato and barley shochu wastewaters (alcoholic distillery wastewaters) while the wild type removed only 30%. Waste sludge was easily separated from effluent wastewater because PAWA cells made large flocks that rapidly settled. Component analysis suggested that PAWA sludge could be used as a protein source for feedstuff and as a phosphorus source for fertilizer. Under anaerobic conditions, denitrification was rapid, resulting in the removal of large amounts of nitrogen from barley shochu wastewater. These results suggest that small shochu manufacturers could benefit from using PAWA to remove phosphorus and organic compounds and then by using a combination of the upflow anaerobic sludge blanket and the downflow hanging sponge method (UASB-DHS method) for nitrification/denitrification.     

Comparison of startup and anaerobic wastewater treatment in UASB, hybrid and baffled reactor

An experimental study was carried out to compare the performance of selected anaerobic high rate reactors operated simultaneously at 37?°C. The three reactors, namely upflow anaerobic sludge bed reactor (UASB), hybrid of UASB reactor and anaerobic filter (anaerobic hybrid reactor – AHR) and anaerobic baffled reactor (ABR), were inoculated with the anaerobic digested sludge from municipal wastewater treatment plant and tested with synthetic wastewater. This wastewater contained sodium acetate and glucose with balanced nutrients and trace elements (COD 6000?mg?·?l^?1). Organic loading rate (B _v ) was increased gradually from an initial 0.5?kg?·?m^?3?·?d^?1 to 15?kg?·?m^?3?·?d^?1 in all the reactors. From the comparison of the reactors' performance, the lowest biomass wash-out resulted from ABR. In the UASB, significant biomass wash-out was observed at the B _v 6?kg?·?m^?3?·?d^?1, and in the AHR at the B _v 12?kg?·?m^?3?·?d^?1. The demand of sodium bicarbonate for pH maintenance in ABR was two times higher as for UASB and AHR. The efficiency of COD removal was comparable for all three reactors – 80–90%. A faster biomass granulation was observed in the ABR than in the other two reactors. This fact is explained by the kinetic selection of filamentous bacteria of the Methanotrix sp. under a high (over 1.5?g?·?l^?1) acetate concentration.     

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.     

Kinetic Modelling and Characterization of Microbial Community Present in a Full-Scale UASB Reactor Treating Brewery Effluent

The performance of a full-scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was investigated by microbial analysis and kinetic modelling. The microbial community present in the granular sludge was detected using fluorescent in situ hybridization (FISH) and further confirmed using polymerase chain reaction. A group of 16S rRNA based fluorescent probes and primers targeting Archaea and Eubacteria were selected for microbial analysis. FISH results indicated the presence and dominance of a significant amount of Eubacteria and diverse group of methanogenic Archaea belonging to the order Methanococcales, Methanobacteriales, and Methanomicrobiales within in the UASB reactor. The influent brewery wastewater had a relatively high amount of volatile fatty acids chemical oxygen demand (COD), 2005 mg/l and the final COD concentration of the reactor was 457 mg/l. The biogas analysis showed 60–69 % of methane, confirming the presence and activities of methanogens within the reactor. Biokinetics of the degradable organic substrate present in the brewery wastewater was further explored using Stover and Kincannon kinetic model, with the aim of predicting the final effluent quality. The maximum utilization rate constant U _max and the saturation constant (K _B) in the model were estimated as 18.51 and 13.64 g/l/day, respectively. The model showed an excellent fit between the predicted and the observed effluent COD concentrations. Applicability of this model to predict the effluent quality of the UASB reactor treating brewery wastewater was evident from the regression analysis (R ²?=?0.957) which could be used for optimizing the reactor performance.     

Liquefaction and gasification during anaerobic digestion of coffee waste by two-phase methane fermentation with slurry-state liquefaction

In order to improve the gas evolution rate during anaerobic digestion of coffee waste by two-phase methane fermentation with slurry-state liquefaction, the liquefaction and gasification processes were separately investigated. In the liquefaction process (including the acidification process), treatment at a pH above 6 had no major effects on the liquefaction and acidification rates. However, the VFA production rates were 880 and 320 mg/l·d during mesophilic (37°C) and thermophilic (53°C) liquefaction, respectively. Mesophilic conditions were superior to thermophilic conditions in the liquefaction. With respect to the gasification process, a high TOC volumetric loading rate of 21 g/l·d was achieved during thermophilic gasification. However, the mesophilic gasification did not yield stable data, even at a low TOC volumetric loading rate of 2 g/l·d. The gas yield was 1.7 l/g TOC consumed during thermophilic gasification. The thermophilic liquefaction and thermophilic gasification reactors were connected in series and a two-phase experiment was conducted with the reactors at various volumetric ratios. The maximum gas evolution rate of 1.43 l/l·d was achieved with a combination of a gasification reactor with a 0.45l working volume and liquefaction reactor with a 2l working volume. This rate was 1.7 times higher than the rate obtained in a previous study.     

Characteristics of methanogenic granules grown on propionate in an upflow anaerobic sludge blanket (UASB) reactor

Granulation of a propionate-degrading consortia was performed with a mesophilic propionate-acclimatized sludge in an upflow anaerobic sludge blanket (UASB) reactor. The granules formed were relatively small, ranging mainly from 0.3 to 0.6 mm in diameter, but had an excellent sedimentation velocity due to a high specific gravity of 1.355 g/cm³ (ash content, 48.2%). The ash consisted mainly of calcium (30.2%), phosphorus (19.7%), and magnesium (3.95%) forming plate crystals in the granules. The populations of three bacterial trophic groups present in the granules, propionate-degraders, hydrogenotrophic and aceticlastic methanogens were 5.6 × 10⁸, 1.6 × 10¹⁰, and 2 × 10⁹ (in most probable number/g mixed-liquor volatile suspended solids [MLVSS]), respectively, while the specific utilization rates of propionate, hydrogen, and acetate of the granules were 9.4, 850, and 20.9 (mmol/g MLVSS·d), respectively. Electron microscopic analysis showed that Methanothrix spp. appeared dominant over the granules. Total granular sludge concentration retained in the UASB reactor during 178 d of operation was 80.0 g mixed-liquor suspended solids (MLSS)/l-reactor, corresponding to 41.4 g MLVSS/l-reactor, which realized a high-rate methanogenic fermentation of propionate of 85 g chemical oxygen demand (COD)/l-reactor·d.     

Accumulation of propionic acid during anaerobic treatment of distillery wastewater from barley-Shochu making

Synthetic wastewater consisting aliphatic acids contained in distillery wastewater from barley-shochu making was treated anaerobically. It was suggested that propionic acid was produced from lactic acid and citric acid via succinic acid. Since it appears to be difficult to treat anaerobically wastewater in which propionic acid is accumulated, we attempted to repress the production of propionic acid during acidification. The amount of propionic acid produced increased with an increase in the hydraulic retention time (HRT) at pH 7. Although the treatment was examined using different pHs at a shorter HRT of 10 h, it was difficult to repress the production of propionic acid.     

Pyridine degradation characteristics of a newly isolated bacterial strain and its application with a novel reactor for the further treatment in pyridine wastewater

A pyridine-degrading strain Gemmobacter sp. ZP-12, isolated from an activated sludge, was able to use pyridine as the sole carbon and nitrogen source for the growth. The strain could effectively degrade pyridine and remove TOC over a wide range of initial pyridine concentrations. The pyridine degradation rate for 100, 500, 1000, 1500 and 2000 mg/L was 2.90 ± 0.17; 13.72 ± 0.21, 20.40 ± 0.24, 31.09 ± 0.26, 27.63 ± 0.17 mg/L/h, respectively. During the pyridine degraded, a large amount of NH₄⁺-N was released and accumulated. The accumulation of NH₄⁺-N increased with the increase of pyridine concentration. For further removing the NH₄⁺-N producing in pyridine degradation, an aerobic-moving bed biofilm reactor coupled with intermittent-aeration membrane biological reactor (a-MBBR-IMBR) was constructed, in which the strain and the aerobic / anoxic mixed sludge combined to remove the pollutants in the wastewater containing 500 mg/L pyridine. After 96 h of operation, the final TOC removal efficiency was 96.5 ± 1.05 %. The average residual concentration of NO₃⁻-N and NH₄⁺-N was respectively 9.09 ± 4.13 mg/L and 7.85 ± 3.88 mg/L. The study provides a viable option for treating pyridine wastewater.     

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%.     

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.     

Removal of PO4 and NH4 from wastewaters by UASB methanogenic fermentation

The removal of PO₄ and NH₄ from wastewaters was investigated by stimulating the formation of precipitates containing PO₄ and NH₄ in granulated sludge in UASB methanogenic fermentation. In a synthetic wastewater containing volatile fatty acids, the removal efficiencies of PO₄ and NH₄ were 85 (influent=406 mg/l) and 60% (influent=100 mg/l), respectively, when 8 mM Ca and 8 mM Mg were added in the influent. In a heat-treated liquor of sewage sludge, a PO₄ removal efficiency of 70% (influent=53 mg/l) was achieved by the addition of 2 mM Ca and 3.5 mM Mg; removal of NH₄, however, was not observed during the experimental periods.     

Enhanced sulfate reduction and trichloroethylene (TCE) biodegradation in a UASB reactor operated with a sludge developed from hydrothermal vents sediments: Process and microbial ecology

Most Trichloroethylene (TCE) biodegradation reports refer to methanogenic conditions, however, in this work, enhanced sulfidogenesis and TCE biodegradation were achieved in an upflow anaerobic sludge blanket (UASB) reactor in which a completely sulfidogenic sludge, from hydrothermal vents sediments, was developed. The work was divided in three stages, (i) sludge development and sulfate reducing activity (SRA) evaluation, (ii) TCE biodegradation and (iii) SRA evaluation after TCE biodegradation. For (i) SR was 98 ± 0.1%, 84% as sulfide (H₂S, 1200 ± 28 mg/L), sulfate reducing activity (SRA) was 188 ± 50 mg COD H₂S/g VSS*d. For (ii) The reactor reached 74% of TCE removal, concentrations of vinyl chloride of 16 ± 0.3 μM (5% of the TCE added) and ethene 202 ± 81 μM (67% of the TCE added), SRA of 161 ± 7 mg COD H₂S/g VSS*d, 68% of sulfide (H₂S) production and 93% of COD removal. For (iii) SRA was of 248 ± 22 mg COD H₂S/g VSS*d demonstrating no adverse effects due to TCE.Among the genera of the microorganisms identified in the sludge during TCE biodegradation were: Dehalobacter, Desulfotomaculum, Sulfospirillum, Desulfitobacterium, Desulfovibrio and Clostridium. To the best of our knowledge, this is the first report using a sulfidogenic UASB reactor to biodegrade TCE. The overall conclusions of this work are that the reactor is efficient on both, sulfate and TCE biodegradation and it could be used to decontaminate wastewater containing organic solvents and relatively high concentrations of sulfate.     

Phototrophic bacterial production of oleic acid in an illuminated upflow anaerobic sludge blanket reactor

Phototrophic bacterial cells in the effluent from a lighted upflow anaerobic sludge blanket reactor supplied with a medium containing 142 mg S (as SO₄ ^2–) l^–1 accumulated a 6.8% w/w oleic acid content in cells and 19 mg cell-bound oleic acid l^–1 in the effluent. Pure cultures of Rhodopseudomonas palustris and Blastochloris sulfoviridis isolated from the effluent also accumulated 5.1 and 6.4% w/w oleic acid contents in cells, respectively. The oleic acid content in the cells recovered from the LUASB reactor effluent was related to the phototrophic bacterial population in the LUASB reactor. The inverse relationship was observed in the LUASB reactor between phototrophic bacterial growth and sulfate concentration in the influent.     

Anammox bacteria enrichment in upflow anaerobic sludge blanket (UASB) reactor

We investigated the anaerobic ammonium oxidation (anammox) reaction in a labscale upflow anaerobic sludge blanket (UASB) reactor. Our aim was to detect and enrich the organisms responsible for the anammox reaction using a synthetic medium that contained low concentrations of substrates (ammonium and nitrite). The reactor was inoculated with granular sludge collected from a full-scale anaerobic digestor used for treating brewery wastewater. The experiment was performed during 260 days under conditions of constant ammonium concentration (50 mg NH₄/⁺-N/L) and different nitrite concentrations (50∼150 mg NO₂-N/L). After 200 days, anammox activity was observed in the system. The microorganisms involved in this anammox reaction were identified as CandidatusB. Anammoxidans andK. Stuttgartiensis using fluorescencein situ hybridization (FISH) method.     

Improvement of the anaerobic treatment of potato processing wastewater in a UASB reactor by co-digestion with glycerol

The effect of three different types of glycerol on the performance of up-flow anaerobic sludge blanket (UASB) reactors treating potato processing wastewater was investigated. High COD removal efficiencies were obtained in both control and supplemented UASB reactors (around 85%). By adding 2 ml glycerol product per liter of raw wastewater, the biogas production could be increased by 0.74 l biogas ml⁻¹ glycerol product, which leads to energy values in the range of 810–1270 kWh_electric per m³ product. Moreover, a better in-reactor biomass yield was observed for the supplemented UASB reactor (0.012 g VSS g⁻¹ COD_removed) compared to the UASB control (0.002 g VSS g⁻¹ COD_removed), which suggests a positive effect of glycerol on the sludge blanket growth.