Granulation of biomass in thermophilic upflow anaerobic sludge blanket reactors treating acidified wastewaters

The development of granular sludge in thermophilic (55 degrees C) upflow anaerobic sludge blanket reactors was investigated. Acetate and a mixture of acetate and butyrate were used as substrates, serving as models for acidified waste-waters. Granular sludge with either Methanothrix or Methanosarcina as the predominant acetate utilizing methanogen was cultivated by allowing the loading rate to increase whenever the acetate concentration in the effluent dropped below 200 and 700 mg COD/L, respectively. The highest methane generation rates, up to 162 kg CH(4)-COD/m(3) day, or 2.53 mole CH(4)/L day, were achieved at hydraulic retention times down to 21 min, with granules consisting of Methanothrix. The formation of Methanothrix granules did not depend on the type of seed material, nor on the addition of inert support particles. The growth of granules proceeded rapidly with adapted seed material, even when the reactors were inoculated with low concentrations. With mesophilic seed materials growth of granules took much longer. Thermophilic Methanothrix granules strongly resemble mesophilic granules of the "filamentous" type. Some factors governing the thermophilic granulation process are discussed.     

Biotransformation of nitrophenols in upflow anaerobic sludge blanket reactors

Four identical bench-scale upflow anaerobic sludge blanket (UASB) reactors, R1, R2, R3 and R4, were used to assess nitrophenols degradation at four different hydraulic retention times (HRT). Reactor R1 was used as control, whereas R2, R3, and R4 were fed with 2-nitrophenol (2-NP), 4-nitrophenol (4-NP), and 2,4-dinitrophenol (2,4-DNP), respectively. The concentration of each nitrophenol was gradually varied from 2 to 30 mg/l during acclimation. After acclimation reactors were operated under steady-state conditions at four different HRTs – 30, 24, 18, and 12 h, to study its effect on the removal of nitrophenols. Overall removal of 2-NP and 4-NP was always more than 99% but 2,4-DNP removal decreased from 96% to 89.7% as HRT was lowered from 30 to 12 h. 2-Aminophenol (2-AP), 4-aminophenol (4-AP) and 2-amino,4-nitrophenol (2-A,4-NP) were found to be the major intermediates during the degradation of 2-NP, 4-NP and 2,4-DNP, respectively. Out of the total input of nitrophenolic concentration (30 mg/l), on molar basis, about 41.2–48.4% of 2-NP, 59.4–68% of 4-NP, 30–26.6% of 2,4-DNP was recovered in the form of their respective amino derivatives at 30–12 h HRT. COD removal was 98–89%, 97–56%, 97–52%, and 94–46% at 30–12 h HRT for R1, R2, R3 and R4, respectively. Average cell growth was observed to be 0.15 g volatile suspended solid (VSS) per g COD consumed. Methanogenic inhibition was observed at lower HRTs (18 and 12 h), however denitrification was always more than 99% with non-detectable level of nitrite. The granules developed inside the reactors were black in color and their average size varied between 1.9 and 2.1 mm.     

Granular sludge formation in upflow anaerobic sludge blanket (UASB) reactors

The state of the art for upflow anaerobic sludge blanket (UASB) reactors is discussed, focusing on the microbiology of immobilized anaerobic bacteria and the mechanism of granule formation. The development of granular sludge is the key factor for successful operation of the UASB reactors. Criteria for determining if granular sludge has developed in a UASB reactor is given based on the densities and diameters of the granular sludge. The shape and composition of granular sludge can vary significantly. Granules typically have a spherical form with a diameter from 0.14 to 5 mm. The inorganic mineral content varies from 10 to 90% of the dry weight of the granules, depending on the wastewater composition etc. The main components of the ash are calcium, potassium, and iron. The extracellular polymers in the granular sludge are important for the structure and maintenance of granules, while the inorganic composition seems to be of less importance. The extracellular polymer content varies between 0.6 and 20% of the volatile suspended solids and consists mainly of protein and polysaccharides. Both Methanosaeta spp. (formerly Methanothrix) and Methanosarcina spp. have been identified as important aceticlastic methanogens for the initial granulation and development of granular sludge. Immunological methods have been used to identify other methanogens in the granules. The results have showed that, besides the aceticlastic methanogens Methanosaeta spp. and Methanosarcina spp., hydrogen and formate utilizing bacteria are also present, e.g., Methanobacterium formicicum, Methanobacterium thermoautotrophicum, and Methanobrevibacter spp. Microcolonies of syntrophic bacteria are often observed in the granules, and the significant electron transfer in these microcolonies occurs through interspecies hydrogen transfer. The internal organization of the various groups of bacteria in the granules depends on the wastewater composition and the dominating metabolic pathways in the granules. Internal organization is observed in granules where such an arrangement is beneficial for an optimal degradation of the wastewater. A four-step model is given for the initial development of granular sludge. (c) 1996 John Wiley & Sons, Inc.     

Granulation in thermophilic upflow anaerobic sludge blanket (UASB) reactors

The state of the art for thermophilic UASB reactors is discussed focusing on the start-up of UASB reactors, the influence of the waste water composition and temperature on the development and maintenance of thermophilic granules, and the microbial composition and structure of thermophilic granules.     

An original procedure for physical simulation of upflow anaerobic sludge blanket reactors

A procedure for sludge blanket reactors physical simulation is presented. To simulate the liquid phase, a solution of 1.0 g/l of polyvinyl alcohol provided the desired rheological behavior. Biological granules presented in UASB reactors were simulated using granules of a water absorbent acrylic polymer capable to expand up to one hundred times in volume when moistened. The biochemical reactions that produces methane and carbon dioxide in anaerobic reactors were simulated reacting nitric acid and sodium bicarbonate. This procedure differs from gas injection since the gas is produced at the reactor core as it happens in the actual situation, thus the simulation is more realistic. Flow visualization and residence time distribution measurements were used to verify if the proposed procedure acceptably simulates UASB reactors treating wastewater. All tests were performed in a 10.5 l bench scale reactor and the results allow to recommend this method whenever flow visualization and reactor physical simulation are useful for reactor design and development.     

High upflow velocity and organic loading rate improve granulation in upflow anaerobic sludge blanket reactors

Five laboratory scale upflow anaerobic sludge blanket (UASB) reactors were seeded with nongranular sewage sludge. Granulation was obtained after 15–35 days when between 0.5 and 2.0m/h upflow liquid velocity was applied, with an organic loading rate (OLR) of 8g COD/l.d (COD is the chemical oxygen demand). Granules had different physical characteristics and specific activity (g COD_REMOVED/g volatile suspended solids) depending on the upflow liquid velocity applied. Granules were obtained in short startup periods (5 and 14 days) when a pilot-scale (180l) UASB reactor with a height of 4.7m was used to study hydraulic effects on the granulation process.     

Hydraulics of laboratory and full-scale upflow anaerobic sludge blanket (UASB) reactors

Laboratory-scale upflow anaerobic sludge blanket (UASB) reactors are often used as test platforms to evaluate full-scale applications. However, for a given volume specific hydraulic loading rate and geometry, the gas and liquid flows increase proportionally with the cube root of volume. In this communication, we demonstrate that a laboratory-scale reactor had plug-flow hydraulics, while a full-scale reactor had mixed flow hydraulics. The laboratory-scale reactor could be modeled using an existing biochemical model, and parameters identified, but because of computational speed with plug-flow hydraulics, mixed systems are instead recommended for parameter identification studies. Because of the scaling issues identified, operational data should not be directly projected from laboratory-scale results to the full-scale design.     

Extracellular polymers in granular sludge from different upflow anaerobic sludge blanket (UASB) reactors

Thermal extraction was used to quantify extracellular polymers (ECP) in granules from anaerobic upflow reactors. The optimal time for extraction was determined as the time needed before the intracellular material gives a significant contribution to the extracted extracellular material due to cell lysis. ECP contents of 41 to 92 mg · g^–1 volatile suspended solids of granules were found depending on the type of granular sludge examined. The content of polysaccharides, protein and lipids in the extracted ECP was quantified. Furthermore, the different methyl esters of the lipids were determined and quantified. Lower amounts of polysaccharides and proteins were found in the extracellular material from granules grown on methanogenic and acetogenic substrates compared to granules grown on more complex substrates. In contrast, the lipid content was lower on complex substrates. Changing the feed of an upflow anaerobic sludge blanket reactor from a sugar-containing waste-water to a synthetic waste-water containing acetate, propionate and butyrate resulted in a decrease in both the protein and polysaccharide content and an increase in the lipid content of the extracellular material. Furthermore, the amount of protein and polysaccharides in the ECP found under mesophilic conditions was significantly higher than under thermophilic conditions, while the lipid content was lower.     

Tetrachloroethene dechlorination kinetics by Dehalospirillum multivorans immobilized in upflow anaerobic sludge blanket reactors

Tetrachloroethene (C₂Cl₄) dechlorination kinetics in upflow anaerobic sludge blanket (UASB) reactors was determined after introducing de novo activities into the granular sludge. These activities were introduced by immobilizing Dehalospirillum multivorans in a test reactor containing unsterile granular sludge, and in a reference reactor, R1, containing sterile granular sludge. A second reference reactor, R2, contained only unsterile granular sludge and served as a control. The kinetic experiments were performed by pulsing the reactors with C₂Cl₄ in a recirculating batch mode. Formate and acetate were added as electron donor and carbon source. Both reactors inoculated with D. multivorans dechlorinated C₂Cl₄ to an equimolar amount of C₂H₂Cl₂ with only traces of C₂HCl₃ in the effluent. In the control reactor, C₂HCl₃ accumulated before C₂H₂Cl₂ was produced. A computer simulation program (AQUASIM) was used to estimate the kinetic parameters. The half-saturation constants (K _s) for C₂Cl₄ and C₂HCl₃ were almost equal in the reactors containing D.␣multivorans (17 μM and 18 μM for C₂Cl₄; 26 μM and 28 μM for C₂HCl₃), indicating no influence of sludge bacteria on the affinity of D. multivorans for C₂Cl₄ and C₂HCl₃. The maximum dechlorination rates (k _m X _B) were about twice as high in the reactor containing D.␣multivorans immobilized in sterile sludge (11 mmol C₂Cl₄ l sludge⁻¹ day⁻¹ and 27 mmol C₂HCl₃ l sludge⁻¹ day⁻¹) than in the test reactor (4.4 mmol C₂Cl₄ l sludge⁻¹ day⁻¹ and 15 mmol C₂HCl₃ l sludge⁻¹ day⁻¹). Compared to other C₂Cl₄-degrading systems, the dechlorination rates of the inoculated reactors and their affinities for C₂Cl₄ and C₂HCl₃ were high. Therefore, introduction of de novo activity is promising for the use of anaerobic reactors to bioremediate C₂Cl₄-polluted water. Received: 5 November 1998 / Received revision: 25 January 1999 / Accepted: 31 January 1999     

Performance of upflow anaerobic sludge blanket (UASB) reactor treating wastewaters containing carbon tetrachloride

The anaerobic biodegradation of carbon tetrachloride (CT) was investigated during the granulation process by reducing the hydraulic retention time, increasing the chemical oxygen demand (COD) and CT loadings in a 2l laboratory-scale upflow anaerobic sludge blanket (UASB) reactor. Anaerobic unacclimated sludge and glucose were used as seed and primary substrate, respectively. Granules were developed 4 weeks after start-up, which grew at an accelerated rate for 8 months, and then became fully grown. The effect of operational parameters such as influent CT concentrations, COD, CT loading, food to biomass ratio and specific methanogenic activity (SMA) were also considered during granulation. The granular sludge cultivated had a maximum diameter of 2.1 mm and SMA of 1.6 g COD/g total suspended solid (TSS) day. COD and CT removal efficiencies of 92 and 88% were achieved when the reactor was firstly operating at CT and COD loading rates of 17.5 mg/l day and 12.5 g/l day, respectively. This corresponds to hydraulic retention time of 0.28 day and food to biomass ratio of 0.5 g COD/g TSS day. Kinetic coefficients of maximum specific substrate utilization rate, half velocity coefficient, growth yield coefficient and decay coefficient were determined to be 2.4 × 10^–3 mg CT/TSS day^–1, 1.37 mg CT/l, 0.69 mg TSS/mg CT and 0.046 day^–1, respectively for CT biotransformation during granulation.     

Feasibility of expanded granular sludge bed reactors for the anaerobic treatment of low-strength soluble wastewaters

The application of the expanded granular sludge bed (EGSB) reactor for the anaerobic treatment of low-strength soluble wastewaters using ethanol as a model substrate was investigated in laboratory-scale reactors at 30oC. Chemical oxygen demand (COD) removal efficiency was above 80% at organic loading rates up to12 g COD/L . d with influent concentrations as low as 100 to 200 mg COD/L. These results demonstrate the suitability of the EGBS reactor for the anaerobic treatment of low-strength wastewaters. The high treatment performance can be attributed to the intense mixing regime obtained by high hydraulic and organic loads. Good mixing of the bulk liquid phase for the substrate-biomass contact and adequate expansion of the substrate-biomass contact and adequate expansion of the sludge bed for the degassing were obtained when the liquid upflow velocity (V(up)) was greater than 2.5 m/h. Under such conditions, an extremely low apparent K(s) value for acetoclastic methanogenesis of 9.8 mg COD/L was observed. The presence of dissolved oxygen in the wastewater had no detrimental effect on the treatment performance. Sludge piston flotation from pockets of biogas accumulating under the sludge bed occurred at V(up) lower than 2.5 m/h due to poor bed expansion. This problem is expected only in small diameter laboratory-scale reactors. A. more important restriction of the EGSB reactor was the sludge washout occurring at V(up) higher than 5.5 m/h and which was intensified at organic loads higher than 7 g COD/L. d due to buoyancy forces from the gas production. To achieve an equilibrium between the mixing intensity and the sludge hold-up, the operation should be limited to an organic loading rate of 7 g COD/L d. and to a liquid up-flow velocity between 2.5 and 5.5 m/h (c) 1994 John Wiley & Sons, Inc.     

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%.     

Improved performance of upflow anaerobic sludge blanket (UASB) reactors by operating alternatives

Summary The efficient operation of UASB reactors treating complex soluble wastewater containing high protein and lipid content was attempted by mixing in different modes. Higher superficial flow rate increased COD removal efficiency, sludge retainment, and methane content in biogas not only in the start-up period but also at high volumetric loading rates. However, formation of sludge particles in larger size was hindered by increased upflow liquid velocity.     

Anaerobic dechlorination of pentachlorophenol in fixed-film and upflow anaerobic sludge blanket reactors using different inocula

Longterm performance and stability of two upflow anaerobic sludge blanket (UASB) reactors inoculated with granular sludge and treating a synthetic waste water containing pentachlorophenol (PCP) and phenol were studied. A similar system consisting of two fixed-film reactors inoculated with anaerobic digested sewage sludge were further studied. One reactor in each series received glucose in addition to the phenols. Dechlorination of PCP proceeded via two different dominating pathways in the respective reactor systems, suggesting that two distinct microbial populations were present, probably originating from the different inocula. Dechlorinating activity was maintained for more than 18 months in the UASB reactors and was generally higher than in the fixed-film reactors. In the fixed-film reactors, dechlorination of PCP suddenly decreased after 15.5 months of operation compared to earlier performance. Since no operational parameters had been changed, this indicated that the enriched culture was unstable on a longterm basis. Addition of yeast extract to the medium restored activity. General process stability in both reactor systems was clearly enhanced by the addition of glucose and was superior in the UASB/granular sludge system. The better performance and the higher stability in the UASB/granular sludge reactor highlights the importance of thorough screening of inocular prior to start-up of processes treating waste waters containing xenobiotic compounds.Abbreviations PCP pentachlorophenol - TeCP tetrachlorophenol - TCP trichlorophenol - DCP dichlorophenol - UASB upflow anaerobic sludge blanket - HRT hydraulic retention time     

Immobilization patterns and dynamics of acetate-utilizing methanogens immobilized in sterile granular sludge in upflow anaerobic sludge blanket reactors

Sterile granular sludge was inoculated with either Methanosarcina mazeii S-6, Methanosaeta concilii GP-6, or both species in acetate-fed upflow anaerobic sludge blanket (UASB) reactors to investigate the immobilization patterns and dynamics of aceticlastic methanogens in granular sludge. After several months of reactor operation, the methanogens were immobilized, either separately or together. The fastest immobilization was observed in the reactor containing M. mazeii S-6. The highest effluent concentration of acetate was observed in the reactor with only M. mazeii S-6 immobilized, while the lowest effluent concentration of acetate was observed in the reactor where both types of methanogens were immobilized together. No changes were observed in the kinetic parameters (Ks and mumax) of immobilized M. concilii GP-6 or M. mazeii S-6 compared with suspended cultures, indicating that immobilization does not affect the growth kinetics of these methanogens. An enzyme-linked immunosorbent assay using polyclonal antibodies against either M. concilii GP-6 or M. mazeii S-6 showed significant variations in the two methanogenic populations in the different reactors. Polyclonal antibodies were further used to study the spatial distribution of the two methanogens. M. concilii GP-6 was immobilized only on existing support material without any specific pattern. M. mazeii S-6, however, showed a different immobilization pattern: large clumps were formed when the concentration of acetate was high, but where the acetate concentration was low this strain was immobilized on support material as single cells or small clumps. The data clearly show that the two aceticlastic methanogens immobilize differently in UASB systems, depending on the conditions found throughout the UASB reactor.     

Kinetics of anaerobic treatment of slaughterhouse wastewater in batch and upflow anaerobic sludge blanket reactor

The kinetics of anaerobic treatment of slaughterhouse wastewater in batch and upflow anaerobic sludge blanket (UASB) reactors was investigated. Different concentrations of organic matter in slaughterhouse wastewater did not change the first order kinetics of the reaction. In batch digesters, methane and nitrogen production stopped after 30-40, 20-30 h, respectively, and in UASB reactors it was terminated after 30-40 days. The constant of velocity was 3.93 and 0.23 h(-1) respectively, for methane and nitrogen production. The yield coefficient, Yp was 343 and 349 ml CH4 per g of chemical oxygen demand at standard temperature and pressure conditions for batch reactors and UASB reactor, respectively.     

Two-stage anaerobic digestion of tomato, cucumber, common reed and grass silage in leach-bed reactors and upflow anaerobic sludge blanket reactors

Anaerobic digestion of tomato, cucumber, common reed and grass silage was studied in four separate two-stage reactor configuration consisting of leach bed reactor (LBR) and upflow anaerobic sludge blanket reactor (UASB). LBR studies showed that COD solubilization for cucumber and grass silage was higher (50%) than tomato (35%) and common reed (15%). Results also showed that 31-39% of initial TKN present in tomato and cucumber was solubilized in the leachates and 47-54% of the solubilized TKN was converted to NH₄-N. The corresponding values for common reed and grass silage were 38-50% and 18-36%, respectively. Biomethanation of the leachates in UASB reactors resulted in methane yields of 0.03-0.14 m³ CH₄ kg⁻¹VS_fed for the studied crop materials. Thus, high COD solubilization, high nitrogen mineralization and solubilization rates were feasible during anaerobic digestion of lignocellulosic materials in a two-stage LBR-UASB reactor system.     

Recycle in upflow anaerobic sludge blanket reactor on treatment of real textile dye effluent

The discharge of textile wastewater containing dye in the environment is varying for both toxicology and esthetical reasons as dyes impede light penetration, damage the quality of the receiving streams. Upflow anaerobic sludge blanket reactor with anaerobic digester sludge treating starch wastewater has been used to investigate the removal efficiency of chemical oxygen demand (COD) and colour of textile dye wastewater. In this study, the starch and textile dye wastewater was mixed at 70 and 30%, respectively, and the experiments were carried out with recycle of treated wastewater at different percentage as 10, 20, 30 and 40. Maximum removal of COD and colour was 96% and 93.3%, respectively, at 30% recycle. At various OLR and HRT, the maximum removal of COD, colour was 95.9%, 93% at 6.81 kg COD/m³d and 96%, 93% with 24 h of HRT. The maximum production of biogas at 24 h of HRT with 30% recycle was about 355 l/d. The Volatile fatty acid/Alkalinity ratio of methanogenic reactor was found to be 0.049–0.053. The result provided evidence, the starch and dye wastewater have wide variation in their characteristics was treated on combination, this new technology supports the effective utilization of starch waste in destruction of dye.