Effect of pH on phase separated anaerobic digestion

A pilot scale experiment was performed for a year to develop a two-phase anaerobic process for piggery wastewater treatment (COD: 6,000 mg/L, BOD: 4,000 mg/L, SS: 500 gm/L, pH 8.4, alkalinity 6,000 mg/L). The acidogenic reactor had a total volume of 3 m³, and the methanogenic reactor, an, anaerobic up-flow sludge filter, combining a filter and a sludge bed, was also of total volume 3 m³ (1.5 m³ of upper packing material). Temperatures of the acidogenic and methanogenic reactors kept at 20°C and 35°C., respectively. When the pH of the acidogenic reactor was controlled at 6.0–7.0 with HCl, the COD removal efficiency increased from 50 to 80% over a period of six months, and as a result, the COD of the final effluent fell in the range of 1,000–1,500 mg/L. BOD removal efficiency over the same period was above 90%, and 300 to 400 mg/L was maintained in the final effluent. The average SS in the final effluent was 270 mg/L. The methane production was 0.32 m³ CH₄/kg COD_removed and methane content of the methanogenic reactor was high value at 80–90%., When the pH of the acidogenic reactor was not controlled over the final two months, the pH reached 8.2 and acid conversion decreased compared with that of pH controlled, while COD removal was similar to the pH controlled operation. Without pH control, the methane content in the gas from methanogenic reactor improved to 90%, compared to 80% with pH control.     

Effects of key operational parameters on biohydrogen production via anaerobic fermentation in a sequencing batch reactor

In this study, a series of tests were conducted in a 6 L anaerobic sequencing batch reactor (ASBR) to investigate the effect of pH, hydraulic retention time (HRT) and organic loading rate on biohydrogen production at 28 °C. Sucrose was used as the main substrate to mimic carbohydrate-rich wastewater and inoculum was prepared from anaerobic digested sludge without pretreatment. The reactor was operated initially with nitrogen sparging to form anaerobic condition. Results showed that methanogens were effectively suppressed. The optimum pH value would vary depending on the HRT. Maximum hydrogen production rate and yield of 3.04 L H₂/L reactor d and 2.16 mol H₂/mol hexose respectively were achieved at pH 4.5, HRT 30 h, and OLR 11.0 kg/m³ d. Two relationships involving the propionic acid/acetic acid ratio and ethanol/acetic acid ratio were derived from the analysis of the metabolites of fermentation. Ethanol/acetic acid ratio of 1.25 was found to be a threshold value for higher hydrogen production.     

Development of an immobilized cell reactor for the production of 1,3-propanediol by Citrobacter freundii

Citrobacter freundii DSM 30040 immobilized on modified polyurethane carrier particles PUR 90/16 was used for continuous glycerol fermentation in an anaerobic fixed bed reactor with effluent recycle and pH control (fixed bed loop reactor). The fermentor was run with buffered mineral medium under growth conditions resulting in the permanent renewal of active biomass. The effects of glycerol concentration in the feed, dilution rate (D), pH and temperature (T) were investigated to optimize the process. With 400 mm glycerol in the feed, pH 6.9, T = 36°C and D = 0.5 h^–1 the maximum productivity could be determined as 8.2 g/l per hour of 1,3-propanediol.     

Anaerobic Immobilized Yeast Cell Fermentation and Anaerobic Remediation in Hybrid Reactor for Mineralization of Dicarboxylic Acid Solid Waste

Summary The solid resinous product (SRP) containing unsaturated/saturated dicarboxylic acid residues, phthalic acid and maleic acid is discharged as a solid waste during cracking of benzene over vanadium at temperatures above 500°C in the dicarboxylic acid manufacturing industry. In the present study the solid waste was diluted with water to a concentration of 0.5% w/v for microbial degradation. The waste was fermented in a reactor containing mesoporous activated carbon on which was immobilized Saccharomyces cerevisiae at an optimum residence time of 24 h at pH 6.5. The immobilized-yeast-treated samples were further treated in an upflow anaerobic reactor at an hydraulic retention time (HRT) of 0.1038 days at a hydraulic flow rate of 7.34 × 10⁻³ m³/day and chemical oxygen demand (COD) loading rate of 2.19 kg/m³/day. The pathway followed in the degradation of dicarboxylic acid into end products by anaerobic metabolism in the yeast cell fermentor and in the upflow anaerobic reactor was confirmed through HPLC, Fourier transform infra red spectroscopy and proton and ¹³C NMR spectroscopy.     

Anaerobic biodegradation of triphenylmethane dyes in a hybrid UASFB reactor for wastewater remediation

Anaerobic digestions have been proved more successful than aerobic systems for the degradation and destruction of dye-containing wastewaters. The performance of a hybrid up flow anaerobic sludge-filter bed (UASFB) reactor was tested with a synthetic wastewater containing Crystal violet (CV) as a carbon source and sodium acetate as a co-substrate. Continuous feeding of the reactor started with an initial OLR of 0.9 g COD/l-d and then it was increased step wise to 4 g COD l⁻¹ d⁻¹, while maintaining constant HRT (24 h). The optimum pH value and temperature for decolorization of crystal violet by this mixed culture species under anaerobic conditions were found to be 8–9 and 30–35°C respectively. N,N-dimethylaminophenol and N,N-bis (dimethylamino) benzophenone (Michler’s Ketone) were detected as the degradative metabolites of Crystal Violet. Subsequently, N,N-dimethylaminophenol was further degraded to aniline in the reactor whereas Michler’s ketone was not degraded under anaerobic conditions. The UASFB bioreactor was able to remove the CV completely up to a loading rate of 100 mg CV l⁻¹d⁻¹.     

Fermentative production of 1,3-propanediol from glycerol by Clostridium butyricum up to a scale of 2m3

Summary The conversion of glycerol to 1,3-propanediol (PD) by Clostridium butyricum DSM 5431 was studied in anaerobic culture. Growth and product formation were optimal at pH = 7.0 and T = 35° C, while aeration rate and stirrer speed were found to have no significant influence. As increasing amounts of initial glycerol led to inhibition of growth, cultivations were done in fed-batch operation. Comparative cultivations were carried out in an air-lift (ALR) and a stirred-tank reactor (STR) having equal working volumes (V _L = 30 l) and no difference in product formation was found. The process was scaled up to reactor sizes of 1.2 m³ (ALR) and 2.0 m³ (STR). The same results were obtained irrespective of reactor volume as well as reactor type (STR/ALR). PD concentrations of approximately 50–58 g·l^–1 and overall productivities of 2.3–2.9 g·l^–1 ·h^–1 could be reached. Offprint requests to: W.-D. Deckwer     

Effect of starch addition on the biological conversion and microbial community in a methanol-fed UASB reactor during long-term continuous operation

The effect of starch addition on the microbial composition and the biological conversion was investigated using two upflow anaerobic sludge bracket (UASB) reactors treating methanolic wastewater: one reactor was operated with starch addition, and another reactor was operated without starch addition. Approximately 300 days of operation were performed at 30 kg COD/m³/d, and then, the organic load of the reactors was gradually increased to 120 kg COD/m³/d. Successful operation was achieved at 30 kg COD/m³/d in both reactors; however, the methanol-fed reactor did not perform well at 120 kg COD/m³/d while the methanol-starch-fed reactor did. The granule analysis revealed the granule developed further only in the methanol-starch-fed reactor. The results of the microbial community analysis revealed more Methanosaeta cells were present in the methanol-starch-fed reactor, suggesting the degradation of starch produced acetate as an intermediate, which stimulated the growth of Methanosaeta cells responsible for the extension of granules.     

Anaerobic digestion of a petrochemical effluent using an anaerobic hybrid digester

Summary An anaerobic hybrid reactor was used in the anaerobic treatment of an acidic petrochemical effluent. An organic loading rate of 20.04 kg COD/(m³d) at a HRT of 17 hours was obtained with a volatile fatty acid removal of 91%, and COD removal of 84%. A final reactor effluent containing 44 mg/l ammonia nitrogen and 12.3 mg/l PO₄-P was produced.     

High-rate nitrogen removal from livestock manure digester liquor by combined partial nitritation–anammox process

In this study, combination of a partial nitritation reactor, using immobilized polyethylene glycol (PEG) gel carriers, and a continuous stirred granular anammox reactor was investigated for nitrogen removal from livestock manure digester liquor. Successful nitrite accumulation in the partial nitritation reactor was observed as the nitrite production rate reached 2.1 kg-N/m³/day under aerobic nitrogen loading rate of 3.8 kg-N/m³/day. Simultaneously, relatively high free ammonia concentrations (average 50 mg-NH₃/l) depressed the activity of nitrite oxidizing bacteria with nitrate concentration never exceeding 3% of TN concentration in the effluent of the partial nitritation reactor (maximum 35.2 mg/l). High nitrogen removal rates were achieved in the granular anammox reactor with the highest removal rate being 3.12 kg-N/m³/day under anaerobic nitrogen loading rate of 4.1 kg-N/m³/day. Recalcitrant organic compounds in the digester liquor did not impair anammox reaction and the SS accumulation in the granular anammox reactor was minimal. The results of this study demonstrated that partial nitritation–anammox combination has the potential to successfully remove nitrogen from livestock manure digester liquor.     

Identification of bacteria coexisting with anammox bacteria in an upflow column type reactor

Anammox process has attracted considerable attention in the recent years as an alternative to conventional nitrogen removal technologies. In this study, a column type reactor using a novel net type acrylic fiber (Biofix) support material was used for anammox treatment. The Biofix reactor was operated at a temperature of 25°C (peak summer temperature, 31.5°C). During more than 340 days of operation for synthetic wastewater treatment, the nitrogen loading rates of the reactor were increased to 3.6 kg-N/m³/d with TN removal efficiencies reaching 81.3%. When the reactor was used for raw anaerobic sludge digester liquor treatment, an average TN removal efficiency of 72% was obtained with highest removal efficiency of 81.6% at a nitrogen loading rate of 2.2 kg-N/m³/d. Results of extracellular polymeric substances (EPS) quantification revealed that protein was the most abundant component in the granular sludge and was found to be almost twice than that in the sludge attached to the biomass carriers. The anammox granules in the Biofix reactor illustrated a dense morphology substantiated by scanning electron microscopy and EPS results. The results of DNA analyses indicated that the anammox strain KSU-1 might prefer relatively low nutrient levels, while the anammox strain KU2 strain might be better suited at high nutrient concentration. Other types of bacteria were also identified with the potential of consuming dissolved oxygen in the influent and facilitating survival of anammox bacteria under aerobic conditions.     

Optimization of two-phase thermophilic anaerobic digestion of biowaste for hydrogen and methane production through reject water recirculation

The optimization of a two-phase thermophilic anaerobic process treating biowaste for hydrogen and methane production was carried out at pilot scale using two stirred reactors (CSTRs) and without any physical/chemical pre-treatment of inoculum. During the experiment the hydrogen production at low hydraulic retention time (3d) was tested, both with and without reject water recirculation and at two organic loading rate (16 and 21 kgTVS/m³d). The better yields were obtained with recirculation where the pH reached an optimal value (5.5) thanks to the buffering capacity of the recycle stream. The specific gas production of the first reactor was 51 l/kgVS_fed and H₂ content in biogas 37%. The mixture of gas obtained from the two reactors met the standards for the biohythane mix only when lower loading rate were applied to the first reactor, with a composition of 6.7% H₂, 40.1% CO₂ and 52.3% CH₄ the overall SGP being 0.78 m³/kgVS_fed.     

Mesophilic batch anaerobic co-digestion of pulp and paper sludge and monosodium glutamate waste liquor for methane production in a bench-scale digester

This paper presented results from anaerobic co-digestion of pulp and paper sludge (PPS) and monosodium glutamate waste liquor (MGWL). A bench-scale anaerobic digester, 10 L in volume was developed, to operate under mesophilic (37 ± 2 °C) batch condition. Under versatile and reliable anaerobic conduct, high efficiency for bioconversion of PPS and MGWL were obtained in the system. The accumulative methane yield attained to 200 mL g⁻¹ VS_added and the peak value of methane daily production was 0.5 m³/(m³ d). No inhibitions of volatile fatty acids (VFAs) and ammonia on anaerobic co-digestion were found. pH 6.0-8.0 and alkalinity 1000-4000 mg CaCO₃/L were got without adjustment. This work showed that there was a good potential to the use of PPS and MGWL to anaerobic co-digestion for methane production.     

The influence of different substrate pH values on the performance of a downflow anaerobic fixed bed reactor treating a petrochemical effluent

Summary Neutralizing requirements for the anaerobic treatment of an acidic petrochemical effluent in a downflow anaerobic fixed bed reactor were examined. Neutralization (pH 6.0 with NaOH) of the effluent prior to digestion resulted in a Na⁺ concentration of over 3 g/l which was detrimental. Decreasing the Na⁺ concentration and subsequent replacement of NaOH by a mixture of Ca(OH)₂, NaOH and KOH resulted in an increase in reactor performance. The addition of different alkalines resulted in the best loading rates thusfar applied in the anaerobic treatment of this petrochemical effluent. During the final stages of this study, the effluent (pH 3.95) was treated at a loading rate of 10.37 kg COD/m³. d (HRT=1.35 d) with more than 94% fatty acid removal.     

Two-stage anaerobic treatment of dairy wastewater using HUASB with PUF and PVC carrier

In the present study, an attempt has been made to treat dairy wastewater entirely via anaerobic treatment over a period of 215 days, using two-stage Hybrid Upflow Anaerobic Sludge Blanket (HUASB) reactors, which offer the advantages associated both with fixed film and upflow sludge blanket treatments. A HUASB with polyurethane foam cubes was used for stage I, and a HUASB utilizing PVC-cut rings was used for stage II. The output from stage I was used as the input for stage II. The two-stage reactor was operated at an organic loading rate that varied from 10.7 to 21.4 kg COD m³/d for a period of 215 days, including the start-up period. The ideal organic loading rate for the two-stage reactor was 19.2 kg COD/m³/d. A further 21.4 kg COD m³/d increase in the organic loading rate resulted in the souring of the reactor function in stage I, which consequently reduced the overall reactor performance. Combined COD removal during the stable operation period (10.7 to 19.2 kg COD m³/d) occurred in a range between 97 and 99%. The methane content in the biogas varied from 65 to 70% in stage I, and from 63 to 66% in stage II. The two-stage anaerobic treatment using HUASB with PUF and PVC described in this work is expected to constitute a better alternative for the complete treatment of dairy wastewater than high-rate anaerobic, anaerobic/aerobic, and two-phase anaerobic treatment methods.     

A comparison study on the high-rate co-digestion of sewage sludge and food waste using a temperature-phased anaerobic sequencing batch reactor system

Assessing contemporary anaerobic biotechnologies requires proofs on reliable performance in terms of renewable bioenergy recovery such as methane (CH₄) production rate, CH₄ yield while removing volatile solid (VS) effectively. This study, therefore, aims to evaluate temperature-phased anaerobic sequencing batch reactor (TPASBR) system that is a promising approach for the sustainable treatment of organic fraction of municipal solid wastes (OFMSW). TPASBR system is compared with a conventional system, mesophilic two-stage anaerobic sequencing batch reactor system, which differs in operating temperature of 1st-stage. Results demonstrate that TPASBR system can obtain 44% VS removal from co-substrate of sewage sludge and food waste while producing 1.2 m³CH₄/m³_system/d (0.2 m³CH₄/kgVS_added) at organic loading rate of 6.1 gVS/L/d through the synergy of sequencing-batch operation, co-digestion, and temperature-phasing. Consequently, the rapid and balanced anaerobic metabolism at thermophilic stage makes TPASBR system to afford high organic loading rate showing superior performance on OFMSW stabilization.     

Treatment of food processing wastewater in a full-scale jet biogas internal loop anaerobic fluidized bed reactor

A full-scale jet biogas internal loop anaerobic fluidized bed (JBILAFB) reactor, which requires low energy input and allows enhanced mass transfer, was constructed for the treatment of food processing wastewater. This reactor has an active volume of 798 m³ and can treat 33.3 m³ wastewater per hour. After pre-treating the raw wastewater by settling, oil separating and coagulation-air floating processes, the reactor was operated with a relatively shorter start-up time (55 days). Samples for the influent and effluent of the JBILAFB reactor were taken and analyzed daily for the whole process including both the start-up and stable running periods. When the volumetric COD loading fluctuated in the range of 1.6–5.6 kg COD m⁻³ day⁻¹, the COD removal efficiency, the volatile fatty acid(VFA)/alkalinity ratio, the maximum biogas production and the content of CH₄ in total biogas of the reactor were found to be 80.1 ± 5%, 0.2–0.5, 348.5 m³ day⁻¹ and 94.5 ± 2.5%, respectively. Furthermore, the scanning electron microscope (SEM) results showed that anaerobic granular sludge and microorganism particles with biofilm coexisted in the reactor, and that the bacteria mainly in bacilli and cocci were observed as predominant species. All the data demonstrated that the enhanced mass transfer for gas, liquid and solid phases was achieved, and that the formation of microorganism granules and the removal of inhibitors increased the stability of the system.     

Separation and simultaneous determination of nalidixic acid, hydroxynalidixic acid and carboxynalidixic acid in serum and urine by micellar electrokinetic capillary chromatography

Separation in capillary electrophoresis is governed by various factors, including buffer type, buffer concentration, pH, temperature, voltage and micelles. Through proper adjustment of these parameters, nalidixic acid and its two major metabolites, 7-hydroxynalidixic and 7-carboxynalidixic, could be separated by micellar electrokinetic capillary chromatography using an electrophoretic electrolyte consisting of 50 mM borate buffer (pH 9) containing 25 mM sodium dodecyl sulphate and 10% acetonitrile. A linear relationship between concentration and peak area for each compound was obtained in the concentration range 0.15–100 μg ml⁻¹, with a correlation coefficient greater than 0.999 and detection limits in the 0.2–0.7 ng ml⁻¹ range. Intra- and inter-day precision values of about 0.8–1.2% RSD (n=11) and 1.3–2.0% RSD (n=30), respectively, were obtained. The method has been applied to the analysis of nalidixic acid and its two major metabolites in serum and urine with limits of sensitivity lower than 0.8 ng ml⁻¹.     

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.     

Anaerobic treatment of synthetic medium-strength wastewater using a multistage biofilm reactor

A laboratory-scale multistage anaerobic biofilm reactor of three compartments with a working volume of 54-L was used for treating a synthetic medium-strength wastewater containing molasses as a carbon source at different influent conditions. The start-up period, stability and performance of this reactor were assessed at mesophilic temperature (35 °C). During the start-up period, pH fluctuations were observed because there was no microbial selection or zoning, but as the experiment progressed, results showed that phase separation had occurred inside the reactor. COD removal percentages of 91.6, 91.6, 90.0 and 88.3 were achieved at organic loading rates of 3.0, 4.5, 6.75 and 9.0 kg COD/m³ day, respectively. A decrease in HRT from 24 to 16 h had no effect on COD removal efficiency. When HRT decreased to 8 h, COD removal efficiency was still 84.9%. Recirculation ratios of 0.5 and 1.0 had no effect on COD removal but other factors such as the volatile fatty acid (VFA) content were affected. The effect of toxic shock was also investigated and results showed that the main advantage of using this bioreactor lies in its compartmentalized structure.     

Partial nitritation and anammox of a livestock manure digester liquor and analysis of its microbial community

A swim-bed reactor for partial nitritation with polymeric coagulant treatment and an UASB reactor for anammox were applied to the treatment of livestock manure digester liquor. The partial nitritation was maintained for 32 days under a 1.6 kg N/m³/d nitrogen loading rate (NLR) with an average conversion efficiency of 51%, and achieved 1.65 kg N/m³/d of the maximum nitrite production rate under 2.58 kg N/m³/d of NLR. Although 200 mg/L of TOC remained in the effluent of the partial nitritation reactor, the anammox nitrogen removal rate was not significantly decreased and a relatively high rate of 2.0 kg N/m³/d was obtained under a NLR of 2.2 kg N/m³/d. 16S rRNA gene analysis showed that Nitrosomonas and KSU-1 were dominant in the partial nitritation and anammox reactor, respectively. The results of this study demonstrated that the partial nitritation-anammox process has possibility of applying to the nitrogen removal of livestock manure digester liquor.