Application of Methanobrevibacter acididurans in anaerobic digestion.

To operate anaerobic digesters successfully under acidic conditions, hydrogen utilizing methanogens which can grow efficiently at low pH and tolerate high volatile fatty acids (VFA) are desirable. An acid tolerant hydrogenotrophic methanogen viz. Methanobrevibacter acididurans isolated from slurry of an anaerobic digester running on alcohol distillery wastewater has been described earlier by this lab. This organism could grow optimally at pH 6.0. In the experiments reported herein, M. acididurans showed better methanogenesis under acidic conditions with high VFA, particularly acetate, than Methanobacterium bryantii, a common hydrogenotrophic inhabitant of anaerobic digesters. Addition of M. acididurans culture to digesting slurry of acidogenic as well as methanogenic digesters running on distillery wastewater showed increase in methane production and decrease in accumulation of volatile fatty acids. The results proved the feasibility of application of M. acididurans in anaerobic digesters.     

Thermophilic anaerobic digester with ultrafilter for solids stabilization.

A thermophilic anaerobic digester with ultrafilter (TADU) for solids separation offers potential advantages of higher VS destruction, biomass retention, and pathogen removal. However, potential disadvantages include ultrafilter fouling, decreasing flux, and high VFA concentrations. In this study, a thermophilic anaerobic digester coupled to a sintered titanium, cross-flow ultrafilter was operated for over five months. Dairy manure was digested (HRT of 23 days). The filtrate VFA concentration was low (220 mg/L as HAc), average VS destruction was 49%, and a low average effluent fecal coliform concentration of 10(2) MPN/100 mL was observed. The low coliform value may be beneficial if dewatered biosolids are used for livestock bedding since low pathogen counts help prevent mastitis. Ultrafilter fluxes of 40-80 L/m2-hr were maintained by cleaning using caustic (3.5% NaOH) followed by water and acid (3% phosphoric acid). Sand from livestock bedding was found to damage the pump and ultrafilter. If TADU were implemented at full scale, then replacing sand bedding with dewatered biosolids should be considered.     

Anaerobic monodigestion of poultry manure: determination of operational parameters for CSTR

In this work the anaerobic monodigestion for the treatment of turkey manure was evaluated, without its codigestion with another substrate. The effect of the organic loading rate (OLR) and the substrate concentration (high total solids (TS) concentration) or product concentration (high volatile fatty acids (VFA) and/or ammonia (NH(3)-N) concentrations) was studied. The results show that for a continuous stirred tank reactor (CSTR) operation, a maximum of 40 g/L of TS and 4.0 g/L of ammonium (NH(4)(+)) was required. In addition, the maximum organic loading rate (OLR) will not exceed 1.5 kg VS/m(3)d. Higher TS and NH(4)(+) concentrations and OLR lead to a reduction on the methane productivity and volatile solids (VS) removal. During the CSTR operation, a high alkalinity concentration (above 10 g/L CaCO(3)) was found; this situation allowed maintaining a constant and appropriate pH (close to 7.8), despite the VFA accumulation. In this sense, the alkalinity ratio (α) is a more appropriate control and monitoring parameter of the reactor operation compared to pH. Additionally, with this parameter a VS removal of 80% with a methane productivity of 0.50 m(3)(CH4)/m(3)(R)d is achieved.     

The effect of temperature on the performance and stability of thermophilic anaerobic digestion.

Sustainable operation of an anaerobic sewage sludge digester requires the effective shuttling of carbon from complex organic material to methane gas. The accumulation of intermediates and metabolic products such as volatile fatty acids and hydrogen gas not only reveal inefficiency within the digestion process, but can be detrimental to reactor operation at sufficiently high levels. Eight anaerobic digesters (1 mesophilic and 7 thermophilic) were operated in order to determine the effect of steady-state digestion temperature on the operational stability and performance of the digestion process. Replicate reactors operated at 57.5 degrees C, the highest temperature studied, were prone to accumulation of volatile fatty acids (4052 and 3411 mg/L as acetate) and gaseous hydrogen. Reactors operated at or below 55 degrees C showed no such accumulation of intermediate metabolites. Overall methanogenesis was also greatly reduced at 57.5 degrees C (0.09 L CH4/g VS fed) versus optimal methane formation at 53 degrees C (0.40 L CH4/g VS fed). Microbial community assessment and free energy calculations suggest that the accumulation of fatty acids and hydrogen, and relatively poor methanogenic performance at 57.5 degrees C are likely due to temperature limitations of thermophilic aceticlastic methanogens.     

Odor control during post-digestion processing of biosolids through bioaugmentation of anaerobic digestion.

The effects of bioaugmenting anaerobic biosolids digestion with a commercial product that contained selected strains of bacteria from genera Bacillus, Pseudomonas, and Actinomycetes, along with ancillary organic compounds containing various micronutrients were evaluated. The main objective of the study was to investigate the effects of bioaugmentation specifically on the performance of methanogenesis during anaerobic digestion, as well as on the generation and fate of odor-causing compounds during the storage of the digested biosolids. The bench-scale digester with 5 g/L bioaugment generated 29% more net CH4 than a control during the eight weeks of operation. In addition, the average residual propionic acid concentration in the bioaugmented digester was 46% lower than that in the control. The biosolids digested in the bioaugmented digester generated a negligible amount of methyl mercaptan (CH3SH) during 10 days of post-digestion storage, while CH3SH concentration in the control reached nearly 300 ppmv during the same period. Similarly peak dimethyl sulfide (CH3SCH3) generated by stored biosolids from the bioaugmented digester was only 37% of that from the control. Similar results were obtained in a subsequent short term study designed to confirm the repeatability of the findings.     

Methane conversion efficiency as a simple control parameter for an anaerobic digester at high loading rates

The anaerobic digestion process is globally applied to the treatment of highly concentrated wastes such as industrial and rural effluents, and sewage sludge. However, it is known to be relatively unstable. When loaded with high concentrations of organic material, unwanted volatile fatty acids (VFA) are often produced rather than methane (CH4) gas which can lead to digester acidification and failure. This study investigated digester behaviour under high loading rates, testing the usefulness of stoichiometric methane conversion efficiency as a digester control parameter at high loading rates. Our results show that, in general, the CH4 production rate was proportional to the feed rate (loading rate). However, at very high loading rates, the CH4 production rate was not proportional to the increase in the feeding rate. Consequently, VFA accumulated and the H2 partial pressure increased. The proportionality of the loading rate and gas production rate is stoichiometrically expressed as the conversion efficiency. We found that conversion efficiency was a useful indicator as an early warning of digester imbalance. The digester remained stable at conversion efficiencies above 75%. Dropping below 70% signified the onset of digester failure. As loading rate and methane production data are readily available on-line in most anaerobic digestion plants, the conversion efficiency can be monitored on-line and used as an efficient control technique to maintain safe operation of anaerobic digesters at high loading rates.     

Syntrophic acetate oxidation in two-phase (acid-methane) anaerobic digesters

The microbial processes involved in two-phase anaerobic digestion were investigated by operating a laboratory-scale acid-phase (AP) reactor and analyzing two full-scale, two-phase anaerobic digesters operated under mesophilic (35 °C) conditions. The digesters received a blend of primary sludge and waste activated sludge (WAS). Methane levels of 20% in the laboratory-scale reactor indicated the presence of methanogenic activity in the AP. A phylogenetic analysis of an archaeal 16S rRNA gene clone library of one of the full-scale AP digesters showed that 82% and 5% of the clones were affiliated with the orders Methanobacteriales and Methanosarcinales, respectively. These results indicate that substantial levels of aceticlastic methanogens (order Methanosarcinales) were not maintained at the low solids retention times and acidic conditions (pH 5.2-5.5) of the AP, and that methanogenesis was carried out by hydrogen-utilizing methanogens of the order Methanobacteriales. Approximately 43, 31, and 9% of the archaeal clones from the methanogenic phase (MP) digester were affiliated with the orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales, respectively. A phylogenetic analysis of a bacterial 16S rRNA gene clone library suggested the presence of acetate-oxidizing bacteria (close relatives of Thermacetogenium phaeum, 'Syntrophaceticus schinkii,' and Clostridium ultunense). The high abundance of hydrogen consuming methanogens and the presence of known acetate-oxidizing bacteria suggest that acetate utilization by acetate oxidizing bacteria in syntrophic interaction with hydrogen-utilizing methanogens was an important pathway in the second-stage of the two-phase digestion, which was operated at high ammonium-N concentrations (1.0 and 1.4 g/L). A modified version of the IWA Anaerobic Digestion Model No. 1 (ADM1) with extensions for syntrophic acetate oxidation and weak-acid inhibition adequately described the dynamic profiles of volatile acid production/degradation and methane generation observed in the laboratory-scale AP reactor. The model was validated with historical data from the full-scale digesters.     

Anaerobic digestion of catering wastes: effect of micronutrients and retention time.

Source-separated foodwastes collected from a campus catering facility were processed in bench-scale single-stage anaerobic digesters. The feedstock contained a varied mix of fruits, vegetables, meats and fried foods. A constant organic loading rate (OLR) was maintained with differing hydraulic retention times (HRT). Regular addition of trace elements or prolonged retention time allowed stable digestion at high total volatile fatty acid (TVFA) levels. Reactors on HRT of 25, 50, and 100 days with no micronutrient supplementation exhibited methanogenic failure after approximately 40, 100 and 90 days respectively, while duplicate reactors with micronutrient supplementation maintained stable digestion. An extended HRT of 180 days has so far allowed continued digestion (for reactors with and without micronutrient supplementation) at levels of ammonia nitrogen exceeding 5.7 g l(-1) and volatile fatty acid levels exceeding 15 g l(-1), usually considered inhibitory or toxic.     

Two-stage thermophilic-mesophilic anaerobic digestion of waste activated sludge from a biological nutrient removal plant.

A two-stage thermophilic-mesophilic anaerobic digestion pilot-plant was operated solely on waste activated sludge (WAS) from a biological nutrient removal (BNR) plant. The first-stage thermophilic reactor (HRT 2 days) was operated at 47, 54 and 60 degrees C. The second-stage mesophilic digester (HRT 15 days) was held at a constant temperature of 36-37 degrees C. For comparison with a single-stage mesophilic process, the mesophilic digester was also operated separately with an HRT of 17 days and temperature of 36-37 degrees C. The results showed a truly thermophilic stage (60 degrees C) was essential to achieve good WAS degradation. The lower thermophilic temperatures examined did not offer advantages over single-stage mesophilic treatment in terms of COD and VS removal. At a thermophilic temperature of 60 degrees C, the plant achieved 35% VS reduction, representing a 46% increase compared to the single-stage mesophilic digester. This is a significant level of degradation which could make such a process viable in situations where there is no primary sludge generated. The fate of the biologically stored phosphorus in this BNR sludge was also investigated. Over 80% of the incoming phosphorus remained bound up with the solids and was not released into solution during the WAS digestion. Therefore only a small fraction of phosphorus would be recycled to the main treatment plant with the dewatering stream.     

Optimizing volatile fatty acid production in partial acidogenesis of swine wastewater.

This research has been conducted based on the fact that acetic and butyric acids are favorable substrates for methanogens, and that a low level of propionic acid production during acidogenesis minimizes the inhibition effect on methanogenic growth. Raw swine wastewater was pretreated with ammonia stripping to enhance acidogenesis. The ammonia nitrogen concentration of less than 1.2 g/L did not significantly affect the biochemical acidogenic potential of swine wastewater. For acidogenesis of swine wastewater, a set of experiments were carried out to produce short chain volatile fatty acids (VFA) in laboratory-scale continuously stirred tank reactors. The production of acetic, propionic, and butyric acids associated with simultaneous changes in hydraulic retention time (HRT) and temperature was investigated. Response surface methodology was successfully applied to approximate the responses of the VFA productions. The optimum physiological conditions where the maximum acetic and butyric acids production occurred were 2.4 days HRT at 34 degrees C and 2.1 days HRT at 35 degrees C, respectively. The propionic acid production linearly increased as both HRT and temperature increased.     

Anaerobic digestion of corn ethanol thin stillage in batch and by high-rate down-flow fixed film reactors

Thin stillage (CTS) from a dry-grind corn ethanol plant was evaluated as a carbon source for anaerobic digestion (AD) by batch and high rate semi-continuous down-flow stationary fixed film (DSFF) reactors. Biochemical methane potential (BMP) assays were carried out with CTS concentrations ranging from approximately 2,460-27,172 mg total chemical oxygen demand (TCOD) per litre, achieved by diluting CTS with clean water or a combination of clean water and treated effluent. High TCOD, SCOD and volatile solids (VS) removal efficiencies of 85 ± 2, 94 ± 0 and 82 ± 1% were achieved for CTS diluted with only clean water at an organic concentration of 21,177 mg TCOD per litre, with a methane yield of 0.30 L methane per gram TCOD(removed) at standard temperature and pressure (STP, 0 °C and 1 atmosphere). Batch studies investigating the use of treated effluent for dilution showed promising results. Continuous studies employed two mesophilic DSFF anaerobic digesters treating thin stillage, operated at hydraulic retention times (HRT) of 20, 14.3, 8.7, 6.3, 5 and 4.2 d. Successful digestion was achieved up to an organic loading rate (OLR) of approximately 7.4 g TCOD L(-1)d(-1) at a 5 d HRT with a yield of 2.05 LCH(4) L(-1)d(-1) (at STP) and TCOD and VS removal efficiencies of 89 ± 3 and 85 ± 3%, respectively.     

Treatment of low and medium strength sewage in a lab-scale gradual concentric chambers (GCC) reactor.

One of the major challenges of anaerobic technology is its applicability for low strength wastewaters, such as sewage. The lab-scale design and performance of a novel Gradual Concentric Chambers (GCC) reactor treating low (165+/-24 mg COD/L) and medium strength (550 mg COD/L) domestic wastewaters were studied. Experimental data were collected to evaluate the influence of chemical oxygen demand (COD) concentrations in the influent and the hydraulic retention time (HRT) on the performance of the GCC reactor. Two reactors (R1 and R2), integrating anaerobic and aerobic processes, were studied at ambient (26 degrees C) and mesophilic (35 degrees C) temperature, respectively. The highest COD removal efficiency (94%) was obtained when treating medium strength wastewater at an organic loading rate (OLR) of 1.9 g COD/L.d (HRT = 4 h). The COD levels in the final effluent were around 36 mg/L. For the low strength domestic wastewater, a highest removal efficiency of 85% was observed, producing a final effluent with 22 mg COD/L. Changes in the nutrient concentration levels were followed for both reactors.     

Behavior of cellulose-degrading bacteria in thermophilic anaerobic digestion process.

Previously, we found that the newly isolated Clostridium sp. strain JC3 became the dominant cellulose-degrading bacterium in thermophilic methanogenic sludge. In the present study, the behavior of strain JC3 in the thermophilic anaerobic digestion process was investigated quantitatively by molecular biological techniques. A cellulose-degrading experiment was conducted at 55 degrees C with a 9.5 L of anaerobic baffled reactor having three compartments (Nos. 1, 2, 3). Over 80% of the COD input was converted into methane when 2.5 kgCOD m(-3) d(-1) was loaded for an HRT of 27 days. A FISH probe specific for strain JC3 was applied to sludge samples harvested from the baffled reactor. Consequently, the ratio of JC3 cells to DAPI-stained cells increased from below 0.5% (undetectable) to 9.4% (compartment 1), 13.1% (compartment 2) and 21.6% (compartment 3) at day 84 (2.5 kgCOD m(-3)d(-1)). The strain JC3 cell numbers determined by FISH correlated closely with the cellulose-degrading methanogenic activities of retained sludge. A specific primer set targeting the cellulase gene (cellobiohydrolaseA: cbhA) of strain JC3 was designed and applied to digested sludge for treating solid waste such as coffee grounds, wastepaper, garbage, cellulose and so on. The strain JC3 cell numbers determined by quantitative PCR correlated closely with the cellulose-sludge loading of the thermophilic digester. Strain JC3 is thus important in the anaerobic hydrolysis of cellulose in thermophilic anaerobic digestion processes.     

Strategies for the anaerobic digestion of the organic fraction of municipal solid waste: an overview.

Different process strategies for anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) are reviewed weighing high-solids versus low-solids, mesophilic versus thermophilic and single-stage versus multi-stage processes. The influence of different waste characteristics such as composition of biodegradable fractions, C:N ratio and particle size is described. Generally, source sorting of OFMSW and a high content of food waste leads to higher biogas yields than the use of mechanically sorted OFMSW. Thermophilic processes are more efficient than mesophilic processes in terms of higher biogas yields at different organic loading rates (OLR). Highest biogas yields are achieved by means of wet thermophilic processes at OLRs lower than 6 kg-VS x m(-3) d(-1). High-solids processes appear to be relatively more efficient when OLRs higher than 6 kg-VS x m(-3)d(-1) are applied. Multi-stage systems show in some investigations a higher reduction of recalcitrant organic matter compared to single-stage systems, but they are seldom applied in full-scale. An extended cost-benefit calculation shows that the highest overall benefit of the process is achieved at an OLR that is lower and a hydraulic retention time (HRT) that is longer than those values of OLR and HRT, at which the highest biogas production is achieved.     

Continuous hydrogen and methane production in a two-stage cheese whey fermentation system

The feasibility of integrating biological hydrogen and methane production in a two-stage process using mixed cultures and cheese whey powder (CWP) as substrate was studied. The effect of operational parameters such as hydraulic retention time (HRT) and organic loading rate (OLR) on the volumetric hydrogen (VHPR) and methane (VMPR) production rates was assessed. The highest VHPR was 28 L H2/L/d, obtained during stable operation in a CSTR at HRT and OLR of 6 h and 142 g lactose/ L/d, respectively. Moreover, hydrogen (13 L/L/d) was produced even at HRT as low as 3.5 h and OLR of 163 g lactose/L/d, nonetheless, the reactor operation was not stable. Regarding methane production in an UASB reactor, the acidified effluent from the hydrogen-producing bioreactor was efficiently treated obtaining COD removals above 90% at OLR and HRT of 20 g COD/L/d and 6 h, respectively. The two-stage process for continuous production of hydrogen and methane recovered over 70% of the energy present in the substrate. This study demonstrated that hydrogen production can be efficiently coupled to methane production in a two-stage system and that CWP is an adequate substrate for energy production.     

Comparison of methane production by co-digesting fruit and vegetable waste with first stage and second stage anaerobic digester sludge from a two stage digester

Fruit and vegetable waste (FVW) was co-digested with first stage (FSS) and second stage anaerobic digester sludge (SSS) separately, over the course of 10 days, in batch reactors. Addition of FVW significantly increased the methane production in both sludges. After 10 days of digestion FSS + FVW produced 514 ± 57 L CH(4) kg VS(-1)(added) compared with 392 ± 16 L CH(4) for the SSS + FVW. The increased methane yield was most likely due to the higher inoculum substrate ratio of the FSS. The final VS and COD contents of the sewer sludge and FVW mixtures were not significantly different from the control values suggesting that all of the FVW added was degraded within 10 days. It is recommended that FVW be added to the first stage of the anaerobic digester in order to maximize methane generation.     

Two-stage anaerobic digestion process for complete inactivation of enteric bacterial pathogens in human night soil.

Anaerobic digestion offers a good alternative for human waste treatment. However, the fate of enteric bacterial pathogens present in human night soil (HNS) remains a major concern for hygienic safety of the process. A two-stage anaerobic digestion process, consisting of separate acidogenic and methanogenic digesters, was designed and its efficacy in the inactivation of Salmonella typhi was compared to a single-stage digestion process. In a single-stage digestion, complete pathogen inactivation was achieved only in the digesters with high levels of volatile fatty acids (VFA approximately equal to 18,000 mg/l) and acidic pH (approximately equal to 6.0). These digesters, however, showed drastic reduction in methane yield. In the two-stage digestion process, S. typhi was completely inactivated in the acidogenic digester and the methanogenic digester was free from the pathogen even after receiving a daily dose of the pathogen. The process also achieved complete inactivation of other enteric pathogens, viz., Shigella dysenteriae and Vibrio cholerae. The two-stage process was efficient in biogas generation from HNS. Thus, the two-stage process ensures complete hygienic safety in anaerobic digestion of human night soil.     

Application of ADM1 for the simulation of anaerobic digestion of olive pulp under mesophilic and thermophilic conditions.

The management of the wastewater originating from olive oil producing industries poses a serious environmental problem. Recently, two-phase production of olive oil has been developed, leading to almost complete elimination of the bulk of the generated wastewater and, is thus regarded as an environmentally friendly technology. However, the main waste stream (olive pulp) is a slurry material characterized by high solids concentration (approximately 30%), requiring stabilisation before its final disposal. The anaerobic digestion of olive pulp is studied in this work under mesophilic and thermophilic conditions in CSTR-type digesters. The digesters were fed with water-diluted (1:4) olive pulp at an HRT of 20 days and an OLR of 3.94 kg COD m(-3) d(-1). In order to study the process kinetics, the digesters were subjected to impulse disturbances of different substrates. The IWA anaerobic digestion model was used to simulate the reactors' response. Some key process parameters, such as the specific maximum uptake rate constants and the saturation constants for the volatile fatty acids degradation were estimated and compared with the standard values suggested by the ADM1.     

Development of an ecologically sustainable wastewater treatment system

The present study aimed mainly for the development of a wastewater treatment system incorporating enhanced primary treatment, anaerobic digestion of coagulated organics, biofilm aerobic process for the removal of soluble organics and disinfection of treated water. An attempt was also made to study the reuse potential of treated water for irrigation and use of digested sludge as soil conditioner by growing marigold plants. Ferric chloride dose of 30 mg/l was found to be the optimum dose for enhanced primary treatment with removals of COD and BOD to the extent of 60% and 77%, respectively. Efficient anaerobic digestion of ferric coagulated sludge was performed at 7 days hydraulic retention time (HRT). Upflow aerobic fixed film reactor (UAFFR) was very efficient in removals of COD/BOD in the organic loading rate (OLR) range of 0.25 to 3 kg COD/m(3)/day with COD and BOD removals in the range 65-90 and 82-96, respectively. Photo-oxidation followed by disinfection saved 50% of chlorine dose required for disinfection of treated effluent and treated water was found to be suitable for irrigation. The result also indicated that anaerobically digested sludge may be an excellent soil conditioner. From the results of this study, it is possible to conclude that the developed wastewater treatment system is an attractive ecologically sustainable alternative for sewage treatment from institutional/industrial/residential campuses.     

Anaerobic digestion of blackwater from vacuum toilets and kitchen refuse in a continuous stirred tank reactor (CSTR).

The objective of this research was mesophilic anaerobic digestion of blackwater from vacuum toilets (BW) and kitchen refuse (KR) in a CSTR within an ecological sanitation system. A detailed investigation of the BW characteristics was carried out. Research on anaerobic digestion was performed with CSTR of 101 volume at HRT of 10, 15 and 20 days. The digestion of BW at 20 days HRT showed stable performance without inhibition effects, in spite of relatively high ammonium concentrations. The removal of total and particulate COD was 61% and 53%, respectively, and the methane yield 10/CH4/cap/day. The addition of kitchen refuse (KR) improved the performance of the CSTR in terms of COD removal efficiency and methane yield. At 20 days HRT the removal of total and particulate COD increased up to 71% and 67%, respectively, and the methane yield to 27/CH4/cap/day. The results at 15 days HRT showed similar performance. At HRT of 10 days, the anaerobic treatment was limited but reached steady state conditions at higher VFA concentrations in the effluent, with a decrease of COD removal of 30 to 33% and of methane yields of 19 to 21%.