Thermophilic Anaerobic Reactor Research Articles

Long term operation of a thermophilic anaerobic reactor: Process stability and efficiency at decreasing sludge retention time

The aim of this study was to evaluate the performance of thermophilic sludge digestion at decreasing sludge retention time (SRT) and increasing organic loading rate (OLR), in terms of methane production, effluent stabilisation, hygienisation and dewaterability. Focus was put on determining indicators to help prevent process failure. To this end, a lab-scale reactor was operated for nearly 2 years at 55 °C. Methane production rate was increased (from 0.2 to 0.4–0.6 m CH 4 3 m reactor - 3 d −1) by decreasing the SRT from 30 to 15–10 days, while increasing the OLR from 0.5 to 2.5–3.5 kg VS m reactor - 3 d −1. Sludge dewaterability was worsened at SRT below 15 days; while pathogen destruction was always successful. The following concentrations might be used to prevent process failure: VFA C2–C5 (3.7 g COD L −1), acetate (0.6 g L −1), acetate/propionate (0.5), intermediate alkalinity (1.8 g CaCO 3 L −1), intermediate/partial alkalinity (0.9), intermediate/total alkalinity (0.5), CH 4 in biogas (55%).

Treatment efficiency and VFA composition of a thermophilic anaerobic contact reactor treating food industry wastewater

The objective of this study was to examine the effects of organic loading rate and hydraulic retention time on volatile fatty acid composition and treatment efficiency of high rate thermophilic anaerobic contact reactor (TACR) treating potato-chips wastewaters. The operational performance of TACR was monitored from start-up by assessing chemical oxygen demand (COD) removal efficiencies, volatile fatty acid (VFA) production and biogas composition. The reactor was studied at different organic loading rates (OLRs) ranging from 0.6 to 8 kg COD/m 3 d. The COD removal efficiencies were found to be 86–97% and the methane percentage of the biogas produced was 68–89% during the OLRs studied. The approximate methane yield was found to be 0.42 m 3 CH 4/kg COD removed. The major intermediate products of anaerobic digestion were acetate, propionate, iso-butyrate, butyrate, iso-valerate, valerate, iso-caproate and caproate. The use of thermophilic anaerobic contact reactor offers a sustainable technology for the treatment of potato-chips wastewaters since high COD removal efficiencies and high methane percentage in the biogas produced can be attained even at high OLRs.

Illumination enhances methane production from thermophilic anaerobic digestion

Incandescent lamp illumination enhanced methane production from a thermophilic anaerobic digestion reactor (55 degrees C) supplied with glucose. After 10 days of operation, the volume of methane produced from light reactors was approximately 2.5 times higher than that from dark reactors. A comparison of the carbon balance between light and dark conditions showed that methane produced from hydrogen and carbon dioxide in the light reactors was higher than that from the dark reactors. When hydrogen or acetate was fed into the reactors, methane production with added hydrogen was faster and higher under light conditions than under dark conditions. The use of blue light-emitting diodes also enhanced methane production over that under dark conditions. The 16S rRNA gene copy numbers for Methanothermobacter spp. in the light reactor and in the dark reactor were at the same level. The copy number for Methanosarcina spp. in the light reactors was approximately double than that in the dark reactors. These results suggest that blue light enhances the methanogenic activity of hydrogenotrophic methanogens.

Microbial community in methanogenic packed-bed reactor successfully operating at short hydraulic retention time

The microbial community in a thermophilic anaerobic packed-bed reactor, which had been successfully operated to convert acetic and butyric acids to methane at a short hydraulic retention time (from 24 h to 1.9 h), was investigated. Archaea closely related to known methanogens were detected by 16S rRNA gene analyses of the effluents, together with diverse types of unidentified bacteria.

Microbial community structure in a thermophilic anaerobic hybrid reactor degrading terephthalate.

A thermophilic terephthalate-degrading methanogenic consortium was successfully enriched for 272 days in an anaerobic hybrid reactor, and the microbial structure was characterized using terminal RFLPs, clone libraries and fluorescence in-situ hybridization with rRNA-targeted oligonucleotide probes. All the results suggested that Methanothrix thermophila-related methanogens, Desulfotomaculum-related bacterial populations in the Gram-positive low-G + C group, and OP5-related populations were the key members responsible for terephthalate degradation under thermophilic methanogenic conditions except during periods when the reactor experienced heat shock and pump failure. These perturbations caused a significant shift in bacterial population structure in sludge samples taken from the sludge bed but not from the surface of the packing materials. After system recovery, many other bacterial populations emerged, which belonged mainly to the Gram-positive low-G + C group and Cytophaga-Flexibacter-Bacteroides, as well as beta-Proteobacteria, Planctomycetes and Nitrospira. These newly emerged populations were probably also capable of degrading terephthalate in the hybrid system, but were out-competed by those bacterial populations before perturbations.

Design of A Solar Thermophilic Anaerobic Reactor for Small Farms

A 10 m 3 completely stirred tank reactor has been designed for anaerobic treatment of liquid cow manure under thermophilic conditions (50°C), using a solar heating system mounted on the reactor roof. Simulation models for two systems have been developed. The first system consists of loose components and in the second system the components are integrated. Each of the two systems includes a heat recovery unit to heat the influent by extracting heat from the effluent. The integrated system includes an extra chamber for the pumps and the heat recovery unit, which are in the same insulating cover as the reactor and the solar collector. The control system is based on simple on/off strategy. An auxiliary heater, operated with the produced biogas, can be used during cold months. The measured and calculated design parameters are presented. The simulation results show that the temperature fluctuation of the reactor during the night is less than 1°C, which is too small to harm the microbial activity. The results show that an annual net thermal energy production and overall annual energy efficiency of 100 and 95%, respectively, could be obtained from the integrated system.

Effect of redox mediator, AQDS, on the decolourisation of a reactive azo dye containing triazine group in a thermophilic anaerobic EGSB reactor

The feasibility of thermophilic (55 °C) anaerobic treatment applied to colour removal of a triazine contained reactive azo dye was investigated in two 0.53 l expanded granular sludge blanket (EGSB) reactors in parallel at a hydraulic retention time (HRT) of 10 h. Generally, this group of azo dyes shows the lowest decolourisation rates during mesophilic anaerobic treatment. The impact of the redox mediator addition on colour removal rates was also evaluated. Reactive Red 2 (RR2) and anthraquinone-2,6-disulfonate (AQDS) were selected as model compounds for azo dye and redox mediator, respectively. The reactors achieved excellent colour removal efficiencies with a high stability, even when high loading rates of RR2 were applied (2.7 g RR2 l −1 per day). Although AQDS addition at catalytic concentrations improved the decolourisation rates, the impact of AQDS on colour removal was less apparent than expected. Results show that the AQDS-free reactor R2 achieved excellent colour removal rates with efficiencies around 91%, compared with the efficiencies around 95% for the AQDS-supplied reactor R1. Batch experiments confirmed that the decolourisation rates were co-substrate dependent, in which the volatile fatty acids (VFA) mixture was the least efficient co-substrate. The highest decolourisation rate was achieved in the presence of either hydrogen or formate, although the presence of glucose had a significant impact on the colour removal rates.

Ammonia inhibition on thermophilic anaerobic digestion

This study evaluated both chronic and acute toxicity of ammonia in thermophilic anaerobic digestion of synthetic wastewater over a range of acclimation concentrations. The inhibition effects of ammonia, in terms of total ammonia nitrogen (TAN), under various pH values and acclimation conditions on thermophilic aceticlastic methanogens were investigated. Completely mixed thermophilic anaerobic reactors operated at a chemical oxygen demand (COD) loading rate of 4 g/l day and a solid retention time (SRT) of 7 days were subjected to TAN concentrations of 0.40, 1.20, 3.05, 4.92, and 5.77 g/l. The reactor operations presented a case of chronic inhibition and it was observed that TAN concentrations of 4.92 and 5.77 g/l caused a drop in methane production by as much as 39% and 64%, respectively with respect to control. Batch anaerobic toxicity assays (ATA) were also performed to evaluate the acute toxicity effects of TAN and pH on methanogenesis at thermophilic condition. Modeling based on the results of ATA indicated that aceticlastic methanogens acclimated to high concentrations of TAN were less sensitive to increase in TAN and could tolerate wider pH ranges. TAN concentration causing 100% inhibition occurred in the range of 8–13 g/l, depending on acclimation condition and system pH.

Sodium Inhibition of Thermophilic Methanogens

This study investigated the sodium inhibition of methanogens using two thermophilic (55°C) anaerobic sequencing batch reactors (ASBRs). The ASBRs were operated at a chemical oxygen demand (COD) loading of 4 g/L/day and a hydraulic retention time of 3 days. To evaluate the chronic toxicity of sodium to methanogens, the biomass in one of the ASBRs was acclimated to increasing sodium concentrations of 4.1, 7.1, and 12.0 g/L while the feed to the second ASBR was not supplemented with any additional sodium. The methanogenic activity (mL CH4/g volatile suspended solids/day) decreased by nearly 44% at an acclimation concentration of 12.0 g Na+/L, but the COD removal efficiency and methane production did not vary appreciably at the different acclimation concentrations studied. The acute toxicity of sodium to methanogens was determined by a series of batch anaerobic toxicity assays (ATAs). The biomass acclimated to different concentrations of sodium was collected from the ASBRs and used as inocula for the batch tests, and the sodium concentration was varied up to 17.7 g/L. The methanogens in the biomass acclimated to 0, 4.1, 7.1, and 12.0 g Na+/L were completely inhibited (100% inhibition) at predicted sodium concentrations of 10.6, 12.7, 18.0, and 22.8 g/L, respectively. To simulate the results of batch ATA in the ASBR, 7-day feeding with sodium concentrations in the influent measuring 6.2, 10.6, and 16.0 g/L were introduced into the reactor. Among each feeding, the reactor was operated with no additional sodium in the feed with 2–3 week intervals. Even though the methanogenic activity was not significantly affected at 6.2 and 10.6 g/L of sodium, there was a deterioration in methanogenic activity at 16.0 g/L dosage of sodium.

Thermotoga lettingae sp. nov., a novel thermophilic, methanol-degrading bacterium isolated from a thermophilic anaerobic reactor

Thermotoga lettingae sp. nov., a novel thermophilic, methanol-degrading bacterium isolated from a thermophilic anaerobic reactor

Thermotoga lettingae sp. nov., a novel thermophilic, methanol-degrading bacterium isolated from a thermophilic anaerobic reactor.

A novel, anaerobic, non-spore-forming, mobile, Gram-negative, thermophilic bacterium, strain TMOT, was isolated from a thermophilic sulfate-reducing bioreactor operated at 65 C with methanol as the sole substrate. The G+C content of the DNA of strain TMOT was 39.2 mol%. The optimum pH, NaCl concentration, and temperature for growth were 7.0, 1.0%, and 65 degrees C, respectively. Strain TMOT was able to degrade methanol to CO2 and H2 in syntrophic culture with Methanothermobacter thermautotrophicus AH or Thermodesulfovibrio yellowstonii. Thiosulfate, elemental sulfur, Fe(III) and anthraquinone-2,6-disulfonate were able to serve as electron acceptors during methanol degradation. In the presence of thiosulfate or elemental sulfur, methanol was converted to CO2 and partly to alanine. In pure culture, strain TMOT was also able to ferment methanol to acetate, CO2 and H2. However, this degradation occurred slower than in syntrophic cultures or in the presence of electron acceptors. Yeast extract was required for growth. Besides growing on methanol, strain TMOT grew by fermentation on a variety of carbohydrates including monomeric and oligomeric sugars, starch and xylan. Acetate, alanine, CO2, H2, and traces of ethanol, lactate and alpha-aminobutyrate were produced during glucose fermentation. Comparison of 16S rDNA genes revealed that strain TMOT is related to Thermotoga subterranea (98%) and Thermotoga elfii (98%). The type strain is TMOT (= DSM 14385T = ATCC BAA-301T). On the basis of the fact that these organisms differ physiologically from strain TMOT, it is proposed that strain TMOT be classified as a new species, within the genus Thermotoga, as Thermotoga lettingae.

Ammonia inhibition on thermophilic aceticlastic methanogens

The inhibition effects of total ammonia nitrogen (TAN) on aceticlastic methanogenic activity using biomass from thermophilic anaerobic reactors were investigated in this study. Anaerobic Toxicity Assays (ATA) were conducted after the biomass was acclimated to different levels of TAN. The TAN background concentrations in the reactors were 400, 1,200, and 3,050 mg/L. The results from ATA showed: 1) high TAN concentrations could cause inhibition of aceticlastic methanogens; 2) biomass acclimated to higher TAN concentrations could alleviate the inhibition effect due to the increase of TAN concentration; 3) the lethal TAN concentration for methanogens was approximately 10,000 mg/L regardless of the background TAN concentration; 4) ATA results also revealed the role played by pH. At a given TAN concentration, methanogenic activity varied with the pH values. The highest methanogenic activity was always observed at a pH of 7.0-7.5. 5) It was also observed that acclimation could increase the pH tolerance range, which made methanogens less sensitive to pH changes.

Aerobic waste activated sludge (WAS) for start-up seed of mesophilic and thermophilic anaerobic digestion

Since there are very limited numbers of thermophilic anaerobic digesters being operated, it is often difficult to start up a new one using sludge from an existing reactor as a seed. However, for obvious reasons it seems few attempts have been made to compare the start-up performance of thermophilic anaerobic digestion using different sources of seed sludges. The purpose of this study was to evaluate the start-up performance of anaerobic digestion using aerobic waste activated sludge (WAS) from a plant which has no anaerobic digesters and mesophilic anaerobic digested sludge (ADS) as the seed source at both mesophilic (35°C) and thermophilic (55°C) temperatures. In this study, two experiments were conducted. First, thermophilic anaerobic reactors were seeded with WAS (VSS=4400 mg/L) and ADS (VSS=14,500 mg/L) to investigate start-up performance with a feed of acetate as well as propionate. The results show that WAS started to produce CH 4 soon after acetate feeding without a lag time, while ADS had a lag time of 10 days. When the feed was changed to propionate, WAS removed propionate down to below the detection limit of 10 mg/L, while ADS removed little propionate and produced little CH 4. Second, in order to further compare the methanogenic activity of WAS and ADS, both mesophilic and thermophilic reactors were operated. WAS acclimated to anaerobic conditions shortly (<5 days at both mesophilic and thermophilic) and after acclimating it produced more CH 4 per unit amount of seeded VSS than ADS. WAS at mesophilic temperature biodegraded acetate at the same rate as for thermophilic. However WAS at mesophilic temperature biodegraded propionate at a much faster rate than at thermophilic. WAS as the seed source of anaerobic digestion resulted in much better performance than ADS at both mesophilic and thermophilic temperatures for both acetate and propionate metabolism.

Measurement of microbial numbers and biomass contained in thermophilic anaerobic reactors.

This paper describes the determination of the microbial population in terms of the number, biomass, and composition of single- and two-phase, laboratory-scale thermophilic (55 degrees C) anaerobic reactors under steady-state conditions. Epifluorescence microscopy with 4',6-diamidine-2-phenylindole (DAPI) as fluorochrome was used to determine the total number of microorganisms in the reactors, and autofluorescence microscopy was used to determine the total methanogenic bacteria populations. The results obtained by the direct count methods were compared to the quantity of biomass contained in the system, which was determined by volatile suspended solids. The concentration of acidogenic bacteria was estimated by subtraction of the autofluorescence results from those of the DAPI epifluorescence microscopy. The viable bacterial population was determined by plating techniques using an anaerobic chamber. The total bacterial and methanogenic populations of single-stage digesters increase when the hydraulic retention time decreases; nevertheless, the percentages of the principal bacterial groups (acidogenic and methanogenic) remain constant at 87% and 13%, respectively. In the two-stage reactors, the percentages of the acidogenic and methanogenic groups are 99% and 26% of the total population in the acidogenic and methanogenic reactors, respectively. In the single-stage reactors, biomass determinations can be used to estimate microbial concentrations, and vice versa, as there is a high positive correlation between microorganism concentration and biomass. The syntrophic relationship between the bacteria involved in the anaerobic process is a possible explanation for the low values of viable population obtained in the reactors studied. Nevertheless, there is a high correlation between direct counts by epifluorescence microscopy and viable plate counts for the combined system studied.

Analysis of the methane production in thermophilic anaerobic reactors: use of autofluorescence microscopy

Methanogenic activity in thermophilic, anaerobic reactors was determined by comparing the amount of methane generated in single- and two-stage systems with the size of the methanogenic population, as determined by microscopy. The methanogenic activities were 2.71 × 10−9 ml methane cell−1 d−1 and 1.10 × 10−9 ml methane cell−1 d−1 for 10 and 4 days of the hydraulic retention time (HRT), in the single-stage system. In the two-stage system, 7.49 × 10−9 ml methane cell−1 d−1 in the acidogenic reactor and 1.56 × 10−9 ml methane cell−1 d−1 in the methanogenic reactor for 4 days of the HRT. A high correlation was evident between the methane production and methanogenic population [0.1354 ln(x) − 2.1375](R 2 0.8619).

Cultivation and in situ detection of a thermophilic bacterium capable of oxidizing propionate in syntrophic association with hydrogenotrophic methanogens in a thermophilic methanogenic granular sludge.

The thermophilic, anaerobic, propionate-oxidizing bacterial populations present in the methanogenic granular sludge in a thermophilic (55 degrees C) upflow anaerobic sludge blanket reactor were studied by cultivation and in situ hybridization analysis. For isolation of propionate-degrading microbes, primary enrichment was made with propionate as the sole energy source at 55 degrees C. After several attempts to purify the microbes, a thermophilic, syntrophic, propionate-oxidizing bacterium, designated strain SI, was isolated in both pure culture and coculture with Methanobacterium thermoautotrophicum. Under thermophilic (55 degrees C) conditions, strain SI oxidized propionate, ethanol, and lactate in coculture with M. thermoautotrophicum. In pure culture, the isolate was found to ferment pyruvate. 16S ribosomal DNA sequence analysis revealed that the strain was relatively close to members of the genus Desulfotomaculum, but it was only distantly related to any known species. To elucidate the abundance and spatial distribution of organisms of the strain SI type within the sludge granules, a 16S rRNA-targeted oligonucleotide probe specific for strain SI was developed and applied to thin sections of the granules. Fluorescence in situ hybridization combined with confocal laser scanning microscopy revealed that a number of rod-shaped cells were present in the middle and inner layers of the thermophilic granule sections and that they formed close associations with hydrogenotrophic methanogens. They accounted for approximately 1.1% of the total cells in the sludge. These results demonstrated that strain SI was one of the significant populations in the granular sludge and that it was responsible for propionate oxidation in the methanogenic granular sludge in the reactor.

Monitoring of specific methanogenic activity of granular sludge by confocal laser scanning microscopy during start-up of thermophilic upflow anaerobic sludge blanket reactor

Confocal, laser-scanning microscopy was applied to acquire coenzyme F420-based autofluorescence images of middle sections of sludge granules during start-up of a thermophilic reactor that were seeded with mesophilically-grown microorganisms of granular sludge. Digital images were analyzed to calculate weighted averages of autofluorescence. The values were related (r2=0.97) to specific methanogenic activities of granular sludge as the granules developed to steady state.

Anaerobic Thermophilic (55°C) Treatment of TMP Whitewater in Reactors Based on Biomass Attachment and Entrapment

Thermomechanical pulping (TMP) whitewater was treated in thermophilic (55°C) anaerobic laboratory-scale reactors using three different reactor configurations. In all reactors up to 70% COD removals were achieved. The anaerobic hybrid reactor, composed of an upflow anaerobic sludge blanket (UASB) and a filter, gave degradation rates up to 10 kg COD/m3d at loading rates of 15 kg COD/m3d and hydraulic retention time (HRT) of 3.1 hours. The anaerobic multi-stage reactor, consisting of three compartments, each packed with granular sludge and carrier elements, gave degradation rates up to 9 kg COD/m3d at loading rates of 15-16 kg COD/m3d, and HRT down to 2.6 hours. Clogging and short circuiting eventually became a problem in the multi-stage reactor, probably caused by too high packing of the carriers. The anaerobic moving bed biofilm reactor performed similar to the other reactors at loading rates below 1.4 kg COD/m3d, which was the highest loading rate applied. The use of carriers in the anaerobic reactors allowed short HRT with good treatment efficiencies for TMP whitewater.

Anaerobic thermophilic (55°C) treatment of TMP whitewater in reactors based on biomass attachment and entrapment

Abstract Thermomechanical pulping (TMP) whitewater was treated in thermophilic (55°C) anaerobic laboratory-scale reactors using three different reactor configurations. In all reactors up to 70 % COD removals were achieved. The anaerobic hybrid reactor, composed of an upflow anaerobic sludge blanket (UASB) and a filter, gave degradation rates up to 10 kg COD/m 3 d at loading rates of 15 kg COD/m 3 d and hydraulic retention time (HRT) of 3.1 hours. The anaerobic multi-stage reactor, consisting of three compartments, each packed with granular sludge and carrier elements, gave degradation rates up to 9 kg COD/m 3 d at loading rates of 15–16 kg COD/m 3 d, and HRT down to 2.6 hours. Clogging and short circuiting eventually became a problem in the multi-stage reactor, probably caused by too high packing of the carriers. The anaerobic moving bed biofilm reactor performed similar to the other reactors at loading rates below 1.4 kg COD/m 3 d, which was the highest loading rate applied. The use of carriers in the anaerobic reactors allowed short HRT with good treatment efficiencies for TMP whitewater.

The adsorption of heavy metals in an acidogenic thermophilic anaerobic reactor

A two-phase, thermophilic anaerobic reactor was operated with a starch-based feed. Copper, zinc, nickel and lead (3 mM) were individually added to the feed each for a period of 30 h. The results were analysed to determine the extent of metal binding in the acidogenic, first-stage reactor and the degree of protection that this afforded to the traditionally more sensitive methanogenic phase. The results showed that zinc and nickel were not bound particularly well, that lead was bound very strongly and that copper had binding characteristics that were between these two extremes. When these findings were compared with an earlier study with a mesophilic sludge, zinc was shown to have a very different behaviour. A possible reason for this is given. An examination of the gas production by the methanogenic stage, in relation to the amount of metal reaching this stage, suggested that the phase separation did not offer any real protection from the toxic effects of heavy metals.