Bioaugmentation of a two-stage thermophilic (68 degrees C/55 degrees C) anaerobic digestion concept for improvement of the methane yield from cattle manure

The possibility of improving a two-stage (68 degrees C/55 degrees C) anaerobic digestion concept for treatment of cattle manure was studied. In batch experiments, a 10-24% increase of the specific methane yield from cattle manure and its fractions was obtained, when the substrates were inoculated with bacteria of the genus Caldicellusiruptor and Dictyoglomus. In a reactor experiment inoculation of a 68 degrees C pretreatment reactor with Caldicellusiruptor resulted in a 93% increase in the methane yield of the pretreatment reactor for a period of 18 days, but gave only a slight increase in the overall methane yield of the two-stage setup.     

A novel process configuration for anaerobic digestion of source-sorted household waste using hyper-thermophilic post-treatment

A novel reactor configuration was investigated for anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW). An anaerobic hyper-thermophilic (68 degrees C) reactor R68 was implemented as a post-treatment step for the effluent of a thermophilic reactor R1 (55 degrees C) in order to enhance hydrolysis of recalcitrant organic matter, improve sanitation and ease the stripping of ammonia from the reactor. The efficiency of the combined system was studied in terms of methane yield, volatile solids (VS) reduction, and volatile fatty acid (VFA) production at different hydraulic retention times (HRT). A single-stage thermophilic (55 degrees C) reactor R2 was used as control. VS reduction and biogas yield of the combined system was 78-89% and 640-790 mL/g VS, respectively. While the VS reduction in the combined system was up to 7% higher than in the single-stage treatment, no increase in methane yield was observed. Shifting the HRT of the hyper-thermophilic reactor from 5 days to 3 days resulted in a drop in the methanogenic activity in the hydrolysis reactor to a minimum. Operation of R68 at HRTs of 24-48 h was sufficient to achieve high VS conversion into VFAs. Removal of pathogens was enhanced by the hyper-thermophilic post-treatment. 7% of the ammonia load was removed in the hyper-thermophilic reactor with a flow of headspace gas through the reactor equivalent to four times the biogas flow produced in reactor R1.     

Hydrogenogenic CO conversion in a moderately thermophilic (55 degrees C) sulfate-fed gas lift reactor: competition for CO-derived H(2)

Thermophilic (55 degrees C) sulfate reduction in a gas lift reactor fed with CO gas as the sole electron donor was investigated. The reactor was inoculated with mesophilic granular sludge with a high activity of CO conversion to hydrogen and carbon dioxide at 55 degrees C. Strong competition for H(2) was observed between methanogens and sulfate reducers, while the homoacetogens present consumed only small amounts of H(2). The methanogens appeared to be more sensitive to pH and temperature shocks imposed to the reactor, but could not be completely eliminated. The fast growth rates of the methanogens (generation time of 4.5 h) enabled them to recover fast from shocks, and they rapidly consumed more than 90% of the CO-derived H(2). Nevertheless, steep increases in sulfide production in periods with low methane production suggests that once methanogenesis is eliminated, sulfate reduction with CO-rich gas as electron donor has great potential for thermophilic biodesulfurization.     

Acetate Degradation at 70 degrees C in Upflow Anaerobic Sludge Blanket Reactors and Temperature Response of Granules Grown at 70 degrees C

Anaerobic acetate degradation at 70 degrees C and at 55 degrees C (as a reference) was studied by running laboratory upflow anaerobic sludge blanket (UASB) reactors inoculated with mesophilic granular sludge. In UASB reactors fed with acetate-containing media (3 g of chemical oxygen demand [COD] per liter, corresponding to 47 mM acetate) approximately 50 days was needed at 70 degrees C and less than 15 days was needed at 55 degrees C to achieve an effluent COD of 500 to 700 mg/liter. In the UASB reactors at both 70 and 55 degrees C up to 90% of the COD was removed. Batch assays showed that sludges from two 70 degrees C UASB reactors, one run at a low effluent acetate concentration and the other run at a high effluent acetate concentration, exhibited slightly different responses to temperatures in the range from 37 to 70 degrees C. Both 70 degrees C sludges, as well as the 55 degrees C sludge, produced methane at temperatures of 37 to 73 degrees C. The 55 degrees C sludge exhibited shorter lag phases than the 70 degrees C sludges and higher specific methane production rates between 37 and 65 degrees C.     

Enhancement of bioenergy production from organic wastes by two-stage anaerobic hydrogen and methane production process

The present study investigated a two-stage anaerobic hydrogen and methane process for increasing bioenergy production from organic wastes. A two-stage process with hydraulic retention time (HRT) 3 d for hydrogen reactor and 12 d for methane reactor, obtained 11% higher energy compared to a single-stage methanogenic process (HRT 15 d) under organic loading rate (OLR) 3 gVS/(L d). The two-stage process was still stable when the OLR was increased to 4.5 gVS/(L d), while the single-stage process failed. The study further revealed that by changing the HRT_hydrogen:HRT_methane ratio of the two-stage process from 3:12 to 1:14, 6.7%, more energy could be obtained. Microbial community analysis indicated that the dominant bacterial species were different in the hydrogen reactors (Thermoanaerobacterium thermosaccharolyticum-like species) and methane reactors (Clostridiumthermocellum-like species). The changes of substrates and HRT did not change the dominant species. The archaeal community structures in methane reactors were similar both in single- and two- stage reactors, with acetoclastic methanogens Methanosarcina acetivorans-like organisms as the dominant species.     

Thermophilic (55-65 degrees C) and extreme thermophilic (70-80 degrees C) sulfate reduction in methanol and formate-fed UASB reactors

The feasibility of thermophilic (55-65 degrees C) and extreme thermophilic (70-80 degrees C) sulfate-reducing processes was investigated in three lab-scale upflow anaerobic sludge bed (UASB) reactors fed with either methanol or formate as the sole substrates and inoculated with mesophilic granular sludge previously not exposed to high temperatures. Full methanol and formate degradation at temperatures up to, respectively, 70 and 75 degrees C, were achieved when operating UASB reactors fed with sulfate rich (COD/SO4(2-)=0.5) synthetic wastewater. Methane-producing archaea (MPA) outcompeted sulfate-reducing bacteria (SRB) in the formate-fed UASB reactor at all temperatures tested (65-75 degrees C). In contrast, SRB outcompeted MPA in methanol-fed UASB reactors at temperatures equal to or exceeding 65 degrees C, whereas strong competition between SRB and MPA was observed in these reactors at 55 degrees C. A short-term (5 days) temperature increase from 55 to 65 degrees C was an effective strategy to suppress methanogenesis in methanol-fed sulfidogenic UASB reactors operated at 55 degrees C. Methanol was found to be a suitable electron donor for sulfate-reducing processes at a maximal temperature of 70 degrees C, with sulfide as the sole mineralization product of methanol degradation at that temperature.     

The transformation and toxicity of anthraquinone dyes during thermophilic (55 degrees C) and mesophilic (30 degrees C) anaerobic treatments

We studied in batch assays the transformation and toxicity of anthraquinone dyes during incubations with anaerobic granular sludge under mesophilic (30 degrees C) and thermophilic (55 degrees C) conditions. Additionally, the electron shuttling capacity of the redox mediator anthraquinone-2-sulfonic acid (AQS) and subsequent increase on decolourisation rates was investigated on anthraquinone dyes. Compared with incubations at 30 degrees C, serum bottles at 55 degrees C presented distinctly higher decolourisation rates not only with an industrial wastewater containing anthraquinone dyes, but also with model compounds. Compared with batch assays at 30 degrees C, the first-order rate constant "k" of the Reactive Blue 5 (RB5) was enhanced 11-fold and 6-fold for bottles at 55 degrees C supplemented and free of AQS, respectively. However, the anthraquinone dye Reactive Blue 19 (RB19) demonstrated a very strong toxic effect on volatile fatty acids (VFA) degradation and methanogenesis at both 30 degrees C and 55 degrees C. The apparent inhibitory concentrations of RB19 exerting 50% reduction in methanogenic activity (IC50-value) were 55 mg l(-1) at 30 degrees C and 45 mg l(-1) at 55 degrees C. Further experiments at both temperatures revealed that RB19 was mainly toxic to methanogens, because the glucose oxidizers including acetogens, propionate-forming, butyrate-forming and ethanol-forming microorganisms were not affected by the dye toxicity.     

Methanogenesis in thermophilic biogas reactors

Methanogenesis in thermophilic biogas reactors fed with different wastes is examined. The specific methanogenic activity with acetate or hydrogen as substrate reflected the organic loading of the specific reactor examined. Increasing the loading of thermophilic reactors stabilized the process as indicated by a lower concentration of volatile fatty acids in the effluent from the reactors. The specific methanogenic activity in a thermophilic pilot-plant biogas reactor fed with a mixture of cow and pig manure reflected the stability of the reactor. The numbers of methanogens counted by the most probable number (MPN) technique with acetate or hydrogen as substrate were further found to vary depending on the loading rate and the stability of the reactor. The numbers of methanogens counted with antibody probes in one of the reactor samples was 10 times lower for the hydrogen-utilizing methanogens compared to the counts using the MPN technique, indicating that other non-reacting methanogens were present. Methanogens that reacted with the probe againstMethanobacterium thermoautotrophicum were the most numerous in this reactor. For the acetate-utilizing methanogens, the numbers counted with the antibody probes were more than a factor of 10 higher than the numbers found by MPN. The majority of acetate utilizing methanogens in the reactor wereMethanosarcina spp. single cells, which is a difficult form of the organism to cultivatein vitro. No reactions were observed with antibody probes raised againstMethanothrix soehngenii orMethanothrix CALS-1 in any of the thermophilic biogas reactors examined. Studies using 2-¹⁴C-labeled acetate showed that at high concentrations (more than approx. 1 mM) acetate was metabolized via the aceticlastic pathway, transforming the methyl-group of acetate into methane. When the concentration of acetate was less than approx. 1 mM, most of the acetate was oxidized via a two-step mechanism (syntrophic acetate oxidation) involving one organism oxidizing acetate into hydrogen and carbon dioxide and a hydrogen-utilizing methanogen forming the products of the first microorganism into methane. In thermophilic biogas reactors, acetate oxidizing cultures occupied the niche ofMethanothrix species, aceticlastic methanogens which dominate at low acetate concentrations in mesophilic systems. Normally, thermophilic biogas reactors are operated at temperatures from 52 to 56° C. Experiments using biogas reactors fed with cow manure showed that the same biogas yield found at 55° C could be obtained at 61° C after a long adaptation period. However, propionate degradation was inhibited by increasing the temperature.     

Effect of temperature and temperature fluctuation on thermophilic anaerobic digestion of cattle manure

The influence of temperature, 50 and 60 degrees C, at hydraulic retention times (HRTs) of 20 and 10 days, on the performance of anaerobic digestion of cow manure has been investigated in completely stirred tank reactors (CSTRs). Furthermore, the effect of both daily downward and daily upward temperature fluctuations has been studied. In the daily downward temperature fluctuation regime the temperatures of each reactor was reduced by 10 degrees C for 10 h while in the daily upward fluctuation regime the temperature of each reactor was increased 10 degrees C for 5 h. The results show that the methane production rate at 60 degrees C is lower than that at 50 degrees C at all experimental conditions of imposed HRT except when downward temperature fluctuations were applied at an HRT of 10 days. It also was found that the free ammonia concentration not only affects the acetate-utilising bacteria but also the hydrolysis and acidification process. The upward temperature fluctuation affects the maximum specific methanogenesis activity more severely as compared to imposed downward temperature fluctuations. The results clearly reveal the possibility of using available solar energy at daytime to heat up the reactor(s) without the need of heat storage during nights, especially at an operational temperature of 50 degrees C and at a 20 days HRT, and without the jeopardising of the overheating.     

Illumination enhances methane production from thermophilic anaerobic digestion

Incandescent lamp illumination enhanced methane production from a thermophilic anaerobic digestion reactor (55°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.     

Methane production using whole and screened dairy manure in conventional and fixed-film reactors

The technical feasibility of adopting the fixed-film reactor concept for biogas production from screened dairy manure was investigated. The methane production capability of laboratory-scale 4-L anaerobic reactors (conventional and fixed-film) receiving screened dairy manure and operated at 35 degrees C was compared. Dairy manure filtrate with 4.4% total solids (TS) and 3.4% volatile solids (VS) (average value) was prepared from 1:1 manure-water slurry. The feed material was added intermittently at loading rates ranging from 2.34 to 25 and 2.25 to 785 g VS/L d, respectively, for the conventional and fixed-film reactors. Maximum methane production rate (L CH(4)/L d) for the conventional reactor was 0.63 L CH(4)/L d achieved at a 6-day hydraulic retention time (HRT). For the fixed-film reactor the maximum production rate was 3.53 L CH(4)/L d when operated at a loading rate of 262 g VS/L d (3 h HRT). The fixed-film reactor was capable of sustaining a loading of 785 g VS/L d (1 h HRT). The fixed-film reactor performed much better than the conventional reactors. These results indicate that a large reduction of required reactor volume is possible through application of a fixed-film concept combined with a liquid-solid separation pretreatment of dairy manure.     

Thermophilic (55 degrees C) conversion of methanol in methanogenic-UASB reactors: influence of sulphate on methanol degradation and competition

Two upflow sludge bed reactors (UASB) were operated for 80 days at 55 degrees C with methanol as the substrate with an organic loading rate (OLR) of about 20 g CODl(-1) per day and a hydraulic retention time (HRT) of 10 h. One UASB was operated without sulphate addition (control reactor-R1) whereas the second was fed with sulphate at a COD:SO4(2-) ratio of 10 (sulphate-fed reactor-R2), providing an influent sulphate concentration of 0.6 g l(-1). For both reactors, methanogenesis was the dominant process with no considerable accumulation of acetate. The methanol removal averaged 93% and 83% for R1 and R2, respectively, and total sulphate removal was achieved in the latter. The pathway of methanol conversion for both sludges was investigated by measuring the fate of carbon in the presence and absence of bicarbonate or specific inhibitors for a sludge sample collected at day 72. In both sludges, about 70% of the methanol was syntrophically converted to methane and/or sulphide, via the intermediate H2/CO2. A strong competition between methanogens and sulphidogens took place in the R2 sludge with half of the methanol-COD being used by methane-producing bacteria and the other half by sulphate-reducing bacteria. Acetate was not an important intermediate for both sludges, and played a slightly more important role for the sulphate-adapted sludge (R2), sustained by the higher amount of bicarbonate produced during sulphate-reduction. The pathway study indicates that, although acetate does not represent an important intermediate, the system is susceptible to its accumulation.     

Effects of pH and hydraulic retention time on hydrogen production versus methanogenesis during anaerobic fermentation of organic household solid waste under extreme-thermophilic temperature (70 degrees C)

Two continuously stirred tank reactors were operated with household solid waste at 70 degrees C, for hydrogen and methane production. The individual effect of hydraulic retention time (HRT as 1, 2, 3, 4, and 6 days) at pH 7 or pH (5, 5.5, 6, 6.5, 7) at 3-day HRT was investigated on the hydrogen production versus methanogenesis. It was found that at pH 7, the maximum hydrogen yield was 107 mL-H(2)/g VS(added) (volatile solid added) but no stable hydrogen production was obtained as after some time methanogenesis was initiated at all tested HRTs. This demonstrated that sludge retention time alone was not enough for washing out the methanogens at pH 7 under extreme-thermophilic conditions. Oppositely, we showed that keeping the pH level at 5.5 was enough to inhibit methane and produce hydrogen stably at 3-day HRT. However, the maximum stable hydrogen yield was low at 21 mL-H(2)/g VS(added).     

Comparative performance and microbial diversity of hyperthermophilic and thermophilic co-digestion of kitchen garbage and excess sludge

The objective of this study was to evaluate the performance characteristics of a hyperthermophilic digester system that consists of an acidogenic reactor operated at hyperthermophilic (70 degrees C) conditions in series with a methane reactor operated at mesophilic (35 degrees C), thermophilic (55 degrees C), and hyperthermophilic (65 degrees C) conditions. Lab-scale reactors were operated continuously, and were fed with co-substrates composed of artificial kitchen garbage (TS 9.8%) and excess sludge (TS 0.5%) at the volumetric ratio of 20:80. In the acidification step, COD solubilization was in the range of 22-46% at 70 degrees C, while it was 21-29% at 55 degrees C. The average protein solubilization was 44% at 70 degrees C. The double bond fatty acid removal ratio at 70 degrees C was much higher than at 55 degrees C. These results suggested that the optimal operation conditions for the acidogenic fermenter were about 3.1 days of HRT and 4 days of SRT at 70 degrees C. Methane conversion efficiency and the VS removal percentage in the methanogenic step following acidification was around 65% and 64% on average at 55 degrees C, respectively. The optimal operational conditions for this system are acidogenesis performed at 70 degrees C and methanogenesis at 55 degrees C. The key microbes determined in the hyperthermophilic acidification step were Anaerobic thermophile IC-BH at 6.4 days of HRT and Thermoanaerobacter thermohydrosulfuricus DSM 567 at 2.4 days of HRT. These results indicated that the hyperthermophilic system provides considerable advantages in treating co-substrates containing high concentrations of proteins, lipids, and nonbiodegradable solid matter.     

Thermophilic anaerobic digestion of livestock waste: the effect of ammonia

Ammonia concentrations of 4 g N/l or more inhibited thermophilic digestion of cattle manure. A stable digestion of cattle manure could be maintained with ammonia concentrations up to 6 g N/l after 6 months of operation. However, the methane yield was reduced and the concentration of volatile fatty acids increased from 1 to 3 g/l as acetate, compared to controls with an ammonia concentration of 2.5 g N/l. The temporary strong inhibition following an one-step increase in ammonia concentration was reduced by applying a gradual increase. The specific methanogenic activity of ammonia-inhibited reactors (6 g N/l) with acetate or hydrogen as substrate was reduced by 73 and 52%, respectively. Tests of ammonia toxicity on the acetate- and hydrogen-utilizing populations showed a higher sensitivity of the aceticlastic compared to the hydrogenotrophic methanogens; the specific growth rate for the aceticlastic methanogens was halved at ammonia concentrations of 3.5 g N/l, compared to 7 g N/l for the hydrogenotrophic methanogens. Correspondence to: B. K. Ahring     

Difference in sporogenous bacterial populations in thermophilic (55 degrees C) and mesophilic (35 degrees C) anaerobic sewage digestion

Spores, sporeforming vegetative cells, and asporogenous populations were enumerated in two semicontinuous anaerobic fermentors digesting municipal primary sludge at 35 and 55 degrees C for more than 87 days. In the 35 degrees C fermentor, the anaerobic total population was 312.5 X 10(6)/ml, with 25.0 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 23.1 X 10(6), 59.2 X 10(6), 26.2 X 10(6), and 7.3 X 10(6)/ml, respectively, with 2.8 X 10(6), 6.7 X 10(6), 3.4 X 10(6), and 1.5 X 10(6)/ml being sporogenous, respectively. The sporeformers accounted for 8.0 to 20.0% of each of the respective populations. In the 55 degrees C fermentor, the anaerobic total population was 512.5 X 10(6)/ml, with 336.6 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 97.7 X 10(6), 190.7 X 10(6), 75.8 X 10(6), and 11.2 X 10(6)/ml, respectively, with 47.8 X 10(6), 110.6 X 10(6), 43.3 X 10(6), and 5.1 X 10(6)/ml, respectively, being sporogenous. The sporeformers represented 45.5 to 65.7% of each of the respective populations. The numbers of thermophilic sporeforming vegetative cells in the 55 degrees C fermentor were 9.0 to 19.8 times higher than their counterparts in the 35 degrees C fermentor. Most sporeformers were in the vegetative state in the 35 and 55 degrees C fermentors. After 18 days of fermentation at 55 degrees C, sporeformers carried out most of the digestion; however, the digestion was shared by both sporeformers and asporogenous bacteria after 87 days of fermentation. In the 35 degrees C fermentor, asporogenous bacteria digested most of the sludge. During the 18- and 87-day experimental periods, sporeformers were never predominant.     

Potential for thermophilic (50 degrees C) anaerobic dechlorination of pentachlorophenol in different ecosystems

Thermophilic (50 degrees C) anaerobic biodegradation of pentachlorophenol (PCP) was investigated by using different inocula from natural ecosystems and anaerobic digesters. The inocula tested were three freshwater sediments, four anaerobic sewage sludge samples from digesters treating sludge from wastewater plants with various industrial inputs, and digested manure from an anaerobic reactor. Only one digested-sludge sample and the manure sample were from thermophilic environments. The initial PCP concentration was 7.5 or 37.5 microM. After 8 months, PCP had disappeared from the sediment samples and various, less chlorinated intermediates were present. Additions of extra PCP were degraded within 4 weeks, and a maximal observed dechlorination rate of 1.61 mumol/liter/day in the vials with addition of 7.5 microM PCP and 7.50 mumol/liter/day in the vials with addition of 37.5 microM PCP were measured for a freshwater sediment. In contrast, only 2.8 to 17.5% of the initial PCP added had disappeared from the sludge samples after 8 months of incubation. The complex pattern of intermediates formed indicated that the dechlorination of PCP proceeded via different pathways, involving at least two different populations in the dechlorination processes.     

Effects of mixing on methane production during thermophilic anaerobic digestion of manure: lab-scale and pilot-scale studies

The effect of mixing on anaerobic digestion of manure was evaluated in lab-scale and pilot-scale experiments at 55 degrees C. The effect of continuous (control), minimal (mixing for 10 min prior to extraction/feeding) and intermittent mixing (withholding mixing for 2h prior to extraction/feeding) on methane production was investigated in three lab-scale continuously stirred tank reactors. On comparison to continuous mixing, intermittent and minimal mixing strategies improved methane productions by 1.3% and 12.5%, respectively. Pilot-scale studies also supported the lab-scale results with an average 7% increase in biogas yields during intermittent mixing compared to continuous mixing. The effect of mixing intensities (minimal, gentle or vigorous) in batch assays at 55 degrees C showed that when the process was overloaded by high substrate to inoculum ratio (40/60), gentle (35 times per minute) or minimal mixing (10 min mixing before feeding) was advantageous compared to vigorous mixing (110 times per minute). On the other hand, under low substrate to inoculum ratio (10/90), gentle mixing was the best. The study thus indicated that mixing schemes and intensities have some effect on anaerobic digestion of manures.     

Difference in sporogenous bacterial populations in thermophilic (55 degrees C) and mesophilic (35 degrees C) anaerobic sewage digestion.

Spores, sporeforming vegetative cells, and asporogenous populations were enumerated in two semicontinuous anaerobic fermentors digesting municipal primary sludge at 35 and 55 degrees C for more than 87 days. In the 35 degrees C fermentor, the anaerobic total population was 312.5 X 10(6)/ml, with 25.0 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 23.1 X 10(6), 59.2 X 10(6), 26.2 X 10(6), and 7.3 X 10(6)/ml, respectively, with 2.8 X 10(6), 6.7 X 10(6), 3.4 X 10(6), and 1.5 X 10(6)/ml being sporogenous, respectively. The sporeformers accounted for 8.0 to 20.0% of each of the respective populations. In the 55 degrees C fermentor, the anaerobic total population was 512.5 X 10(6)/ml, with 336.6 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 97.7 X 10(6), 190.7 X 10(6), 75.8 X 10(6), and 11.2 X 10(6)/ml, respectively, with 47.8 X 10(6), 110.6 X 10(6), 43.3 X 10(6), and 5.1 X 10(6)/ml, respectively, being sporogenous. The sporeformers represented 45.5 to 65.7% of each of the respective populations. The numbers of thermophilic sporeforming vegetative cells in the 55 degrees C fermentor were 9.0 to 19.8 times higher than their counterparts in the 35 degrees C fermentor. Most sporeformers were in the vegetative state in the 35 and 55 degrees C fermentors. After 18 days of fermentation at 55 degrees C, sporeformers carried out most of the digestion; however, the digestion was shared by both sporeformers and asporogenous bacteria after 87 days of fermentation. In the 35 degrees C fermentor, asporogenous bacteria digested most of the sludge. During the 18- and 87-day experimental periods, sporeformers were never predominant.     

Optimisation of biogas production from manure through serial digestion: lab-scale and pilot-scale studies

In the present study, the possibility of optimizing biogas production from manure by serial digestion was investigated. In the lab-scale experiments, process performance and biogas production of serial digestion, two methanogenic continuously stirred tank reactors (CSTR) connected in series, was compared to a conventional one-step CSTR process. The one-step process was operated at 55 degrees C with 15d HRT and 5l working volume (control). For serial digestion, the total working volume of 5l was distributed as 70/30%, 50/50%, 30/70% or 13/87% between the two methanogenic reactors, respectively. Results showed that serial digestion improved biogas production from manure compared to one-step process. Among the tested reactor configurations, best results were obtained when serial reactors were operated with 70/30% and 50/50% volume distribution. Serial digestion at 70/30% and 50/50% volume distribution produced 13-17.8% more biogas and methane and, contained low VFA and residual methane potential loss in the effluent compared to the one-step CSTR process. At 30/70% volume distribution, an increase in biogas production was also noticed but the process was very unstable with low methane production. At 13/87% volume distribution, no difference in biogas production was noticed and methane production was much lower than the one-step CSTR process. Pilot-scale experiments also showed that serial digestion with 77/23% volume distribution could improve biogas yields by 1.9-6.1% compared to one-step process. The study thus suggests that the biogas production from manure can be optimized through serial digestion with an optimal volume distribution of 70/30% or 50/50% as the operational fluctuations are typically high during full scale application. However, process temperature between the two methanogenic reactors should be as close as possible in order to derive the benefits of serial coupling.