Effect of aeration rate and waste load on evolution of volatile fatty acids and waste stabilization during thermophilic aerobic digestion of a model high strength agricultural waste

Thermophilic aerobic digestion (TAD) is a relatively new, dynamic and versatile low technology for the economic processing of high strength waste slurries. Waste so treated may be safely disposed of or reused. In this work a model high strength agricultural waste, potato peel, was subjected to TAD to study the effects of oxygen supply at 0.1, 0.25, 0.5 and 1.0 vvm (volume air per volume slurry per minute) under batch conditions at 55 degrees C for 156 h on the process. Process pH was controlled at 7.0 or left unregulated. Effects of waste load, as soluble chemical oxygen demand (COD), on TAD were studied at 4.0, 8.0, 12.0 and 16.0 gl(-1) (soluble COD) at pH 7.0, 0.5 vvm and 55 degrees C. Efficiency of treatment, as degradation of total solids, total suspended solids and soluble solid, as well as soluble COD significantly increased with aeration rate, while acetate production increased as the aeration rate decreased or waste load increased, signifying deterioration in treatment. Negligible acetate, and no other acids were produced at 1.0 vvm. Production of propionate and other acids increased after acetate concentration had started to decrease and, during unregulated reactions coincided with the drop in the pH of the slurry. Acetate production was more closely associated with periods of oxygen limitation than were other acids. Reduction in oxygen availability led to deterioration in treatment efficiency as did increase in waste load. These variables may be manipulated to control treated waste quality.     

Protein enrichment of corn cob heteroxylan waste slurry by thermophilic aerobic digestion using Bacillus stearothermophilus

Thermophilic aerobic digestion (TAD) of heteroxylan waste was implemented at waste load of 30gL(-1) with mineral nitrogen supplementation to study effect of the process on waste degradation, protein accretion and quality. Digestions were carried out at 45 50, 55, 60 and 65 degrees C using Bacillusstearothermophilus in a CSTR under batch conditions at 1.0vvm aeration rate, pH 7.0 for a maximum of 120h. Amylase and xylanase activities appeared rapidly in the digest, while basal protease activity appeared early in the digestion and increased towards end of the processes. Highest degradation of volatile suspended solid, hemicellulose and fibre occurred at 55 degrees C while highest degradation of total suspended solid occurred at 60 degrees C. Highest protein accretion (258.8%) and assimilation of mineral nitrogen and soluble protein occurred at 55 degrees C. The % content of amino acids of digest crude protein increased relative to raw waste and with digestion temperature. Quality of digest protein was comparable to the FAO standard for feed use. TAD has potentials for use in the protein enrichment of waste.     

Effect of process temperature, pH and suspended solids content upon pasteurization of a model agricultural waste during thermophilic aerobic digestion

Thermophilic aerobic digestion(TAD), or liquid composting, is a versatile new process for the treatment and stabilization of high strength wastes of liquid or, perhaps more importantly, slurry consistency. The pattern of inactivation of various pathogenic and indicator organisms was studied using batch digestions under conditions that may be expected to be found in full-scale TAD processes. Rapid inactivation of test populations occurred within the first 10 min from the start of digestion. The inactivation rate was slightly lower when digestions were conducted below 60 degrees C. In some instances, a 'tail' was apparent, possibly indicating the survival of relatively resistant sub-populations particularly in the case of Serratia marcescens and Enterococcus faecalis, or of clumping or attachment of cells to particulate materials. The effect of pH on the inactivation of the test populations depended on the temperature of digestion, but varied with the test population. At 55 degrees C Escherichia coli was more sensitive to temperature effects at pH 7 than at pH 8, but was more sensitive at pH 8, 60 degrees C. The reverse was the case at 60 degrees C for Ent. faecalis. An increase in the solid content of the digesting waste caused a progressive increase in the protection of test organisms from thermal inactivation. Challenging a TAD process with test strains allows (via estimation of D-values) a quantification of the cidal effects of such processes, with a view to manipulating process variables to enhance such effects.     

Yield and protein quality of thermophilic Bacillus spp. biomass related to thermophilic aerobic digestion of agricultural wastes for animal feed supplementation

Bacillus spp. responsible for thermophilic aerobic digestion (TAD) of agricultural wastes were studied for their growth rate, yield and protein quality (amino acid profile) under conditions that approximate full-scale waste digestion as pointers to the capacity of TAD to achieve protein enrichment of wastes for reuse in animal feeding. Specific growth rates of the thermophiles varied with temperature and aeration rates. For Bacillus coagulans, the highest specific growth rate was 1.98 muh(-1); for Bacillus licheniformis 2.56 muh(-1) and for Bacillus stearothermophilus 2.63 muh(-1). Molar yield of B. stearothermophilus on glucose increased with temperature to a peak of 0.404 g g(-1) at 50 degrees C before declining. Peak concentration of overflow metabolite (acetate) increased from 10 mmol at 45 degrees C to 34 mmol at 65 degrees C before declining. Accumulation of biomass in all three isolates decreased with increase in temperature while protein content of biomass increased. Highest biomass protein (79%) was obtained in B. stearothermophilus at 70 degrees C. Content of most essential amino acids of the biomass improved with temperature. Amino acid profile of the biomass was comparable to or superior to the FAO standard for SCP intended for use in animal feeding. Culture condition (waste digestion condition) may be manipulated to optimize protein yield and quality of waste digested by TAD for recycling in animal feed.     

The role of HRT and low temperature on the acid-phase anaerobic digestion of municipal and industrial wastewaters

Two identical 31 completely mixed reactors with solids recycling capabilities were used to investigate the effects of hydraulic retention time (HRT) and low temperatures on volatile fatty acid (VFA) production. One reactor was fed with a 1:1 ratio of diluted primary sludge and a starch-rich industrial wastewater, while the other was fed with diluted primary sludge alone. The VFA and soluble COD concentrations and specific production rates reached their highest values at 30 h HRT and at 25 degrees C. Further increase in HRT (at 25 degrees C) or decrease in temperature (at an HRT of 30 h) resulted in lower amounts of VFA and COD produced. All parameters related to VFA and COD production were significantly higher in the industrial-municipal reactor than in the municipal-only reactor. The VFA:COD ratios were very high, with values ranging from about 0.8 to 1.0 indicating that hydrolysis was the rate-limiting step. Degradation of proteins (measured by ammonia production) was inhibited by the starch-rich wastewater in the industrial-municipal reactor, while no evidence of inhibition was found in the municipal-only reactor. This study revealed that VFA production was feasible at low temperatures (down to 8 degrees C), particularly in the presence of the industrial waste. Ultimately, the amount VFA produced was adequate, in most cases, to support subsequent biological nutrient removal (BNR) processes.     

Diversity of thermophilic populations during thermophilic aerobic digestion of potato peel slurry

Aims:  To study the diversity of thermophiles during thermophilic aerobic digestion (TAD) of agro-food waste slurries under conditions similar to full-scale processes.
Methods and Results:  Population diversity and development in TAD were studied by standard microbiological techniques and the processes monitored by standard fermentation procedures. Facultative thermophiles were identified as Bacillus coagulans and B. licheniformis, while obligate thermophiles were identified as B. stearothermophilus . They developed rapidly to peaks of 10⁷ to 10⁸ in ≤48 h. Stability of obligate thermophiles increased with process temperatures. Thermophiles were unstable at process pH above or below neutral, but developed rapidly at all aeration rates. Peak populations were higher in the median than at extremes of aeration rates. Obligate thermophiles were unstable at low aeration rates. Process self-heating was higher at lower than at higher aeration rate. Beyond 96 h most thermophiles were present as spores.
Conclusions:  Limited range of indigenous thermophiles drives TAD of slurry. They develop rapidly and are stable at most digestion conditions.
Significance and Impact of the Study:  Development and stability of thermophiles in TAD suggest that the process may be operated in a wide range of conditions; and even at short HRT in continuous processes without compromising efficiency.     

Thermophilic anaerobic digestion of sewage sludge: focus on the influence of the start-up. A review

The thermophilic anaerobic digestion (TAD) of sewage sludge has often been found to be less stable than mesophilic treatment. In comparison to mesophilic digesters, thermophilic reactors treating sludge are generally characterized by relatively high concentrations of volatile fatty acids (VFA) in the effluent along with poor effluent quality, indicating a lower level of process stability. However, reviewing the literature related to the procedure for obtaining a thermophilic inoculum, it seems that most of the problems associated with the instability and the accumulation of organic intermediates are the result of the manner in which the thermophilic sludge has been obtained. In this paper, the different options available for obtaining an anaerobic digester operating at thermophilic temperature (55°C) have been reviewed. In this light, rapid heating to the target temperature followed by the development of thermophilic microorganisms, which can be determined by VFA dropping to ≤500?mg acetic acid L^?1 before increasing the organic loading rate (OLR), has been determined the most suitable means of establishing TAD.     

Ultrasonic-pretreated waste activated sludge hydrolysis and volatile fatty acid accumulation under alkaline conditions: Effect of temperature

The effect of temperature on the hydrolysis and acidification of ultrasonic-pretreated waste activated sludge (WAS) under alkaline conditions was investigated in this study. The experiment temperatures were set at 10, 20, 37, and 55°C. Experimental results showed that the hydrolysis of ultrasonic-pretreated WAS under alkaline conditions increased significantly with temperature from 10 to 55°C, while the volatile fatty acid (VFA) accumulation was not augmented as temperature increased. Among the four temperatures tested, 37°C was the point with the highest VFA accumulation after 72h fermentation. VFA accumulation decreased markedly at 55°C compared to 37°C. Mechanism investigation revealed that among all the temperatures tested, 37°C was the temperature at which consumptions of WAS protein and carbohydrate, activities of key enzymes related to VFA formation and ratio of Bacteria to Archaea all reached the maximum. Due to activities of related microorganisms inhibited by higher temperature (55°C), VFA accumulation decreased at 55°C.     

Thermophilic anaerobic digestion of high strength wastewaters

Investigations on the thermophilic anaerobic treatment of high-strength wastewaters (14-65 kg COD/m(3)) are presented. Vinasse, the wastewater of alcohol distilleries, was used as an example of such wastewaters. Semicontinuously fed digestion experiments at high retention times revealed that the effluent quality of digestion at 55 degrees C is comparable with that at 30 degrees C at similar loading rates. The amount of methane formed per kilogram of vinasse drops almost linearly with increasing vinasse concentrations. This can be attributed to increasing concentrations of inhibitory compounds, resulting in increasing volatile fatty acid (VFA) concentrations in the effluent. The treatment of vinasse was also investigated using upflow anaerobic sludge blanket (UASB) reactors. Thermophilic granular sludge, cultivated on sucrose, was used as seed material. The sludge required a 4-month adaptation period, during which the size of the sludge granules decreased significantly. However, the settling characteristics remained satisfactory. After adaptation, high loading and methane generation rates could be accommodated at satisfactory treatment efficiencies, namely, 86.4 kg COD/m(3) day and 26 m(3) CH(4)(STP)/m(3) day, respectively. As in the semicontinuously fed digesters, the effluent VFA concentrations were virtually independent of the loading rates applied, indicating that the toxicity of the vinasse is more important than the loading rate in determining the efficiency of the conversion of vinasse to methane.     

Thermal-alkaline solubilization of waste activated sludge as a pre-treatment stage for anaerobic digestion

This work studied the hydrolysis kinetics and the solubilization of waste activated sludge under a medium range temperature (50-90 degrees C) and pH in the alkaline region (8-11), as a pretreatment stage for anaerobic digestion. The hydrolysis rate for the solubilization of volatile suspended solids (VSS) followed a first-order rate. A linear polynomial hydrolysis model was derived from the experimental results leading to a satisfactory correlation between the hydrolysis rate coefficient, pH, and temperature. At pH 11 and a temperature of 90 degrees C the concentration of the VSS was 6.82%, the VSS reduction reached 45% within ten hours and at the same time the soluble COD was 70.000 mg/l and the total efficiency for methane production 0.28 l of CH4 per g of VSS loading.     

Influence of pH and temperature on soluble substrate generation with primary sludge fermentation

The study was undertaken to evaluate the effect of pH and temperature control on the generation of soluble fermentation products from primary sludge. The effect was tested by running parallel experiments under pH and temperature controlled and uncontrolled conditions. In fermentation experiments conducted at 20 degrees C without pH control, the average soluble COD release was 14 mg per liter of wastewater treated, representing a potential increase of 5% in the biodegradable COD content of the primary sedimentation effluent. The corresponding average VFA generation was 9.2mg COD l(-1). The nutrient release was practically negligible and stayed at 0.4 mg l(-1) for nitrogen and 0.1mg l(-1) for phosphorus. Acetic acid accounted more than 45% of the generated VFA in all experimental runs. The acetic acid content of the VFA decreased with increasing initial VSS concentrations and higher pH levels. VFA generation by fermentation was significantly affected with temperature and pH control. Temperature change between 10 and 24 degrees C induced a five-fold increase in VFA generation, from 610 mg l(-1) at 10 degrees C to 2950 mg l(-1) at 24 degrees C.     

Effect of saponification on the anaerobic digestion of solid fatty residues

In France, fatty residues considered as "non-ultimate" waste will not be accepted in landfill from 2002, in accordance with French legislation. Anaerobic digestion appears as an alternative process to mobilize and profitably use such fermentable waste. In this work, the effect of an alkaline pretreatment on the degradation of hexane extractible matter (HEM) and the production of volatile fatty acids (VFAs) was compared in reactors working at pH 6.5 and 8.5. The results obtained showed that 40% (+/- 0.1) of HEM were degraded at pH 8.5 versus 10% (+/- 0.3) at pH 6.5, regardless of the alkali agent used to saponify the greasy wastes. The highest performances of VFA production (8.45% +/- 0.3) were obtained at pH 8.5 with greases saponified by potassium hydroxide, compared to results (4.25% +/- 0.1) with greasy waste saponified by sodium hydroxide. This difference in VFA production might be attributable to biotoxic molecules generated during the saponification of greases by soda, limiting consequently the VFA production process.     

Kinetic studies on thermal denaturation of C-phycocyanin

The kinetics of thermal denaturation of a biliprotein, C-phycocyanin (C-PC) isolated from Spirulina platensis were studied at different pH values, ranging from 4.0 to 8.0. The denaturation of C-PC follows the first order kinetics and rate constant at pH 5.0 and temperature 55 degrees C is found to be 4.37 x 10(-5) s(-1), which increases to 5.46 x 10(-1) s(-1) at pH 7.0. The denaturation rate is much higher at 65 degrees C and pH 7.0 (7.96 x 10(-4)), as compared to at pH 5.0 (1.46 x 10(-4)). The thermal stability of C-PC is more at pH 5.0, as compared to other pH values. The observed differences in entropy values at pH 5.0, as compared to other pH values indicate a considerably close fit structure of the protein at pH 5.0, which increases the stability of native structure, even at higher temperature (65 degrees C).     

Aerobic and two-stage anaerobic-aerobic sludge digestion with pure oxygen and air aeration

The degradability of excess activated sludge from a wastewater treatment plant was studied. The objective was establishing the degree of degradation using either air or pure oxygen at different temperatures. Sludge treated with pure oxygen was degraded at temperatures from 22 degrees C to 50 degrees C while samples treated with air were degraded between 32 degrees C and 65 degrees C. Using air, sludge is efficiently degraded at 37 degrees C and at 50-55 degrees C. With oxygen, sludge was most effectively degraded at 38 degrees C or at 25-30 degrees C. Two-stage anaerobic-aerobic processes were studied. The first anaerobic stage was always operated for 5 days HRT, and the second stage involved aeration with pure oxygen and an HRT between 5 and 10 days. Under these conditions, there is 53.5% VSS removal and 55.4% COD degradation at 15 days HRT - 5 days anaerobic, 10 days aerobic. Sludge digested with pure oxygen at 25 degrees C in a batch reactor converted 48% of sludge total Kjeldahl nitrogen to nitrate. Addition of an aerobic stage with pure oxygen aeration to the anaerobic digestion enhances ammonium nitrogen removal. In a two-stage anaerobic-aerobic sludge digestion process within 8 days HRT of the aerobic stage, the removal of ammonium nitrogen was 85%.     

The purification and characterization of a dextranase from Lipomyces starkeyi

Dextranase produced by Lipomyces starkeyi was purified 43-fold, by carboxymethyl-Sepharose chromatography followed by agarose gel-filtration chromatography. The purified enzyme showed four bands by SDS/polyacrylamide gel electrophoresis with estimated mass 74 kDa, 71 kDa, 68 kDa and 65 kDa. This preparation exhibited multiple isoelectric points between 5.6 and 6.1. All the isoelectric forms were active and catalytically similar. The dextranase contained a carbohydrate moiety (8%). The physical properties of the enzyme were pH and temperature optima of 5.0 and 55 degrees C, respectively. This dextranase was stable between pH 2.5 and 7.0 at temperatures below 40 degrees C. Lipomyces dextranase was a typical endodextranase with the final product of dextran hydrolysis being isomalto-oligosaccharides from glucose to isomaltotetrose.     

Formation of soluble oligomers and amyloid fibrils with physical properties of the scrapie isoform of the prion protein from the C-terminal domain of recombinant murine prion protein mPrP-(121-231)

Prion diseases are fatal neurodegenerative disorders associated with conformational conversion of the cellular prion protein, PrP(C), into a misfolded, protease-resistant form, PrP(Sc). Here we show, for the first time, the oligomerization and fibrillization of the C-terminal domain of murine PrP, mPrP-(121-231), which lacks the entire unstructured N-terminal domain of the protein. In particular, the construct we used lacks amino acid residues 106-120 from the so-called amyloidogenic core of PrP (residues 106-126). Amyloid formation was accompanied by acquisition of resistance to proteinase K digestion. Aggregation of mPrP-(121-231) was investigated using a combination of biophysical and biochemical techniques at pH 4.0, 5.5, and 7.0 and at 37 and 65 degrees C. Under partially denaturing conditions (65 degrees C), aggregates of different morphologies ranging from soluble oligomers to mature amyloid fibrils of mPrP-(121-231) were formed. Transmission electron microscopy analysis showed that roughly spherical aggregates were readily formed when the protein was incubated at pH 5.5 and 65 degrees C for 1 h, whereas prolonged incubation led to the formation of mature amyloid fibrils. Samples incubated at 65 degrees C at pH 4.0 or 7.0 presented an initial mixture of oligomers and protofibrils or fibrils. Electrophoretic analysis of samples incubated at 65 degrees C revealed formation of sodium dodecyl sulfate-resistant oligomers (dimers, trimers, and tetramers) and higher molecular weight aggregates of mPrP-(121-231). These results demonstrate that formation of an amyloid form with physical properties of PrP(Sc) can be achieved in the absence of the flexible N-terminal domain and, in particular, of residues 106-120 of PrP and does not require other cellular factors or a PrP(Sc) template.     

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.     

The effects of digestion temperature and temperature shock on the biogas yields from the mesophilic anaerobic digestion of swine manure

In order to obtain basic design criteria for anaerobic digesters of swine manure, the effects of different digesting temperatures, temperature shocks and feed loads, on the biogas yields and methane content were evaluated. The digester temperatures were set at 25, 30 and 35 degrees C, with four feed loads of 5%, 10%, 20% and 40% (feed volume/digester volume). At a temperature of 30 degrees C, the methane yield was reduced by only 3% compared to 35 degrees C, while a 17.4% reduction was observed when the digestion was performed at 25 degrees C. Ultimate methane yields of 327, 389 and 403 mL CH(4)/g VS(added) were obtained at 25, 30 and 35 degrees C, respectively; with moderate feed loads from 5% to 20% (V/V). From the elemental analysis of swine manure, the theoretical biogas and methane yields at standard temperature and pressure were 1.12L biogas/g VS(destroyed) and 0.724 L CH(4)/g VS(destroyed), respectively. Also, the methane content increased with increasing digestion temperatures, but only to a small degree. Temperature shocks from 35 to 30 degrees C and again from 30 to 32 degrees C led to a decrease in the biogas production rate, but it rapidly resumed the value of the control reactor. In addition, no lasting damage was observed for the digestion performance, once it had recovered.     

Effect of temperature and pH on the ribulose-1,5-diphosphate carboxylase activity of the thermophilic hydrogen bacterium, Pseudomonas thermophila

The activity of ribulose 1,5-diphosphate (RDP) carboxylase was found in the soluble fraction of the cytoplasm from sonicated Pseudomonas thermophila K-2 cells. The enzyme is relatively thermolabile and completely loses its activity at 80 degrees C. The activity of RDP carboxylase at 60 degrees C increases by 40% during the first 10 min of heating in the presence of Mg2+ ions, bicarbonate and dithiothreitol, and again decreases if the enzyme is heated over 20 min. The optimum temperature of the enzyme is 50--55 degrees C. The specific activity of the enzyme in fresh preparations under these conditions reaches 0.22 unit per 1 mg of protein in the extract. The calculated value of the activation energy for RDP carboxylase is 6.4 kcal-mole-1, but 11.6 kcal-mole-1 in frozen preparations. The optimal pH is 7.0--7.3 depending on the buffer. The temperature optimum for the enzyme action does not depend on pH within the range of 7.3 to 8.8. Therefore, RDP carboxylase of Ps. thermophila K-2 differs from RDP carboxylases of mesophilic cultures studied earlier by a higher susceptibility to a decrease in temperature (the enzyme activity is negligible at 30 degrees C), by a lower value of the activation energy at suboptimal temperatures, and by a lower pH optimum of the enzyme action.     

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.