Comparative effect of different aerobic pretreatments on the kinetics and macroenergetic parameters of anaerobic digestion of olive mill wastewater in continuous mode

A comparative kinetic study was carried out on the anaerobic digestion of olive mill wastewater (OMW) and OMW that was previously fermented with Geotrichum candidum, Azotobacter chroococcum and Aspergillus terreus. The reactors used were continuously fed and contained sepiolite as support for the mediating bacteria. A kinetic model for multicomponent substrate removal by anaerobic digestion has been used. The model is based on the linear removal concept which is a special case of the broader Monod equation. The second-order kinetic constant, k _{2( s )}, was found to be influenced by the pretreatment carried out, and was 4.2, 4.0 and 2.5 times higher for Aspergillus, Azotobacter and Geotrichum-pretreated OMWs than that obtained in the anaerobic digestion of untreated OMW. This was significant at 95% confidence level. This behaviour is believed to be due to the lower levels of phenolic compounds and biotoxicity present in the pretreated OMWs. In fact, the kinetic constant increased when the phenolic compound content and biotoxicity of the pretreated OMWs decreased. In addition, the macroenergetic parameters of the anaerobic digestion of OMW, i.e. the specific rate of substrate uptake for cell maintenance, m, and the yield coefficient for the biomass, Y, decreased by a factor of 2.4, 3.6 and 5.1 and increased by a factor of 1.9, 2.2 and 2.4 respectively, for the OMWs previously treated with Geotrichum candidum, Azotobacter chroococcum and Aspergillus terreus in relation to the observed values for the untreated OMW.     

Kinetic behaviour of waste tyre rubber as microorganism support in an anaerobic digester treating cane molasses distillery slops

The kinetics of anaerobic digestion of cane molasses distillery slops was investigated using a continuous-flow bioreactor which contained waste tyre rubber as support, to which the microorganisms became immobilized. Hydraulic retention times (HRT) ranging from 1 to 10 days were investigated at an average influent chemical oxygen demand (COD) concentration of 47.7?g/l. The maximum substrate utilization rate, k, and half saturation coefficient, K _L, were determined to be 1.82?kg COD_removed/kg VSS day and 0.33?kg COD/kg VSS day. The yield coefficient, Y, and sludge decay rate coefficient, K _d, were also determined to be 0.06?kg VSS/kg COD_removed and 0.05?day^-1, respectively. Methane production was maximum (6.75?l/l day) at a 2 day HRT corresponding to a biomass loading rate of 2.578?kg COD/kg VSS day. Biogas yield ranged between 0.51?l/g COD (HRT=2 days) and 0.25?l/g COD (HRT=1?day). In addition, the methane percentage in the biogas varied between 70.5% (HRT=10?days) and 47.5% (HRT=1?day). The close relationship between biomass loading rate and specific substrate utilization rate supported the use of Monod equations. Finally, the experimental values of effluent substrate concentration were reproduced with deviations equal to or less than 10% in every case.     

Anaerobic digestion of slaughterhouse waste: main process limitations and microbial community interactions

Fresh pig/cattle slaughterhouse waste mixtures, with different lipid-protein ratios, were characterized and their anaerobic biodegradability assessed in batch tests. The resultant methane potentials were high (270-300 L_CH4 kg⁻¹_COD) making them interesting substrates for the anaerobic digestion process. However, when increasing substrate concentrations in consecutive batch tests, up to 15 g_COD kg⁻¹, a clear inhibitory process was monitored. Despite the reported severe inhibition, related to lipid content, the system was able to recover activity and successfully degrade the substrate. Furthermore, 16SrRNA gene-based DGGE results showed an enrichment of specialized microbial populations, such as β-oxidizing/proteolitic bacteria (Syntrophomonas sp., Coprothermobacter sp. and Anaerobaculum sp.), and syntrophic methanogens (Methanosarcina sp.). Consequently, the lipid concentration of substrate and the structure of the microbial community are the main limiting factors for a successful anaerobic treatment of fresh slaughterhouse waste.     

Influence of dilution and phase separation on the anaerobic digestion of olive mill wastewaters

The high organic content of Olive Mill Wastewaters (OMW) causes some difficulties in maintaining the anaerobic process efficiency at high level. The two phase anaerobic system was used to treat olive mill wastewaters, diluted with tap water. Phase separation was accomplished through control of the hydraulic retention time and initial COD removal in two reactors operated in series. The effect of substrate concentration and phase separation on removal efficiency has been investigated. Experimental results indicated that yield of 0.322 to 0.335 litre biogas/g COD removal were obtained with two phase anaerobic treatment and space loading rate of 2.3 and 2.4 gCOD/l.day. The maximum methane production rate near to the theoretical value and corresponded to 360 ml of CH₄ for 1g COD removal.Abbreviations OMW Olive Mill Wastewaters - VFA Volatile Fatty Acid - COD Chemical Oxygen Demand - HRT High Retention Time - TKN Total Kjeldahl Nitrogen     

Enhanced Methane Production from Anaerobic Digestion of Disintegrated and Deproteinized Excess Sludge

To improve biogas yield and methane content in anaerobic digestion of excess sludge from the wastewater treatment plant, the sludge was disintegrated by using various methods (sonication, alkaline and thermal treatments). Since disintegrated sludge contains a high concentration of soluble proteins, the resulting metabolite, ammonia, may inhibit methane generation. Therefore, the effects of protein removal from disintegrated sludge on methane production were also studied. As a result, an obvious enhancement of biogas generation was observed by digesting disintegrated sludge (biogas yield increased from 15 to 36 ml/g COD_added·day for the raw excess sludge and the sonicated sludge, respectively). The quality of biogas was also improved by removing proteins from the disintegrated sludge. About 50% (w/w) of soluble proteins were removed from the suspension of disintegrated sludge by salting out using 35 g MgCl₂·6H₂O/l and also by isoelectric point precipitation at pH 3.3. For deproteinized sludge, methane production increased by 19%, and its yield increased from 145 ml/g COD_removed to 325 ml/g COD_removed. Therefore, the yield and quality of biogas produced from digestion of excess sludge can be enhanced by disintegrating the sludge and subsequent protein removal. Revisions requested 14 November 2005; Revisions received 13 January 2006     

Impact of milling,enzyme addition,and steam explosion on the solid waste biomethanation of an olive oil production plant

Anaerobic digestion is a consolidated bioprocess which can be further enhanced by incorporating an upstream pretreatment unit. The olive oil production produces a large amount of solid waste which needs to be properly managed and disposed. Three different pretreatment techniques were evaluated in regard to their impact on the anaerobic biodegradability: manual milling of olive pomace (OP), enzyme maceration, direct enzyme addition, and thermal hydrolysis of two-phase olive mill waste. The Gompertz equation was used to obtain parameters for comparison purposes. A substrate/inoculum ratio 0.5 was found to be the best to be used in anaerobic batch test with olive pomace as substrate. Mechanical pretreatment of OP by milling increases the methane production rate while keeping the maximum methane yield. The enzymatic pretreatment showed different results depending on the chosen pretreatment strategies. After the enzymatic maceration pretreatment, a methane production of 274 ml CH₄ g VS _added ^?1 was achieved, which represents an improvement of 32 and 71 % compared to the blank and control, respectively. The direct enzyme addition pretreatment showed no improvement in both the rate and the maximum methane production. Steam explosion showed no improvement on the anaerobic degradability of two-phase olive mill waste; however, thermal hydrolysis with no rapid depressurization enhanced notoriously both the maximum rate (50 %) and methane yield (70 %).     

Comparative study of anaerobic digestion of olive mill wastewater (OMW) and OMW previously fermented with Aspergillus terreus

A comparative kinetic study was carried out on the anaerobic digestion of olive mill wastewater (OMW) and OMW that was previously fermented with Aspergillus terreus. Two completely-mixed bioreactors were used for the study. The results obtained were evaluated using the Chen-Hashimoto methane production model to obtain values for the maximum specific growth rate ( _max) and kinetic constant (K) of the process for each case studied. The kinetic parameters were found to be influenced by the pretreatment carried out, and were 4.5 and 3.8 times higher for pretreated OMW than that obtained in the anaerobic digestion of untreated OMW. This was significant at 95 % confidence level. This behaviour is believed to be due to the lower levels of phenolic compounds and biotoxicity present in the pretreated OMW, as compared to untreated OMW, resulting in an improved process performance and stability. Finally, the experimental values of methane production were reproduced with deviations equal to or less than 10% in every case.The authors wish to thank the Comisión Interministerial de Ciencia y Technología-CICYT-(project AMB-92723C03-01) and Junta de Andalucia for providing financial support.     

Two-stage biological treatment of olive mill wastewater with whey as co-substrate

Olive mill wastewater (OMW) is a highly polluting wastewater, caused by a high organic load and phenol content. These characteristics suggest that it may be suitable for aerobic treatment and anaerobic bacterial digestion. Aerobic treatment coupled with anaerobic bacterial digestion may be economically feasible as the methane produced is a valuable energy source while simultaneously purifying the OMW. In an attempt to improve the overall performance of the process, the addition of a co-substrate such as whey to the aerobic treatment pre-treatment of OMW by the yeast Candida tropicalis was studied.The two-stage system operated satisfactorily up to an organic loading rate (OLR) of 3.0 kg COD L⁻¹ day⁻¹ with a biogas production rate of 1.25 L_biogas L_reactor⁻¹ day⁻¹ and a total COD reduction in excess of 93% (62% COD reduction in aerobic pretreatment and 83% COD reduction in anaerobic digestion). Fifty-four percent of the phenol was biodegraded during the aerobic treatment stage, and biogas with between 68% and 75% methane was produced during anaerobic digestion.     

Influence of lignin on the methanization of lignocellulosic wastes

The effect on anaerobic digestion of reducing the lignin content of vine shoots to 1% (w/w), by treatment with sodium chlorite in an acid medium at 80°C, is reported. The yields of methane obtained were 240 ml of CH₄ g⁻¹ of VS (volatile solids) fed for untreated vine shoots, and 370 ml of CH₄ g⁻¹ of VS fed for treated vine shoots. A mathematical model was used to calculate the kinetic parameters H and μ, and the increased biodegradability of the substrate in which lignin had been removed was confirmed. A study of the mass balances of the process under optimum conditions (temperature = 35°C; loading rate of 1 g litre⁻¹ digester day⁻¹) enabled the percentage of degraded cellulose to be calculated (35·5% for untreated vine shoots, 81·5% for the treated vine shoots), as were the volumes of biogas and methane produced per gram of VS introduced (VS₁) and degraded. The blocking effect of lignin on the methanization process was confirmed.     

Anaerobic digestion of the vegetable fraction of municipal refuses: mesophilic versus thermophilic conditions

The phenomena limiting the anaerobic digestion of vegetable refuses are studied through batch tests carried out using anaerobic sludge previously selected under either mesophilic (37v°C) or thermophilic (55v°C) conditions. The compositions of the hydrolysed cellulosic and hemicellulosic fractions of these materials are simulated by starch and hemicellulose hydrolysates, respectively. Non-hydrolysed mixtures of vegetable waste with sewage sludge are used to ascertain whether the hydrolysis of these polymeric materials is the limiting step of the digestion process or not. The experimental data of methane production are then worked out by a first-order equation derived from the Monod's model to estimate the kinetic rate constant and methane production yield for each material. Comparison of these results shows that passing from mesophilic to thermophilic conditions is responsible for a slight deceleration of methane production but remarkably enhances both methanation yield and methane content of biogas. The final part of the study deals with the fed-batch digestion of the same residues in static digester. Working under thermophilic conditions at a loading rate threshold of 6.0 g_COD/l · d, the hemicellulose hydrolysate ensures the highest methane productivity (60 mmol_CH4/l · d) and methane content of biogas (60%), while unbalance towards the acidogenic phase takes place under the same conditions for the starch hydrolysate. The intermediate behaviour of the non-hydrolysed mixture of vegetable waste with sewage sludge demonstrates that hemicellulose hydrolysis is the limiting step of digestion and suggests the occurrence of ligninic by products inhibition on methane productivity.     

One-stage and two-stage anaerobic digestion of lipid-extracted algae

Waste-grown microalgae are a potentially important biomass for wastewater treatment. The lipid accumulated in microalgae could be utilized as feedstocks for biodiesel production. The algal residues, as major by-products derived from lipid extraction, mainly consist of carbohydrate and protein, making anaerobic digestion an efficient way to recover energy. The conversion of lipid-extracted algal residues into methane plays dual role in renewable energy production and sustainable development of microalgal biodiesel industry. Therefore, an anaerobic fermentation process for investigation of the methane production potential of algal residues was conducted in this paper. The effect of inoculum to substrate ratios (ISRs) on the methane production by anaerobic digestion of Chlorella sp. residue in a single stage was evaluated. The maximum methane yield of 195.6 ml CH₄/g volatile solid (VS) was obtained at an ISR of 1:1. The stability and progress of the reaction from algal residues to methane were monitored by measuring the pH, volatile fatty acids (VFAs), total ammoniacal nitrogen (TAN), and methane volume. Based on the results of one-stage experiments, two-stage technology was proposed and was found to be more suitable for high organic load. The optimum conditions for acidogenesis and methanogenesis are indicated in this paper.     

Anaerobic digestion of glycerol derived from biodiesel manufacturing

The anaerobic digestion of glycerol derived from biodiesel manufacturing, in which COD was found to be 1010 g/kg, was studied in batch laboratory-scale reactors at mesophilic temperature using granular and non-granular sludge. Due to the high KOH concentration of this by-product, H₃PO₄ was added to recover this alkaline catalyst as agricultural fertilizer (potassium phosphates). Although it would not be economically viable, a volume of glycerol was distilled and utilised as reference substrate. The anaerobic revalorisation of glycerol using granular sludge achieved a biodegradability of around 100%, while the methane yield coefficient was 0.306 m³ CH₄/kg acidified glycerol. Anaerobic digestion could be a good option for revalorising this available, impure and low priced by-product derived from the surplus of biodiesel companies. The organic loading rate studied was 0.21–0.38 g COD/g VSS d, although an inhibition phenomenon was observed at the highest load.     

Biochemical methane potential (BMP) test for thickened sludge using anaerobic granular sludge at different inoculum/substrate ratios

The biochemical methane potential (BMP) test for thickened sludge was evaluated at three different inoculum/substrate (I:S) ratios. The cumulative methane yield was 51.4 mL CH₄/g VS_added at an I:S ratio of 1:1, 76.3 mL CH₄/g VS_added at an I:S ratio of 1:3, and 21.9 mL CH₄/g VS_added at an I:S ratio of 1:8. The greatest ultimate methane yield and methane production rate constant were achieved at an I:S ratio of 1:3, whereas the least was obtained at an I:S ratio of 1:8. The maximum methane production rate constant was 0.38/day and the minimum methane production rate constant was 0.0016/day. For the case of a lower I:S ratio, the biomass activity may be affected due to the low substrate concentration. On the other hand, for the case of higher I:S ratios, anaerobic digestion of thickened sludge was inhibited by higher concentrations of volatile fatty acids and lower pH.     

Enhancement of the ozonation of wine distillery wastewaters by an aerobic pretreatment

The decomposition of the organic substrate present in wine distillery wastewaters (WDW) is studied in batch reactors, by an ozonation process, by an aerobic degradation and by another ozonation of the aerobically pretreated wastewaters. In the ozonation process, the effects on the substrate removal obtained of the temperature, pH and the presence of H₂O₂ and UV radiation are established, and an approximate kinetic study is conducted which leads to the evaluation of the apparent kinetic constants for the substrate reduction. In the aerobic degradation treatment, the evolution of the substrate, biomass and total phenolic compounds are followed during the process, and a kinetic study is performed by using the Contois model, which applied to the experimental data provides the specific kinetic parameters q_max and K₁. Finally, in the ozonation of the pretreated wastewaters, the?influence of the operating variables is established, and the effect of this aerobic pretreatment on the substrate removal and kinetic constants obtained in the ozonation stage is also discussed.     

Effect of the solid content on anaerobic digestion of meat and bone meal

The effect of the solid content on anaerobic digestion of meat and bone meal (MBM) was investigated in batch reactors at MBM solid contents of 1%, 2%, 5% and 10%. There was no significant difference in the specific methane (CH₄) production potential with respect to the total volatile MBM solids (TVS) applied at these solid contents, which ranged from 351 to 381 ml CH₄/g TVS. However, the highest CH₄ yield with respect to the removed volatile MBM solids (RVS) was 482 ml CH₄/g RVS at the MBM solid content of 5%; the CH₄ yields were 384–448 ml CH₄/g RVS at the other MBM solid contents. The lag time of CH₄ production rose with the increase in the solid content. The longer lag time at MBM solid contents of 5% and 10% was due to inhibition caused by high concentrations of volatile fatty acids (VFAs) and free ammonia in the reactors, but the inhibition was reversible. The production of VFAs during the digestion varied with solid contents: at the solid content of 1%, only acetic acid was detected; at 2%, both acetic and propionic acids were detected; and at 5% and 10%, acetic, propionic, butyric and valeric acids were detected. After 93-day digestion, the volatile MBM solid reduction was 92%, 91%, 79% and 80% at MBM solid contents of 1%, 2%, 5% and 10%, respectively.     

Fate of organic carbon in UAFB treating raw sewage: impact of moderate to low temperature

The anaerobic biological treatment of raw sewage under low ambient temperatures was investigated in an up-flow anaerobic fixed bed (UAFB) reactor, over a 220-day trial period. The operating temperature was lowered stepwisely from 35 to 25, 20 and 15 °C. Obvious accumulation of acetic acid was observed in the effluent as the temperature dropped, which almost tripled at 15 °C compared to that at 35 °C. An interesting psychrophilic-forward-shift trend of methane production rate following the decrease of temperature was discovered. The methane yield at each trial was 169, 201, 256 and 269 L_STP CH₄/kg COD_removed. For each temperature trial, about 32.6%, 28.3%, 34.4% and 34.9% were removed in form of methane. Based on PCR-DGGE results, the band corresponded to Methanosphaera spp. only showed up on 15 °C profile. Analysis of both methane production and DGGE results indicated that psychro-tolerant microorganisms dominated the archaeal populations in the UAFB under low-temperature.     

A review of anaerobic digestion bio-kinetics

Anaerobic digestion is the key to sustainable wastewater management and bioenergy production. Kinetics plays an important role in the design of bioreactors, processes, and process scale-up in anaerobic digestion. This article focuses on a state-of-the-art literature review on the experimental kinetic studies of conventional anaerobic bioreactors and anaerobic membrane bioreactors. Various kinetic models that were used to fit the experimental data and derive the kinetic parameters are summarized and discussed in the literature. The values of the maximum specific growth rate µ_max, half saturation constant K_s, decay co-efficient k_d, sludge yield Y, and methane yield Y_CH4 from experimental studies are summarized for each model. This paper can serve as an updated comprehensive source of anaerobic bio-kinetic studies and digester design.     

Comparison of Seven Chemical Pretreatments of Corn Straw for Improving Methane Yield by Anaerobic Digestion

Agriculture straw is considered a renewable resource that has the potential to contribute greatly to bioenergy supplies. Chemical pretreatment prior to anaerobic digestion can increase the anaerobic digestibility of agriculture straw. The present study investigated the effects of seven chemical pretreatments on the composition and methane yield of corn straw to assess their effectiveness of digestibility. Four acid reagents (H₂SO₄, HCl, H₂O₂, and CH₃COOH) at concentrations of 1%, 2%, 3%, and 4% (w/w) and three alkaline reagents (NaOH, Ca(OH)₂, and NH₃·H₂O) at concentrations of 4%, 6%, 8%, and 10% (w/w) were used for the pretreatments. All pretreatments were effective in the biodegradation of the lignocellulosic straw structure. The straw, pretreated with 3% H₂O₂ and 8% Ca(OH)₂, acquired the highest methane yield of 216.7 and 206.6 mL CH₄ g VS ⁻¹ in the acid and alkaline pretreatments, which are 115.4% and 105.3% greater than the untreated straw. H₂O₂ and Ca(OH)₂ can be considered as the most favorable pretreatment methods for improving the methane yield of straw because of their effectiveness and low cost.     

Miscanthus for biogas production: Influence of harvest date and ensiling on digestibility and methane hectare yield

The 8,000 biogas plants currently in operation in Germany are mainly fed with biomass from annual crops. However, feedstock from perennial crops such as miscanthus is expected to be more environmentally benign. If miscanthus is to be used in greater amounts as a substrate for anaerobic digestion, storage will become a relevant topic, as a continuous supply of biomass throughout the year is necessary. The objective of this study was to identify the miscanthus harvest time that best balances the simultaneous achievement of high silage quality, high digestibility and high methane hectare yields. For this purpose, biomass from four miscanthus genotypes with varying senescence characteristics was harvested on three different dates in autumn 2017. Part of the biomass was ensiled, and the methane yield of both ensiled and non‐ensiled biomass was analysed in a biogas batch test to assess the effect of ensiling on the methane hectare yield and digestion velocity. The ensiled biomass was found to have an up to 7% higher substrate‐specific methane yield and also showed a higher digestion velocity than the non‐ensiled biomass. The silage quality was best when miscanthus was harvested in mid‐October, due to highest lactic acid content (average: 3.0% of DM) and lowest pH (average: 4.39) compared to the harvests in mid‐September and beginning of October. Mass losses during ensiling (as high as 7.6% of fresh matter for the M. sinensis genotype Sin55) were compensated for by a higher substrate‐specific methane yield (up to 353 Nml CH₄ (g oDM)^?1) in ensiled miscanthus. This resulted in non‐significantly different methane hectare yields for non‐ensiled (average: 4.635 Nm³ CH₄/ha) and ensiled miscanthus biomass (4.803 Nm³ CH₄/ha). A comparison of the four genotypes suggests that Miscanthus x giganteus is the most suitable genotype for ensiling as it had the best silage quality.     

Production and characterization of β-glucosidase from Aspergillus niger fermentation: Application for organic fraction of municipal solid waste hydrolysis and methane enhancement

The anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) is currently an attractive treatment process with energy production in the form of biogas. Hydrolysis is the rate-limiting step for the anaerobic digestion of solid wastes. Thus, in the present study fungal enzymatic pretreatment of OFMSW was applied to enhance biogas production. Two enzyme cocktails rich on β-glucosidase were produced from submerged fermentation of Aspergillus niger on basal medium using OFMSW as carbon source and urea (Urea cocktail) and Ulva rigida as nitrogen source (Ulva cocktail). Ulva cocktail displayed an important effect on OFMSW solubilization. Therefore, an increase of reducing sugar concentration about 60% was obtained which was in correlation with chemical oxygen demand (COD) increase. The performance of enzymatic pretreatment on anaerobic digestion of OFMSW was studied by conducting biochemical methane potential tests. Results showed that the enzymatic pretreatment improved methane yield of OFMSW even at high solid concentration. High methane yield about 500 ml/g total volatile solid was obtained, which corresponds up to 68% enhancement over the control.