Effect of feed to inoculum ratios on biogas yields of food and green wastes

Biogas and methane yields of food and green wastes and their mixture were determined using batch anaerobic digesters at mesophilic (35 ± 2 °C) and thermophilic (50 ± 2 °C) temperatures. The mixture was composed of 50% food waste and 50% green waste, based on the volatile solids (VS) initially added to the reactors. The thermophilic digestion tests were performed with four different feed to inoculum (F/I) ratios (i.e., 1.6, 3.1, 4.0 and 5.0) and the mesophilic digestion was conducted at one F/I (3.1). The results showed that the F/I significantly affected the biogas production rate. At four F/Is tested, after 25 days of thermophilic digestion, the biogas yield was determined to be 778, 742, 784 and 396 mL/g VS for food waste, respectively; 631, 529, 524 and 407 mL/g VS for green waste, respectively; and 716, 613, 671 and 555 mL/g VS for the mixture, respectively. About 80% of the biogas production was obtained during the first 10 days of digestion. At the F/I of 3.1, the biogas and methane yields from mesophilic digestion of food waste, green waste and their mixture were lower than the yields obtained at thermophilic temperature. The biogas yields were 430, 372 and 358 mL/g VS, respectively, and the methane yields were 245, 206, and 185 mL/g VS, respectively.     

Feasibility of biogas production from anaerobic co-digestion of herbal-extraction residues with swine manure

The objective of this work was to examine the feasibility of biogas production from the anaerobic co-digestion of herbal-extraction residues with swine manure. Batch and semi-continuous experiments were carried out under mesophilic anaerobic conditions. Batch experiments revealed that the highest specific biogas yield was 294 mL CH₄ g⁻¹ volatile solids added, obtained at 50% of herbal-extraction residues and 3.50 g volatile solids g⁻¹ mixed liquor suspended solids. Specific methane yield from swine manure alone was 207 mL CH₄ g⁻¹ volatile solid added d⁻¹ at 3.50 g volatile solids g⁻¹ mixed liquor suspended solids. Furthermore, specific methane yields were 162, 180 and 220 mL CH₄ g⁻¹ volatile solids added d⁻¹ for the reactors co-digesting mixtures with 10%, 25% and 50% herbal-extraction residues, respectively. These results suggested that biogas production could be enhanced efficiently by the anaerobic co-digestion of herbal-extraction residues with swine manure.     

Potential for methane production from anaerobic co-digestion of swine manure with winery wastewater

This work examines the methane production potential for the anaerobic co-digestion of swine manure (SM) with winery wastewater (WW). Batch and semi-continuous experiments were carried out under mesophilic conditions. Batch experiments revealed that the highest specific methane yield was 348 mL CH₄ g⁻¹ COD added, obtained at 85.4% of WW and 0.7 g COD g⁻¹ VS. Specific methane yield from SM alone was 27 mL CH₄ g⁻¹ COD added d⁻¹. Furthermore, specific methane yields were 49, 87 and 107 mL CH₄ g⁻¹ COD added d⁻¹ for the reactors co-digesting mixtures with 10% WW, 25% WW and 40% WW, respectively. Co-digestion with 40% WW improved the removal efficiencies up to 52% (TCOD), 132% (SCOD) and 61% (VSS) compared to SM alone. These results suggest that methane can be produced very efficiently by the co-digestion of swine manure with winery wastewater.     

Strategies for optimizing recovery of the biogas process following ammonia inhibition

Strategies for recovery of ammonia-inhibited thermophilic biogas process, were evaluated in batch and lab-scale reactors. Active methane producing biomass (digested cattle manure) was inhibited with NH(4)Cl and subsequently, 3-5 days later, diluted with 50% of water, or with 50% digested manure, or with 50% fresh manure or kept undiluted. Dilution with fresh cattle manure resulted in the highest methane production rate during the recovery period while dilution with digested cattle manure gave a more balanced recovery according to the fluctuations in volatile fatty acids. Furthermore, the process recovery of a 7600m(3) biogas plant suffering from ammonia inhibition was observed. The ammonia concentration was only gradually lowered via the daily feeding with cattle manure, as is the normal procedure at Danish full-scale biogas plants. Recovery took 31 days with a 40% methane loss and illustrates the need for development of efficient process recovery strategies.     

Anaerobic co-digestion of desugared molasses with cow manure; focusing on sodium and potassium inhibition

Desugared molasses (DM), a syrup residue from beet-molasses, was investigated for biogas production in both batch and in continuously-stirred tank reactor (CSTR) experiments. DM contained 2-3 times higher concentration of ions than normal molasses, which could inhibit the biogas process. The effect of sodium and potassium concentration on biogas production from manure was also investigated. Fifty percent inhibition occurred at sodium and potassium concentration of 11 and 28 g/L, respectively. The reactor experiments were carried out to investigate the biogas production from DM under different dilutions with water and co-digestion with manure. Stable operation at maximum methane yield of 300 mL-CH₄/gVS-added was obtained at a mixture of 5% DM in cow manure. The biogas process was inhibited at DM concentrations higher than 15%. Manure was a good base substrate for co-digestion, and a stable anaerobic digestion could be achieved by co-digesting DM with manure at the concentration below 15% DM.     

Enhanced methane and hydrogen production from municipal solid waste and agro-industrial by-products co-digested with crude glycerol

The effects of crude glycerol on the performance of single-stage anaerobic reactors treating different types of organic waste were examined. A reactor treating the organic fraction of municipal solid waste produced 1400 mL CH₄/d before the addition of glycerol and 2094 mL CH₄/d after the addition of glycerol. An enhanced methane production rate was also observed when a 1:4 mixture of olive mill wastewater and slaughterhouse wastewater was supplemented with crude glycerol. Specifically, by adding 1% v/v crude glycerol to the feed, the methane production rate increased from 479 mL/d to 1210 mL/d. The extra glycerol-COD added to the feed did not have a negative effect on the reactor performance in either case. Supplementation of the feed with crude glycerol also had a significant positive effect on anaerobic fermentation reactors. Hydrogen yield was 26 mmole H₂/g VS added and 15 mmole H₂/g VS added in a reactor treating the organic fraction of municipal solid waste and a 1:4 mixture of olive mill and slaughterhouse wastewater. The addition of crude glycerol to the feed enhanced hydrogen yield at 2.9 mmole H₂/g glycerol added and 0.7 mmole H₂/g glycerol added.     

Effect of temperature on the performance of an anaerobic tubular reactor treating fruit and vegetable waste

This study compares the performance of anaerobic digestion of fruit and vegetable waste (FVW) in the thermophilic (55 °C) process with those under psychrophilic (20 °C) and mesophilic (35 °C) conditions in a tubular anaerobic digesters on a laboratory scale. The hydraulic retention time (HRT) ranged from 10 to 20 days, and raw fruit and vegetable waste was supplied in a semi-continuous mode at various concentrations of total solids (TS) (4, 6, 8 and 10% on dry weight). Biogas production from the experimental thermophilic digester was higher on average than from psychrophilic and mesophilic digesters by 144 and 41%, respectively. The net energy production in the thermophilic digester was 195.7 and 49.07 kJ per day higher than that for the psychrophilic and mesophilic digesters, respectively. The relation between the daily production of biogas and the temperature indicates that for the same produced quantity of biogas, the size of the thermophilic digester can be reduced with regard to that of the psychrophilic and the mesophilic digesters.     

Anaerobic digestion of whole stillage from dry-grind corn ethanol plant under mesophilic and thermophilic conditions

Anaerobic digestion of whole stillage from a dry-grind corn-based ethanol plant was evaluated by batch and continuous-flow digesters under thermophilic and mesophilic conditions. At whole corn stillage concentrations of 6348 to 50,786 mg total chemical oxygen demand (TCOD)/L, at standard temperature (0 °C) and pressure (1 atm), preliminary biochemical methane potential assays produced 88 ± 8 L (49 ± 5 L CH₄) and 96 ± 19 L (65 ± 14 L CH₄) biogas per L stillage from mesophilic and thermophilic digesters, respectively. Continuous-flow studies for the full-strength stillage (TCOD = 254 g/L) at organic loadings of 4.25, 6.30 and 9.05 g TCOD/L days indicated unstable performance for the thermophilic digester. Among the sludge retention times (SRTs) of 60, 45 and 30 days tested, the mesophilic digestion was successful only at 60 days-SRT which does not represent a practical operation time for a large scale bioethanol plant. Future laboratory studies will focus on different reactor configurations to reduce the SRT needed in the digesters.     

Evaluation of dairy cattle manure as a supplement to improve net energy gain in fermentative hydrogen production from sucrose

This study evaluated fermentative biohydrogen production from sucrose supplemented with dairy cattle manure at different sucrose:manure ratios. Hydrogen yields found in this study (2.9-5.3 M hydrogen/M sucrose) at ambient temperature are higher than literature results obtained at mesophilic temperatures. This study demonstrated that dairy cattle manure could serve as a buffering agent to maintain recommended pH levels; as a nutrient source to provide the required nutrients for hydrogen production; as a seed to produce hydrogen from sucrose; and as a co-substrate to improve the hydrogen yield. Based on an analysis of the net energy gain, it is concluded that positive net energy gains can be realized with non-thermal pretreatment and/or by combining dark fermentation with anaerobic digestion or microbial fuel cells to extract additional energy from the aqueous products of dark fermentation.     

Effect of initial COD concentration, nutrient addition, temperature and microbial acclimation on anaerobic treatability of broiler and cattle manure

In this study, anaerobic treatability and biogas generation potential of broiler and cattle manure were investigated. For this purpose, seven sets of anaerobic batch reactor experiments were performed using broiler and cattle manure and their mixtures in five different ratios (100% broiler; 75% broiler, 25% cattle; 50% broiler, 50% cattle; 25% broiler, 75% cattle; 100% cattle). These manure mixtures had two different initial chemical oxygen demand (COD) (12,000 and 53,500 mg/l) concentrations. The effects of initial COD concentration, nutrient and trace metal supplementation, microbial acclimation and digestion temperature were investigated. Results revealed that the efficiency of total COD removal was 32.0-43.3% and 37.9-50% for initial COD concentrations of 12,000 and 53,500 mg/l, respectively. The biogas yields observed for initial COD concentrations of 12,000 and 53,500 mg/l were 180-270 and 223-368 ml gas/g COD added, respectively. A decrease in biogas yield was observed as the fraction of broiler manure increased in mixture of broiler and cattle manure at initial COD values of 53,500 mg/l.     

Evaluation of Integrated Anaerobic Digestion and Hydrothermal Carbonization for Bioenergy Production

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal carbonization (HTC) are promising technologies for bioenergy production from biomass in terms of biogas and HTC biochar, respectively. In this study, the combination of AD and HTC is proposed to increase overall bioenergy production. Wheat straw was anaerobically digested in a novel upflow anaerobic solid state reactor (UASS) in both mesophilic (37 °C) and thermophilic (55 °C) conditions. Wet digested from thermophilic AD was hydrothermally carbonized at 230 °C for 6 hr for HTC biochar production. At thermophilic temperature, the UASS system yields an average of 165 L_CH4/kg_VS (VS: volatile solids) and 121 L _CH4/kg_VS at mesophilic AD over the continuous operation of 200 days. Meanwhile, 43.4 g of HTC biochar with 29.6 MJ/kg_{dry_biochar} was obtained from HTC of 1 kg digestate (dry basis) from mesophilic AD. The combination of AD and HTC, in this particular set of experiment yield 13.2 MJ of energy per 1 kg of dry wheat straw, which is at least 20% higher than HTC alone and 60.2% higher than AD only.     

Methanogenic population dynamics during start-up of anaerobic digesters treating municipal solid waste and biosolids

An aggressive start-up strategy was used to initiate codigestion in two anaerobic, continuously mixed bench-top reactors at mesophilic (37 degrees C) and thermophilic (55 degrees C) conditions. The digesters were inoculated with mesophilic anaerobic sewage sludge and cattle manure and were fed a mixture of simulated municipal solid waste and biosolids in proportions that reflect U.S. production rates. The design organic loading rate was 3.1 kg volatile solids/m3/day and the retention time was 20 days. Ribosomal RNA-targeted oligonucleotide probes were used to determine the methanogenic community structure in the inocula and the digesters. Chemical analyses were performed to evaluate digester performance. The aggressive start-up strategy was successful for the thermophilic reactor, despite the use of a mesophilic inoculum. After a short start-up period (20 days), stable performance was observed with high gas production rates (1.52 m3/m3/day), high levels of methane in the biogas (59%), and substantial volatile solids (54%) and cellulose (58%) removals. In contrast, the mesophilic digester did not respond favorably to the start-up method. The concentrations of volatile fatty acids increased dramatically and pH control was difficult. After several weeks of operation, the mesophilic digester became more stable, but propionate levels remained very high. Methanogenic population dynamics correlated well with performance measures. Large fluctuations were observed in methanogenic population levels during the start-up period as volatile fatty acids accumulated and were subsequently consumed. Methanosaeta species were the most abundant methanogens in the inoculum, but their levels decreased rapidly as acetate built up. The increase in acetate levels was paralleled by an increase in Methanosarcina species abundance (up to 11.6 and 4.8% of total ribosomal RNA consisted of Methanosarcina species ribosomal RNA in mesophilic and thermophilic digesters, respectively). Methanobacteriaceae were the most abundant hydrogenotrophic methanogens in both digesters, but their levels were higher in the thermophilic digester.     

Anaerobic digestion of cattle waste at mesophilic and thermophilic temperatures

Methanogenesis was studied using stirred, bench-top fermentors of 3-1 working volume fed on a semi-continuous basis with waste obtained from cattle fed a high grain, finishing diet. Digestion was carried out at 40 and 60°C. CH₄ production was 11.8, 18.3, 61.9 and 84.5% higher in the thermophilic than the mesophilic digestor at the 3, 6, 9 and 12 g volatile solids (VS) l^–1 reactor volume loading rates, respectively. When compared on an energetic basis CH₄ production was 7.4, 18.3, 72.9 and 107.3 kJ day higher in the thermophilic than the mesophilic digestor. CH₄ production decreased more rapidly with each increase in VS loading rate and decrease in retention time (RT) in the mesophilic than the thermophilic digestor. When expressed as l g^–1 VS fed or as kJ kJ^–1 fed, the amount of CH₄ was 49% less at the highest compared to the lowest loading rate in the mesophilic digestor. In the thermophilic digestor the decrease was only 16%. Propionate accumulated in the mesophilic digestor at the two highest loading rates, reaching concentrations of about 50 mM, but were only about 13 mM in the thermophilic digestor. Isobutyrate, isovalerate plus 2-methylbutyrate, and valerate also accumulated at the higher loading rates.     

Biogas production from different substrates in an experimental Continuously Stirred Tank Reactor anaerobic digester

Different mixtures were digested in a single-stage, batch, mixed, laboratory scale mesophilic anaerobic digester at the Biomass Research Centre Laboratory (University of Perugia). The yield and the composition of biogas from the different substrates were evaluated and the cumulative curves were estimated. Two experimental campaigns were carried out, the first on three mixtures (chicken, pig and bovine manures), the second on animal and vegetal biomasses (chicken and cow manure, olive husk) with different inocula (rumen fluid and digested sludge). In the first campaign pig manure mixture showed the maximum biogas production (0.35 Nm³/kg) and energy content (1.35 kWh/kg VS); in the second one the differences in produced biogas from the different inocula were analyzed: olive husk with piggery manure anaerobically digested as inoculum showed the higher biogas (0.28 Nm³/kg VS) and methane yield (0.11 Nm³/kg VS), corresponding to an energetic content of 1.07 kWh/kg VS. All data obtained from the laboratory scale anaerobic digester are comparable to the values in literature for several biomass and in particular for olive husk, dairy manure and chicken manure.     

Methane recovery from the anaerobic codigestion of municipal sludge and FOG

The anaerobic biodegradability of a mix of municipal primary sludge (PS), thickened waste activated sludge (TWAS) and fat, oil, and grease (FOG) was assessed using semi-continuous feed, laboratory-scale anaerobic digesters operated at mesophilic (35 °C) and thermophilic (52 °C) temperature. Addition of a large FOG fraction (48% of the total VS load) to a PS + TWAS mix, resulted in 2.95 times larger methane yield, 152 vs. 449 mL methane @ STP/g VS added at 35 °C and 2.6 times larger methane yield, 197 vs. 512 mL methane @ STP/g VS added at 52 °C. The high FOG organic load fraction was not inhibitory to the process. The results of this study demonstrate the benefit of sludge and FOG codigestion.     

Biotechnological intensification of biogas production

The importance of syntrophic relationships among microorganisms participating in biogas formation has been emphasized, and the regulatory role of in situ hydrogen production has been recognized. It was assumed that the availability of hydrogen may be a limiting factor for hydrogenotrophic methanogens. This hypothesis was tested under laboratory and field conditions by adding a mesophilic (Enterobacter cloacae) or thermophilic hydrogen-producing (Caldicellulosyruptor saccharolyticus) strain to natural biogas-producing consortia. The substrates were waste water sludge, dried plant biomass from Jerusalem artichoke, and pig manure. In all cases, a significant intensification of biogas production was observed. The composition of the generated biogas did not noticeably change. In addition to being a good hydrogen producer, C. saccharolyticus has cellulolytic activity; hence, it is particularly suitable when cellulose-containing biomass is fermented. The process was tested in a 5-m³ thermophilic biogas digester using pig manure slurry as a substrate. Biogas formation increased at least 160–170% upon addition of the hydrogen-producing bacteria as compared to the biogas production of the spontaneously formed microbial consortium. Using the hydrogenase-minus control strain provided evidence that the observed enhancement was due to interspecies hydrogen transfer. The on-going presence of C. saccharolyticus was demonstrated after several months of semicontinuous operation.     

Improving pig manure conversion into biogas by thermal and thermo-chemical pretreatments

Thermal (70–190 °C) and thermo-chemical (pH = 10 and 12, 25 °C and 90–190 °C) treatments were investigated in order to maximise the production of methane from pig manure. Methane production from treated and raw manure was assessed from batch mesophilic biochemical methane potential tests. Methane potential of manure soluble fraction increased with the temperature of thermal treatments whereas temperatures higher than 135 °C were necessary to improve the methane potential of the total fraction. The best results were obtained with the highest temperature (190 °C).     

Strategies for recovering inhibition caused by long chain fatty acids on anaerobic thermophilic biogas reactors

Long chain fatty acids (LCFA) concentrations over 1.0 g L⁻¹ were inhibiting manure thermophilic digestion, in batch and semi-continuous experiments, resulting in a temporary cease of the biogas production. The aim of the work was to test and evaluate several recovery actions, such as reactor feeding patterns, dilution and addition of adsorbents, in order to determine the most appropriate strategy for fast recovery of the reactor activity in manure based plants inhibited by LCFA. Dilution with active inoculum for increasing the biomass/LCFA ratio, or addition of adsorbents for adsorbing the LCFA and reducing the bioavailable LCFA concentration, were found to be the best recovery strategies, improving the recovery time from 10 to 2 days, in semi-continuously fed systems. Moreover, acclimatization was introduced by repeated inhibition and process recovery. The subsequent exposure of the anaerobic biomass to an inhibitory concentration of LCFA improved the recovery ability of the system, indicated as increasing degradation rates from 0.04 to 0.16 g COD_CH₄/g VS day. The incubation time between subsequent pulses, or discontinuous LCFA pulses, seems to be a decisive process parameter to tackle LCFA inhibition in manure anaerobic co-digestion.     

Production of an acidic and thermostable lipase of the mesophilic fungus Penicillium simplicissimum by solid-state fermentation

The production of a lipase by a wild-type Brazilian strain of Penicillium simplicissimum in solid-state fermentation of babassu cake, an abundant residue of the oil industry, was studied. The enzyme production reached about 90 U/g in 72 h, with a specific activity of 4.5 U/mg of total proteins. The crude lipase showed high activities at 35–60 °C and pH 4.0–6.0, with a maximum activity at 50 °C and pH 4.0–5.0. Enzyme stability was enhanced at pH 5.0 and 6.0, with a maximum half-life of 5.02 h at 50 °C and pH 5.0. Thus, this lipase shows a thermophilic and thermostable behavior, what is not common among lipases from mesophilic filamentous fungi. The crude enzyme catalysed the hydrolysis of triglycerides and p-nitrophenyl esters (C4:0–C18:0), preferably acting on substrates with medium-chain fatty acids. This non-purified lipase in addition to interesting properties showed a reduced production cost making feasible its applicability in many fields.     

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO₂, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic‐ and thermophilic anaerobic cultures were enriched to convert CO₂ to CH₄ by addition of H₂. Enrichment at thermophilic temperature (55°C) resulted in CO₂ and H₂ bioconversion rate of 320 mL CH₄/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic‐ and thermophilic‐enriched cultures, as revealed by PCR–DGGE. Nonetheless, they all belonged to the order Methanobacteriales, which can mediate hydrogenotrophic methanogenesis. Biogas upgrading was then tested in a thermophilic anaerobic reactor under various operation conditions. By continuous addition of hydrogen in the biogas reactor, high degree of biogas upgrading was achieved. The produced biogas had a CH₄ content, around 95% at steady‐state, at gas (mixture of biogas and hydrogen) injection rate of 6 L/(L day). The increase of gas injection rate to 12 L/(L day) resulted in the decrease of CH₄ content to around 90%. Further study showed that by decreasing the gas–liquid mass transfer by increasing the stirring speed of the mixture the CH₄ content was increased to around 95%. Finally, the CH₄ content around 90% was achieved in this study with the gas injection rate as high as 24 L/(L day). Biotechnol. Bioeng. 2012; 109: 2729–2736. © 2012 Wiley Periodicals, Inc.