Effects of Temperature and Carbon-Nitrogen (C/N) Ratio on the Performance of Anaerobic Co-Digestion of Dairy Manure,Chicken Manure and Rice Straw: Focusing on Ammonia Inhibition

Anaerobic digestion is a promising alternative to disposal organic waste and co-digestion of mixed organic wastes has recently attracted more interest. This study investigated the effects of temperature and carbon-nitrogen (C/N) ratio on the performance of anaerobic co-digestion of dairy manure (DM), chicken manure (CM) and rice straw (RS). We found that increased temperature improved the methane potential, but the rate was reduced from mesophilic (30∼40°C) to thermophilic conditions (50∼60°C), due to the accumulation of ammonium nitrogen and free ammonia and the occurrence of ammonia inhibition. Significant ammonia inhibition was observed with a C/N ratio of 15 at 35°C and at a C/N ratio of 20 at 55°C. The increase of C/N ratios reduced the negative effects of ammonia and maximum methane potentials were achieved with C/N ratios of 25 and 30 at 35°C and 55°C, respectively. When temperature increased, an increase was required in the feed C/N ratio, in order to reduce the risk of ammonia inhibition. Our results revealed an interactive effect between temperature and C/N on digestion performance.     

Improvement of activated sludge stabilisation and filterability during anaerobic digestion by fruit and vegetable waste addition

Anaerobic co-digestion of fruit and vegetable waste (FVW) and activated sludge (AS) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of AS:FVW ratio and the organic loading rate (OLR) on digesters performances were examined. The mixtures having AS:FVW ratios of 100:00, 65:35, 35:65, by a total solid (TS) basis were operated at an hydraulic retention time (HRT) of 20d. However, 30:70, 20:80, 15:85, 10:90 and 0:100 ratios were tested at an HRT of 10d. To investigate effects of aerobic and anaerobic digestion on the sludge filterability, specific resistance to filtration (R) was also determined. Increasing FVW proportions in the feedstock significantly improved the biogas production yield. The reactor that was fed with a 30:70 ratio showed the highest VS removal and biogas production yield of 88% and 0.57 L g(-1) VS added, respectively. The filterability results showed that the anaerobic effluent was characterised by a slightly better filterability efficiency of 1.6 x 10(16) m kg(-1) than 1.74 x 10(16) m kg(-1) of aerobic effluent. However, FVW addition improved the anaerobic co-digestion effluent filterability (5.52 x 10(14) m kg(-1)).     

Volatile fatty acids productivity by anaerobic co-digesting waste activated sludge and corn straw: Effect of feedstock proportion

Volatile fatty acids (VFAs) are the most suitable and biodegradable carbon substrates for many bioprocesses. This study explored a new approach to improve the VFAs production from anaerobic co-digesting waste activated sludge (WAS) with corn straw (CS). The effect of feedstock proportion on the acidification efficiency was investigated. The maximum VFAs yield (corresponding fermentation time) was substantially increased 69% (96 h), 45% (72 h), 13% (120 h) and 12% (120 h) with 50%, 35%, 25% and 20% CS proportion of feedstock, respectively. HAc (acetic acid) was consistently the most abundant, followed by HPr (propionic acid) and n-HBu (butyric acid) in the co-digesting tests. The increase of CS in feedstock led to more production of HAc and HPr. Moreover, the consumption of protein and carbohydrate were also improved remarkably from 2955 and 249 mg COD/L (individual WAS fermentation) to 6575 and 815 mg COD/L (50%_WAS:50%_CS co-digestion) from 120 onward, respectively. The highest contribution of CS to additional VFAs production was1113 mg VFAs (as COD)/g CS/L in the 65%_WAS:35%_CS co-digesting test. Our study indicated a valuable method to improve VFAs production from anaerobic co-digesting WAS and CS.     

Biohydrogen and methane production by co-digestion of cassava stillage and excess sludge under thermophilic condition

Thermophilic anaerobic hydrogen and methane production by co-digestion of cassava stillage (CS) and excess sludge (ES) was investigated in this study. The improved hydrogen and subsequent methane production were observed by co-digestion of CS with certain amount of ES in batch experiments. Compared with one phase anaerobic digestion, two phase anaerobic digestion offered an attractive alternative with more abundant biogas production and energy yield, e.g., the total energy yield in two phase obtained at VS_CS/VS_ES of 3:1 was 25% higher than the value of one phase. Results from continuous experiments further demonstrated that VS_CS/VS_ES of 3:1 was optimal for hydrogen production with the highest hydrogen yield of 74 mL/g total VS added, the balanced nutrient condition with C/N ratio of 1.5 g carbohydrate-COD/g protein-COD or 11.9 g C/g N might be the main reason for such enhancement. VS_CS/VS_ES of 3:1 was also optimal for continuous methane production considering the higher methane yield of 350 mL/g total VS added and the lower propionate concentration in the effluent.     

牲畜粪便与麦秆混合厌氧发酵的产气量、发酵时间及最优温度

探索牲畜粪便与作物秸秆混合发酵的产气量和发酵时间与发酵温度之间的关系,是解决农村户用沼气原料选择、确定最优发酵温度和提高农作物秸秆资源化利用效率的关键.采用可控型恒温发酵装置,以猪粪、牛粪和麦秆作为发酵原料,以常温厌氧发酵池的底物为接种物,在总固体（total solid,TS）质量分数为8%的条件下进行批量试验,研究了混合发酵的产气量、发酵时间及最优温度.结果表明：粪便与麦秆混合发酵明显提高了原料的产气效率,其中猪粪与麦秆混合发酵的累积产气量比猪粪作为单一发酵原料高24倍,而牛粪与麦杆混合发酵的累积产气量与单一牛粪无显著差异.猪粪、牛粪与麦秆混合发酵的最优温度均在30 ℃以上,发酵时间在60 d左右.厌氧发酵的发酵时间不总是随着温度的升高而缩短,单一以温度来断定厌氧发酵时间的长短是不可行的.     

Bioconversion of wheat straw cellulose/hemicellulose to ethanol by Saccharomyces uvarum and Pachysolen tannophilus

The information presented in this publication represents current research findings on the production of glucose and xylose from straw and subsequent direct fermentation of both sugars to ethanol. Agricultural straw was subjected to thermal or alkali pulping prior to enzymatic saccharification. When wheat straw (WS) was treated at 170 degrees C for 30-60 min at a water-to-solids ratio of 7:1, the yield of cellulosic pulp was 70-82%. A sodium hydroxide extration yielded a 60% cellulosic pulp and a hemicellulosic fraction available for fermentation to ethanol. The cellulosic pulps were subjected to cellulase hydrolysis at 55 degrees C for production of sugars to support a 6-C fermentation. Hemicellulose was recovered from the liquor filtrates by acid/alcohol precipitation followed by acid hydrolysis to xylose for fermentation. Subsequent experiments have involved the fermentation of cellulosic and hemicelluosic hydrolysates to ethanol. Apparently these fermentations were inhibited by substances introduced by thermal and alkali treatment of the straws, because ethanol efficiencies of only 40-60% were achieved. Xylose from hydrolysis of wheat straw pentosans supported an ethanol fermentation by Pachysolen tannophilus strain NRRL 2460. This unusual yeast is capable of producing ethanol from both glucose and xylose. Ethanol yields were not maximal due to deleterious substances in the WS hydrolysates.     

Effect of reactor configuration on biogas production from wheat straw hydrolysate

The potential of wheat straw hydrolysate for biogas production was investigated in continuous stirred tank reactor (CSTR) and up-flow anaerobic sludge bed (UASB) reactors. The hydrolysate originated as a side stream from a pilot plant pretreating wheat straw hydrothermally (195 °C for 10–12 min) for producing 2nd generation bioethanol [Kaparaju, P., Serrano, M., Thomsen, A.B., Kongjan, P., Angelidaki, I., 2009. Bioethanol, biohydrogen and biogas production from wheat straw in a biorefinery concept. Bioresource Technology 100 (9), 2562–2568]. Results from batch assays showed that hydrolysate had a methane potential of 384 ml/g-volatile solids (VS)_added. Process performance in CTSR and UASB reactors was investigated by varying hydrolysate concentration and/or organic loading rate (OLR). In CSTR, methane yields increased with increase in hydrolysate concentration and maximum yield of 297 ml/g-COD was obtained at an OLR of 1.9 g-COD/l d and 100% (v/v) hydrolysate. On the other hand, process performance and methane yields in UASB were affected by OLR and/or substrate concentration. Maximum methane yields of 267 ml/g-COD (COD removal of 72%) was obtained in UASB reactor when operated at an OLR of 2.8 g-COD/l d but with only 10% (v/v) hydrolysate. However, co-digestion of hydrolysate with pig manure (1:3 v/v ratio) improved the process performance and resulted in methane yield of 219 ml/g-COD (COD removal of 72%). Thus, anaerobic digestion of hydrolysate for biogas production was feasible in both CSTR and UASB reactor types. However, biogas process was affected by the reactor type and operating conditions.     

Optimization of biogas production from co-digestion of whey and cow manure

Anaerobic co-digestion is effective and environmentally attractive technology for energy recovery from organic waste. Organic, agricultural and industrial wastes are good substrates for anaerobic co-digestion because they contain high levels of easily biodegradable materials. In this paper enhancement of biogas production from codigestion of whey and cow manure was investigated in a series of batch experiments. The influence of whey ratio on specific biogas production in a mixture with cow manure was analyzed at 35 and 55°C, for different initial pH values and for different concentrations of supplemental bicarbonate in experiments carried out over 12 days. Good biogas production (6.6 dm³/dm³), methane content (79.4%) in a biogas mixture and removal efficiencies for total solids (16%) were achieved at optimum process conditions (temperature of 55°C, 10% v/v of whey and 5 g/dm³ NaHCO₃ in the initial mixture). In order to validate optimized conditions for co-digestion of whey and cow manure in the one-stage batch process, the experiments were performed within 45 days. The high biogas production (21.8 dm³/dm³), a good methane content (78.7%) in a biogas mixture as well as maximum removal efficiencies for total solids (32.3%), and chemical oxygen demand (56.3%), respectively indicate that whey could be efficiently degraded to biogas in a onestage batch process when co-digested with cow manure.     

Anaerobic co-digestion of swine manure with energy crop residues

Anaerobic co-digestion involves the treatment of different substrates with the aim of improving the production of biogas and the stability of the process. In this research, co-digestion of swine manure (SM) and energy crop residues (ECRs) was studied. The mixtures evaluated contained SM combined with maize (Mz), rapeseed (Rs) or sunflower (Sf) residues. Batch and semi-continuous experiments were performed to determine methane (CH₄) yields and the behavior of reactors while co-digesting agricultural wastes. Three different proportions of ECRs were tested in batch experiments for co-digestion with SM: 25, 50, and 75% volatile solids (VS). On the basis of the results obtained from batch tests, a mixture with a 50% ECR content was selected for the second stage of the study. Mesophilic reactors with a 3 L working volume were used for semi-continuous experiments. The hydraulic retention time (HRT) was set at 30 days and the reactors were kept under these operational conditions over four HRTs. The addition of ECR to the co-digestion system resulted in a major increase in the amount of biogas produced daily. The highest biogas yield was obtained when co-digesting Rs (3.5 L/day), although no improvement was observed in specific gas production from the addition of the co-substrate.     

Effect of ferrous chloride on biogas production and enzymatic activities during anaerobic fermentation of cow dung and <Emphasis Type="Italic">Phragmites</Emphasis> straw

The effect of ferrous (added as FeCl₂) on the anaerobic co-digestion of Phragmites straw and cow dung was studied by investigating the biogas properties, pH values, organic matter degradation (COD) and enzyme activities (cellulase, protease and dehydrogenase) at different stages of mesophilic fermentation. The results showed that Fe²⁺ addition increased the cumulative biogas yields by 18.1 % by extending the peak period with high daily biogas yields. Meanwhile, the methane (CH₄) contents in the Fe²⁺ added groups were generally higher than the control group before the 15th day. The pH values were not significantly impacted by Fe²⁺ concentrations during the fermentation process. The COD concentrations, cellulase, protease and dehydrogenase activities varied with the added Fe²⁺ concentrations and the stages of the fermentation process. At the beginning stage of fermentation (4th day), Fe²⁺ addition increased the biogas production by improving the cellulase and dehydrogenase activities which caused a decline in COD. At the peak stage of fermentation (8th day), Fe²⁺ addition enhanced the cellulase and protease activities, and resulted in lower COD contents than the control group. When the biogas yields decreased again (13th day), the COD contents varied similar with the protease and dehydrogenase activities, whilst cellulase activities were not sensitive to Fe²⁺ concentrations. At the end of fermentation (26th day), Fe²⁺ addition decreased the cellulase activities, led to lower COD contents and finally resulted the lower biogas yields than the control group. Taking the whole fermentation process into account, the promoting effect of Fe²⁺ addition on biogas yields was mainly attributed to the extension of the gas production peak stage and the improvement of cellulase activities.     

Pilot-scale semisolid fermentation of straw.

Semisolid fermentation of ryegrass straw to increase its animal feed value was successfully performed on a pilot scale. The pilot plant, which could handle 100 kg of straw per batch, was designed so that all major operations could take place in one vessel. The straw was hydrolyzed at 121 degrees C for 30 min with 0.5 N H2SO4 (7:3 liquid:solid), treated with ammonia to raise the pH to 5.0, inoculated with Candida utilis, and fermented in a semisolid state (70% moisture). During fermentation the straw was held stationary with air blown up through it. Batch fermentation times were 12 to 29 h. Semisolid fermentation did not require agitation and supported abundant growth at 20 to 40 degrees C even at near zero oxygen tensions. Fermentation increased the protein content, crude fat content, and in vitro rumen digestibility of the straw.     

Hazelnut husk as a substrate for the cultivation of shiitake mushroom (Lentinula edodes)

The possibility of using hazelnut husk (HH) as a new basal ingredient for substrate preparation in Lentinula edodes cultivation was investigated. Some chemical properties of the substrates prepared by HH alone and its mixtures with wheat straw (WS), beech wood-chip (BWC) and wheat bran (WB) in different ratios were compared, and their effects on spawn run time, days to first harvest (earliness), yield and biological efficiency (BE) were determined. The N content of the substrate prepared from HH alone was very high (0.82%), and thus the C:N ratio of substrates decreased with an increase in the rate of HH in the mixtures. Yield and BE in the HH alone substrate was considerably low compared with the controls (80BWC:10WS:10M and 60BWC:20WS:20WB), and decreased with an increase in the rate of HH in the mixtures. However, when the HH content in the mixtures was kept below 50%, the yield was relatively high (50HH:50WS and 50HH:50BWC). Even when the HH content increased to 75% in the mixture, the comparable yield and BE to the controls could be obtained by adding 10% of WB as nutrients (75HH:15WS:10WB and 75HH:15BWC:10WB). The results revealed that HH could be used as a new basal ingredient for substrate preparation in L. edodes cultivation.     

Pilot-scale semisolid fermentation of straw

Semisolid fermentation of ryegrass straw to increase its animal feed value was successfully performed on a pilot scale. The pilot plant, which could handle 100 kg of straw per batch, was designed so that all major operations could take place in one vessel. The straw was hydrolyzed at 121 degrees C for 30 min with 0.5 N H2SO4 (7:3 liquid:solid), treated with ammonia to raise the pH to 5.0, inoculated with Candida utilis, and fermented in a semisolid state (70% moisture). During fermentation the straw was held stationary with air blown up through it. Batch fermentation times were 12 to 29 h. Semisolid fermentation did not require agitation and supported abundant growth at 20 to 40 degrees C even at near zero oxygen tensions. Fermentation increased the protein content, crude fat content, and in vitro rumen digestibility of the straw.     

Effect of ruminally protected fat on in vitro fermentation and apparent nutrient digestibility in buffaloes (Bubalus bubalis)

Experiments were conducted to evaluate effects of supplementation of calcium salts of long chain fatty acids (Ca-LCFA) as a rumen inert fat (PF) on in vitro fermentation and apparent nutrient digestion in adult buffaloes fed wheat straw based diets. For the in vitro fermentation study, five total mixed rations (TMR) consisting of a concentrate mixture (CM), green Sorghum bicolor, WS and supplemented without (C) or with 30 g/kg dry matter (DM) rice bran fatty acid oil (RBO) (30 RBO) or 20 g/kg RBO + 10 g/kg PF (20 RBO/10 PF) or 10 g/kg RBO + 20 g/kg PF (10 RBO/20 PF) or 30 g/kg PF in the DM in the ratio of 340:50:580:30 were prepared. The in vitro DM degradability (IVDMD), TN, trichloro acetic acid precipitable N (TCA-N), non-protein N (NPN) and ammonia N (NH₃-N) were similar among groups. Within the fat supplemented groups, total volatile fatty acid (TVFA) concentration increased linearly (P=0.025) with PF supplementation. Apparent nutrient digestibility was determined on 20 adult buffaloes divided into five equal groups fed CM supplemented without (C) or with 300 g RBO (30 RBO) or 200 g RBO + 100 g PF (20 RBO/10 PF) or 100 g RBO + 200 g PF (10 RBO/20 PF) or 300 g PF (30 PF) along with limited green S. bicolor and WS maintaining forage: concentrate ratio of 650:350. Fat supplementation had no effect on the DM intake and apparent digestibilities of DM, organic matter (OM), crude protein (CP), total carbohydrate (TCHO) and neutral detergent fiber (aNDF). Within fat supplemented groups, inclusion of PF increased digestibilities of DM, OM, ether extract (EE), TCHO, aNDF and ADF. Supplemental fat also increased the digestible energy (DE) and metabolizable energy (ME) content of the diet, which also increased linearly with PF supplementation. All buffaloes were in positive N, Ca and P balances. We conclude that 200–300 g supplemental PF in the form of Ca-LCFA can be included in straw based diets fed to buffaloes to increase its energy density without adversely affecting DM intake and digestibility.     

Metabolomics reveals stage-specific metabolic pathways of microbial communities in two-stage anaerobic fermentation of corn-stalk

Analysis of intracellular metabolites is essential to delineate metabolic pathways of microbial communities for evaluation and optimization of anaerobic fermentation processes. The metabolomics are reported for a microbial community during two stages of anaerobic fermentation of corn stalk in a biogas digester using GC–MS. Acetonitrile/methanol/water (2:2:1, by vol) was the best extraction solvent for microbial community analysis because it yielded the largest number of peaks (>200), the highest mean summed value of identified metabolites (23) and the best reproducibility with a coefficient of variation of 30 % among four different extraction methods. Inter-stage comparison of metabolite profiles showed increased levels of sugars and sugar alcohols during methanogenesis and fatty acids during acidogenesis. Identification of stage-specific metabolic pathways using metabolomics can therefore assist in monitoring and optimization of the microbial community for increased biogas production during anaerobic fermentation.     

Sulphur fate and anaerobic biodegradation potential during co-digestion of seaweed biomass (Ulva sp.) with pig slurry

Seaweed (Ulva sp.) stranded on beaches were utilized as co-substrate for anaerobic digestion of pig slurry in three-month co-digestion tests in pilot scale anaerobic digesters in the laboratory. The methanogenic potential of Ulva sp. was low compared to that of other potential co-substrates available for use by farmers: 148 N m3CH4/t of volatile solids or 19 N m3CH4/t of crude product. When used as a co-substrate with pig manure (48%/52% w/w), Ulva sp. seaweed did not notably disrupt the process of digestion; however, after pilot stabilisation, biogas produced contained 3.5% H2S, making it unsuitable for energy recovery without treatment. Sequentially addition of the sulphate reduction inhibitor, potassium molybdate, to a final concentration of 3mM, temporarily reduced H2S emissions, but was unable to sustain this reduction over the three-month period. According to these pilot tests, the use of seaweed stranded on beaches as co-substrate in farm-based biogas plants shows some limitations.     

Copper stressed anaerobic fermentation: biogas properties,process stability,biodegradation and enzyme responses

The effect of copper (added as CuCl₂) on the anaerobic co-digestion of Phragmites straw and cow dung was studied in pilot experiments by investigating the biogas properties, process stability, substrate degradation and enzyme activities at different stages of mesophilic fermentation. The results showed that 30 and 100 mg/L Cu²⁺ addition increased the cumulative biogas yields by up to 43.62 and 20.77% respectively, and brought forward the daily biogas yield peak, while 500 mg/L Cu²⁺ addition inhibited biogas production. Meanwhile, the CH₄ content in the 30 and 100 mg/L Cu²⁺-added groups was higher than that in the control group. Higher pH values (close to pH 7) and lower oxidation–reduction potential (ORP) values in the Cu²⁺-added groups after the 8th day indicated better process stability compared to the control group. In the presence of Cu²⁺, the degradation of volatile fatty acids (VFAs) and other organic molecules (represented by chemical oxygen demand, COD) generated from hydrolysis was enhanced, and the ammonia nitrogen (NH₄ ⁺-N) concentrations were more stable than in the control group. The contents of lignin and hemicellulose in the substrate declined in the Cu²⁺-added groups while the cellulose contents did not. Neither the cellulase nor the coenzyme F₄₂₀ activities could determine the biogas producing efficiency. Taking the whole fermentation process into account, the promoting effect of Cu²⁺ addition on biogas yields was mainly attributable to better process stability, the enhanced degradation of lignin and hemicellulose, the transformation of intermediates into VFA, and the generation of CH₄ from VFA.     

Effect of C/N ratio and moisture content on the composting of poplar wood

Summary Milled poplar wood (1.7 mm mesh size) was composted in lab-scale reactors. Initial C/N ratios were adjusted to 10:1, 30:1, and 50:1 using urea as the nitrogen source. At each C/N ratio, three moisture levels (30, 50, and 70%) were tested. C/N ratios of 50:1 or 30:1 and moisture content of 70% favored more effective composting as indicated by higher levels of mineralization of the poplar wood to CO₂.     

Productivity of agricultural residues used for the cultivation of the medicinal fungus Lentinula edodes

Lentinula edodes mushrooms were produced by solid-state fermentation, using as substrates different mixtures of wheat straw (WS), corn-cobs (CC), and oak-wood sawdust (OS). Studies were conducted for evaluating their bio-transformation efficiency with respect to substrate colonization, time of sporophore production, biological efficiency, and mushroom nitrogen content, as well as basidiocarp number and size. First, the potential of residue-substrates to support early fructification was evaluated in glass-tubes. The mushrooms appeared 50–60 days after inoculation, with WS and CC promoting earlier sporophore initiation than did OS. A second experiment, where L. edodes mushrooms were cultivated in ‘bag-logs’, revealed high productivity on CC and high mushroom protein content on OS-based substrates. However, WS appeared to promote early fructification and mushroom quality. Finally, mushroom production characteristics in tubes and bags were correlated with nitrogen content and C/N ratio of substrates. Early fructification was positively related to nitrogen content. Substrate mixtures with lower C/N ratio favoured earlier sporophore induction.     

Comparison of characterization and microbial communities in rice straw- and wheat straw-based compost for <Emphasis Type="Italic">Agaricus bisporus</Emphasis> production

Rice straw (RS) is an important raw material for the preparation of Agaricus bisporus compost in China. In this study, the characterization of composting process from RS and wheat straw (WS) was compared for mushroom production. The results showed that the temperature in RS compost increased rapidly compared with WS compost, and the carbon (C)/nitrogen (N) ratio decreased quickly. The microbial changes during the Phase I and Phase II composting process were monitored using denaturing gradient gel electrophoresis (DGGE) and phospholipid fatty acid (PLFA) analysis. Bacteria were the dominant species during the process of composting and the bacterial community structure dramatically changed during heap composting according to the DGGE results. The bacterial community diversity of RS compost was abundant compared with WS compost at stages 4–5, but no distinct difference was observed after the controlled tunnel Phase II process. The total amount of PLFAs of RS compost, as an indicator of microbial biomass, was higher than that of WS. Clustering by DGGE and principal component analysis of the PLFA compositions revealed that there were differences in both the microbial population and community structure between RS- and WS-based composts. Our data indicated that composting of RS resulted in improved degradation and assimilation of breakdown products by A. bisporus, and suggested that the RS compost was effective for sustaining A. bisporus mushroom growth as well as conventional WS compost.