High-Rate two-phase process for the anaerobic degradation of cellulose, employing rumen microorganisms for an efficient acidogenesis

A novel two-stage anaerobic process for the microbial conversion of cellulose into biogas has been developed. In the first phase, a mixed population of rumen bacteria and ciliates was used in the hydrolysis and fermentation of cellulose. The volatile fatty acids (VFA) produced in this acidogenic reactor were subsequently converted into biogas in a UASB-type methanogenic reactor.A stepwise increase of the loading rate from 11.9 to 25.8 g volatile solids/L reactor volume/day (g VS/L/day) did not affect the degradation efficiency in the acidogenic reactor, whereas the methanogenic reactor appeared to be overloaded at the highest loading rate. Cellulose digestion was almost complete at all loading rates applied. The two-stage anaerobic process was also tested with a closed fluid circuit. In this instance total methane production was 0.438 L CH(4)g VS added, which is equivalent to 98% of the theoretical value. The application of rumen microorganisms in combination with a high-rate methane reactor is proposed as a means of efficient anaerobic degradation of cellulosic residues to methane. Because this newly developed two-phase system is based on processes and microorganisms from the ruminant, it will be referred to as "Rumen Derived Anaerobic Digestion" (RUDAD-) process.     

Production of biogas from water hyacinth (Eichhornia crassipes) (Mart) (Solms) in a two-stage bioreactor

A two-stage rumen-derived anaerobic digestion process was tested for the conversion of water hyacinth shoots and a mixture of the shoots with cowdung (7:3) into biogas. Under conditions similar to those of the rumen and loading rates (LR) in the range of 11.6–19.3g volatile solids (VS) l–1d–1 in the rumen reactor, the degradation efficiencies were 38% for the shoots and 43% for the mixture. The major fermentation products were volatile fatty acids (VFA) with a maximum yield of 7.92mmolg–1 VS digested, and biogas with a yield of 0.2lg–1 VS digested. The effect of varying LR, solid retention time (SRT) and dilution rates on the extent of degradation of the water hyacinth–cowdung mixture was examined. Overall conversion of the substrate was highest at the loading rate of 15.4gVS.l–1d–1. Varying the retention times between 60 and 120h had no effect on the degradation efficiency, but a decrease was observed at retention times below 60h. The overall performance of the reactor was depressed by changing the dilution rate from 0.5 to 0.34h–1. By applying a LR of 15.4VS. l–1d–1, a SRT of 90h and a dilution rate of 0.5h–1 in the rumen reactor, and connecting it to a methanogenic reactor of the upflow anaerobic sludge blanket type, 100% conversion efficiency of the VFA into biogas with a methane content of 80% was achieved. The average methane gas yield was 0.44lg–1 VS digested.     

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

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

Microbial Community Dynamics of a Continuous Mesophilic Anaerobic Biogas Digester Fed with Sugar Beet Silage

The aim of the study was to investigate the long‐term fermentation of an extremely sour substrate without any addition of manure. In the future, the limitation of manure and therefore the anaerobic digestion of silage with a very low buffering capacity will be an increasing general bottleneck for energy production from renewable biomass. During the mesophilic anaerobic digestion of sugar beet silage (without top and leaves) as the sole substrate (without any addition of manure), which had an extreme low pH of around 3.3, the highest specific gas production rate (spec. GPR) of 0.72 L/g volatile solids (VS) d was achieved at a hydraulic retention time (HRT) of 25 days compared to an organic loading rate (OLR) of 3.97 g VS/L d at a pH of around 6.80. The methane (CH₄) content of the digester ranged between 58 and 67 %, with an average of 63 %. The use of a new charge of substrate (a new harvest of the same substrate) with higher phosphate content improved the performance of the biogas digester significantly. The change of the substrate charge also seemed to affect the methanogenic population dynamics positively, thus improving the reactor performance. Using a new substrate charge, a further decrease in the HRT from 25 to 15 days did not influence the digester performance and did not seem to affect the structure of the methanogenic population significantly. However, a decrease in the HRT affected the size of the methanogenic population adversely. The lower spec. GPR of 0.54 L/g VS d attained on day 15 of the HRT could be attributed to a lower size of methanogenic population present in the anaerobic digester during this stage of the process. Furthermore, since sugar beet silage is a relatively poor substrate, in terms of the buffering capacity and the availability of nutrients, an external supply of buffering agents and nutrients is a prerequisite for a safe and stable digester operation.     

Digestion of sand-laden manure slurry in an upflow anaerobic solids removal (UASR) digester

Studies on the performance of a laboratory scale upflow anaerobic solids removal (UASR) digester were carried out using sand-laden cow manure slurries having total solids (TS) concentration as 50 and 100 g/l. Hydraulic retention time (HRT) was maintained as 32.4 days, which resulted in the volatile solids (VS) loading rates of 1 and 1.64 g/l d. The UASR system was designed to remove sand from the manure slurry, while anaerobically digesting biodegradable solids inside a single reactor. To enhance the contact of microorganisms and substrate, the liquor from the top of the digester was recirculated through the bed of settled solids at its bottom. Volatile solids reduction through this process was observed to be 62% and 68% in the case of feed slurries having TS concentration as 50 and 100 g/l (referred in the text as 5% and 10% feed slurries), respectively. The methane production rates were observed to be 0.22 and 0.38 l/l d, while methane yield was 0.21 and 0.27 l CH₄/g VS loaded, for 5% and 10% feed slurries, respectively. This indicates that the increase in the VS loading had a positive impact on methane production rate and methane yield. It would be of interest to study the performance of a UASR digester at higher solids loadings and with longer solids retention times. Nonetheless, the presented study showed that sand-laden manure slurries can be successfully digested in a UASR digester producing methane energy equivalent to 4 kW h per m³ of digester volume per day.     

Characteristics of carbohydrate degradation and the rate-limiting step in anaerobic digestion

The characteristics of the degradation of cellulose, soluble starch, and glucose in the acidogenic phase and the effects of the substrate loading rate and biological solids retention time on the methanogenic phase of anaerobic digestion were investigated. The results obtained from continuous experiments using laboratory-scale anaerobic chemostat reactors elucidated the true rate-limiting step of anaerobic digestion. The specific rate of substrate utilization decreased in the following order: glucose, soluble starch, acetic acid, and cellulose. The rate of the hydrolysis of cellulose was so low that this was shown to be the rate-limiting step in overall anaerobic digestion. Among methanogenic bacteria Methanosarcina would provide a higher substrate utilization rate than Methanothrix, and the maximum allowable substrate loading rate in the methanogenic phase was 11.2 g acetic acid/L day.     

Methane Yields and Digestion Dynamics of Press Fluids from Mechanically Dehydrated Maize Silages Using Different Types of Digesters

The mechanical dehydration of ensiled agricultural crops results in two major products: a fibrous press cake and a press fluid containing mainly easily digestible constituents. This study is aimed at the investigation on methane yields and digestion dynamics of the press fluids from maize silages using different types of digesters. Methane yields investigated in batch experiments account for 390?C506?l_N CH₄/kg?volatile solids (VS) with a degree of degradation of the organic matter in the fluid of more than 90%. The investigation of digestion dynamics in a continuously working stirrer tank digester at different levels of retention time and volume load suggests that a stable fermentation of press fluids can only be achieved with retention times of more than 20?days and with volume loads below 2?g VS/l/day. In a continuously working fixed bed digester a steady fermentation could be achieved at a retention time of 8?days and a volume load of 3?g VS/l/day.     

Characterisation of solid and liquid fractions of dairy manure with regard to their component distribution and methane production

Dairy manure with a total solids content of 77.2g TS/l was separated by means of screening and coagulation-flocculation treatments, using CaO as coagulant and a cationic polyacrylamide as flocculant, obtaining liquid and solid fractions. The solid fraction separated contained 33.4% of the initial total mass of dairy manure plus chemical solutions, containing also 75.2% of the TS, 80.4% of the VS, 58.5% of the total Kjeldahl nitrogen (TKN) and 87.4% of the total phosphorus (P(T)) present in the initial dairy manure. 83.7% of the liquid fraction chemical oxygen demand (COD) was anaerobically biodegradable (COD(BD)). Methane production for the separated liquid fraction was 0.604l CH4 NCTP/g VS added, being 0.307 and 0.371l CH4 NCTP/g VS added for dairy manure and screened dairy manure, respectively. The characteristics of this liquid fraction would allow its treatment in high loading anaerobic reactors having shorter hydraulic retention times, smaller reactor size and a higher methane volumetric production rate than conventional anaerobic reactors treating either manure or screened manure.     

Anaerobic digestion of processed municipal solid waste using a novel high solids reactor: Maximum solids levels and mixing requirements

Summary Novel, laboratory-scale, high solids reactors operated under mesophilic conditions were used to study the anaerobic fermentation of processed municipal solid waste (MSW) to methane. The anaerobic digestion consortium was introduced to high solids levels through gradual adaptation. The maximum sludge solids level for stable anaerobic fermentation performance was identified as approximately 36% wt/wt. Recovery of the anaerobic consortium, following dilution of inhibitory high solids levels, was swift. Reactor mixing requirements were also studied. No significant difference in fermentation performance was observed between agitator speeds of 1 and 25 rpm. Preliminary fermentation performance tests showed that solids loading rates as high as 9.5 g VS (volatile solids) feed/L sludge^.d, at 32% solids within the reactor, were possible. Under these conditions, operation was stable with an average pH of 7.8–8.0, total volatile fatty acid pools of <20 mM, and a biogas composition of 55%–60% methane.     

Integrating sugarcane molasses into sequential cellulosic biofuel production based on SSF process of high solid loading

Background

Sugarcane bagasse (SCB) is one of the most promising lignocellulosic biomasses for use in the production of biofuels. However, bioethanol production from pure SCB fermentation is still limited by its high process cost and low fermentation efficiency. Sugarcane molasses, as a carbohydrate-rich biomass, can provide fermentable sugars for ethanol production. Herein, to reduce high processing costs, molasses was integrated into lignocellulosic ethanol production in batch modes to improve the fermentation system and to boost the final ethanol concentration and yield.

Results

The co-fermentation of pretreated SCB and molasses at ratios of 3:1 (mixture A) and 1:1 (mixture B) were conducted at solid loadings of 12% to 32%, and the fermentation of pretreated SCB alone at the same solid loading was also compared. At a solid loading of 32%, the ethanol concentrations of 64.10 g/L, 74.69 g/L, and 75.64 g/L were obtained from pure SCB, mixture A, and mixture B, respectively. To further boost the ethanol concentration, the fermentation of mixture B (1:1), with higher solid loading from 36 to 48%, was also implemented. The highest ethanol concentration of 94.20 g/L was generated at a high solid loading of 44%, with an ethanol yield of 72.37%. In addition, after evaporation, the wastewater could be converted to biogas by anaerobic digestion. The final methane production of 312.14 mL/g volatile solids (VS) was obtained, and the final chemical oxygen demand removal and VS degradation efficiency was 85.9% and 95.9%, respectively.

Conclusions

Molasses could provide a good environment for the growth of yeast and inoculum. Integrating sugarcane molasses into sequential cellulosic biofuel production could improve the utilization of biomass resources.

     

Anaerobic Digestion of Dairy Manure in a Hybrid Reactor with Biogas Recirculation

Summary A novel anaerobic hybrid reactor (AHR) configuration incorporating floating support media for biomass immobilization and biogas recirculation for enhanced mixing was used for anaerobic digestion of dairy manure. No pretreatment or solid liquid separation was applied. The reactor was operated at high influent volatile solids (VS) and organic loading rates (OLR) of up to 9.87% and 7.30 g VS/l day, respectively. After 149 days of continuous operation the results revealed that a high amount (38.1 g VSS) of biomass was able to attach itself to the support medium being used. The investigated AHR configuration achieved COD, BOD, TS, and VS removal efficiencies of 48–63, 64–78, 55–65, and 59–68%, respectively, at a hydraulic retention time (HRT) of 15 days. The corresponding average methane production value obtained in this study was 0.191 l/g VS added.     

Comparative evaluation of biogas production from dairy manure and co‐digestion with maize silage by CSTR and new anaerobic hybrid reactor

This study aimed to investigate potential methane production through anaerobic digestion of dairy manure and co‐digestion with maize silage. Two different anaerobic reactor configurations (single‐stage continuously stirred tank reactor [CSTR] and hybrid anaerobic digester) were used and biogas production performances for each reactor were compared. The HR was planned to enable phase separation in order to improve process stability and biogas production under higher total solids loadings (≥4%). The systems were tested under six different organic loading rates increased steadily from 1.1 to 5.4 g VS/L.d. The CSTR exhibited lower system stability and biomass conversion efficiency than the HR. The specific biogas production of the hybrid system was between 440 and 320 mL/gVS with 81–65% volatile solids (VS) destruction. The hybrid system provided 116% increase in specific biogas production and VS destruction improved by more than 14%. When MS was co‐digested together with dairy manure, specific biogas production rates increased about 1.2‐fold. Co‐digestion was more beneficial than mono‐material digestion. The hybrid system allowed for generating methane enriched biogas (>75% methane) by enabling phase separation in the reactor. It was observed that acidogenic conditions prevailed in the first two compartments and the following two segments as methanogenic conditions were observed. The pH of the acidogenic part ranged between 4.7 and 5.5 and the methanogenic part was between 6.8 and 7.2.     

Effects of thermo-chemical pre-treatment of grass silage on methane production by anaerobic digestion

Dried grass silage (GS) was pre-treated at different NaOH loading rates (1%, 2.5%, 5% and 7.5% by volatile solids (VS) mass in grass silage) and temperatures (20 °C, 60 °C, 100 °C and 150 °C) to determine effects on its bio-degradability in terms of the hydrolysis yield and degradation of ligno-cellulosic materials for biogas production. At 100 °C and the four NaOH loadings, up to 45% of the total COD was solubilised and up to 65.6%, 36.1% and 21.2% of lignin, hemicellulose and cellulose were removed, respectively; biological methane production potentials obtained were 359.5, 401.8, 449.5 and 452.5 ml CH?/g VS added, respectively, being improved by 10-38.9% in comparison with untreated GS. VS removals following anaerobic digestion were 67.6%, 76.9%, 85.3%, 95.2% and 96.7% for untreated GS and GS treated at the four NaOH loadings, respectively. 100 °C and the NaOH loading rate of 5% is recommended as a proper GS pre-treatment condition.     

Generation of soluble lignin-derived compounds during degradation of barley straw in an artificial rumen reactor

Summary The supernatants of effluents from an artificial rumen reactor degrading barley straw have been shown to contain lignin-derived compounds by UV spectral characteristics and pyrolysis mass spectrometry (PYMS). Most of these compounds were shown to be released by the action of rumen microorganisms. The compounds were quantified by measuring absorbance at 280 nm using bamboo-milled wood lignin as a standard. The concentration of the compounds rose from 0.5 mg·ml^–1 at solid and liquid retention times (SRT and HRT) of 60 and 12 h, respectively, and a loading rate (LR) of 25 g total solids (TS)·l^–1 per day to 3.5 mg·ml^–1 at a SRT of 144 h, an HRT of 20 days and an LR of 15 g TS·1^–1 per day. The highest concentration was below the level known to be toxic to rumen microorganisms in vitro. No indications were found for anaerobic lignin degradation in the rumen reactor. Offprint requests to: H. J. M. Op den Camp     

Anaerobic digestion of alfalfa silage with recirculation of process liquid

Process liquid recirculation initially stimulated one-phase anaerobic digestion of alfalfa silage in two semi-continuously fed and stirred tank reactors. Thus, with increased pH, alkalinity and stability it was possible to increase the organic loading rate to 3 g VS L(-1) d(-1), as compared to 2.25 g VS L(-1) d(-1) in a control reactor without recirculation. However, the recirculation of liquid eventually caused an accumulation of organic and inorganic substances, leading to an inhibition of hydrolysis and methanogenesis. This inhibition of microbial activity was prevented in one of the processes by replacing 50% of the recirculated process liquid with water during the second half of the operation period. A multiple linear regression model of principal components using seven input variables explained the variance in output variables nearly as well as the original model using all 23 measured input variables. The results show that it is necessary to adjust the degree of liquid recirculation to reach an optimal process.     

Conversion of cereal residues into biogas in a rumen-derived process

A recently developed high-rate, two-phase process, which employs rumen microorganisms for efficient acidogenesis, was tested for anaerobic degradation of barley straw, rye straw, and maize stover. Under conditions similar to those of the rumen and loading rates varying between 9.8 and 26.0 g of organic matter/I/day in the first phase (acidogenic reactor), total fibre degradation efficiencies ranged between 42% and 57%, irrespective of the loading rate applied. Average specific production of volatile fatty acids and biogas/g volatile solid digested in the acidogenic reactor varied between 6.9 and 11.2 mmol and 0.10 and 0.25 l, respectively.The effect of varying solid retention times on the extent of degradation of barley straw was examined. Changing of retention times in the range of 60 to 156 h had no effect on degradation efficiency, but a decrease in efficiency was observed at retention times below 60 h.By connecting the acidogenic reactor in series to an Upflow Anaerobic Sludge Blanket (UASB) methanogenic reactor the volatile fatty acids were converted into biogas. Average methane contents of the gases produced in the acidogenic reactor and in the UASB reactor were 30±3% and 78±3%, respectively.     

Polyhydroxyalkanoate production using waste vegetable oil and filtered digestate liquor of chicken manure

Abstract

The production of polyhydroxyalkanoates (PHA) using digestate of chicken manure combined with waste sunflower oil as no-cost feedstocks in a multi-stage process was investigated. Using Cupriavidus necator H16 in combined culture media, a maximum PHA accumulation of 4.6?±?0.2?g/L at 75.1?±?1.4% of cell dry matter and a residual cell matter yield of 1.5?±?0.1?g/L were obtained after 96?hr of cultivation (30?°C, 160?rpm, pH 7.0) in flask-based experiments. Manure was acidogenically fermented in a continuous stirring tank reactor in fed-batch mode. The bioreactor was operated at varying organic loading rates (OLR) and hydraulic retention times (HRT) ranging from 1–4?g volatile solids (VS)/L/d and 4–8?days, respectively. Optimal operation was observed at an OLR of 4?g VS/L/d and an HRT of 4?days. Analysis showed the presence of significant amounts of ammonia, potassium, magnesium, calcium, and trace elements, i.e. Fe, Cu, Ni, Mn, Co, Zn, Cr in the digestate. The micro-filtered digestate was utilized as a complex culture media base while waste oil served as an additional carbon source and supplemented for effective PHA production. The total volatile fatty acid content of digestate greatly affected the growth performance of the PHA-producing microorganism Cupriavidus necator H16.     

Characteristics and biogas production potential of municipal solid wastes pretreated with a rotary drum reactor

This study was conducted to determine the characteristics and biogas production potential of organic materials separated from municipal solid wastes using a rotary drum reactor (RDR) process. Four different types of wastes were first pretreated with a commercial RDR system at different retention times (1, 2 and 3 d) and the organic fractions were tested with batch anaerobic digesters with 2.6 g VS L(-1) initial loading. The four types of waste were: municipal solid waste (MSW), a mixture of MSW and paper waste, a mixture of MSW and biosolids, and a mixture of paper and biosolids. After 20 d of thermophilic digestion (50+/-1 degrees C), it was found that the biogas yields of the above materials were in the range of 457-557 mL g VS(-1) and the biogas contained 57.3-60.6% methane. The total solid and volatile solid reductions ranged from 50.2% to 65.0% and 51.8% to 66.8%, respectively. For each material, the change of retention time in the RDR from 1 to 3d did not show significant (alpha=0.05) influence on the biogas yields of the recovered organic materials. Further studies are needed to determine the minimum retention time requirements in the RDR system to achieve effective separation of organic from inorganic materials and produce suitable feedstock for anaerobic digesters.     

The effect of organic loading rate and retention time on hydrogen production from a methanogenic CSTR

The possibility of shifting a methanogenic process for hydrogen production by changing the process parameters viz., organic loading rate (OLR) and hydraulic retention time (HRT) was evaluated. At first, two parallel semi-continuously fed continuously stirred tank reactors (CSTR) were operated as methanogenic reactors (M1 and M2) for 78 days. Results showed that a methane yield of 198-218 L/kg volatile solids fed (VS(fed)) was obtained when fed with grass silage at an OLR of 2 kgVS/m3/d and HRT of 30 days. After 78 days of operation, hydrogen production was induced in M2 by increasing the OLR from 2 to 10 kgVS/m3/d and shortening the HRT from 30 to 6 days. The highest H? yield of 42 L/kgVS(fed) was obtained with a maximum H? content of 24%. The present results thus demonstrate that methanogenic process can be shifted towards hydrogen production by increasing the OLR and decreasing HRT.     

Anaerobic organic acid production of food waste in once-a-day feeding and drawing-off bioreactor

Acidogenesis of food waste was studied in a 2-L reactor with semi-continuous mode operation (once-a-day feeding and draw-off) for maximum 65 days to examine optimal volatile acid compositions for biological nitrogen removal (BNR) and enhanced biological phosphorus removal (ENPR). Various operational parameters of hydraulic retention time (HRT), organic loading rate (ORL), pH and temperature were investigated for soluble chemical oxygen demand (SCOD), volatile fatty acid composition, nitrogen and phosphate. The yields (gTVFA/g VS) and the volumetric productivity (gTVFA/d L) increased with HRT from 0.26-0.32, 1.25-1.50 (at 4 days) to 0.36-0.39, 1.71-1.83 (at 12 days). However, the acetate fraction (%) decreased with HRT from 35.7-37.5 at 4 days to 23.5-25 at 12 days. The yields decreased with increase of organic loading from 0.34-0.37 at 5 g/L d to 0.29-0.30 at 13 g/L d and the productivity increased from 1.63-1.65 to 3.61-3.75. The yield and productivity were highest at 35 degrees C among 25, 35 and 45 degrees C. The yield and productivity at pH 5.5 and 6.0 were best and very similar to each other. The condition of 35 degrees C, pH 6.0, HRT 8 days, ORL 9 g/L d resulted in TVFA, SCOD, acetate and butyrate of 25, 39.5, 12 and 5.25 g/L, respectively.