Thermophilic methane production from cattle waste.

Methane production from waste of cattle fed a finishing diet was investigated, using four 3-liter-working volume anaerobic digestors at 60 degrees C. At 55 degrees C a start-up culture, in which waste was the only source of bacteria, was generated within 8 days and readily adapted to 60 degrees C, where efficiency of methanogenesis was greater. Increasing the temperature from 60 to 65 degrees C tended to drastically lower efficiency. When feed concentrations of volatile solids (VS, organic matter) were increased in steps of 2% after holding for 1 months at a given concentration, the maximum concentrations for efficient fermentation were 8.2, 10.0, 11.6, and 11.6% for the retention times (RT) of 3, 6, 9, and 12 days, respectively. The VS destructions for these and lower feed concentrations were 31 to 37, 36 to 40, 47 to 49 and 51 to 53% for the 3-, 6-, 9-, and 12-day RT digestors, respectively, and the corresponding methane production rates were about 0.16, 0.18, 0.20, and 0.22 liters/day per g of VS in the feed. Gas contained 52 to 57% methane. At the above RT and feed concentrations, alkalinity rose to 5,000 to 7,700 mg of CaCo3 per liter (pH to 7.5 to 7.8), NH3 plus NH4+ to 64 to 90 mM, and total volatile acids to 850 to 2,050 mg/liter as acetate. The 3-day RT digestor was quite stable up to 8.2% feed VS and at this feed concentration produced methane at the very high rate of 4.5 liters/day per liter of digestor. Increasing the percentage of feed VS beyond those values indicated above resulted in greatly decreased organic matter destruction and methane production, variable decrease in pH, and increased alkalinity, ammonia, and total volatile acid concentrations, with propionate being the first to accumulate in large amounts. In a second experiment with another lot of waste, the results were similar. These studies indicate that loading rates can be much higher than those previously thought useful for maximizing methanogenesis from cattle waste.     

Thermophilic methane production from cattle waste

Methane production from waste of cattle fed a finishing diet was investigated, using four 3-liter-working volume anaerobic digestors at 60 degrees C. At 55 degrees C a start-up culture, in which waste was the only source of bacteria, was generated within 8 days and readily adapted to 60 degrees C, where efficiency of methanogenesis was greater. Increasing the temperature from 60 to 65 degrees C tended to drastically lower efficiency. When feed concentrations of volatile solids (VS, organic matter) were increased in steps of 2% after holding for 1 months at a given concentration, the maximum concentrations for efficient fermentation were 8.2, 10.0, 11.6, and 11.6% for the retention times (RT) of 3, 6, 9, and 12 days, respectively. The VS destructions for these and lower feed concentrations were 31 to 37, 36 to 40, 47 to 49 and 51 to 53% for the 3-, 6-, 9-, and 12-day RT digestors, respectively, and the corresponding methane production rates were about 0.16, 0.18, 0.20, and 0.22 liters/day per g of VS in the feed. Gas contained 52 to 57% methane. At the above RT and feed concentrations, alkalinity rose to 5,000 to 7,700 mg of CaCo3 per liter (pH to 7.5 to 7.8), NH3 plus NH4+ to 64 to 90 mM, and total volatile acids to 850 to 2,050 mg/liter as acetate. The 3-day RT digestor was quite stable up to 8.2% feed VS and at this feed concentration produced methane at the very high rate of 4.5 liters/day per liter of digestor. Increasing the percentage of feed VS beyond those values indicated above resulted in greatly decreased organic matter destruction and methane production, variable decrease in pH, and increased alkalinity, ammonia, and total volatile acid concentrations, with propionate being the first to accumulate in large amounts. In a second experiment with another lot of waste, the results were similar. These studies indicate that loading rates can be much higher than those previously thought useful for maximizing methanogenesis from cattle waste.     

Detailed study of anaerobic digestion of Spirulina maxima algal biomass

Biomass of the blue-green alga Spirulina maxima was converted to methane using continuous stirred tank digesters with an energy conversion efficiency of 59%. Digesters were operated using once-a-day feeding with a retention time (theta) between 5 and 40 days, volatile solid concentrations (S(to)) between 20 and 100 kg VS/m(3), and temperatures between 15 and 52 degrees C. The results indicated a maximum methane yield of 0.35 m(3) (STP)/kg VS added at theta 30 days and S(to) 20 kg VS/m(3). Under such conditions, the energy conversion of the algal biomass to methane was 59%. The maximum methane production rate of 0.80 m(3) (STP)/m(3) day was obtained with theta= 20 days and S = 100 kg VS/m(3). The mesophilic condition at 35 degrees C produced the maximum methane yield and production rate. The process was stable and characterized by a high production of volatile acids (up to 23, 200 mg/L), alkalinity (up to 20, 000 mg/L), and ammonia (up to 7000 mg/L), and the high protein content of the biomass produced a well buffered environment which reduced inhibitory effects. At higher loading rates, the inhibition of methanogenic bacteria was observed, but there was no clear-cut evidence that such a phenomenon was due to nonionized volatile acids or gaseous ammonia. The kinetic analysis using the model proposed by Chen and Hashimoto indicated that the minimum retention time was seven days. The optimum retention time increased gradually from 11 to 16 days with an increase in the initial volatile solid concentration. The kinetic constant K decreased with the improvement in the digester performance and increased in parallel with the ammonia concentration in the culture media.     

Methane recovery from water hyacinth through whole-cell immobilization technology

The concepts of feed pretreatment, phase separation, and whole-cell immobilization technology have been incorporated in this investigation for the development of rational and cost-effective two- and three-stage methane recovery systems from water hyacinth (WH)Analyses of laboratory data reveal that a three-stage system could be designed with an alkali pretreatment stage [3.6% Na(2)CO(3) + 2.5% Ca(OH)(2) W/W, 24 h HRT] followed by an open acid reactor (2.1 days HRT) and closed immobilized methane reactor (12 h HRT), providing steady-state COD conversion of 62-65%, TVA conversion of 91-95%, and gas productivity of 4.08-5.36 L/L reactor volume/day with 82% methane. A gas yield of 50 L/kg WH/day (dry wt basis) at 35-37 degrees C is possible with this system. Insulation bricks, with particle size distribution of 500-3000 mum, were used as support material in the reactors at organic loading rate of 20 kg COD/m(3) day. The reactors matured in 15-18 weeksSubstantial reduction in retention time for the conversion of volatile acids in immobilized methane reactors prompted further research on the combined immobilized reactor to make possible an additional reduction in the cost of a WH-based biogas system. Evaluation of laboratory data reveals that a two-stage system could be designed with an open alkali pretreatment stage and a combined immobilized reactor (12 h HRT), providing steady-state COD conversion of 53% and gas productivity of 3.1 L/L reactor volume/day with 86% methane. A gas yield of 44 L/kg WH/day (dry wt basis) at 35-37 degrees C could be obtained from this system. Insulation bricks, with 500-1000 mum particle size distribution, was used as support material at an organic loading rate of 15 kg COD/m(3) day. Notwithstanding the fact that the technology in this study has been developed with water hyacinth as substrate, the implicit principles could be extended to any other organic substrate.     

Effect of Temperature and Retention Time on Methane Production from Beef Cattle Waste

The effect of temperature and retention time on the rate of methane production from waste of beef cattle fed a finishing diet was investigated by using continuously mixed 3-liter working volume anaerobic fermentors. The temperatures ranged from 30 to 65°C with 5°C increments between fermentors. The fermentors were fed once per day with 6% volatile solids (organic matter). Retention time for each temperature was varied from 18 to 2.5 days. After 3-volume turnovers, samples were obtained on 4 consecutive days. The highest methane production rate (liters/liter of fermentor per day) and methane yield at that rate (liters/gram of volatile solids) were 1.27 and 0.19 at 9 days and 30°C, 1.60 and 0.16 at 6 days and 35°C, 2.28 and 0.23 at 6 days and 40°C, 2.42 and 0.24 at 6 days and 45°C, 2.83 and 0.14 at 3 days and 50°C, 2.75 and 0.14 at 3 days and 55°C, 3.18 and 0.14 at 2.5 days and 60°C, and 1.69 and 0.17 at 6 days and 65°C. Volatile solids degradation at these retention times and temperatures was between 46 and 54%. The concentrations of volatile acids in the 30 to 55°C fermentors were generally below 2,000 mg/liter, with the exception of the 3-day retention time. The 60 and 65°C fermentors were usually above this level for all retention times. These studies indicate potential rates of methane production from the fermentation of untreated waste of beef cattle fed high-grain finishing diets. This information should serve as preliminary guidelines for various kinetic analyses and aid in economic evaluations of the potential feasibility of fermenting beef cattle waste to methane.     

Microstructure and molecular microbiology of rapidly formed hydrogen‐ and methane‐producing granules in a phase‐separated anaerobic environment

Hydrogen and methane were simultaneously produced in a two‐phase reactor, operated to separate the reactions of hydrogen and methanogen production. Each reactor was inoculated with a seed enriched with different microbial consortia. The first phase was operated with a hydraulic retention time of 7 days and at an organic loading rate of 7.7 g VS L^?1 d^?1 that produced a stable pH of 5.5. This suppressed the growth of methanogens and as a result, the off gas contained up to 27% hydrogen. The second phase was operated with a hydraulic retention time of 12 days and at an organic loading rate of 3.6 g VS L^?1 d^?1. This permitted the growth of hydrogenotrophs and methanogens to produce methane at a concentration of 60%. Examination of the microbial population of the two reactors both microscopically and using PCR, showed an effective separation of hydrogen‐ and methane‐producing microbial communities. The study revealed that the suppression of hydrogentrophs and methanogens can be achieved by adopting rapid method that leads the growth of hydrogen‐ and methane‐producing granules in phase‐separated anaerobic environment.     

Effect of substrate concentration on dry mesophilic anaerobic digestion of organic fraction of municipal solid waste (OFMSW)

The influence of total solid contents during anaerobic mesophilic treatment of the organic fraction of municipal solid waste (MSW) has been studied in this work. The work was performed in batch reactors of 1.7L capacity, during a period of 85-95 days. Two different organic substrate concentrations were studied: 931.1 mgDOC/L (20% TS) and 1423.4 mgDOC/L (30% TS). Experimental results showed that the reactor with 20% total solids content had significantly higher performance. Thus, the startup phase ended at 14 days and the total DOC removal was 67.53%. The startup in reactor R30 ended at 28 days obtaining 49.18% DOC removal. Also, the initial substrate concentration contributed substantially to the amount of methane in the biogas. Hence, the total methane production in the methanogenic phase was 7.01 L and 5.53 L at the end of the experiments for R20 and R30, respectively.     

Methane emission,digestive characteristics and faecal archaeol in heifers fed diets based on silage from brown midrib maize as compared to conventional maize

The aim of the present experiment was to compare silage prepared from maize having a brown midrib (BMR) mutation with control (CTR) maize to identify their effects on enteric methane emission, digesta mean retention time (MRT), ruminal fermentation and digestibility. In addition, the utility of archaeol present in faecal samples was validated as a proxy for methane production. Seven German Holstein heifers were fed total mixed rations with a maize-silage proportion (either BMR or CTR) of 920 g/kg dry matter (DM) in a change-over design. Heifers were fed boluses with markers to measure MRT; faeces were collected for 7 days and rumen fluid was collected on the penultimate day. Methane emission was measured in respiration chambers on one day. Data were analysed by t-test and regression analysis. DM intake did not differ between the two diets. The apparent digestibility of DM and most nutrients was unaffected by diet type, but apparent digestibility of neutral and acid detergent-fibre was higher in those heifers fed BMR than in those fed CTR. Comparisons between diets revealed no difference in particle or solute MRT in the gastro-intestinal tract and the reticulorumen. Concentrations of short-chain fatty acid and ammonia in rumen fluid and its pH were not affected by silage type. Independent of the mode of expression [l/d, l/kg DM intake, l/kg digested organic matter], methane emissions were not affected by maize-silage type, but with BMR, there was a trend towards lower methane production per unit of digested neutral detergent fibre than there was with CTR silage. Results of the present study show that feeding heifers BMR silage does not increase methane emissions despite a higher fibre digestibility as compared to CTR silage. Therefore, it is assumed that improvements in animal productivity achieved by feeding BMR silage, as some studies have reported, can be obtained without extra environmental cost per unit of milk or meat. Neither faecal archaeol content [µg/g] nor daily amount excreted [mg/d] is suitable to predict methane production in absolute terms [l per day]. However, faecal archaeol content has a certain potential for predicting the methane yield [l per kg DM intake] of individual animals.     

Effect of mixing duration and vacuum on methane production rate from beef cattle waste

The effects of mixing duration and vacuum on methane production rates from anaerobically fermented beef cattle wastes were discussed. The results showed that continuously mixed fermentors produced significantly (P < 0.05) higher methane production rates than fermentors mixed two hours per day. However, the rates from the continuously mixed fermentors were only 8-11% higher than the intermittently mixed fermentors at six and four days hydraulic retention time (HRT), respectively. There was no significant difference between the vacuum and conventional fermentors at six days HRT, but there was a significant difference at four days HRT. The CH(4) production rate of the vacuum fermentors was 5% higher than the conventional fermentors at four days HRT. The results of these experiments compared well with predicted CH(4) production rates. These results suggest that there is little potential for increasing the fermentation rates of livestock wastes by increased mixing or vacuum.     

Methane Production by Fermentor Cultures Acclimated to Waste from Cattle Fed Monensin, Lasalocid, Salinomycin, or Avoparcin

The ability of microorganisms to ferment waste from cattle fed monensin, lasalocid, or salinomycin to methane was determined. Continuously mixed anaerobic fermentors with 3-liter working volumes at 55°C were used; fermentors were fed once per day. Initially, all fermentors were fed waste without antibiotics at 6% volatile solids (VSs, organic matter) and a 20-day retention time (RT) for 60 days. Waste from animals fed monensin, lasalocid, or salinomycin at 29, 20, and 16.5 mg per kg of feed, respectively, was added to duplicate fermentors at the above VSs, and RT. Avoparcin (5 to 45 mg/liter) was not fed to animals but was added directly to duplicate fermentors. Lasalocid and salinomycin had minimal effects on the rate of methane production at RTs of 20 days and later at 6.5 days. Avoparcin caused an increase in organic acids from 599 to 1,672 mg/liter (as acetate) after 4 weeks, but by 6 weeks, acid concentrations declined and the rate of methane production was similar to controls at a 6.5-day RT. The monensin fermentors stopped producing methane 3 weeks after antibiotic addition. However, after a 6-month acclimation period, the microorganisms apparently adapted, and methane production rates of 1.65 and 2.51 liters per liter of fermentor volume per day were obtained with 6% VSs, and RTs of 10 and 6.5 days, respectively. This compares with 1.78 and 2.62 liters/liter per day for controls (P > 0.05). All fermentors that were fed waste containing antibiotics had lower pH values and ammonia and alkalinity concentrations, suggesting less buffering capacity and protein catabolism than in controls. Acclimation results obtained with fermentors at 35°C were similar to those for fermentors at 55°C. These studies indicate that waste from cattle fed these selected growth-promoting antibiotics can be thermophilically fermented to methane at RTs of 6.5 days or longer and VS concentrations of 6%, at rates comparable to waste without antibiotics.     

Methane fermentation of coastal mud sediment by a two-stage upflow anaerobic sludge blanket (UASB) reactor system

The removal of organic matter from a coastal mud sediment was carried out by a methane fermentation process under anaerobic conditions. In a batch acidogenic fermentation, the addition of vitamins containing thiamine, nicotinic acid and biotin dramatically enhanced acetate production from the mud sediment (200 g wet wt l(-1) artificial sea water), yielding 77 mM acetate after 6 days, which corresponded to 77% of the organic matter in the mud sediment, measured on the basis of chemical oxygen demand. Thereafter, the two-fold diluted, post-acidogenic fermentation liquor (PAF liquor) was continuously treated at 2.4x original dilution rate day(-1) for 30 days, using an upflow anaerobic sludge blanket methanogenic reactor containing the acclimated methanogenic sludge from the mud sediment. Acetate, 42 mM in the PAF liquor, was converted to methane at a maximum methane production rate of 96 mmol l(-1) day(-1); and 87.5% of the acetate and 88.7% of the total organic carbon in the PAF liquor were removed. Moreover, an efficient treatment of the mud sediment was carried out by a semi-continuous, two-stage reactor system, where the culture broth was circulated between acidogenic and methanogenic reactors. This two-stage reactor system gave a stable operation at 4-day intervals for one treatment period, yielding 112 mmol methane from the wet mud in the PAF liquor (278 g l(-1)).     

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     

Dissolved methane oxidation and competition for oxygen in down-flow hanging sponge reactor for post-treatment of anaerobic wastewater treatment

Post-treatment of anaerobic wastewater was undertaken to biologically oxidize dissolved methane, with the aim of preventing methane emission. The performance of dissolved methane oxidation and competition for oxygen among methane, ammonium, organic matter, and sulfide oxidizing bacteria were investigated using a lab-scale closed-type down-flow hanging sponge (DHS) reactor. Under the oxygen abundant condition of a hydraulic retention time of 2h and volumetric air supply rate of 12.95m(3)-airm(-3)day(-1), greater than 90% oxidation of dissolved methane, ammonium, sulfide, and organic matter was achieved. With reduction in the air supply rate, ammonium oxidation first ceased, after which methane oxidation deteriorated. Sulfide oxidation was disrupted in the final step, indicating that COD and sulfide oxidation occurred prior to methane oxidation. A microbial community analysis revealed that peculiar methanotrophic communities dominating the Methylocaldum species were formed in the DHS reactor operation.     

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.     

Biomass retention and performance of anaerobic fixed-film reactors treating acetic acid wastewater

An acetic-acid-based synthetic wastewater of different organic concentrations was successfully treated at 35 degrees C in anaerobic downflow fixed-film reactors operated at high organic loading rates and short hydraulic retention times (HRTs). Substrate removal and methane production rates close to theoretical values of complete volumetric chemical oxygen demand (COD) removal and maximum methane conversion were obtained. A high concentration of biofilm biomass was retained in the reactor. Steady-state biofilm concentration increased with increased organic loading rate and decreased HRTs, reaching a maximum of 8.3 kg VFS/m(3) at a loading rate of 17 kg COD/m(3) day. Biofilm substrate utilization rates of up to 1.6 kg COD/kg VFS day were achieved. Soluble COD utilization rates at various COD concentrations can be described by half-order reaction kinetics.     

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.     

Dry-thermophilic anaerobic digestion of simulated organic fraction of Municipal Solid Waste: Process modeling

Solid retention time (SRT) is a very important operational variable in continuous and semicontinuous waste treatment processes since the organic matter removal efficiency - expressed in terms of percentage of Dissolved Organic Carbon (% DOC) or Volatile Solids (% VS) removed - and the biogas or methane production are closely related with the SRT imposed. Optimum SRT is depending on the waste characteristics and the microorganisms involved in the process and, hence, it should be determined specifically in each case.In this work a series of experiments were carried out to determine the effect of SRT, from 40 to 8 days, on the performance of the dry (30% Total Solids) thermophilic (55 °C) anaerobic digestion of organic fraction of Municipal Solid Wastes (OFMSW) operating at semicontinuous regime of feeding.The experimental results show than 15 days is the optimum SRT (the best between all proved) for this process. Besides, data of organic matter concentration and methane production versus SRT have been used to obtain the kinetic parameters of the kinetic model of Romero García (1991): the maximum specific growth rate of the microorganisms (μ_max = 0.580 days⁻¹) and the fraction of substrate non-biodegradable (α = 0.268).     

Ammonia–methane two-stage anaerobic digestion of dehydrated waste-activated sludge

The study investigated methane production from dehydrated waste-activated sludge (DWAS) with approximately 80% water content under thermophilic conditions. The repeated batch-wise treatment of DWAS using methanogenic sludge unacclimated to high concentrations of ammonia, increased the ammonia production up to 7,600 mg N per kilogram total wet sludge of total ammonia concentration, and stopped the methane production. Investigation revealed that the loading ratio of DWAS for methanogenic sludge influences anaerobic digestion. Methane production significantly decreased and ammonia concentration increased with the increase in loading ratio of DWAS. Since the semicontinuous culture revealed that approximately 50% of organic nitrogen in DWAS converted to ammonia at sludge retention time (SRT) after 4 days at 37 degrees C and 1.33 days at 55 degrees C, the previous stripping of the ammonia produced from DWAS was carried out. The stripping of ammonia increased methane production significantly. This ammonia-methane two-stage anaerobic digestion demonstrated a successful methane production at SRT 20 days in the semicontinuous operation using a laboratory-scale reactor system.     

Granule development and performance in sucrose fed anaerobic baffled reactors

Two 90 L anaerobic baffled reactors were used to study the granulation of sludge and the effect of the organic loading rate and NaHCO3/COD ratios on reactor performance. Furthermore, it was determined whether an anaerobic baffled reactor would promote phase separation and if additive of bentonite or granular active carbon was capable of enhancing granule formation. In order to minimize feed variations, and have a totally biodegradable substrate, a synthetic sucrose substrate was used. Granulation was achieved in both reactors within 75 days. However, the granules from the granular active carbon amended reactor appeared earlier and were larger and more compact. The reactors were maintained at a hydraulic retention time of 20 h during performance study stage. The results showed that when organic loading rate were changed from 2.15 to 6.29 kg COD m(-3)day(-1), chemical oxygen demand (COD) removal was not decreased (91-93%), but a slight increase in effluent COD was observed. It was found that the COD removals were generally good (87-92%) and had not obviously change with the decreasing NaHCO3/COD ratios. From the bacterial distribution and the concentration of volatile fatty acids in four compartments, it was concluded that a separation of phases occurred within the anaerobic baffled reactors.     

Bioconversion of industrial hemp to ethanol and methane: the benefits of steam pretreatment and co-production

Several scenarios for ethanol production, methane production (by anaerobic digestion) and co-production of these, using autumn harvested hemp as substrate, were investigated and compared in terms of gross energy output. Steam pretreatment improved the methane production rate compared with mechanical grinding. The methane yield of steam pretreated stems was similar both with and without pre-hydrolysis with cellulolytic enzymes. Co-production of ethanol and methane from steam pretreated stems gave a high yield of transportation fuel, 11.1-11.7 MJ/kg processed stem dry matter (DM); more than twice that of ethanol production alone from hexoses, 4.4-5.1 MJ/kg processed stem DM. Co-production from the whole hemp plant would give 2600-3000 L ethanol and 2800-2900 m(3) methane, in total 171-180 GJ per 10,000 m(2) of agricultural land, based on a biomass yield of 16 Mg DM. Of this, the yeast and enzymes from ethanol production were estimated to contribute 700 m(3) (27 GJ) of methane.