Effect of Dietary Monensin or Chlortetracycline on Methane Production from Cattle Waste

Wastes from feedlot cattle fed finishing diets containing either monensin, chlortetracycline, or no antibiotic were investigated as substrates for methane production. We used continuously mixed anaerobic fermentors with 3-liter working volumes at 35 and 55°C; these fermentors were fed once per day. Within a few days after waste from animals fed monensin was added, the volume of methane produced began to decrease in the 55°C fermentors. After 9 days of daily feeding, methane production was severely inhibited, the pH dropped from 7.6 to 5.9, and the concentration of volatile acids increased from 543 to 6,300 mg/liter (as acetate). Although additions of waste from cattle fed monensin were discontinued after 9 days, the fermentors did not resume gas production within 8 weeks. The addition of waste from cattle which had been fed chlortetracycline reduced the methane production rate approximately 20%; however, pH and volatile acid values were comparable to control fermentor values after 40 days. Similar effects were observed with the 35°C fermentors. In a batch fermentation experiment in which 50-g portions of volatile solids from waste of animals fed monensin, chlortetracycline, or no antibiotics were added to fermentors, monensin delayed the onset of methane production for about 40 days, but then these fermentors began to produce methane at a rate comparable to the control rate. The ultimate methane yields from the three types of waste after 180 days were not significantly different. These studies indicate that monensin has a detrimental effect on the conversion of feedlot wastes to methane, unless microorganisms can be adapted to the levels that are present in these wastes.     

Development of a two-phase combination fermenter for the conversion of cellulose to methane

Cellulose degradation to methane under continuous fermentation conditions was compared using fully mixed, fully mixed with solids return, sludge-blanket, and fixed-film fermenters. In fully mixed fermenters, a decrease in hydraulic retention time (HRT) of two weeks or less caused the wash out of anaerobes capable of converting volatile fatty acids to methane, while at increased feeding rates over 1 g/L day the rapid growth of cellulolytic anaerobes upset the balance between acid formation and its conversion to CH(4). Circulation of cellulose and difficulty in settling of cellulose with attached bacteria imposed problems in the use of other types of fermenters. On the basis of information obtained from this study, a fermenter which combined a fully mixed phase for cellulose degradation and a fixed-film phase with pre-immobilized bacteria for converting fatty acids to CH(4) in one vessel, was devised. Using this fermenter, a mixed culture converted cellulose to CH(4) at 4.8 g/L day at a HRT of six days as compared to 0.7 g/L day at a HRT of 28 days in the fully mixed fermenter.     

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 from cattle waste: Effects of temperature, hydraulic retention time, and influent substrate concentration on kinetic parameter (k)

The effects of temperature (35 and 55 degrees C), influent volatile solids (VS) concentration (S(0) = 43, 64, 82, 100, and 128 kg VS/m(3)) and hydraulic retention time (HRT = 4, 5, 8, 10, 15, and 25 days) on methane (CH(4)) production from cattle waste were evaluated using 3-dm(3) laboratoryscale fermentors. The highest CH(4) production rate achieved was 6.11 m(3) CH(4) m(-3) fermentor day(-1) at 55 degrees C, four days HRT, and S(0) = 100 kg VS/m(3). Batch fermentations showed an ultimate CH(4) yield (B(0)) of 0.42 m(3) CH(4)/kg VS fed. The maximum loading rates for unstressed fermentation were 7 kg VS m(-3) day(-1) at 35 degrees C and 20 kg VS m(-3) day(-1) at 55 degrees C. The kinetic parameter (K, an increasing K indicates inhibition of fermentation) increased exponentially as S(0) increased, and was described by: K = 0.8 + 0.0016 e(0.06S(0) ). Temperature had no significant effect on K for S(0) between 40 and 100 kg VS/m(3). The above equation predicted published K values for cattle waste within a mean standard error of 7%.     

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.     

Novel membrane bioreactor: Able to cope with fluctuating loads, poorly water soluble VOCs, and biomass accumulation

We determined the effect of different mixing intensities on the performance, methanogenic population dynamics, and juxtaposition of syntrophic microbes in anaerobic digesters treating cow manure from a dairy farm. Computer automated radioactive particle tracking in conjunction with computational fluid dynamics was performed to quantify the shear levels locally. Four continuously stirred anaerobic digesters were operated at different mixing intensities of 1,500, 500, 250, and 50 revolutions per min (RPM) over a 260-day period at a temperature of 34 +/- 1 degrees C. Animal manure at a volatile solids (VS) concentration of 50 g/L was fed into the digesters daily at five different organic loading rates between 0.6 and 3.5 g VS/L day. The different mixing intensities had no effect on the biogas production rates and yields at steady-state conditions. A methane yield of 0.241 +/- 0.007 L CH(4)/g VS fed was obtained by pooling the data of all four digesters during steady-state periods. However, digester performance was affected negatively by mixing intensity during startup of the digesters, with lower biogas production rates and higher volatile fatty acids concentrations observed for the 1,500-RPM digester. Despite similar methane production yields and rates, the acetoclastic methanogenic populations were different for the high- and low-intensity mixed digesters with Methanosarcina spp. and Methanosaeta concilii as the predominant methanogens, respectively. For all four digesters, epifluorescence microscopy revealed decreasing microbial floc sizes beginning at week 4 and continuing through week 26 after which no microbial flocs remained. This decrease in size, and subsequent loss of microbial flocs did not, however, produce any long-term upsets in digester performance.     

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.     

Influence of temperature on the performance of anaerobic fixed-film reactors utilizing screened dairy manure

Thermophilic (55°C) and mesophilic (35°C and 22°C) anaerobic digestions in laboratory scale (4 litre) fixed-film reactors fed with screened dairy manure were successfully operated over a range of hydraulic retention times, from 1 to 20 days. Maximum methane production rates of 1·82, 1·68 and 1·28 litres CH₄ litre⁻¹ day⁻¹ occurred at 1, 1·5 and 1 days HRT for the respective 55°C, 35°C and 22°C reactors. Both thermophilic and mesophilic digestions achieved maximum biodegradation efficiency at 10 days HRT. The thermophilic fixed-film reactor performed better than completely-mixed reactors in terms of methane production at HRTs shorter than 2 days. From the results, mesophilic fixed-film reactor operated at 35°C provided optimum methane production and net energy output between 1 and 5 days HRT.     

Hydrogen and methane production through two-stage mesophilic anaerobic digestion of olive pulp

The present study focused on the anaerobic biohydrogen production from olive pulp (two phase olive mill wastes, TPOMW) and the subsequent anaerobic treatment of the effluent for methane production under mesophilic conditions in a two-stage process. Biohydrogen production from water-diluted (1:4) olive pulp was investigated at hydraulic retention times (HRT) of 30 h, 14.5 h and 7.5 h while methane production from the effluent of hydrogenogenic reactor was studied at 20 d, 15 d, 10 d and 5 d HRT. In comparison with previous studies, it has been shown that the thermophilic hydrogen production process was more efficient than the mesophilic one in both hydrogen production rate and yield. The methanogenic reactor was successfully operated at 20, 15 and 10 days HRT while it failed when an HRT of 5 days was applied. Methane productivity reached the maximum value of 1.13 ± 0.08 L/L/d at 10 days HRT whereas the methane yield increased with the HRT. The Anaerobic Digestion Model no. 1 (ADM1) was applied to the obtained experimental data from the methanogenic reactor to simulate the digester response at all HRT tested. The ability of the model to predict the experimental results was evident even in the case of the process failure, thus implying that the ADM1 could be a valuable tool for process design even in the case of a complex feedstock. In general, the two-stage anaerobic digestion proved to be a stable, reliable and effective process for energy recovery and stabilization treatment of olive pulp.     

Continuous methane fermentation and the production of vitamin B12 in a fixed-bed reactor packed with loofah

A fixed-bed reactor with acclimated methanogens immobilized on a loofah support was studied on a laboratory scale to evaluate the system producing methane from the mixture of CO(2) and H(2) gas, with the production of vitamin B(12) as a by-product. Fermentation using CO(2)/H(2) acclimated methanogens was conducted in a jar fermentor with hydraulic retention times (HRTs) of three and six days. The performance of the reactor was mainly dependent on the HRT. With an HRT of three days, the methane production rate and the vitamin B(12) concentration in the culture broth were 6.18 l/l-reactor/h and 2.88 mg/l-culture liquid; these values were 11.96 l/l-reactor/h and 37.54 mg/l-culture liquid for an HRT of six days. A higher total cell mass of methanogens retained 42.5 g dry cell/l-culture liquid was achieved in the HRT of six days. The loofah carrier immobilized almost 95% of the methanogens, which led to a more effective bio-reaction. It was also observed that the fermentation system had a better ability to buffer pH, especially for an HRT of six days.     

Emission of methane and carbon dioxide and earthworm survival during composting of pharmaceutical sludge and spent mycelia

Emissions of methane (CH4) and carbon dioxide (CO2) from spent mycelia of the mold Penicilium notatum and sludge from the effluent treatment facility (ETPS) of a pharmaceutical industry were estimated twice during a two-week composting before vermicomposting. These wastes are dumped in landfills or sometimes used in agricultural fields and no reports are available on their greenhouse gas producing potentials. The solid wastes contained appreciable organic carbon and nitrogen while very high Fe, Mn and Zn were found in ETPS only. Pure wastes did not support germination of Vigna radiata L. while mixing soil with ETPS and spent mycelia at the ratios of 12:1 and 14:1 led to 80% and 50% germination, respectively. The wastes were mixed with cowdung at the ratios of 1:1, 1:3 and 3:1 for composting. Carbon dioxide emissions were always significantly higher than CH4 emissions from all the treatments due to prevalence of aerobic condition during composting. From some treatments, CH4 emissions increased with time, indicating increasing activity of anaerobic bacteria in the waste mixtures. Methane emissions ranged from 21.6 to 231.7 microg m(-2) day(-1) while CO2 emissions were greater than thousand times at 39.8-894.8 mg m(-2) day(-1). The amount of C emitted as CH4-C and CO2-C from ranged from 0.007% to 0.081% of total C composted. Cowdung emitted highest CH4 followed by spent mycelia and ETPS while ETPS emitted more CO2 than spent mycelia but lesser than cowdung. Global warming potential of emitted CH4 was found to be in the range of 10.6-27.7 mg-CO2-equivalent on a 20-year time horizon. The results suggest that pharmaceutical wastes can be an important source of CH4 and CO2 during composting or any other stockpiling under suitable moisture conditions. The waste mixtures were found not suitable for vermicomposting after two weeks composting and earthworms did not survive long in the mixtures.     

Anaerobic digestion of animal waste: effect of mixing

Six laboratory scale biogas mixed anaerobic digesters were operated to study the effect of biogas recycling rates and draft tube height on their performance. The digesters produced methane at 0.40-0.45 L per liter of digester volume per day. A higher methane production rate was observed in unmixed digesters, while increased biogas circulation rate reduced methane production. However, different draft tube heights caused no difference in the methane production rate. Air infiltration (up to 15% oxygen in the biogas) was observed in the digesters mixed by biogas recirculation. Slight air permeability of tubing or leakage on the vacuum side of the air pump may have caused the observed air infiltration. The similar performance of the mixed and unmixed digesters might be the result of the low solids concentration (50 g dry solids per liter of slurry) in the fed animal slurry, which could be sufficiently mixed by the naturally produced biogas.     

Biofuels generation from sweet sorghum: fermentative hydrogen production and anaerobic digestion of the remaining biomass

The present study focuses on the exploitation of sweet sorghum biomass as a source for hydrogen and methane. Fermentative hydrogen production from the sugars of sweet sorghum extract was investigated at different hydraulic retention times (HRT). The subsequent methane production from the effluent of the hydrogenogenic process and the methane potential of the remaining solids after the extraction process were assessed as well. The highest hydrogen production rate (2550 ml H(2)/d) was obtained at the HRT of 6h while the highest yield of hydrogen produced per kg of sorghum biomass was achieved at the HRT of 12h (10.4l H(2)/kg sweet sorghum). It has been proved that the effluent from the hydrogenogenic reactor is an ideal substrate for methane production with approximately 29l CH(4)/kg of sweet sorghum. Anaerobic digestion of the solid residues after the extraction process yielded 78l CH(4)/kg of sweet sorghum. This work demonstrated that biohydrogen production can be very efficiently coupled with a subsequent step of methane production and that sweet sorghum could be an ideal substrate for a combined gaseous biofuels production.     

Conversion of straw-manure mixtures to methane at mesophilic and thermophilic temperatures

The effects of temperature (35 and 55 degrees C) and straw-manure mixtures (90, 75, 50, and 0% straw) on methane (CH(4)) production were studied using laboratory-scale fermentors. Batch fermentations showed that the ultimate CH(4) yield (B(0)) of straw-manure mixtures was directly proportional to the relative proportion and B(0) of the individual components. Also, hammer or ball milling did not increase the B(0) of straw. Kinetic analysis showed that fermentation stress occurred when the straw content of straw-manure mixtures was higher than 40% at 35 degrees C, or higher than 75% at 55 degrees C. This fermentation stress was observed to occur when the free ammonia concentration was below 10 g/m(3).     

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.     

Pile mixing increases greenhouse gas emissions during composting of dairy manure

The effect of pile mixing on greenhouse gas (GHG) emissions during dairy manure composting was determined using large flux chambers designed to completely cover replicate pilot-scale compost piles. GHG emissions from compost piles that were mixed four times during the 80 day trial were approximately 20% higher than emissions from unmixed (static) piles. For both treatments, carbon dioxide (CO(2)), methane (CH(4)), and nitrous oxide (N(2)O) accounted for 75-80%, 18-21%, and 2-4% of GHG emissions, respectively. Seventy percent of CO(2) emissions and 95% of CH(4) emissions from all piles occurred within first 23 days. By contrast, 80-95% of N(2)O emissions occurred after this period. Mixed and static piles released 2 and 1.6 kg GHG (CO(2)-Eq.) for each kg of degraded volatile solids (VS), respectively. Our results suggest that to minimize GHG emissions, farmers should store manure in undisturbed piles or delay the first mixing of compost piles for approximately 4 weeks.     

Two-stage biogas production by co-digesting molasses wastewater and sewage sludge

We evaluated the feasibility of co-digesting molasses wastewater and sewage sludge in a two-stage hydrogen- and methane-producing system. The highest energy was recovered at the 21-h hydraulic retention time (HRT) of the first hydrogenic reactor and at 56-h HRT of the secondary methanogenic reactor. Hence, the two-stage system recovered 1,822 kJ from 1 L of the mixed wastes (19.7: hydrogenic reactor plus, 1,802 kJ L^?1: methanogenic reactor). Despite the overloaded VFA-run with a short HRT of 56 h, the GAC-CH₄ reactor increased methane production rate and yields due to enhanced pH buffer capacity. An RNA-based community analysis showed that the Ethanoligenens and Methanosaeta dominated the hydrogen and methane bioreactor, respectively. The two-stage system of co-digesting molasses and sewage sludge is particularly cost-effective due to non-pretreatment of sewage sludge.     

Methane consumption in two temperate forest soils

Forest soils are thought to be an important sink for atmospheric methane. To evaluate methane consumption,¹⁴C-labeled methane was added to the headspace of intact soil cores collected from a mixed mesophytic forest and from a red spruce forest located in the central Appalachian Mountains. Both soils consumed the added methane at initially high rates that decreased as the methane mixing ratio of the air decreased. The mixed mesophytic forest soil consumed an average of 2 mg CH₄ m^–2 d^–1 versus 1 mg CH, m^–2 d^–1 for the spruce forest soil. The addition of acetylene to the headspace completely suppressed methane consumption by the soils, suggesting that an aerobic methane-consuming microorganism mediated the process. At both forest sites, methane mixing ratios in soil air spaces were greater than that in the air overlying the soil surface, indicating that these soils had the ability to produce methane. Models of methane emission from forest soils to the atmosphere must represent methane flux as the balance between production and consumption of methane, which are controlled by very different factors     

The potential of biological methane generation from chicken manure

The potential for biological methane generating from the manure of laying hens was investigated in the laboratory. Fresh manure was collected, analyzed, and used to prepare medium for bacterial growth. At 55°C and under anaerobic conditions, methanogenic cultures were initiated by incubating the medium with different inoculations from various natural environments. Since there were no significant differences in gas production among these initiated cultures after 40 days of acclimation, they were mixed to maintain a genetic pool. The mixed culture was then challenged with different retention times (RT) and different volatile solid (VS) concentrations for the selection of optimal conditions and cultures. The conditions were finally selected to be 4-day RT and 6% VS for the maximal rate of gas production. The optimal pH and temperature were determined to be 7.5 and 50°C, respectively. Under such conditions the selected culture produced total gas at a rate 4.5 L/L day and methane (70% of total gas) 3.2 L/L day. The chicken manure therefore was able to support the methane yield at 270 L/kg of VS, a value comparably higher than other kinds of livestock wastes.     

Single-stage, batch, leach-bed, thermophilic anaerobic digestion of spent sugar beet pulp

Spent sugar beet pulp as received was digested in a single-stage, batch, unmixed, leach-bed, laboratory scale thermophilic anaerobic digester. Biogasification of each 0.450 kg (wet weight) batch of spent pulp was initiated by inoculating with anaerobically digested liquor from previous run. The average methane yield was 0.336 m3 CH4 at STP (kgVS)(-1), the maximum methane production rate was 0.087 m3 CH4 at STP (kgVS)(-1)d(-1), average lag time to initiate methanogenesis was only 0.44 days and time required to achieve 95% methane yield was 8 days. The pH in the digesters ranged between 8.0 and 9.5. High rates of methane generation were sustained even at high pH values. The equivalent organic loading rate in the batch digesters was 4 kgCODm(-3)d(-1). The digestion process used here offers significant improvements over one-stage and two-stage systems reported in the literature with comparable performance as it is a single-stage system where the feedstock does not require size reduction, and mixing is not required in the digester.