In developing countries, biogas energy production is seen as a technology that can provide clean energy in poor regions and reduce pollution caused by animal manure. Laboratories in these countries have little access to advanced gas measuring equipment, which may limit research aimed at improving local adapted biogas production. They may also be unable to produce valid estimates of an international standard that can be used for articles published in international peer-reviewed science journals. This study tested and validated methods for measuring total biogas and methane ($CH_4$) production using batch fermentation and for characterizing the biomass. The biochemical methane potential (BMP) ($CH_4$ NL $kg^{-1}$ VS) of pig manure, cow manure and cellulose determined with the Moller and VDI methods was not significantly different in this test (p>0.05). The biodegradability using a ratio of BMP and theoretical BMP (TBMP) was slightly higher using the Hansen method, but differences were not significant. Degradation rate assessed by methane formation rate showed wide variation within the batch method tested. The first-order kinetics constant k for the cumulative methane production curve was highest when two animal manures were fermented using the VDI 4630 method, indicating that this method was able to reach steady conditions in a shorter time, reducing fermentation duration. In precision tests, the repeatability of the relative standard deviation (RSDr) for all batch methods was very low (4.8 to 8.1%), while the reproducibility of the relative standard deviation (RSDR) varied widely, from 7.3 to 19.8%. In determination of biomethane concentration, the values obtained using the liquid replacement method (LRM) were comparable to those obtained using gas chromatography (GC). This indicates that the LRM method could be used to determine biomethane concentration in biogas in laboratories with limited access to GC.
This study focuses on the feasibility of bio-gas production using anaerobic digestion by measuring methane generation and biodegradability through the BMP test of industrial organic wastes. Organic wastes consist of entrails of pigs and organic residues of rumen generated from slaughter houses, wastewater sludge from slaughter waste water, fish offal and residues of vegetables from public wholesale markets, and wastewater sludge from the process of wastewater treatment in paper mill. The cumulative methane production by BMP test ranges from 149.3 ml/g-VS to 406.6 ml/g-VS and this is similar to methane generation of the normal wastewater sludge and food waste. As a result of measurement of biodegradability, wastewater sludge (S1 ~ S4) is low, ranging from 27.1% to 58.9 % and organic residues of rumen (G1) is low at 49.6 %. In conclusion, it turned out that raising the hydrolysis by various pre-treatments is necessary in order to produce bio-gas by using industrial organic wastes.
Kim, E.T.;Park, C.G.;Lim, D.H.;Kwon, E.G.;Ki, K.S.;Kim, S.B.;Moon, Y.H.;Shin, N.H.;Lee, S.S.
Asian-Australasian Journal of Animal Sciences
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v.27
no.12
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pp.1721-1725
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2014
The objective of this study was to evaluate the in vitro effects of coconut materials on ruminal methanogenesis and fermentation characteristics, in particular their effectiveness for mitigating ruminal methanogenesis. Fistulated Holstein cows were used as the donor of rumen fluid. Coconut materials were added to an in vitro fermentation incubated with rumen fluid-buffer mixture and timothy substrate for 24 h incubation. Total gas production, gas profiles, total volatile fatty acids (tVFAs) and the ruminal methanogens diversity were measured. Although gas profiles in added coconut oil and coconut powder were not significantly different, in vitro ruminal methane production was decreased with the level of reduction between 15% and 19% as compared to control, respectively. Coconut oil and coconut powder also inhibited gas production. The tVFAs concentration was increased by coconut materials, but was not affected significantly as compared to control. Acetate concentration was significantly lower (p<0.05), while propionate was significantly higher (p<0.05) by addition of the coconut materials than that of the control. The acetate:propionate ratio was significantly lowered with addition of coconut oil and coconut powder (p<0.05). The methanogens and ciliate-associated methanogens in all added coconut materials were shown to decrease as compared with control. This study showed that ciliate-associated methanogens diversity was reduced by more than 50% in both coconut oil and coconut powder treatments. In conclusion, these results indicate that coconut powder is a potential agent for decreasing in vitro ruminal methane production and as effective as coconut oil.
Chung, Chong Min;Hong, Seok Won;Park, Chul Hee;Kim, Young O;Lee, Sang Hyup
Journal of Korean Society of Water and Wastewater
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v.22
no.5
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pp.571-580
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2008
The influence of bacterial stress on anaerobic hydrogen-producing microorganisms was investigated in batch tests using serum bottles. Several physical and chemical stresses (i.e., heating, adding methane producing inhibitor and chemical acidification) were adapted as a pretreament of the seed sludge. In this experiment, the cultivation temperature were set at mesophilic ($35^{\circ}C$) and thermophilic conditions ($55^{\circ}C$) with adjusting pH at 5, 6, and 7 when using the mixture of food waste and activated sludge as a substrate. In conjunction with the pretreatment, hydrogen production was significantly enhanced as compared with that from untreated sludge. However, less biogas (hydrogen and methane) was produced without the pH control, resulted from the decrease of pH to below 4, mainly due to the formation of VFAs. Hydrogen and carbon dioxide gas were analyzed as main components of the biogas while methane not detected. With an application of chemical acidification, the highest hydrogen production value of 248 ml/l/day achieved at pH 7 and $35^{\circ}C$. In addition, more hydrogen gas produced when the ratio of butyric/acetic acid ratio increased. The optimum pH and temperature for hydrogen production were found to be 7 and $35^{\circ}C$, respectively.
Purpose: The kinetic evaluation was performed for swine manure (SM) degradation and biogas generation. Methods: The SM was anaerobically digested using batch digesters at feed to inoculum ratio (F/I) of 1.0 under mesophilic conditions ($36.5^{\circ}C$). The specific gas yield was expressed in terms of gram total chemical oxygen demand (mL/g TCOD added) and gram volatile solids added (mL/g VS added) and their effectiveness was discussed. The biogas and methane production were predicted using first order kinetic model and the modified Gompertz model. The critical hydraulic retention time for biomass washout was determined using Chen and Hashimoto model. Results: The biogas and methane yield from SM was 346 and 274 mL/ TCOD added, respectively after 100 days of digestion. The average methane content in the biogas produced from SM was 79% and $H_2S$ concentration was in the range of 3000-4108 ppm. It took around 32-47 days for 80-90% of biogas recovery and the TCOD removal from SM was calculated to be 85%. When the specific biogas and methane yield from SM (with very high TVFA concentration) was expressed in terms of oven dried volatile solids (VS) basis, the gas yield was found to be over estimated. The difference in the measured and predicted gas yield was in the range of 1.2-1.5% when using first order kinetic model and 0.1% when using modified Gompertz model. The effective time for biogas production ($T_{Ef}$) from SM was calculated to be in the range of 30-45 days and the critical hydraulic retention time ($HRT_{Critical}$) for biomass wash out was found to be 9.5 days. Conclusions: The modified Gompertz model could be better in predicting biogas and methane production from SM. The HRT greater than 10 days is recommended for continuous digesters using SM as feedstock.
Eight feeds (mixture of wheat straw and oil seed cakes in 3:1 ratio) were evaluated for methane emission and fermentation pattern with buffalo rumen liquor as inoculum in an in vitro gas production test. The cakes tested were groundnut cake (GNC), soybean cake (SBC), mustard seed cake (MSC), cotton seed cake (CSC), karanj seed cake expeller extracted (KCEE), karanj seed cake solvent extracted (KCSE), caster bean cake expeller extracted (CBCEE) and caster bean cake solvent extracted (CBCSE). The gas production (ml/g dry matter) was significantly higher with SBC and MSC followed by CSC, GNC, KCSE, KCEE, CBCSE and was the lowest with CBCEE. Methane emission was significantly lower with KCEE, KCSE, CBCEE, CBCSE (20.32- 22.43 ml/g DM) than that with SBC, GNC, CSC (27.34-31.14 ml/g DM). Mustard seed cake was in-between the two groups of oil cakes in methane production. In vitro true digestibility was highest with SBC followed by GNC, CSC, MSC, KCSE, KCEE, CBCSE and CECEE. Ammonia nitrogen level was positively correlated with the amount of protein present in the cake. Total holotrich protozoa were significantly higher with SBC, whereas, large spirotrich protozoa tended to be lower than with other cakes. The counts of small spirotrich and total protozoa were similar with all the cakes. Total volatile fatty acid production and acetate to propionate ratio were significantly higher with SBC and significantly lower with KCEE as compared to the other cakes. Among the conventional oil cakes tested in the present experiment (GNC, SBC, MSC and CSC), mustard seed cake-based feed produced the minimum methane without affecting other fermentation characteristics adversely.
Lee, S.Y.;Yang, S.H.;Lee, W.S.;Kim, H.S.;Shin, D.E.;Ha, Jong K.
Asian-Australasian Journal of Animal Sciences
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v.22
no.1
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pp.42-48
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2009
An in vitro incubation study was conducted to investigate effects of 2-bromoethanesulfonic acid (BES) on ruminal fermentation characteristics and methanogen population. BES at the final concentration of 0, 1 and 5 mM with two different substrates having a different ratio of timothy and concentrate (100% timothy vs. 40% timothy-60% concentrate) was incubated for 0, 24, 48 and 72 h in a $39^{\circ}C$ incubator. Total DNA extracted from culture fluid was used as a template for real-time PCR to measure the population of methanogens. Four different primer sets were used for amplification of total bacteria, total methanogens, the order Methanobacteriales and the order Methanomicrobiales. BES reduced (p<0.01) total gas and methane production in a dose-dependent manner. BES at 5 mM inhibited methane production by more than 95% compared to the control. An interaction between substrate and level of BES in total gas and methane was detected (p<0.01). The decrease of methane production with increasing BES level was more pronounced on mixed substrate than on timothy alone. However, hydrogen production was increased by BES treatment (p<0.01). Total VFA concentration was not affected, but molar percentage of propionate and butyrate was increased and acetate to propionate ratio was reduced by BES treatment (p<0.01). BES did not affect the population density of total bacteria but reduced (p<0.01) the population of total methanogens, the order Methanobacteriales and the order Methanomicrobiales in a dose-dependent manner. The type of substrate did not influence the trend, although the magnitude of response was different between all-roughage and 40% roughage substrate.
Objective: The present study explored the effects of grape seed procyanidin extract (GSPE) on rumen fermentation, methane production and archaeal communities in vitro. Methods: A completely randomized experiment was conducted with in vitro incubation in a control group (CON, no GSPE addition; n = 9) and the treatment group (GSPE, 1 mg/bottle GSPE, 2 g/kg dry matter; n = 9). The methane and volatile fatty acid concentrations were determined using gas chromatography. To explore methane inhibition after fermentation and the response of the ruminal microbiota to GSPE, archaeal 16S rRNA genes were sequenced by MiSeq high-throughput sequencing. Results: The results showed that supplementation with GSPE could significantly inhibit gas production and methane production. In addition, GSPE treatment significantly increased the proportion of propionate, while the acetate/propionate ratio was significantly decreased. At the genus level, the relative abundance of Methanomassiliicoccus was significantly increased, while the relative abundance of Methanobrevibacter decreased significantly in the GSPE group. Conclusion: In conclusion, GSPE is a plant extract that can reduce methane production by affecting the structures of archaeal communities, which was achieved by a substitution of Methanobrevibacter with Methanomassiliicoccus.
Journal of the Korea Organic Resources Recycling Association
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v.27
no.3
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pp.5-13
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2019
This study is conducted to investigate the effects of anaerobic treatments of swine manure slurry alone and combination of livestock manure slurry and fruit pomace on biogas production. Anaerobic co-digestion was evaluated in mesophilic tank reactors for 96 day-incubation period. The organic matter loading of anaerobic digestion was 1 kg of volatile solids(VS) per $1m^3{\cdot}day$. The highest methane production was achieved from the combination of swine manure slury and mandarin pomace(70:30) treatment, whereas the lowest daily and cumulative methane yields was observed in swine manure slurry alone treatment. More than two-fold increase in bio-gas and methane production was obtained by combination of livestock manure slurry and mandarin pomace treatment, compared to the swine manure slurry alone treatment. The co-digestion of livestock manure and fruits pomace has advantages to enhance the production of methane gas, compared to digestion of swine manure slurry alone.
An in vitro gas production technique was used in this study to elucidate the effect of two strains of active live yeast on methane ($CH_4$) production in the large intestinal content of pigs to provide an insight to whether active live yeast could suppress $CH_4$ production in the hindgut of pigs. Treatments used in this study include blank (no substrate and no live yeast cells), control (no live yeast cells) and yeast (YST) supplementation groups (supplemented with live yeast cells, YST1 or YST2). The yeast cultures contained $1.8{\times}10^{10}$ cells per g, which were added at the rates of 0.2 mg and 0.4 mg per ml of the fermented inoculum. Large intestinal contents were collected from 2 Duroc${\times}$Landrace${\times}$Yorkshire pigs, mixed with a phosphate buffer (1:2), and incubated anaerobically at $39^{\circ}C$ for 24 h using 500 mg substrate (dry matter (DM) basis). Total gas and $CH_4$ production decreased (p<0.05) with supplementation of yeast. The methane production reduction potential (MRP) was calculated by assuming net methane concentration for the control as 100%. The MRP of yeast 2 was more than 25%. Compared with the control group, in vitro DM digestibility (IVDMD) and total volatile fatty acids (VFA) concentration increased (p<0.05) in 0.4 mg/ml YST1 and 0.2 mg/ml YST2 supplementation groups. Proportion of propionate, butyrate and valerate increased (p<0.05), but that of acetate decreased (p<0.05), which led to a decreased (p<0.05) acetate: propionate (A: P) ratio in the both YST2 treatments and the 0.4 mg/ml YST 1 supplementation groups. Hydrogen recovery decreased (p<0.05) with yeast supplementation. Quantity of methanogenic archaea per milliliter of inoculum decreased (p<0.05) with yeast supplementation after 24 h of incubation. Our results suggest that live yeast cells suppressed in vitro $CH_4$ production when inoculated into the large intestinal contents of pigs and shifted the fermentation pattern to favor propionate production together with an increased population of acetogenic bacteria, both of which serve as a competitive pathway for the available H2 resulting in the reduction of methanogenic archaea.
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