• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.86-91
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• 2012

An in vitro rumen batch culture study was completed to compare effects of common grasses, leguminous shrubs and non-leguminous shrubs used for livestock grazing in Australia and Ghana on $CH_4$ production and fermentation characteristics. Grass species included Andropodon gayanus, Brachiaria ruziziensis and Pennisetum purpureum. Leguminous shrub species included Cajanus cajan, Cratylia argentea, Gliricidia sepium, Leucaena leucocephala and Stylosanthes guianensis and non-leguminous shrub species included Annona senegalensis, Moringa oleifera, Securinega virosa and Vitellaria paradoxa. Leaves were harvested, dried at $55^{\circ}C$ and ground through a 1 mm screen. Serum bottles containing 500 mg of forage, modified McDougall's buffer and rumen fluid were incubated under anaerobic conditions at $39^{\circ}C$ for 24 h. Samples of each forage type were removed after 0, 2, 6, 12 and 24 h of incubation for determination of cumulative gas production. Methane production, ammonia concentration and proportions of VFA were measured at 24 h. Concentration of aNDF (g/kg DM) ranged from 671 to 713 (grasses), 377 to 590 (leguminous shrubs) and 288 to 517 (non-leguminous shrubs). After 24 h of in vitro incubation, cumulative gas, $CH_4$ production, ammonia concentration, proportion of propionate in VFA and IVDMD differed (p<0.05) within each forage type. B. ruziziensis and G. sepium produced the highest cumulative gas, IVDMD, total VFA, proportion of propionate in VFA and the lowest A:P ratios within their forage types. Consequently, these two species produced moderate $CH_4$ emissions without compromising digestion. Grazing of these two species may be a strategy to reduce $CH_4$ emissions however further assessment in in vivo trials and at different stages of maturity is recommended.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.06a
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• pp.394-398
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• 2005

This study aimed to develop a novel anaerobic two-stage process converting food waste to $H_2$ and $CH_2$. The anaerobic two-stage process was devised by combining hydrogen fermentation with methane fermentation. At the high loading rate of 12.3 kg $Vs/m^3/d$, it could remove 72.5%of VS and convert $VS_{removed}$ to $H_2$ (28.2%) and $CH_4$ (69.9%) on COD basis in 8 days.

• Journal of Animal Environmental Science
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• v.17 no.3
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• pp.171-180
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• 2011

The researchers have tried to reduce ruminal methane gas ($CH_4$) and to convert it into beneficial nutrient for several decades. This study was conducted to screen the methane-reducing vegetables among lettuce, hot pepper, spring onion, onion, turmeric, sesame leaf, garlic, radish sprout, leek and ginger nutritiously on the in vitro ruminal fermentation. The heat-treated vegetables at the 10% of substrate (timothy) were used to reduce methane production on the in vitro anaerobic experiment of 0, 6, 12, 24 and 48 h incubation time. Total gas production, pH, ammonia, $H_2$, $CO_2$, $CH_4$, and volatile fatty acid (VFA) were measured as indicators of in vitro fermentation product containing methane gas. All treatments except garlic showed a tendency to increase in total gas production. The result of ammonia showed that garlic and hot pepper affected rumen bacteria concerned protein metabolism and that lettuce and spring onion increased ammonia production. Garlic decreased $CH_4$ production in inverse proportion to $H_2$. Lettuce, spring onion, onion, garlic, radish sprout, leek and ginger increased propionate of VFA. Garlic balanced the ruminal fermentation in the pH, $H_2$, $CH_4$, acetate and propionate. This results showed that methane production at in vitro study was inhibited by heat-treated garlic supplementation. In conclusion, this study suggests that ruminal fermentation covering methane production might be controled by proper vegetables.

• Korean Journal of Soil Science and Fertilizer
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• v.25 no.1
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• pp.62-69
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• 1992

A series of laboratory experiment was conducted to find out the effects of physico-chemical, microbiological and commercial inhibitors on the odorous gas evolution in the fermentation of livestock feces. The results obtained were summarized as follows. 1. The rate and amount of evolution of gas were the highest at 7 days after incubation, thereafter gradually decreased until 24 days after incubation. 2. The rate and amount of gases were evolved in order of $CO_2>N_2O>CH_4>NH_3>N_2S$, respectively. 3. The highest amount of methane gas was evolved from the poultry feces, those of carbon dioxide and nitrous oxide were evolved from the pig feces, and that of hydrogen sulfide was dominantly evolved from the cattle feces. 4. Negative correlation were obtained between the total amounts of $NH_3$ and $CH_4$, $CO_2$ and $CH_4$, $N_2O$ and $CH_4$, $N_2O$ and $CH_4$, while positive correlations were obtained between the amounts of $CO_2$ and $N_2O$, $CO_2$ and $NH_3$, and $NH_3$ and $N_2O$, respectively. 5. There was no significantly inhibiting effect obtained that the application of commercial gas inhibitor as VK 88. On the other hand there was significantly inhibiting effect obtained when application of fertile paddy soil and photosynthetic bacteria to the fermentation of livestock feces.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.6
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• pp.700-707
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• 2010

Effects of protease-resistant antimicrobial substances (PRA) produced by Lactobacillus plantarum and Leuconostoc citreum on rumen methanogenesis were examined using the in vitro continuous methane quantification system. Four different strains of lactic acid bacteria, i) Lactococcus lactis ATCC19435 (Control, non-antibacterial substances), ii) Lactococcus lactis NCIMB702054 (Nisin-Z), iii) Lactobacillus plantarum TUA1490L (PRA-1), and iv) Leuconostoc citreum JCM9698 (PRA-2) were individually cultured in GYEKP medium. An 80 ml aliquot of each supernatant was inoculated into phosphate-buffered rumen fluid. PRA-1 remarkably decreased cumulative methane production, though propionate, butyrate and ammonia N decreased. For PRA-2, there were no effects on $CH_4$ and $CO_2$ production and fermentation characteristics in mixed rumen cultures. The results suggested that PRA-1 reduced the number of methanogens or inhibited utilization of hydrogen in rumen fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.8
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• pp.1242-1251
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• 2020

Objective: The 3×3 factorial arrangement was used to investigate if either high water-soluble carbohydrates (WSC) cultivars or suitable time of day that the grass cut could improve nutrient values and in vitro fermentation characteristics. Methods: The 3 cultivars were mowed at 3 diurnal time points and included a benchmark WSC ryegrass cultivar 'Premium', and 2 high WSC cultivars AberAvon and AberMagic, which contained, on average, 157, 173, and 193 g/kg dry matter (DM) of WSC, and 36.0, 36.5, and 34.1 g/kg DM of N during 7th regrowth stage, respectively. The fermentation jars were run at 39℃ with gas production recorded and sampled at 2, 5, 8, 11, 14, 17, 22, 28, 36, and 48 h. The rumen liquid was collected from 3 rumen fistulated cows grazing on ryegrass pasture. Results: High WSC cultivars had significantly greater WSC content, in vitro DM digestibility (IVDMD) and total gas production (TGP), and lower lag time than Premium cultivar. Methane production for AberMagic cultivar containing lower N concentration was marginally lower than that for AberAvon and Premium cultivars. Grass cut at Noon or PM contained greater WSC concentration, IVDMD and TGP, and lower N and neutral detergent fiber (NDF) contents, but CH₄ production was also increased, compared to grass cut in AM. Meanwhile, the effects of diurnal cutting time were influenced by cultivars, such as in vitro CH₄ production for AberMagic was not affected by cutting time. The IVDMD and gas production per unit of DM incubated were positively related to WSC concentration, WSC/N and WSC/NDF, respectively, and negatively related to N and NDF concentrations. Conclusion: These results imply either grass cut in Noon or PM or high WSC cultivars could improve nutrient values, IVDMD and in vitro TGP, and that AberMagic cultivar has a slightly lower CH₄ production compared to AberAvon and Premium. Further study is necessary to determine whether the increase of CH₄ production response incurred by shifting from AM cutting to Noon and/or PM cutting could be compensated for by high daily gain from increased WSC concentration and DM digestibility.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.4
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• pp.471-478
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• 2011

The objectives were to compare the ability of various rumen microbial fractions to reduce nitrate and to assess the effect of nitrate on in vitro fermentation characteristics. Physical and chemical methods were used to differentiate the rumen microbial population into the following fractions: whole rumen fluid (WRF), protozoa (Pr), bacteria (Ba), and fungi (Fu). The three nitrogen substrate treatments were as follows: no supplemental nitrogen source, nitrate or urea, with the latter two being isonitrogenous additions. The results showed that during 24 h incubation, WRF, Pr and Ba fractions had an ability to reduce nitrate, and the rate of nitrate disappearance for the Pr fraction was similar to the WRF fraction, while the Ba fraction needed an adaptation period of 12 h before rapid nitrate disappearance. The WRF fraction had the greatest methane ($CH_4$) production and the Pr fraction had the greatest prevailing $H_2$ concentration (p<0.05). Compared to the urea treatment, nitrate diminished net gas and $CH_4$ production during incubation (p<0.05), and ammonia-N ($NH_3$-N) concentration (p<0.01). Nitrate also increased acetate, decreased propionate and decreased butyrate molar proportions (p<0.05). The Pr fraction had the highest acetate to propionate ratio (p<0.05). The Pr fraction as well as the Ba fraction appears to have an important role in nitrate reduction. Nitrate did not consistently alter total VFA concentration, but it did shift the VFA profile to higher acetate, lower propionate and lower butyrate molar proportions, consistent with less $CH_4$ production by all microbial fractions.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.3
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• pp.330-341
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• 2014

Andropogon gayanus is an important grass due to its high biomass production, drought tolerance and favorable growth on low fertility acidic soils. Currently, there is little research on the impact of growth stage on the nutritional quality or the degree of $CH_4$ production that may arise from this forage during ruminal fermentation. The objectives of this study were to determine the effects of regrowth stage of A. gayanus on its chemical composition, in vitro production of gas and CH4, as well as in vitro dry matter (DM) digestibility when grown under tropical Brazilian conditions and conserved as hay or as silage. The nutritional value of A. gayanus grass declined with increasing maturity; however digestible DM yield linearly increased. After 112 d of regrowth, A. gayanus produced higher quality silage (higher lactate and lower pH and butyrate content) and higher DM yield. However, the low levels of crude protein at this time would make protein supplementation a necessity for proper rumen fermentation. No differences in $CH_4$ kinetic parameters were found with advancing maturity or preservation method (hay or silage).

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.3
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• pp.197-205
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• 2013

An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.6
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• pp.864-873
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• 2013

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.