• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.54-62
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• 2018

Objective: Due to the threat of global warming, the livestock industry is increasingly interested in exploring how feed additives may reduce anthropogenic greenhouse gas emissions, especially from ruminants. This study investigated the effect of Rhodophyta supplemented bovine diets on in vitro rumen fermentation and rumen microbial diversity. Methods: Cannulated Holstein cows were used as rumen fluid donors. Rumen fluid:buffer (1:2; 15 mL) solution was incubated for up to 72 h in six treatments: a control (timothy hay only), along with substrates containing 5% extracts from five Rhodophyta species (Grateloupia lanceolata [Okamura] Kawaguchi, Hypnea japonica Tanaka, Pterocladia capillacea [Gmelin] Bornet, Chondria crassicaulis Harvey, or Gelidium amansii [Lam.] Lamouroux). Results: Compared with control, Rhodophyta extracts increased cumulative gas production after 24 and 72 h (p = 0.0297 and p = 0.0047). The extracts reduced methane emission at 12 and 24 h (p<0.05). In particular, real-time polymerase chain reaction analysis indicated that at 24 h, ciliate-associated methanogens, Ruminococcus albus and Ruminococcus flavefaciens decreased at 24 h (p = 0.0002, p<0.0001, and p<0.0001), while Fibrobacter succinogenes (F. succinogenes) increased (p = 0.0004). Additionally, Rhodophyta extracts improved acetate concentration at 12 and 24 h (p = 0.0766 and p = 0.0132), as well as acetate/propionate (A/P) ratio at 6 and 12 h (p = 0.0106 and p = 0.0278). Conclusion: Rhodophyta extracts are a viable additive that can improve ruminant growth performance (higher total gas production, lower A/P ratio) and methane abatement (less ciliateassociated methanogens, Ruminococcus albus and Ruminococcus flavefaciens and more F. succinogenes.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.917-930
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• 2017

This study was conducted to evaluate Raphanus sativus extracts to methane reduction in rumen. Five different levels of R. sativus extracts were used to investigate the most effective dosing level for the decrease of methane production in the rumen. The rumen fluid was collected from a cannulated one Hanwoo cow ($BW=450{\pm}30kg$) consuming 600 g/kg timothy and 400 g/kg concentrate. On fermentation day, rumen fluid was collected at 2 hr postfeeding R. sativus extracts was dosed to achieve final concentration of 0, 1, 3, 5, 7, and 9% respectively, to fermentation bottles containing the mixture of rumen fluid and McDougall's buffer and 300 mg of timothy was added as a substrate. The fermentation was conducted for 3, 6, 9, 12, 24, 48 and 72 hr incubation time at $39^{\circ}C$ with shaking. In vitro ruminal pH values were measured normal range for ruminal fermentation. Dry matter disappearance was significantly higher (p<0.05) at 3 hr incubation time 1, 3 and 5% doses than that of control. The highest methane reduction was observed in 12 hr incubation time 5, 7 and 9%. The carbon dioxide emission was also significantly (p<0.05) lower than that of control at 12 hr incubation time 5, 7 and 9%. The total volatile fatty acid was no significant difference between control and all doses level at 12 and 24 hr incubation time. At 24 hr incubation time, the result of real-time PCR were indicated that M. archea was significantly lower (p<0.05) at all doses level comparing to that of control. In conclusion, R. sativus extracts were significantly decreased methane emission. R. sativus extracts were significantly lower (p<0.05) than that of control at 12 hr incubation time 5, 7 and 9% and no adversely effect in rumen pH, dry matter disappearance and total VFA.

• Journal of agriculture & life science
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• v.51 no.4
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• pp.97-109
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• 2017

This study was conducted to investigate the effects of anti-inflammatory plant extracts on the in vitro rumen fermentation characteristics and methane emission. Anti-inflammatory plant extracts from Morus bombycis Koidz, Mallotus japonicus L., Morus alba L., Paulownia coreana Uyeki, Isodon japonicus Hara and Ginkgo biloba L. were used in the study. The ruminal fluid(5 mL), McDougall buffer(10 mL), timothy as a substrate(0.3 g) and each anti-inflammatory plant extract(5% of substrate) were dispensed anaerobically into 50mL serum bottle. The mixtures were incubated for 3, 9, 12, 24, 48 and 72h at $39^{\circ}C$ without shaking. Supplementation of the anti-inflammatory plant extracts did not effects characteristics(pH, digestibility of dry matter, glucose concentration, ammonia concentration, protein concentration, VFA) on rumen fermentation. Total gas was showed a different pattern depending on treatments. Carbon dioxide was significantly(p<0.05) higher in Morus alba and Isodon japonicus than in control at 48h. Methane was significantly(p<0.05) lower in treatment than in control at initial fermentation. However the more incubation time was increased, the more methane emission was higher in treatment than in control. The concentrations of polyphenol and flavonoid were higher in Ginkgo biloba. In conclusion, supplementation of the anti-inflammatory plant extracts did not effect on rumen fermentation and methane emission was decreased in initial fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.12
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• pp.1728-1735
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• 2008

The present study was conducted with four ruminally canulated Holstein cows to observe the effects of monensin or fish oil on diet fermentation and production of conjugated linoleic acids (CLAs) in the rumen when fed diets supplemented with soybean oil and sodium bicarbonate. Cows of the control treatment were fed a basal diet (CON) consisting of 60% commercial concentrate and 40% chopped rye grass hay. Cows of other treatments were fed the same diet as CON, but the concentrate was supplemented with 7% of soybean oil and 0.5% of sodium bicarbonate (SO-B), SO-B supplemented with monensin (30 ppm, SO-BM) or concentrate supplemented with 6.3% of soybean oil, 0.5% of sodium-bicarbonate, 30 ppm of monensin and 0.7% of fish oil (SO-BMF). Dry matter (DM) intake of the cows was significantly (p<0.011) reduced by feeding the SO-BMF diet compared to the other diets which did not differ in DM intake. Whole tract digestibility of major dietary components was significantly (p<0.004-0.027) higher for SO-BMF than the other supplement-containing diets. Dietary supplements did not clearly affect rumen pH and ammonia concentrations compared to the CON diet. Significantly reduced (p<0.05) total VFA concentration was obtained by the addition of fish oil to the diet (SO-BMF) compared to other diets. No differences, however, were obtained in major VFA proportions as well as in total VFA between the supplemented diets. The SO-BM diet increased (p<0.01-0.05) the concentrations of trans-11 $C_{18:1}$ and linoleic acid in rumen fluid. Total CLA concentration was also increased by the feeding of SO-B and SO-BM diets during early fermentation times (up to 3 h) post-feeding. Cis-9, trans-11 CLA concentration in rumen fluid was highest (p<0.05) for SO-B up to 1 h while the highest (p<0.01) value for SO-BM occurred at 3 h post-feeding. An increased trans-10, cis-12 CLA concentration was obtained from the SO-B and SO-BM diets at 1 and 3 h post feeding compared to the other diets. Supplementation of oils with monensin and sodium bicarbonate increased (p<0.05) the proportions of $C_{18:1}$ and CLA in the plasma of cows, but the effect of monensin and/or fish oil was limited to trans-10, cis-12 CLA.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.901-916
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• 2017

This study was conducted to evaluate natural plant extracts for methane gas reduction in ruminants. Rumen fluid was collected from cannulated Hanwoo cow ($450{\pm}30kg$) consuming 400 g/kg concentrate and 600 g/kg timothy. The 15 ml of mixture comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically into 50 ml serum bottles. Rumen fluid contents were collected and in vitro fermentation prepared control (timothy, 300 mg), ginseng, balloon flower, yucca plant, camellia, tea plant and ogapi extracts were added at the level of 5% against 300 mg of timothy as a substrate (v/w) and incubated for 3, 6, 9, 12, 24, 48, and 72 h. In vitro pH values range 6.55~7.41, this range include rumen titration. The dry matter digestibility was not differ between all treatments and control. Total gas emission was significantly higher (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Carbon dioxide emission was not differ all treatments on 9 h than in control and significantly higher (p<0.05) yucca plant, camellia and tea plant treatments on 12 h than control. Methane emission was not differ all treatments on 6 h than in control. The rumen microbial growth rate was significantly higher (p<0.05) in ginseng, balloon flower on 12 h and significantly higher (p<0.05) in ginseng, yucca plant, tea plant and ogapi treatments on 24 h than in control. Total VFA was significantly higher (p<0.05) in tea plant and ogapi treatments on 12 h than in control and significantly higher (p<0.05) in ginseng, balloon flower treatments on 48 h than in control. Acetic acid was significantly lower (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Propionic acid was significantly higher (p<0.05) in ginseng and balloon flower treatments on 48 h than in control. As a results, sixth natural plant extracts had no significant effect dry matter digestibility and negative on rumen fermentation, but not effect methane reduction.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.12
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• pp.1726-1735
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• 2014

This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (p<0.05) the total gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (p<0.05) the ratio of acetate to propionate, the concentrations of isovalerate, valerate and ammonia-nitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.977-987
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• 2019

Objective: The use of tannin extract and other phytochemicals as dietary additives in ruminants is becoming more popular due to their wide biological actions such as in methane mitigation, bypass of dietary protein, intestinal nematode control, among other uses. Unfortunately, some have strong astringency, low stability and bioavailability, and negatively affecting dry matter intake and digestibility. To circumvent these drawbacks, an effective delivery system may offer a promising approach to administer these extracts to the site where they are required. The objectives of this study were to encapsulate acacia tannin extract (ATE) with native starch and maltodextrin-gum arabic and to test the effect of encapsulation parameters on encapsulation efficiency, yield and morphology of the microparticles obtained as well as the effect on rumen in vitro gas production. Methods: The ATE was encapsulated with the wall materials, and the morphological features of freeze-dried microparticles were evaluated by scanning electron microscopy. The in vitro release pattern of microparticles in acetate buffer, simulating the rumen, and its effect on in vitro gas production was evaluated. Results: The morphological features revealed that maltodextrin/gum-arabic microparticles were irregular shaped, glossy and smaller, compared with those encapsulated with native starch, which were bigger, and more homogenous. Maltodextrin-gum arabic could be used up to 30% loading concentration compared with starch, which could not hold the core material beyond 15% loading capacity. Encapsulation efficiency ranged from $27.7%{\pm}6.4%$ to $48.8%{\pm}5.5%$ in starch and $56.1%{\pm}4.9%$ to $64.8%{\pm}2.8%$ in maltodextrin-gum arabic microparticles. Only a slight reduction in methane emission was recorded in encapsulated microparticles when compared with the samples containing only wall materials. Conclusion: Both encapsulated products exhibited the burst release pattern under the pH conditions and methane reduction associated with tannin was marginal. This is attributable to small loading percentages and therefore, other wall materials or encapsulation methods should be investigated.

• Journal of agriculture & life science
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• v.50 no.2
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• pp.95-105
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• 2016

This study was conducted to evaluate the effects of nitrate-rich plants extracts on the in vitro rumen fermentation characteristics and rumen methane production. The extracts of nitrate-rich plants, as potato, carrot, chinese cabbage, lettuce and spinach were used in this study. The ruminal fluid was collected from a cannulated Hanwoo cow fed concentrate and timothy in the ratio of 6 to 4. The 20mL of mixture, comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically 50mL serum bottles containing 0.3g of timothy substrate and extracts of nitrogen-rich plants. The serum bottles were incubated 39℃ for 9, 12, 24, 48 hours. The pH value was decreased by increased incubation times and normal range to 6.31 to 6.96. The dry matter digestibility was significantly(p<0.05) lower in chinese cabbage than in control at 9h incubation time. Ammonia concentration was significantly(p<0.05) lower in potato, chinese cabbage, lettuce than in control and the rumen microbial growth rate was significantly(p<0.05) higher in carrot than in control at 24h incubation time. The concentrations of acetate and propionate was significantly(p<0.05) lower in treatment than in control. The concentration of butyrate was showed a different pattern depending on treatments. Total gas emissions was significantly(p<0.05) lower in chinese cabbage, lettuce, spinach than in control at 12h, 24h incubation time. Methane production was significantly(p<0.05) lower in potato, chinese cabbage, spinach than in control, carbon dioxide production was significantly(p<0.05) lower in treatment than in control. In conclusion, supplementation of the nitrate-rich plant extracts in ruminal fermentation in vitro resulted in decreasing the methane production without adversely affecting the fermentation characteristics. Particularly the chinese cabbage extract was regard as a potential candidate for reducing the methane emission in ruminants.

• 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.

• 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.