• Journal of Animal Science and Technology
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• v.53 no.2
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• pp.147-154
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• 2011

This study was conducted to evaluate the effects of saponin contained plant extracts on in vitro rumen fermentation characteristics and methane production. Ruminal fluid was collected from rumen cannulated Hanwoo steers fed rice straw and concentrate (5:5). Collected rumen fluids, corn starch and buffer including saponin contained plant extracts (ginseng, Ogapi, soapwort, tea plant and yucca; 0.5%/15 ml) were incubated at $39^{\circ}C$ for 24 h. All incubations were repeated five times. Rumen pH in all treatments was lower (p<0.05) compared with that of the control (no addition) during incubation time. The concentration of total VFA in all treatments was higher (p<0.05) than that of the control after 12h incubation. Compared with the control, the concentration of acetate and propionate in all treatments was lower and higher after 6h incubation, respectively. The concentration of $NH_3$-N in all treatments was lower (p<0.05) than that of the control except for Ogapi or yucca extracts supplementation. The number of protozoa in all treatments was significantly (p<0.05) lower than that of the control except for soapwort extract supplementation. The total gas production and methane production in all treatments was higher (p<0.05) and lower (p<0.05) compared with the control, except for ogapi or soapwort extracts supplementation after 12h incubation, respectively. Therefore, reduction in methane production by saponins may could be results from decreased protozoal population without any negative in vitro fermentation.

• Journal of Animal Science and Technology
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• v.58 no.1
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• pp.4.1-4.7
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• 2016

Background: This study was conducted to evaluate the effects of adding Korean rice wine residue (RWR) in total mixed ration (TMR) on in vitro ruminal fermentation and growth performance of growing Hanwoo steers. Methods: For in vitro fermentation, the experimental treatments were Control (Con: 0 % RWR + TMR), Treatment 1 (T1: 10 % RWR + TMR), and Treatment 2 (T2: 15 % RWR + TMR). The rumen fluid was collected from three Hanwoo steers and mixed with buffer solution, after which buffered rumen fluid was transferred into serum bottles containing 2 g dry matter (DM) of TMR added with or without RWR. The samples were then incubated for 0 h, 12 h, 24 h, or 48 h at $39^{\circ}C$ and 100 rpm. For the in vivo experiment, 27 Hanwoo steers (6 months old) with an average weight of $196{\pm}8.66kg$ were subjected to a 24-week feeding trial. The animals were randomly selected and equally distributed into three groups. After which the body weight, feed intake and blood characteristics of each group were investigated. Results: The pH of the treatments decreased significantly relative to the control during the 12 h of incubation. Total gas production and ammonia nitrogen ($NH_3-N$) was not affected by RWR addition. The total volatile fatty acid (VFA) was lower after 24 h of incubation but at other incubation times, the concentration was not affected by treatments. Feed cost was 8 % and 15 % lower in T1 and T2 compared to control. Blood alcohol was not detected and a significant increase in total weight gain and average daily gain were observed in Hanwoo steers fed with RWR. Conclusion: Overall, the results of this study suggest that TMR amended with 15 % RWR can be used as an alternative feed resource for ruminants to reduce feed cost.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.6
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• pp.806-811
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• 2012

This study was conducted to evaluate effects of plant extracts on methanogenesis and rumen microbial diversity in in vitro. Plant extracts (Artemisia princeps var. Orientalis; Wormwood, Allium sativum for. Pekinense; Garlic, Allium cepa; Onion, Zingiber officinale; Ginger, Citrus unshiu; Mandarin orange, Lonicera japonica; Honeysuckle) were obtained from the Plant Extract Bank at Korea Research Institute of Bioscience and Biotechnology. The rumen fluid was collected before morning feeding from a fistulated Holstein cow fed timothy and commercial concentrate (TDN; 73.5%, crude protein; 19%, crude fat; 3%, crude fiber; 12%, crude ash; 10%, Ca; 0.8%, P; 1.2%) in the ratio of 3 to 2. The 30 ml of mixture, comprising McDougall buffer and rumen liquor in the ratio of 4 to 1, was dispensed anaerobically into serum bottles containing 0.3 g of timothy substrate and plant extracts (1% of total volume, respectively) filled with $O_2$-free $N_2$ gas and capped with a rubber stopper. The serum bottles were held in a shaking incubator at $39^{\circ}C$ for 24 h. Total gas production in all plant extracts was higher (p<0.05) than that of the control, and total gas production of ginger extract was highest (p<0.05). The methane emission was highest (p<0.05) at control, but lowest (p<0.05) at garlic extract which was reduced to about 20% of methane emission (40.2 vs 32.5 ml/g DM). Other plant extracts also resulted in a decrease in methane emissions (wormwood; 8%, onion; 16%, ginger; 16.7%, mandarin orange; 12%, honeysuckle; 12.2%). Total VFAs concentration and pH were not influenced by the addition of plant extracts. Acetate to propionate ratios from garlic and ginger extracts addition samples were lower (p<0.05, 3.36 and 3.38 vs 3.53) than that of the control. Real-time PCR indicted that the ciliate-associated methanogen population in all added plant extracts decreased more than that of the control, while the fibrolytic bacteria population increased. In particular, the F. succinogens community in added wormwood, garlic, mandarin orange and honeysuckle extracts increased more than that of the others. The addition of onion extract increased R. albus diversity, while other extracts did not influence the R. albus community. The R. flavefaciens population in added wormwood and garlic extracts decreased, while other extracts increased its abundance compared to the control. In conclusion, the results indicated that the plant extracts used in the experiment could be promising feed additives to decrease methane gas emission from ruminant animals while improving ruminal fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.3
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• pp.376-380
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• 2006

An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.

• Journal of Life Science
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• v.31 no.5
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• pp.520-526
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• 2021

Microbial protein is one of the sources of protein in the rumen and can also be the source of glutamate production. Glutamic acid is used as fuel in the metabolic reaction in the body and the synthesis of all proteins for muscle and other cell components, and it is essential for proper immune function. Moreover, it is used as a surfactant, buffer, chelating agent, flavor enhancer, and culture medium, as well as in agriculture for such things as growth supplements. Glutamic acid is a substrate in the bioproduction of gamma-aminobutyric acid (GABA). This review provides insights into the role of glutamic acid and glutamic acid-producing microorganisms that contain the glutamate decarboxylase gene. These glutamic acid-producing microorganisms could be used in producing GABA, which has been known to regulate body temperature, increase DM intake and milk production, and improve milk composition. Most of these glutamic acid and GABA-producing microorganisms are lactic acid-producing bacteria (LAB), such as the Lactococcus, Lactobacillus, Enterococcus, and Streptococcus species. Through GABA synthesis, succinate can be produced. With the help of succinate dehydrogenase, propionate, and other metabolites can be produced from succinate. Furthermore, clostridia, such as Clostridium tetanomorphum and anaerobic micrococci, ferment glutamate and form acetate and butyrate during fermentation. Propionate and other metabolites can provide energy through conversion to blood glucose in the liver that is needed for the mammary system to produce lactose and live weight gain. Hence, health status and growth rates in ruminants can be improved through the use of these glutamic acid and/or GABA-producing microorganisms.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.2
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• pp.61-72
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• 2022

The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39℃ for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybean-supplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.

• Journal of Animal Science and Technology
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• v.51 no.5
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• pp.369-378
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• 2009

Protein fractionation was evaluated from whole crop silages of rye (RS), wheat (WS), triticale (TS), oat (OS), barley (BS), and rice straw silage (RSS), and in vitro trial was carried out to examine the effect of silage and extraction of soluble protein on fermentation characteristics, total gas production and degradation. Soluble protein of silages was extracted with borate-phosphate buffer, and fermentation characteristics, gas production and degradation of silages were estimated by incubating anaerobically the mixed solution of strained rumen fluid and artificial saliva (1:1, v/v) containing dried and ground silages placed in nylon bag at $39^{\circ}C$ up to 48h. Soluble protein (SP) content was lowest for RSS as 2.11% in total CP compared to those for other silages. Highest A fraction (NPN) was observed from RS (74.33% of total CP) while those from TS and RSS were relatively low (48%). B2 fraction was relatively higher for RS, RSS and WS than for TS and BS. $B_3$ fraction was lowest in WS among silages. C fraction (27.07) in RSS was higher than in other silages (1.40~9.93%). pH in incubation solution was increased (P<0.01~P<0.001) for extracted silages up to 12h but decreased (P<0.01) at 48h for non-extracted ones. Contents of ammonia-N (P<0.001) and total VFA (P<0.01~P<0.001) were higher for non-extracted silages than for extracted ones. Acetate proportion was increased (P<0.001) in buffer extracted silages while those of propionate and butyrate were decreased (P<0.001) up to 24h incubation. Increased (P<0.001) total gas production was obtained from non-extracted silages up to 12h while gas production was increased (P<0.01) in extracted ones thereafter. In vitro degradation of dry matter and CP was increased (P<0.001) in non-extracted silages but that of neutral detergent fiber was increased (P<0.001) in extracted ones without difference among silages. Difference in mean values of degradability for each silage prior to- and post extraction with borate buffer, however, was not found among silages. It may be concluded that high NPN content of silages may reduce the protein availability in silages and borate buffer soluble components in silages can stimulate the early stage of fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.2
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• pp.211-218
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• 2016

This study evaluated the effect of Candida norvegensis (C. norvegensis) viable yeast culture on in vitro ruminal fermentation of oat straw. Ruminal fluid was mixed with buffer solution (1:2) and anaerobically incubated with or without yeast at $39^{\circ}C$ for 0, 4, 8, 16, and 24 h. A fully randomized design was used. There was a decrease in lactic acid (quadratic, p = 0.01), pH, (quadratic, p = 0.02), and yeasts counts (linear, p<0.01) across fermentation times. However, in vitro dry matter disappearance (IVDMD) and ammonia-N increased across fermentation times (quadratic; p<0.01 and p<0.02, respectively). Addition of yeast cells caused a decrease in pH values compared over all fermentation times (p<0.01), and lactic acid decreased at 12 h (p = 0.05). Meanwhile, yeast counts increased (p = 0.01) at 12 h. C. norvegensis increased ammonia-N at 4, 8, 12, and 24 h (p<0.01), and IVDMD of oat straw increased at 8, 12, and 24 h (p<0.01) of fermentation. Yeast cells increased acetate (p<0.01), propionate (p<0.03), and butyrate (p<0.03) at 8 h, while valeriate and isovaleriate increased at 8, 12, and 24 h (p<0.01). The yeast did not affect cellulolytic bacteria (p = 0.05), but cellulolytic fungi increased at 4 and 8 h (p<0.01), whereas production of methane decreased (p<0.01) at 8 h. It is concluded that addition of C. norvegensis to in vitro oat straw fermentation increased ruminal fermentation parameters as well as microbial growth with reduction of methane production. Additionally, yeast inoculum also improved IVDMD.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.71-79
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• 2018

Objective: Gelidium amansii (Lamouroux) is a red alga belonging to the family Gelidaceae and is commonly found in the shallow coasts of many East Asian countries, including Korea, China, and Japan. G. amansii has traditionally been utilized as an edible alga, and has various biological activities. The objective of this study was to determine whether dietary supplementation of G. amansii could be useful for improving ruminal fermentation. Methods: As assessed by in vitro fermentation parameters such as pH, total gas, volatile fatty acid (VFA) production, gas profile (methane, carbon dioxide, hydrogen, and ammonia), and microbial growth rate was compared to a basal diet with timothy hay. Cannulated Holstein cows were used as rumen fluid donors and 15 mL rumen fluid: buffer (1:2) was incubated for up to 72 h with four treatments with three replicates. The treatments were: control (timothy only), basal diet with 1% G. amansii extract, basal diet with 3% G. amansii extract, and basal diet with 5% G. amansii extract. Results: Overall, the results of our study indicate that G. amansii supplementation is potentially useful for improving ruminant growth performance, via increased total gas and VFA production, but does come with some undesirable effects, such as increasing pH, ammonia concentration, and methane production. In particular, real-time polymerase chain reaction indicated that the methanogenic archaea and Fibrobacter succinogenes populations were significantly reduced, while the Ruminococcus flavefaciens populations were significantly increased at 24 h, when supplemented with G. amansii extracts as compared with controls. Conclusion: More research is required to elucidate what G. amansii supplementation can do to improve growth performance, and its effect on methane production in ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.3
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• pp.395-404
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• 2019

Objective: Wheat bran (WB) and rice bran (RB) are the agricultural by-products used as poultry feed in many developing countries. However, their use for poultry feed is limited due to high fiber and the presence of anti-nutritional substances (e.g. ${\beta}-glucans$). The objective of this study was to develop a method to improve the quality of those brans by reducing the fiber content. Methods: A two-step fermentation method was developed where the second fermentation of first fermented dry bran was carried out. Fermentation was performed at a controlled environment for 3 h and 6 h (n = 6). The composition of brans, buffer solution and rumen liquor was maintained in a ratio of 1:2:3, respectively. Brans were analyzed for dry matter, crude fiber (CF), acid detergent fiber (ADF), neutral detergent fiber (NDF), and acid detergent lignin (ADL) content. Celluloses and hemicelluloses were calculated from the difference of ADF-ADL and NDF-ADF, respectively. Samples were compared by two-factor analysis of variance followed by Tukey's multiple comparison tests (p<0.05). Results: CF %, ADF % and cellulose tended to decrease and NDF % and hemicellulose content was reduced significantly (p<0.05). After the 1st fermentation step, NDF decreased $10.7%{\pm}0.55%$ after 3 h vs $17.0%{\pm}0.78%$ after 6 h in case of WB. Whereas, these values were $2.3%{\pm}0.30%$ (3 h) and $7.5%{\pm}0.69%$ (6 h) in case of RB. However, after the 2nd fermentation step, the decrease in the NDF content amounted to $9.1%{\pm}0.72%$ (3 h), $17.4%{\pm}1.13%$ (6 h) and $9.3%{\pm}0.46%$ (3 h), $10.0%{\pm}0.68%$ (6 h) in WB and RB, respectively. Cellulose and hemicellulose content was reduced up to $15.6%{\pm}0.85%$ (WB), $15.8%{\pm}2.20%$ (RB) and $36.6%{\pm}2.42%$ (WB), $15.9%{\pm}3.53%$ (RB), respectively after 2nd fermentation of 6 h. Conclusion: Two-step fermentation process improved the quality of the brans for their use in poultry feed.