• 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.

• Korean Journal of Poultry Science
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• v.31 no.3
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• pp.177-185
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• 2004

This experiment was conducted to examine the dietary effects of local granular ark shell(GAS) replacing limestone on egg shell quality in aged hens. A total of 200 Hy-Line Brown layers of 54-weeks-old were allotted into four treatments with five replications each. They were fed control(limestone only) 1% GAS, 2% GAS and 3% GAS diets for 6 weeks. GAS was substituted for coarse-type limestone at 0, 1, 2 or 3% of diet on weight basis. Particle sizes of limestone and GAS were within the ranges of 2~4 mm in diameters. In vitro HCl solubility of GAS was significantly lower(6.27 vs. 7.90%) compared to that of limestone(p < 0.05). The dietary treatment did not affect feed intake and egg production rate of the birds. Egg shell strength and thickness tended to be improved for birds fed the diets containing higher levels of GAS. The average egg shell strength and thickness of 2 or 3% GAS groups were significantly greater than those of the control (P < 0.05). The Tibial ash content of birds from 3% GAS group increased significantly as compared to that of the control. Tibial breaking strength, however, was not affected by the dietary treatment. Serum Ca and P contents of the birds were also not different among the dietary treatments. In conclusion, locally prepared GAS can be used as a calcium source in aged layer diet.

• Journal of agriculture & life science
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• v.45 no.3
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• pp.69-79
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• 2011

This study was conducted to estimate effects of by-products of medical herbs replacing rice straw on in vitro fermentation characteristics. Each trial was composed of five treatments including medical herbs : rice straw (%) = 20 : 80 (T1), 40 : 60 (T2), 50 : 50 (T3), 100 : 0 (T4) and the control. Each treatment had eight fermentation times (3, 6, 9, 12, 24, 36, 48 and 72 hours) with three replications. The gas production and DM degradation were significantly (P<0.05) increased by supplementation, especially T4, during the whole fermentation periods. Methane production increased along with addition of by-products similar to the gas production and DM degradation. The pH values ranged from 5.39 to 6.80 and were significantly (P<0.05) decreased by supplementation of by-products of medical herbs. Microbial growth rates reached the peak at between 36 and 48h, thereafter tended to decrease. Although there were no significant differences in the enzyme activities, there was a tendency of increase in T4 treatment. From above results, the replacement levels, particularly 100% replacement of rice straw by by-products of medical herbs, resulted in improving the in vitro fermentation characteristics such as increasing gas production, microbial growth and DM degradation. Also it may help digestion by increasing enzyme activities.

• Animal Bioscience
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• v.35 no.9
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• pp.1379-1389
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• 2022

Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×10⁶ cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.2
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• pp.230-235
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• 2020

Objective: This study was realized to evaluate the nutritional value of rice grains as a replacement for corn grains in the diet of growing Hanwoo steers. Methods: Two experimental diets were prepared: i) Corn total mixed ration (TMR) consisting of 20% corn grains and ii) Rice TMR consisting of 20% rice grains, in a dry matter (DM) basis. These treatments were used for in vitro rumen fermentation and in vivo growth trials. In the rumen fermentation experiment, the in vitro DM digestibility (IVDMD), in vitro crude protein digestibility (IVCPD), in vitro neutral detergent fiber digestibility, pH, ammonia nitrogen, and volatile fatty acids (VFA) were estimated at 48 h, and the gas production was measured at 3, 6, 12, 24, and 48 h. Twenty four growing Hanwoo steers (9 months old; body weight [BW]: 259±13 kg) were randomly divided into two treatment groups and the BW, dry matter intake (DMI), average daily gain (ADG), and feed conversion ratio (FCR) were measured. Results: The in vitro experiment showed that the IVDMD, IVCPD, and VFA production of the Rice TMR were higher than those of the Corn TMR (p<0.05). The growth trial showed no differences (p>0.05) in the final BW, ADG, DMI, and FCR between the two TMRs. Conclusion: The use of rice grains instead of corn grains did not exhibit any negative effects on the rumen fermentation or growth performance, thereby rice grains with a DM of less than 20% could be used as a starch source in the diet of growing steers.

• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.1
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• pp.59-74
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• 2012

Buffer solubility and protein fractionation were evaluated from the hays (timothy, alfalfa and klein) and straws (tall fescue and rice), and $In$ $vitro$ trial was conducted to examine the effect of buffer extraction on fermentation characteristics, degradability and gas ($CO_2$ and $CH_4$) production. Buffer soluble protein (SP) content and A fraction in total protein were highest in alfalfa hay as 61% and 41.77%, respectively while lowest in rice straw (42.8% and 19.78%, respectively). No difference was observed in B1 fraction among forages but B2 fraction was slightly increased in klein hay (12.34%) and tall fescue straw (10.05%) compared with other forages (6.34~8.85%). B3 fraction of tall fescue was highest as 38.49% without difference among other forages while C fraction was highest in rice straw. pH in incubation solution was higher in all forages after extraction than before extraction at 3h (P<0.01) and 6h (P<0.05), and pH from hays of timothy and alfalfa was higher than the other forages at 6h (P<0.05) and 12h (P<0.001). Regardless of extraction, ammonia-N concentration from alfalfa hay was increased at all incubation times and extraction effect was appeared only at 3h incubation time (P<0.01). Total VFA concentration from alfalfa hay was highest up to 24h incubation while those from tall fescue straw and rice straw were lowest. Buffer extraction decreased (P<0.01~P<0.001) the total VFA concentration. Acetic acid proportion was increased (P<0.001) before extraction of forages but no difference was found between forages. Propionic acid($C_3$) proportion was also increased(P<0.001) before extraction in all forages than in straws at 3h, 24h and 48h incubations, and $C_3$ from hays were mostly higher (P<0.05) than from straws. Butyric acid proportion, however, was not affected by extraction at most incubation times. Parameter 'a' regarding to the dry matter (DM) degradation was increase (P<0.001) in all forages before extraction, and was decreased (P<0.05) in tall fescue straw and rice straw compared with hays. Parameter 'b' was also increased (P<0.001) before extraction but no difference was found between forages. Effective degradability of DM (EDDM) was higher (P<0.001) before extraction in most forages except for rice straw. Buffer extraction decreased (P<0.05) all parameters (a, b, and c) regrading to the crude protein (CP) degradation but no difference was found between forages. Effective degradation of CP (EDCP) was lower (P<0.05) in straws than in hays. Parameters 'a' and 'b' regarding to the NDF degradation (P<0.01) and effective degradability of NDF (EDNDF, P<0.001) were also higher in forages before extraction than after extraction but no difference was found between forages. Buffer extraction reduced (P<0.05~P<0.001) $CO_2$ production from all the forages uo to 24h incubation and its production was greater (P<0.05~P<0.01) from hays than straws. Methane ($CH_4$) production was also greater (P<0.01~P<0.001) in all forages at all incubation times, and its production was greater (P<0.05) from hays than from straws at most incubation times. Based on the results of the current study, it can be concluded that buffer solubility and CP fractionation might be closely related with $In$ $vitro$ VFA concentration, degradability and gas ($CO_2$ and $CH_4$) production. Thus, measurement of buffer solubility and protein fractionation of forages might be useful to improve TMR availability in the ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.2
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• pp.266-274
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• 2013

The effect of different phytogenic feed additives on reducing odorous compounds in swine was investigated using in vitro fermentation and analyzed their microbial communities. Soybean meal (1%) added with 0.1% different phytogenic feed additives (FA) were in vitro fermented using swine fecal slurries and anaerobically incubated for 12 and 24 h. The phytogenic FAs used were red ginseng barn powder (Panax ginseng C. A. Meyer, FA1), persimmon leaf powder (Diospyros virginiana L., FA2), ginkgo leaf powder (Ginkgo biloba L., FA3), and oregano lippia seed oil extract (Lippia graveolens Kunth, OL, FA4). Total gas production, pH, ammonianitrogen ($NH_3$-N), hydrogen sulfide ($H_2S$), nitrite-nitrogen ($NO_2{^-}$-N), nitrate-nitrogen ($NO_3{^-}$-N), sulfate (${SO_4}^{--}$), volatile fatty acids (VFA) and other metabolites concentration were determined. Microbial communities were also analyzed using 16S rRNA DGGE. Results showed that the pH values on all treatments increased as incubation time became longer except for FA4 where it decreased. Moreover, FA4 incubated for 12 and 24 h was not detected in $NH_3$-N and $H_2S$. Addition of FAs decreased (p<0.05) propionate production but increased (p<0.05) the total VFA production. Ten 16S rRNA DGGE bands were identified which ranged from 96 to 100% identity which were mostly isolated from the intestine. Similarity index showed three clearly different clusters: I (FA2 and FA3), II (Con and FA1), and III (FA4). Dominant bands which were identified closest to Eubacterium limosum (ATCC 8486T), Uncultured bacterium clone PF6641 and Streptococcus lutetiensis (CIP 106849T) were present only in the FA4 treatment group and were not found in other groups. FA4 had a different bacterial diversity compared to control and other treatments and thus explains having lowest odorous compounds. Addition of FA4 to an enriched protein feed source for growing swine may effectively reduce odorous compounds which are typically associated with swine production.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.11
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• pp.1562-1570
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• 2014

The effects of Lactobacillus mucosae (L. mucosae), a potential direct fed microbial previously isolated from the rumen of Korean native goat, on the rumen fermentation profile of brewers grain were evaluated. Fermentation was conducted in serum bottles each containing 1% dry matter (DM) of the test substrate and either no L. mucosae (control), 1% 24 h broth culture of L. mucosae (T1), or 1% inoculation with the cell-free culture supernatant (T2). Each serum bottle was filled anaerobically with 100 mL of buffered rumen fluid and sealed prior to incubation for 0, 6, 12, 24, and 48 h from which fermentation parameters were monitored and the microbial diversity was evaluated. The results revealed that T1 had higher total gas production (65.00 mL) than the control (61.33 mL) and T2 (62.00 mL) (p<0.05) at 48 h. Consequently, T1 had significantly lower pH values (p<0.05) than the other groups at 48 h. Ammonia nitrogen ($NH_3$-N), individual and total volatile fatty acids (VFA) concentration and acetate:propionate ratio were higher in T1 and T2 than the control, but T1 and T2 were comparable for these parameters. Total methane ($CH_4$) production and carbon dioxide ($CO_2$) were highest in T1. The percent DM and organic matter digestibilities were comparable between all groups at all times of incubation. The total bacterial population was significantly higher in T1 (p<0.05) at 24 h, but then decreased to levels comparable to the control and T2 at 48 h. The denaturing gradient gel electrophoresis profile of the total bacterial 16s rRNA showed higher similarity between T1 and T2 at 24 h and between the control and T1 at 48 h. Overall, these results suggest that addition of L. mucosae and cell-free supernatant during the in vitro fermentation of dried brewers grain increases the VFA production, but has no effect on digestibility. The addition of L. mucosae can also increase the total bacterial population, but has no significant effect on the total microbial diversity. However, inoculation of the bacterium may increase $CH_4$ and $CO_2$ in vitro.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.2
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• pp.177-190
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• 2011

This study was conducted to determine effects of dietary herbaceous peat on in vitro fermentation and milk production in dairy cows. Ruminal pH, gas production, VFA (volatile fatty acid), Ammonia-N, and rumen degradability were examined by the addition of three times over 0, 1, and 5% herbaceous peat with substrate of timothy hay, and the change of rumen fermentation characteristics were evaluated. In 0, 3, 12 and 24 hours cultivation, all treatments did not show a significant difference but the control at 6 hours appeared significantly lower pH compared to 1 and 5% treatments (p<0.05). The gas production of the treatments significantly increased until 12 hours of cultivation compared to control (p<0.05), the rumen ammonia concentration showed a tendency to increase until 24 hours in all treatment groups, and there was no significant difference between treatments. About the rumen degradability, 5% treatment showed higher rumen degradability in all hours than control and 1% treatment (p<0.05). Meanwhile, for in vivo trial, 16 heads of Holstein lactation dairy cows were selected for experiment for four weeks in order to research the change of milk yield, milk compositions and change of somatic cell counts of lactation dairy cows by herbaceous peat feeding. The milk yield of vitamin C and herbaceous peat treatments (T3) was 25.0 kg but the control was 23.2 kg, herbaceous peat treatment (T1) was 23.1 kg, and vitamin C treatment (T2) was 23.4 kg, so there was linear increase effect of milk yield by T3. The partial significance of the milk (fat, milk protein, lactose, MUN and SNF) and change of somatic cell count before and after experiment by the control and treatments about change of milk and somatic cell counts (p<0.05) were recognized. About change of milk in the first half (1~2 weeks) and latter half (3~4 weeks) during four weeks of experiments period, the herbaceous peat supplement treatments showed a tendency of significant decrease of quality of milk protein and SNF. The control and treatments did not show significant change of blood nutrients (total protein, cholesterol, NEFA, BUN), liver function component (AST, GGT) and minerals (Ca, P, Mg) before and after experiment. In summary, it is judged that herbaceous peat feeding for lactation dairy cows would be recommendable based on the results of milk, somatic cell count physiologically.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.6
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• pp.851-858
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• 2002

This study was conducted to determine effects of whole (intact), coarsely-ground (4 mm), finely-ground (1 mm), steam-flaked and steam-flaked-ground (1 mm) corns on in vitro and in situ DM digestibilities and also in vitro fermentation characteristics. After 48 h incubation, in vitro dry matter digestibilities of whole, steam-flaked, coarsely-ground, steam-flaked-ground, and finely-ground corns were 6.79, 61.68, 76.48, 85.72 and 90.31%, respectively. Steam-flaked-ground corn showed the highest digestibility until 24 h incubation (p<0.01). After 48 h incubation, pH of whole corn decreased with a small range. However the values of pH of other media significantly decreased (p<0.01). The gas productions of finely-ground and steam-flaked-ground corns were higher than those of the other corns (p<0.01). After 24 h incubation, $NH_3$-N concentration of finely-ground and steam-flaked-ground corns increased rapidly. Total VFA was the highest in finely-ground corn, followed by steam-flaked-ground, steam-flaked, coarsely-ground and whole corns. Incorporating steam-flaked corn resulted in the highest propionate concentration (p<0.01) and the lowest acetate : propionate value (p<0.05). Finely-ground corn showed the highest in situ DM digestibility throughout the incubation period (p<0.01), followed by coarsely-ground, steam-flaked and whole corns, respectively. Overall, DM of whole corn was merely digested in vitro as well as in situ.