• Journal of agriculture & life science
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• v.51 no.5
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• pp.91-101
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• 2017

The in vitro experiment was conducted to ensure the supplemental level of spent Flammulina velutipes mushroom substrates(SMS) as an energy source in manufacturing of rye silage. Rye harvested at heading stage was ensiled with spent mushroom substrates of 0%(Control), 20%(R-20), 40%(R-40) and 60%(R-60) as fresh matter basis for 6week. The rumen fluid for preparation of in vitro solution was collected from two cannulated Holstein bulls fed a 40:60 concentrate:timothy diet. The experiment was conducted by 3, 6, 9, 12, 24, and 48 hrs of ncubation time with 3 replications. The silages were evaluated fermentation characteristics and dry matter digestibility(DMD) in vitro. The pH of in vitro solution was inclined to decrease with elapsing the incubation time, and that of the R-60 was significantly(p<0.05) lower than the other treatment at 48 hr of incubation. The microbial growth in vitro was inclined to increase with elapsing the incubation time, and that of the R-20 was significantly(p<0.05) greater than the Control at 48 hr of incubation. Gas production was greater(p<0.05) in the Control than the other treatments at 48 hr of incubation. In vitro dry matter digestibility(IVDMD) was higher with increasing the supplemental level of SMS, and was significantly(p<0.05) lower in the Control compared with other treatments throughout whole incubation time. The IVDMD for R-60 was the highest(p<0.05) among treatments at 24 hr and 48 hr of incubation. Considering of above results and the availability of SMS, SMS could be supplemented by 60% in fresh matter basis for rye silage fermentation.

• Journal of Animal Science and Technology
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• v.45 no.1
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• pp.131-142
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• 2003

We evaluated the effects of adding fibrolytic enzyme into ruminant diets on ruminal fermentation (in vitro) and lactational performances of dairy cows (in vivo). Through the in vitro experiment that was carried out with different contents of NDF (34, 38, 43%) in diets, digestibilities of NDF in the rumen appeared not significantly different by the addition of enzyme but were different by NDF content in diets showing higher digestibility in NDF 43% diet. It could be attributed by the relatively higher amount of hemicellulose in the current experimental diets than in conventional diets that might have been digested easily by the addition of fibrolytic enzyme in the rumen. The addition of fibrolytic enzyme tended to increase NDF digestibilities to a little extent both in 0.05 and 0.1% enzyme levels. Ruminal pH, NH3-N concentrations and VFA production in the rumen were not affected by the addition of fibrolytic enzyme. Activities of CMCase and xylanase were higher in enzyme treated diets of both NDF 34 and 38%. In particular, the activities of xylanase that slowly decreased from 0 to 12 hr but rapidly after 24 hr indicates that the major action of the enzyme in the rumen occurs in early period of incubation. Through an in vivo experiment, fibrolytic enzyme addition into the diets of dairy cows indeed affected lactational performance of milk yield. The cows fed enzyme treated diets produced 8% (1.9kg/d) more amounts of milk than with no enzyme addition. Milk composition of milk fat and protein was not affected by enzyme addition. Overall, the results of this in vivo study indicates that fibrolytic enzyme can be used to improve milk production in lactating cows. In respect that animals in different treatments of this study had the same amounts of intake, the increased milk yield with enzyme addition may be attributed to the improved utilization of nutrients in the digestive tract.

• 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.

• Journal of Animal Science and Technology
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• v.51 no.5
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• pp.379-386
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• 2009

The aim of this study was tofind out the effects of supplementation of vitamin A to the diets of high or low amounts of concentrates for ruminants. In the first experiment, ruminal fermentation patterns with the data of pH, VFA production and cellulose disappearance rates in the rumen in vitro were investigated. In the second experiment, enzyme activities, gas production and dry matter degradabilities using cellulolytic bacteria, Ruminococcus flavefaciens were investigated. Ruminal pH was higher in low amounts of concentrates than in high amounts of concentrates as expected, however, no significant differences were found. Cellulose disappearance rates improved in vitamin A addition particularly in early incubation time (before 24h) and also the production of volatile fatty acids increased in vitamin A addition. These trends were more evident in diets containing high amounts of concentrates than in low amounts of concentrates and it may indicate that vitamin A is more required in the diets of high amounts of concentrates. In the second experiment, gas production, enzyme activities and dry matter degradabilities using cellulolytic bacteria, Ruminococcus flavefaciens were not different between vitamin A added and non-added diets. Ruminococcus flavefaciens may not require additional vitamin A for its own growth.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.801-813
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• 2014

A study was conducted to improve the biological activity of two medicinal plants, Eucommia ulmoides Oliv. and Glycyrrhiza uralensis, by fermentation. The biological activity was assessed by determining antibacterial, antioxidant and antimethanogenic properties. Fermentation was achieved by adding the plant materials in MRS broth at 10% (w/v) and different starter cultures at 1% (v/v). Condition for fermentation were incubation temperature of $30^{\circ}C$ and agitation at 150 rpm for 48 h. Six starter cultures, Weissella confusa NJ28 (Genbank accession number KJ914897), Weissella cibaria NJ33 (Genbank accession number KJ914898), Lactobacillus curvatus NJ40 (Genbank accession number KJ914899), Lactobacillus brevis NJ42 (Genbank accession number KJ914900), Lactobacillus plantarum NJ45 (Genbank accession number KJ914901) and Lactobacillus sakei NJ48 (Genbank accession number KJ914902) were used. Antibacterial activity was observed in L. curvatus NJ40 and L. plantarum NJ45 only as opposed to other treatments, including the non-fermented groups, which showed no antibacterial activity. Both plants showed antioxidant activity, although E. ulmoides Oliv. had lower activity than G. uralensis. However, fermentation by all strains significantly improved (p<0.05), antioxidant activity in both plants compared to non-fermented treatment. Six treatments were based on antibacterial activity results, selected for in vitro rumen fermentation; 1) non-fermented E. ulmoides, 2) fermented E. ulmoides NJ40, 3) fermented E. ulmoides NJ45, 4) non-fermented G. uralensis, 5) fermented G. uralensis NJ40, 6) fermented G. uralensis NJ45. A negative control was also added, making a total of 7 treatments for the in vitro experiment. Medicinal plant-based treatments significantly improved (p<0.05) total volatile fatty acid (VFA) concentration. Significant methane reduction per mol of VFA were observed in G. uralensis (p<0.05). Based on the present study, fermentation improves the biological activity of E. ulmoides Oliv. and G. uralensis. Fermented G. uralensis could also be applied as an enteric methane mitigating agent in ruminant animals.

• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.987-996
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• 2003

This study was conducted to determine the effects of the artificial culture medium of wild-ginsengs on in vitro fermentation characteristics. NH$_3$-N concentration was showed the highest in 3% WGM treatment among all treatments and control. In addition, microbial protein synthesis was significantly different in all treatments throughout the incubation time, and WGM 3% treatment was the highest at the 9 h incubation(P〈0.05). Protozoa numbers within rumen were decreased in all WGM treatments at 9 h incubation time, whereas WGM 3% treatment was always decreased throughout the incubation(P〈0.05). NDF and ADF digestibility were proportionally increased as the incubation time in both control and treatments. NDF digestibility showed no significantly difference between control and the 3% treatment, and ADF digestibility was similar in all. Total volatile fatty acid(VFA) concentrations of WGM treatments without 5% were significantly higher than control (P〈0.05). No differences were observed in total VFA, acetate, propionate and butyrate concentration among the WGM treatments. Acetate/Propionate ratio of WGM treatments was higher than control after 12 h incubation(P〈0.05). As a result of the artificial culture medium of wild-ginseng on rumen fermentation characteristics in vitro, microbial protein synthesis of WGM treatment was higher than control, and WGM 3% was the highest in all treatments(P〈0.05). The effect of saponin in artificial culture medium of wild-ginseng tended to decrease NH$_3$-N concentration, while it increases the microbial synthesis in early incubation. Therefore, artificial cultures medium of wild-ginseng can increase utilization of feed by microbial and anti-protozoal effects of saponin, which may enhance microbial synthesis capacity in early fermentation period in rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.6
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• pp.804-811
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• 2013

The effects and significance of ${\gamma}$-amino butyric acid (GABA) producing bacteria (GPB) on in vitro rumen fermentation and reduction of biogenic amines (histamine, methylamine, ethylamine, and tyramine) using corn meal as a substrate were determined. Ruminal samples collected from ruminally fistulated Holstein cows served as inoculum and corn was used as substrate at 2% dry matter (DM). Different inclusion rates of GPB and GABA were evaluated. After incubation, addition of GPB had no significant effect on in vitro fermentation pH and total gas production, but significantly increased the ammonia nitrogen ($NH_3$-N) concentration and reduced the total biogenic amines production (p<0.05). Furthermore, antioxidation activity was improved as indicated by the significantly higher concentration of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) among treated samples when compared to the control (p<0.05). Additionally, 0.2% GPB was established as the optimum inclusion level. Taken together, these results suggest the potential of utilizing GPB as feed additives to improve growth performance in ruminants by reducing biogenic amines and increasing anti-oxidation.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.1
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• pp.51-61
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• 2016

In order to provide recommendations on the most useful forage species to smallholder farmers, eleven grass and eleven legume forages grown in Abomey-Calavi in Republic of Benin were investigated for nutritive value (i.e. chemical composition and energy content) and fermentation characteristics (i.e. gas and volatile fatty acid production, organic matter degradability). The in vitro gas production technique was used, incubating the forages for 120 h under anaerobic condition with buffalo rumen fluid. Compared to legume, tropical grass forages showed lower energy (8.07 vs 10.57 MJ/kg dry matter [DM]) and crude protein level (16.10% vs 19.91% DM) and higher cell wall content (neutral detergent fiber: 63.8% vs 40.45% DM), respectively. In grass forages, the chemical composition showed a quite high crude protein content; the in vitro degradability was slightly lower than the range of tropical pasture. The woody legumes were richer in protein and energy and lower in structural carbohydrates than herbaceous plants, however, their in vitro results are influenced by the presence of complex compounds (i.e. tannins). Significant correlations were found between chemical composition and in vitro fermentation characteristics. The in vitro gas production method appears to be a suitable technique for the evaluation of the nutritive value of forages in developing countries.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.364-370
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• 2008

Chemical analysis and in vitro studies were conducted to investigate the nutritive value for ruminants of cell mass from lysine production (CMLP) which is a by-product of the lysine manufacturing process. Proximate analysis, protein fractionation, and in vitro protein degradation using protease from Streptomyces griseus and strained ruminal fluid were carried out to estimate ruminal protein degradability of CMLP with two reference feedstuffs-soybean meal (SBM) and fish meal (FM). Amino acid composition and pepsin-HCl degradability were also determined to evaluate postruminal availability. CMLP contained 67.8% crude protein with a major portion being soluble form (45.4% CP) which was composed of mainly ammonium nitrogen (81.8% soluble CP). The amount of nucleic acids was low (1.15% DM). The total amount of amino acids contained in CMLP was 40.60% DM, which was lower than SBM (47.69% DM) or FM (54.08% DM). CMLP was composed of mainly fraction A and fraction B2, while the protein fraction in SBM was mostly B2 and FM contained high proportions of B2 and B3 fractions. The proportion of B3 fraction, slowly degradable protein, in CP was the highest in fish meal (23.34%), followed by CMLP (7.68%) and SBM (1.46%). CMLP was degraded up to 51.40% at 18 h of incubation with Streptomyces protease, which was low compared to FM (55.23%) and SBM (83.01%). This may be due to the insoluble portion of CMLP protein being hardly degradable by the protease. The in vitro fermentation by strained ruminal fluid showed that the amount of soluble fraction was larger in CMLP (40.6%) than in SBM (17.8%). However, because the degradation rate constant of the potentially degradable fraction of CMLP (2.0%/h) was lower than that of SBM (5.8%/h), the effective ruminal protein degradability of CMLP (46.95%) was slightly lower than SBM (53.77%). Unavailable fraction in the rumen was higher in CMLP (34.0%) compared to SBM (8.8%). In vitro CP degradability of CMLP by pepsin was 80.37%, which was lower than SBM (94.42%) and FM (89.04%). The evaluation of protein degradability using different approaches indicated that soluble protein in CMLP may supply a large amount of ammonia in the rumen while insoluble protein can be by-passed from microbial attacks due to its low degradability. The results from this study suggest that CMLP can be used as a protein supplement to ruminants for supplying both non-protein nitrogen to rumen microbes and rumen undegradable protein to the host animal.

• Animal Bioscience
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• v.35 no.9
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• pp.1367-1378
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• 2022

Objective: The present study was conducted to examine the gas production, fermentation characteristics, nutrient degradation, and methanogenic community composition of a rumen fluid culture with Broussonetia papyrifera (B. papyrifera) subjected to ensiling or steam explosion (SE) pretreatment. Methods: Fresh B. papyrifera was collected and pretreated by ensiling or SE, which was then fermented with ruminal fluids as ensiled B. papyrifera group, steam-exploded B. papyrifera group, and untreated B. papyrifera group. The gas and methane production, fermentation characteristics, nutrient degradation, and methanogenic community were determined during the fermentation. Results: Cumulative methane production was significantly improved with SE pretreatment compared with ensiled or untreated biomass accompanied with more volatile fatty acids production. After 72 h incubation, SE and ensiling pretreatments decreased the acid detergent fiber contents by 39.4% and 22.9%, and neutral detergent fiber contents by 10.6% and 47.2%, respectively. Changes of methanogenic diversity and abundance of methanogenic archaea corresponded to the variations in fermentation pattern and methane production. Conclusion: Compared with ensiling pretreatment, SE can be a promising technique for the efficient utilization of B. papyrifera, which would contribute to sustainable livestock production systems.