• 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.

• Journal of Nutrition and Health
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• v.28 no.9
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• pp.834-845
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• 1995

This study was conducted in order to establish an in vitro method simulating the physiological function of fibers along the large intestine of humans. Commercial fibers including guar gum, apple pectin, citrus pectin, CM-cellulose, alginic acid and $\alpha$-cellulose, and dietary fiber residues obtained from rice bran, barley, soybean, Korea cabbage, apple, tangerine and sea mustard were employed to determine the water-holding capacity, weight of hydrated residue and fiber content after anaerobic fermentation using human fecal inoculum for 24 hours, followed by dialysis under osmotic suction pressure. The weight of hydrated residue in commercial fibers was in the decreasing order of CM-cellulose > alginic acid, $\alpha$-cellulose > apple pectin, citrus pectin > guar gum and that in food fiber residues was in the decreasing order of rice bran, sea mustard > soybean > tangerine, Korean cabbage > barley > apple. It was demonstrated that the larger the weight of hydrated residue was, the more the weight of human stool increased. Consequently this in vitro method can be used as a preceeding test before undertaking animal or human experiment to predict the physiological effects of fiber residues from diverse food samples as well as commercially refined fibers.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1314-1321
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• 2006

The objective of this work was to study the effect of mixed culture solid substrate fermentation of C. tropicalis ZD-3 with A. niger ZD-8 on detoxification of cottonseed meal (CSM), and to investigate the effect of fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment on the reduction of free gossypol levels during mixed culture solid substrate fermentation of CSM. Experiment 1: Three groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. tropicalis ZD-3, A. niger ZD-8 or mixed culture (C. tropicalis ZD-3 with A. niger ZD-8). One non-inoculated group was used as the control. Levels of initial and final free gossypol (FG), CP and in vitro CP digestibility were assayed. The results indicated that mixed culture fermentation was far more effective than single strain fermentation, which not only had higher detoxification rate, but also had higher CP content and in vitro digestibility. Experiment 2: CSM substrates were treated according to experimental variables including fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment, Then, the treated CSM substrates were inoculated with mixed culture (C. tropicalis ZD-3 with A. niger ZD-8) and incubated at $30^{\circ}C$ for 36 h in a 95% relative humidity chamber. After fermentation ended, FG and CP content of fermented CSM substrate was assayed. The results showed that the appropriate fermentation period was 36 h, and the optimal proportion of CSM in substrate was 70%. Addition of sodium carbonate to CSM substrate was beneficial for fermentative detoxification. Heat treatment could facilitate fermentative detoxification, and supplementation with minerals was instrumental in reducing gossypol levels during mixed culture solid substrate fermentation of CSM.

• Animal Bioscience
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• v.35 no.2
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• pp.184-195
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• 2022

Objective: In this study we aimed to evaluate the effect of dietary live yeast supplementation on ruminal pH pattern, fermentation characteristics and associated bacteria in beef cattle. Methods: This work comprised of in vitro and in vivo experiments. In vitro fermentation was conducted by incubating 0%, 0.05%, 0.075%, 0.1%, 0.125%, and 0.15% active dried yeast (Saccharomyces cerevisiae, ADY) with total mixed ration substrate to determine its dose effect. According to in vitro results, 0.1% ADY inclusion level was assigned in in vivo study for continuously monitoring ruminal fermentation characteristics and microbes. Six ruminally cannulated steers were randomly assigned to 2 treatments (Control and ADY supplementation) as two-period crossover design (30-day). Blood samples were harvested before-feeding and rumen fluid was sampled at 0, 3, 6, 9, and 12 h post-feeding on 30 d. Results: After 24 h in vitro fermentation, pH and gas production were increased at 0.1% ADY where ammonia nitrogen and microbial crude protein also displayed lowest and peak values, respectively. Acetate, butyrate and total volatile fatty acids concentrations heightened with increasing ADY doses and plateaued at high levels, while acetate to propionate ratio was decreased accordingly. In in vivo study, ruminal pH was increased with ADY supplementation that also elevated acetate and propionate. Conversely, ADY reduced lactate level by dampening Streptococcus bovis and inducing greater Selenomonas ruminantium and Megasphaera elsdenii populations involved in lactate utilization. The serum urea nitrogen decreased, whereas glucose, albumin and total protein concentrations were increased with ADY supplementation. Conclusion: The results demonstrated dietary ADY improved ruminal fermentation dose-dependently. The ruminal lactate reduction through modification of lactate metabolic bacteria could be an important reason for rumen pH stabilization induced by ADY. ADY supplementation offered a complementary probiotics strategy in improving gluconeogenesis and nitrogen metabolism of beef cattle, potentially resulted from optimized rumen pH and fermentation.

• 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 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.

• Journal of Animal Science and Technology
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• v.66 no.3
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• pp.523-542
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• 2024

Hanwoo and Jeju Black cattle (Jeju Black) are native breeds of Korean cattle. Jeju Black cattle are recognized as natural monuments and are known to exhibit slower growth rates compared to Hanwoo. While several studies have analyzed the genetic characteristics of these cattle, there has been limited research on the differences in their microbiome. In this study, rumen fluid was obtained from three Hanwoo steers and three Jeju Black steers, and three different diets (total mixed rations [TMRs] for growing, early fattening, and late fattening periods) were used as substrates for in vitro fermentation. The in vitro incubation was conducted for 3 h and 24 h following a 2 × 3 factorial arrangement. After both incubation periods, fermentation characteristics were analyzed, and ruminal microbiome analysis was performed using 16S rRNA gene sequencing, employing both QIIME2 and PICRUSt2. The results revealed significant differences in the ruminal microbiota due to the inoculum effect. At the phylum level, Patescibacteria and Synergistota were found to be enriched in the Jeju Black inoculum-treated group. Additionally, using different inocula also affected the relative abundance of major taxa, including Ruminococcus, Pseudoramibacter, Ruminococcaceae CAG-352, and the [Eubacterium] ruminantium group. These microbial differences induced by the inoculum may have originated from varying levels of domestication between the two subspecies of donor animals, which mainly influenced the fermentation and microbiome features in the early incubation stages, although this was only partially offset afterward. Furthermore, predicted commission numbers of microbial enzymes, some of which are involved in the biosynthesis of secondary metabolites, fatty acids, and alpha amylase, differed based on the inoculum effect. However, these differences may account for only a small proportion of the overall metabolic pathway. Conversely, diets were found to affect protein biosynthesis and its related metabolism, which showed differential abundance in the growing diet and were potentially linked to the growth-promoting effects in beef cattle during the growing period. In conclusion, this study demonstrated that using different inocula significantly affected in vitro fermentation characteristics and microbiome features, mainly in the early stages of incubation, with some effects persisting up to 24 h of incubation.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.491-500
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• 2010

Three experiments were conducted to investigate the effects of polyurethane coated urea on in vitro ruminal fermentation, ammonia release dynamics and lactating performance of Holstein dairy cows fed a steam-flaked corn-based diet. In Exp. 1, a dual-flow continuous culture was run to investigate the effect of polyurethane coated urea on nutrient digestibility, rumen fermentation parameters and microbial efficiency. Three treatment diets with isonitrogenous contents (13.0% CP) were prepared: i) feedgrade urea (FGU) diet; ii) polyurethane coated urea (PCU) diet; and iii) isolated soy protein (ISP) diet. Each of the diets consisted of 40% steam-flaked corn meal, 58.5% forages and 1.5% different sources of nitrogen. PCU and FGU diets had significantly lower digestibility of NDF and ADF (p<0.01) than the ISP diet. Nitrogen source had no significant effect (p = 0.62) on CP digestibility. The microbial efficiency (expressed as grams of microbial N/kg organic matter truly digested (OMTD)) in vitro of the PCU diet (13.0 g N/kg OMTD) was significantly higher than the FGU diet (11.3 g N/kg OMTD), but comparable with the ISP diet (14.7 g N/kg OMTD). Exp. 2, an in vitro ruminal fermentation experiment, was conducted to determine the ammonia release dynamics during an 8 h ruminal fermentation. Three treatment diets were based on steam-flaked corn diets commonly fed to lactating cows in China, in which FGU, PCU or soybean meal (SBM) was added to provide 10% of total dietary N. In vitro $NH_3-N$ concentrations were lower (p<0.05) for the PCU diet than the FGU diet, but similar to that for the SBM diet at all time points. In Exp. 3, a lactation trial was performed using 24 lactating Holstein cows to compare the lactating performance and blood urea nitrogen (BUN) concentrations when cows were fed PCU, FGU and SBM diets. Cows consuming the PCU diet had approximately 12.8% more (p = 0.02) dietary dry matter intake than those consuming the FGU diet. Cows fed the PCU diet had higher milk protein content (3.16% vs. 2.94%) and lower milk urea nitrogen (MUN) concentration (13.0 mg/dl vs. 14.4 mg/dl) than those fed the FGU diet. Blood urea nitrogen (BUN) concentration was significantly lower for cows fed the PCU (16.7 mg/dl) and SBM (16.4 mg/dl) diets than the FGU (18.7 mg/dl) diet. Cows fed the PCU diet had less surplus ruminal N than those fed the FGU diet and produced a comparable lactation performance to the SBM diet, suggesting that polyurethane coated urea can partially substitute soybean meal in the dairy cow diet without impairing lactation performance.

• Journal of The Korean Society of Grassland and Forage Science
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• v.37 no.3
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• pp.216-222
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• 2017

This study was conducted to examine the effect of corn (Zea mays L.) - soybean (Glycine max L.) silage prepared by intercropping method on the nutritive value of the silage, in vitro rumen fermentation characteristics, dry matter degradability, as well as milk yield and milk composition of dairy cows. In a couple of experiments intercropped corn-soybean silage (CSBS) was compared with corn silage (CS) and/or Italian ryegrass hay (IRG). Numerically, CSBS had higher crude protein, ether extract, and lactic acid contents compared to CS. In vitro rumen fermentation analysis demonstrated that up to a 24-h incubation period, both CS and CSBS showed higher total gas production, ammonia N concentration, and dry matter degradability compared to IRG (p<0.05). The investigation on animals was conducted in a commercial dairy farm located in Gyeongju, South Korea, employing 42 Holstein cows that were divided into 2 group treatments: CS and CSBS in a completely randomized design. Although no significant difference was observed in milk yield, animals fed on CSBS showed significantly higher milk protein (p<0.05) and milk fat content (p<0.01), compared to animals fed on CS. Taken together, our findings indicate that corn-soybean silage that is cultivated, harvested, and prepared through intercropping can improve the protein content of the silage, and can also enhance in vitro rumen fermentation, dry matter degradability, and performance of dairy cattle.

• 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.