• Asian-Australasian Journal of Animal Sciences
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• v.12 no.4
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• pp.544-547
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• 1999

The object of this study was to determine the influence of monensin and virginiamycin (VM) on in vitro ruminal fermentation of rice straw or ammoniated rice straw. Rumen fluid was collected from 4 wethers fed 200 g of concentrate supplement with 400 g of untreated (U) or ammoniated (A) rice straw once daily for 28 days. Mixed ruminal microorganisms were incubated in anaerobic media that contained 20% (vol/vol) ruminal fluid and 0.3 g of either U or A rice straw. Monensin and/or VM, dissolved in ethanol, were added in centrifuge tubes at final concentrations of 0, 15, 30, 15+15 and 30+30 ppm of culture fluid. The addition of monensin and VM combination to A rice straw fermentation decreased (p<0.05) the acetate to propionate ratio, total VFA and lactate production, but increased (p<0.05) pH. Total gas production tended to be decreased by the addition of monensin plus VM. Antimicrobial agents decreased $NH_3$ N concentration and dry matter digestibility.

• Reproductive and Developmental Biology
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• v.31 no.4
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• pp.235-240
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• 2007

This study was conducted to examine the viability of Korean native striped cattle (Bos namadicus Falconer, Chikso) clone embryos after embryo transfer. Chikso somatic cell nuclear transfer (SCNT) embryos were produced by fusion of ear skin cells derived from a female Chikso with enucleated oocytes matured in vitro for 18-24 hr. After in vitro culture of SCNT embryos for 7 to 8 days, fresh or vitrified blastocysts derived from SCNT were transferred into a uterine horn of recipient cows. Fifteen of total 43 recipients were pregnant at Day 50 and 4 recipients were maintained to term. Three IVF-derived calves and 1 clone Chikso calf were born. Pregnancy rate was higher when fresh embryos were transferred to recipients compared to vitrified embryos, but development to term was not different between both groups. The clone Chikso calf died at 5 days after birth due to the fullness of amniotic fluid in rumen and the infection of umbilical cord. The result of the present study shows that clone Chikso calf can produced from the embryo transfer of SCNT embryos, however, solution of abortion problem is necessary to improve the cloning efficiency.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.4
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• pp.501-506
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• 2001

This experiment was conducted to evaluate the effects of supplemental unsaturated fatty acids (UFA) on fermentation characteristics, especially on gas production, cellulose degradation and volatile fatty acid (VFA) concentration by mixed ruminal microorganisms. In order to attain this objective, unsaturated fatty acids including oleic acid (C 18:1), linoleic acid (C18:2) and arachidonic acid (C22:4) were added at varying level. Mixed ruminal microbes used in this experiment were obtained from the rumen of a cannulated Holstein cow. Medium pH values after 7 d incubation were significantly affected by type and level of unsaturated fatty acids (p<0.01). All of UFA inhibited total gas production, and especially treatment of arachidonic acid at the levels of 0.01% gave the lowest gas. production after 7 d incubation (p<0.01). Comparison of the population of protozoa revealed that UFA did not have any significant effect on the total protozoa number. The addition of UFA did not effect dry matter degradation. Volatile fatty acid (VFA) composition of the culture was influenced little by UFA, although the considerable amount of iso-type VFA were detected in UFA supplemented incubations. The ratio of acetic acids to propionic acids, however, was lower than control in all the treatments after 7 d incubation (p<0.01).

• Journal of Animal Science and Technology
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• v.51 no.1
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• pp.45-52
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• 2009

This experiment was conducted to observe the effects of anaerobic cellulolytic bacteria culture (Ruminococcus flavefaciens H-20 and Fibrobactor succinogenes H-23) on in vivo ruminal fermentation characteristics in Hanwoo heifers. Four ruminally cannulated Hanwoo heifers ($221\pm7.5kg$) receiving a basal diet containing 3 kg of mixture hay (tall fescue and ochardgrass) and 2 kg of concentrate per day were in a $4\times4$ Latin square with 21-day periods. Treatments were the basal diet without the culture additive (control), the basal diet plus 50 ml/day of bacteria culture of H-20 and H-23 (1%), 150 ml/day of H-20 and H-23 (3%), and 250 ml/day of H-20 and H-23 (5%). In the whole experimental periods, ruminal pH did not differ between treatments. However, the concentration of ruminal ammonia-N was increased in the 3% treatment relative to control and the 1% treatment at 1 hr post-feeding (p<0.05). Avicelase and CMCase (carboxymethyl cellulase) activities in rumen fluid showed no significant difference among treatments. However, xylanase activity was higher in the 5% (119.49, xylose ${\mu}mol$/ml/min) than the 3% treatment (71.02, xylose ${\mu}mol$/ml/min) at 0 hr post-feeding (p<0.05). Concentrations of ruminal total VFA, acetate, propionate and valerate were unaffected by treatments, while butyrate was higher in the 3% treatment (24.48 mM) than control (15.71 mM) at 1 hr post-feeding (p<0.05). Results indicate that minimum 3% inclusion of cellulolytic bacteria cultures improved ruminal fermentation, especially ammonia-N concentration and butyric acid production.

• Journal of Animal Science and Technology
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• v.55 no.1
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• pp.41-49
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• 2013

In vitro and in situ incubation studies were conducted to determine effects of yeast culture supplements (Saccharomyces cerevisiae) on cellulolytic bacterial function and fiber digestion in rice straw. In vitro dry matter digestibility of rice straw gradually increased according to supplemental levels of yeast culture (0.0, 0.2, 0.4, 0.6, 0.8 and 1.0%). Digestibility of rice straw started to increase apparently when yeast culture was added more than 0.6% level (p<0.05). Also, we reconfirmed that in vitro dry matter digestibility was significantly increased by 0.6% of yeast culture addition in 4% NaOH treated and non-treated rice straws (p<0.05). When in situ dry matter digestibility was tested in Korean native goats fed basal diet or experimental diet which contained 1.0% of yeast culture, the yeast culture feeding improved in situ dry matter digestibility in both 4% NaOH treated and non-treated rice straws (p<0.05). In case of real-time PCR monitoring cellulolytic bacterial function, the bacterial population attached on rice straw showed the increasing trends with higher level of yeast culture spraying on rice straw. F. succinogenes and R. flavefaciens were significantly increased in accordance to spraying levels of yeast culture (0.0, 0.1 and 0.3%) at both 12 and 24 hrs of in situ incubation (p<0.05). R. albus was significantly higher population in yeast culture spraying than non-soraying at 12 hrs of in situ incubation (p<0.05). These bacterial populations were showed the increasing trends with digestibility enhancement of rice straw according to the higher levels of yeast culture supplement. Overall, these results clearly suggest that the presence of yeast culture result in noticeable increase of rice straw digestion, which is modulated via good effect on cellulolytic bacterial attachment to fiber substrates.

• 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.

• Korean Journal of Organic Agriculture
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• v.24 no.3
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• pp.453-463
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• 2016

This study was undertaken to investigate the effects of dicarboxylic acid supplementation, as replacement antibiotics, of on in vitro ruminal parameters and milk yield and milk composition in lactating cows. in vitro treatments were 1) Con (4 g of basal diet), 2) CM (4 g of basal diet + 0.05 ml of monensin), 3) CR (4 g of basal diet + 0.1 ml of dicarboxylic acid) and in vivo treatments were 1) Con (25 kg of basal diet/head/day), and 2) CR (25 kg of basal diet + 5 g of dicarboxylic acid/head/day), respectively. A total 10 lactating dairy cows ($649{\pm}19kg$ average body weight, $99{\pm}65$ average milking days) were divided in to two groups according to mean milk yield and number of days of postpartum. The cows fed a basal diet during adaptation (2 wk) and experimental diets during the treatment periods (4 wk). In the first in vitro experiment, there were no statistical differences between treatments in pH, gas production, and ammonia-N and lactic acid concentration during incubation. However, dry matter digestibility was significantly higher in CR treatment compared to control or CM treatment (P<0.05). Total VFA was tended to higher in CR treatment than those of control and CM treatment (P>0.05). In the second experiment, milk yield was significantly higher in treatment (40.39 kg) compared to control (35.19 kg), (P<0.05). Milk composition and MUN were not changed by dietary supplementing dicarboxylic acid. Therefore the present results reporting that supplementation of dicarboxylic acid might enhance the stabilization of ruminal fermentation and increase the milk yield of lactating cows.

• 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 The Korean Society of Grassland and Forage Science
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• v.34 no.3
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• pp.193-201
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• 2014

This study was conducted to examine the effects of defaunation (removal of live protozoa) on fermentation characteristics, degradation of ryegrass hay and $CH_4$ (methane) production by rumen microbes when incubated with plant oils (SO, sunflower oil and LO, linseed oil) in vitro. Sodium lauryl sulfate (0.000375 g/ml) as a defaunation reagent was added into the culture solution and incubated anaerobically up to 24 h at $39^{\circ}C$. pH from defaunation was increased for all treatments from 6 h incubation times (p<0.01-0.001) compared with those from fauantion. Concentration of ammonia-N from defaunation is higher than that from faunation at 3 h (p<0.001), 12 h (p<0.05) and 24 h (p<0.001) incubation times. Defaunation decreased (p<0.01-0.001) total volatile fatty acid concentration at all incubation times. Molar proportions of $C_2$ (acetate, p<0.05-0.001) and butyrate (p<0.01-0.001) were also decreased by defaunation at all incubation times. Molar proportion of $C_3$ (propionate), however, was increased by defaunation at all incubation times (p<0.001). Thus the rate of $C_2$ to $C_3$ was decreased by defaunation at all incubation times (p<0.001). Defaunation decreased ED (effective degradability) of dry matter (p<0.001) and ED of neutral detergent fiber (p<0.001) of ryegrass hay. Defaunation decreased total gas, $CH_4$ production, $CH_4$ % in total gas and $CH_4/CO_2$ at all incubation times (p<0.001). Oil supplementation decreased total gas (p<0.05-0.001), $CH_4$ production (p<0.001) and $CH_4$ % in total gas (p<0.001) compared with control at all incubation times. The result of this study showed that defaunation combined with oil supplementation may cause an alteration of microbial communities and further medicate the fermentation pattern, resulting in both reduction of degradation of ryegrass hay and $CH_4$ production. No difference, however, was observed in all the examinations between SO and LO.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.763-772
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• 2004

The patterns of fungal growth and fiber digestion under the microscope, and tile productions of fibrolytic enzyme were studied in an in vitro culture with Neocallimastix frontalis SA when either filter paper or rice straw was provided as sole energy source. Under the microscopic observation, active zoospores attachment, sporangium development and complex rhizoidal system were founded on the surface and at the edge of filter paper. After 7 days of incubation, a reduced fiber mass, a decreased fiber cohesion and a weakened fiber structure by fungal digestion were clearly observed. Similar fungal development was observed with rice straw, but fungal growth and digestion took place mostly on the damaged and exposed portion of rice straw. Although there were some differences in absolute concentration and pattern, the concentration of both cellulase and xylanase increased with incubation time with the higher activity being obtained with filter paper. Their differences were large especially after 48 and 96hr of incubation(P< 0.05). The filter paper was more good inducer of cellulolytic and xylanolytic enzymes compared with complex substrate, rice straw. These findings suggest that the filter paper is the better energy source for N frontalis than the complex substrate, and structural disintegration by physical process is able to help rumen fungal growth on the lignified roughage although anaerobic rumen fungi have mechanical and enzymatic functions for fiber digestion.