• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.3
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• pp.206-212
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• 2013

This study was conducted to investigate the effect of biological membrane transfer modifier, lysophospholipd (LPLs) on the parameters from in vitro rumen simulated fermentation. Commercially available LPLs product (Lipidol$^{TM}$) was supplemented into experimental diets which consisted of orchard grass and concentrate diet (60:40) in different levels (0.1%, 0.3% and 0.5%). Then in vitro rumen simulated fermentation was performed. Although, a declining trend of pH was found in treatments, all pH values were detected in a range relevant to normal rumen fermentation. Gas production, ammonia nitrogen and total VFA production were greatly influenced by the supplementation of LPLs. All parameters were increased along with increased levels of LPLs in diet. As a result, 0.1% of Lipidol$^{TM}$ is recommended based on the determined in vitro rumen fermentative parameters in this study.

• Asian-Australasian Journal of Animal Sciences
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• v.3 no.4
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• pp.269-279
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• 1990

A 98 d feeding trial carried out to study liveweight change and rumen metabolites in heifers weighing initially 275 kg and given either untreated or ammoniated rice straw supplemented with 0, 0.4, 0.8 or 1.2 kg protein meal consisting of cottonseed meal (60). All 32 animals received 0.6 kg rice polishings/hd/d and had continuous access to molasses/urea block-licks containing 15% urea. The effects on growth rates of treatment of the straw with ammonia and of supplementation with bypass protein were additive. The heifers fed ammoniated straw grew 267 g/hd/d (p<0.001) faster and consumed 11% (p<0.05) more straw than the heifers on untreated straw. The mean growth response to bypass protein was 0.37 kg gain/kg protein meal supplied. Supplementation with protein meal tended (p=0.06) to depress intake of straw, but straw intakes of the unsupplemented groups were high. Small changes in the composition of the block-licks that were fed throughout the feeding trial led to changes in block intake and in intake of untreated straw. Increasing quantities of protein meal fed were associated with linear increase in concentrations of ammonia (p<0.05) and in molar percentages of iso-butyrate (p<0.01), iso-valerate (p<0.01) and valerate (p<0.01) in the rumen fluid of the heifers on a basal diet of untreated straw. However, in the rumen fluid of the heifers given ammoniated straw, the levels of these metabolities were not affected by the quantity of protein meal given.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.113-128
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• 1999

Protozoa can represent as half of the total rumen microbial biomass. Around 10 genera are generally present on the same time in the rumen. Based on nutritional aspects they can be divided in large entodiniomorphs, small entodiniomorphs and isotrichs. Their feeding behaviour and their enzymatic activities differ considerably. Many comparisons between defaunated and refaunated animals were carried out during the last two decades to explain the global role of protozoa at the ruminal or animal levels. It is now generally considered that a presence of an abundant protozoal population in the rumen has a negative effect on the amino acid (AA) supply to ruminants and contribute to generate more methane but, nevertheless, protozoa must not be considered as parasites. They are useful for numerous reasons. They stabilise rumen pH when animal are fed diets rich in available starch and decrease the redox potential of rumen digesta. Because cellulolytic bacteria are very sensitive to these two parameters, protozoa indirectly stimulate the bacterial cellulolytic activity and supply their own activity to the rumen microbial ecosystem. They could also supply some peptides in the rumen medium which can stimulate the growth of the rumen microbiota, but this aspect has never been considered in the past. Their high contribution to ammonia production has bad consequences on the urinary nitrogen excretion but means also that less dietary soluble nitrogen is necessary when protozoa are present. Changes in the molar percentages of VFA and gases from rumen fermentations are not so large that they could alter significantly the use of energy by animals. The answer of animals to elimination of protozoa (defaunation) depends on the balance between energy and protein needs of animals and the supply of nutrients supplied through the diet. Defaunation is useful in case of diets short in protein nitrogen but not limited in energy supply for animals having high needs of proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.11
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• pp.1577-1583
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• 2014

Encapsulation is a method used to protect material from certain undesirable environments, for controlled release at a more favorable time and place. Animal productivity would be enhanced if feed additives are delivered to be utilized at their site of action, bypassing the rumen where they are likely to be degraded by microbial action. A novel method of encapsulation with sesame gum was used to coat nitrate, a known enteric methane mitigating agent, and tested for the effect on methane reduction and other in vitro fermentation parameters using rumen fluid from cannulated Hanwoo steers. Orchard grass was used as basal diet for fermentation. The treatments were matrix (1.1 g sesame gum+0.4 g sesame oil cake) only, encapsulated nitrate (matrix+nitrate [21 mM]), free nitrate (21 mM), and a control that contained no additive. Analyses of fermentation parameters were done at 0, 3, 6, 9, 12, 24, and 48 h time periods. In comparison to control, both free and encapsulated nitrate produced significantly reduced (p<0.01) methane (76% less) and also the total volatile fatty acids were reduced. A significantly higher (p<0.01) concentration of ammonia nitrogen was obtained with the encapsulated nitrate treatment (44%) compared to the free form (28%) and matrix only (20%) (p = 0.014). This might suggest slow release of encapsulated nitrate so that it is fully reduced to ammonia. Thus, this pioneering study found a significant reduction in methane production following the use of sesame gum encapsulated nitrate that shows the potential of a controlled release system in enhancing sustainability of ruminant production while reducing/eliminating the risk of nitrite toxicity.

• Korean Journal of Agricultural Science
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• v.45 no.3
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• pp.419-427
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• 2018

The present study was conducted to investigate the effect of different levels of Korean corn grain on in vitro ruminal fermentation with total mixed ration (TMR) as a basal feed. Three ruminal cannulated Holstein steers (Body Weight $479{\pm}33.0kg$) were used as rumen fluid donors. Treatments for in vitro fermentation were TMR only (control, 3.0 g), TMR substituted partially with high level (HC, TMR 1.5 and corn 1.5 g), and with low level of Korean corn grain (LC, TMR 2.25 and corn 0.75 g), respectively. To measure in vitro ruminal pH, gas production, ammonia N and volatile fatty acids (VFA), the in vitro fermentation incubation was triplicated at $39^{\circ}C$, 120 rpm for 0, 1, 3, 6, 12, 24 and 48 h, respectively. Mean ruminal pH was significantly lower (p < 0.05) for HC than control. Changes in rumen pH was rather similar between the groups till 6 h after incubation, but the lowest pH for HC (pH 5.10) appeared at 48 h compared with control and LC. Total gas production was tended (p < 0.09) to be higher and ammonia N was significantly lower (p < 0.05) for HC than control and LC. Total VFA was higher (p < 0.05) for HC and LC than control but no differences appeared between HC and LC. Overall, the present data indicate that feeding different levels of Korean domestic corn grain may lead to high and sustainable starch degradation in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.6
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• pp.821-827
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• 2002

The purpose of this study was to evaluate the effect of protected fat and heat-extruded soybean meal on the lactation performance of Holstein cows. Twenty-four cows, consisting of 20 lactating cows and 4 rumen-fistulated dry cows, were randomly allocated into four groups with 5 lactating cows and 1 fistulated cow in each group. A replicated 4${\times}$4 Latin square design with four 21 day periods, including 14 days of adaptation and 7 collection days within each period was employed. The experiment was a 2${\times}$2 arrangement, with or without heat-extruded soybean meal and protected fat inclusion. The dietary treatments consisted of supplements of (a) soybean meal (the control), (b) heat-extruded soybean meal, (c) protected fat, and (d) heat-extruded soybean meal and protected fat. The results showed that there were no significant differences in feed intake, milk yield, milk protein content, milk lactose content and body weight change between the dietary treatments. However, cows supplemented with protected fat showed a significantly increased (p<0.05) milk fat yield, 3.5% FCM and total solid yield. The increase in undegradable intake protein (UIP) via heat extruded soybean meal supplement significantly decreased the urea nitrogen concentration in the blood (p<0.05). Dietary fat inclusion significantly increased the blood cholesterol concentration (p<0.01) and decreased the ruminal pH value (p<0.01). Increased dietary UIP significantly decreased the ammonia nitrogen concentration in the rumen (p<0.01), but did not significantly influence the pH and VFA molar percentage in the rumen. It appears that dietary protected fat inclusion could improve milk fat and solid content. Increased dietary undegradable intake protein through heat extruded soybean meal did not improve milk yield. But it could alleviate the adverse effect of decreased milk protein due to dietary fat supplementation. Increased UIP could also decrease the ammonia nitrogen concentration in the rumen and plasma urea nitrogen concentration in the blood.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.6
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• pp.832-840
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• 2014

This study aimed to investigate the effects of storage duration and temperature on the characteristics of wet brewers grains (WBG) as feeds for ruminant animals. Four storage temperatures ($5^{\circ}C$, $15^{\circ}C$, $25^{\circ}C$, and $35^{\circ}C$) and four durations (0, 1, 2, and 3 d) were arranged in a $4{\times}4$ factorial design. Surface spoilage, chemical composition and microorganism density were analyzed. An in vitro gas test was also conducted to determine the pH, ammonia-nitrogen and volatile fatty acid (VFA) concentrations after 24 h incubation. Surface spoilage was apparent at higher temperatures such as $25^{\circ}C$ and $35^{\circ}C$. Nutrients contents decreased concomitantly with prolonged storage times (p<0.01) and increasing temperatures (p<0.01). The amount of yeast and mold increased (p<0.05) with increasing storage times and temperatures. As storage temperature increased, gas production, in vitro disappearance of organic matter, pH, ammonia nitrogen and total VFA from the WBG in the rumen decreased (p<0.01). Our results indicate that lower storage temperature promotes longer beneficial use period. However, when storage temperature exceeds $35^{\circ}C$, WBG should be used within a day to prevent impairment of rumen fermentation in the subtropics such as Southeast China, where the temperature is typically above $35^{\circ}C$ during summer.

• Korean Journal of Organic Agriculture
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• v.28 no.4
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• pp.605-614
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• 2020

This study was conducted to investigate the effects of TMR on in vitro rumen fermentation and methane production of goat with different forage sources. The experiment was arranged 4×2 factorial design. The different forage sources were rice straw (RS), Italian rye grass (IR), timothy (TI) and alfalfa (AL), respectively. There were two different forage : concentrate ratios such as 20:80 (20) and 50:50 (50), respectively. Therefore, totally 8 treatments were used: 1) RS20, 2) RS50, 3) IR20, 4) IR50, 5) TI20, 6) TI50, 7) AL20, and 8) AL50, respectively. The rumen fluid of goat was collected from the slaughterhouse. For fermentation parameters, ruminal pH, total gas, methane, hydrogen, ammonia nitrogen, and volatile fatty acid were determined. The pH values were within an optimal range across all treatments. Total gas productions at TI20 and AL50 were significantly greater than others (p<0.05). Methane production was significantly lower in TI and AL compared with other treatments (p<0.05). The relatively high dietary NDF content in treatments showed significantly lower methane production (p<0.05). Significant alterations treatments were detected at ammonia nitrogen concentration according to the ratio of forage : concentrate (p<0.05). AL treatment showed greater total volatile fatty acid production compared with other treatments (p<0.05). Therefore, the present study suggests that both Timothy and Alfalfa could be recommendable forage sources for goat based on results with volatile fatty acid as an energy source and methane as an index for energy loss and environmental issues. Also, the 50:50 (forage : concentrate) ratio would prefer to 20:80.

• Animal Bioscience
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• v.36 no.8
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.6
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• pp.776-782
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• 2019

Objective: Fasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods. Methods: Three ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40. Results: Fasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility. Conclusion: When in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.