• Asian-Australasian Journal of Animal Sciences
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• 제22권3호
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• pp.356-364
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• 2009

The current ruminant feeding models require parameterization of the digestion kinetics of carbohydrate fractions in feed ingredients to estimate the supply of nutrients from a ration. Using an automated gas production technique, statistically welldefined digestion rate of carbohydrate, including soluble carbohydrate, can be estimated in a relatively easy way. In this study, the gas production during in vitro fermentation was measured and recorded by an automated gas production system to investigate degradation kinetics of carbohydrate fractions of a wide range of ruminant feeds: corn silage, rice straw, corn, soybean hull, soybean meal, and cell mass from lysine production (CMLP). The gas production from un-fractionated, ethanol insoluble residue and neutral detergent insoluble residue of the feed samples were obtained. The gas profiles of carbohydrate fractions on the basis of the carbohydrate scheme of the Cornell Net Carbohydrate and Protein System (A, B1, B2, B3 and C) were generated using a subtraction approach. After the gas profiles were plotted with time, a curve was fitted with a single-pool exponential equation with a discrete lag to obtain kinetic parameters that can be used as inputs for modern nutritional models. The fractional degradation rate constants (Kd) of corn silage were 11.6, 25.7, 14.8 and 0.8%/h for un-fractioned, A, B1 and B2 fractions, respectively. The values were statistically well estimated, assessed by high t-value (>12.9). The Kd of carbohydrate fractions in rice straw were 4.8, 21.1, 5.7 and 0.5%/h for un-fractioned, A, B1 and B2 fractions, respectively. Although the Kd of B2 fraction was poorly defined with a t-value of 4.4, the Kd of the other fractions showed tvalues higher than 21.9. The un-fractioned corn showed the highest Kd (18.2%/h) among the feeds tested, and the Kd of A plus B1 fraction was 18.7%/h. Soybean hull had a Kd of 6.0, 29.0, 3.8 and 13.8%/h for un-fractioned, A, B1 and B2, respectively. The large Kd of fraction B2 indicated that NDF in soybean hull was easily degradable. The t-values were higher than 20 except for the B1 fraction (5.7). The estimated Kd of soybean meal was 9.6, 24.3, 5.0%/h for un-fractioned, A and B1 fractions, respectively. A small amount of gas (5.6 ml at 48 ho of incubation) was produced from fermentation of CMLP which contained little carbohydrate. In summary, the automated gas production system was satisfactory for the estimation of well defined (t-value >12) kinetic parameters and Kd of soluble carbohydrate fractions of various feedstuffs that supply mainly carbohydrate. The subtraction approach, however, should be applied with caution for some concentrates, especially those which contain a high level of crude protein since nitrogen-containing compounds can interfere with gas production.

• Asian-Australasian Journal of Animal Sciences
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• 제29권2호
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• pp.230-240
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• 2016

Information on the effects of different yeast species on ruminal fermentation is limited. This experiment was conducted in a $3{\times}4$ factorial arrangement to explore and compare the effects of addition of three different live yeast species (Candida utilis 1314, Saccharomyces cerevisiae 1355, and Candida tropicalis 1254) at four doses (0, $0.25{\times}10^7$, $0.50{\times}10^7$, and $0.75{\times}10^7$ colony-forming unit [cfu]) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover, and rice straw by mixed rumen microorganisms in dairy cows. The maximum gas production (Vf), dry matter disappearance (IVDMD), neutral detergent fiber disappearance (IVNDFD), and methane production in C. utilis group were less (p<0.01) than other two live yeast supplemented groups. The inclusion of S. cerevisiae reduced (p<0.01) the concentrations of ammonia nitrogen ($NH_3$-N), isobutyrate, and isovalerate compared to the other two yeast groups. C. tropicalis addition generally enhanced (p<0.05) IVDMD and IVNDFD. The $NH_3$-N concentration and $CH_4$ production were increased (p<0.05) by the addition of S. cerevisiae and C. tropicalis compared with the control. Supplementation of three yeast species decreased (p<0.05) or numerically decreased the ratio of acetate to propionate. The current results indicate that C. tropicalis is more preferred as yeast culture supplements, and its optimal dose should be $0.25{\times}10^7$ cfu/500 mg substrates in vitro.

• Asian-Australasian Journal of Animal Sciences
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• 제27권3호
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• pp.330-341
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• 2014

Andropogon gayanus is an important grass due to its high biomass production, drought tolerance and favorable growth on low fertility acidic soils. Currently, there is little research on the impact of growth stage on the nutritional quality or the degree of $CH_4$ production that may arise from this forage during ruminal fermentation. The objectives of this study were to determine the effects of regrowth stage of A. gayanus on its chemical composition, in vitro production of gas and CH4, as well as in vitro dry matter (DM) digestibility when grown under tropical Brazilian conditions and conserved as hay or as silage. The nutritional value of A. gayanus grass declined with increasing maturity; however digestible DM yield linearly increased. After 112 d of regrowth, A. gayanus produced higher quality silage (higher lactate and lower pH and butyrate content) and higher DM yield. However, the low levels of crude protein at this time would make protein supplementation a necessity for proper rumen fermentation. No differences in $CH_4$ kinetic parameters were found with advancing maturity or preservation method (hay or silage).

• Animal Bioscience
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• 제35권10호
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Asian-Australasian Journal of Animal Sciences
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• 제31권3호
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• pp.354-362
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• 2018

Objective: Wheat is an alternative to corn silage for ruminant feeding in northern China. This study examined the effects of harvest time and added molasses on nutritional content, ensiling characteristics and in vitro degradation of whole crop wheat (WCW). Methods: Fresh WCW at the milk-ripe stage was harvested at 0700 h (i.e., in the morning [Mo]) and 1700 h (i.e., in the afternoon [Af]), and then immediately used to prepare silage and make hay. Commercial molasses was added to Af WCW at 0%, 2%, 4%, and 6% (fresh weight) proportions. The WCW treated with molasses was mixed thoroughly prior to ensiling. Results: Dry matter (DM), neutral detergent fiber, water soluble carbohydrate (WSC) content (p<0.01), accumulative gas production in 72 h ($GP_{72h}$, 77.46 mL/g vs 95.15 mL/g) and dry matter disappearance in vitro (69.15% vs 76.77%) were lower (p<0.05), while crude protein (CP) content was higher for WCW silage (WCWS) compared to WCW (p<0.01). The propionic acid and butyric acid concentrations in WCWS from Mo WCW were 1.47% and 0.26%, respectively. However, the propionic and butyric acid concentrations were negligible, while the ammonia nitrogen/total nitrogen ($NH_3-N/TN$, p<0.01) concentration was lower and the rate of gas production at 50% of the maximum (17.05 mL/h vs 13.94 mL/h, p<0.05) was higher for Af WCWS compared to Mo WCWS. The incubation fluid's $NH_3-N$ concentration was lower in WCWS and Af WCW compared to Mo WCW (p<0.05). The CP and WSC content increased with increasing molasses levels (p<0.05). Furthermore, the pH (p<0.01) and time when gas production was 50% of the maximum (2.78 h vs 3.05 h, p<0.05) were lower in silage treated with 4% molasses than silage without molasses. Conclusion: Harvesting wheat crops in the afternoon and adding molasses at 4% level to WCW optimally improved ensiling characteristics, leading to well-preserved silage.

• 한국초지조사료학회지
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• 제32권4호
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• pp.379-386
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• 2012

In this study, the in vitro fermentation parameters of whole crop barley (WCBS-TMR) and Italian ryegrass (IRGS-TMR) silage total mixed rations were compared. A rice straw based diet (RSBD), which was a mixture of rice straw and concentrate (60:40), was used as the control. The feeds were incubated in buffered rumen fluid for 3, 6, 9, 12, 24, 48 and 72 hours at $39^{\circ}C$. At the end of each incubation period the following parameters were determined, total gas, pH, ammonia nitrogen ($NH_3$-N), volatile fatty acids (VFA) and then the acetate to propionate ratio (A/P) was calculated. The dietary treatments did not affect (p>0.05) the overall production of $NH_3$-N, gas, total VFA and all the individual VFA, with the exception of n-butyrate (p<0.001). The treatment diets significantly affected the A/P ratio (p<0.01). The control diet resulted in the lowest A/P ratios, followed by WCBS-TMR and lastly IRGS-TMR had the highest ratios. Gas production was not different between treatments, suggesting a probable similar level of digestibility when treatments are fed to animals. It can therefore be concluded from the present study that WCBS and IRGS are of almost an equivalent nutritional value when incubated in a TMR form. WCBS-TMR however resulted in lower A/P ratios than IRGS-TMR, which is indicative of a more energy efficient diet.

• Asian-Australasian Journal of Animal Sciences
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• 제27권2호
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• pp.169-178
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• 2014

Samples of Clitoria ternatea L. (Cunh$\tilde{a}$) were harvested at 35, 50, 70, and 90 d after a uniformity harvest in a field study designed as a completely randomized design with a total of 18 experimental plots. The dry matter yield of the whole plant was separated quantitatively into leaves, stems, and pods at each harvesting age. Chemical analyses and in vitro gas production kinetics were performed to assess the quality of the plant parts. Yields, chemical composition, and estimates of gas production parameters were analyzed by fitting a mixed statistical model with two types of covariance structures as follows: variance components and an unrestricted structure with heterogeneous variances. Fast and slow gas yielding pools were detected for both leaves and stems, but only a single pool was detected for pods. The homoscedasticity assumption was more likely for all variables, except for some parameters of the gas production kinetics of leaves and stems. There was no presence of typical pods at 35 and 50 d. In the leaves, the fibrous fractions were affected, whereas the non-fibrous fractions were unaffected by the harvesting age. The harvesting age affected the majority of the chemical constituents and gas kinetic parameters related to the stems. The leaves of this legume were the least affected part by the aging process.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1518-1523
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• 2004

This study was conducted to evaluate nutritive value of forages commonly used as ruminant feeds in North Sumatra, Indonesia. Seven species of grasses and five species of legumes were collected during the rainy season. The results showed that chemical composition, in vitro digestibility of dry matter (DMD), organic matter (OMD) and crude protein (CPD), in vitro gas production and metabolizable energy (ME) content greatly varied among the species of grass and legume forages. The CP content ranged from 6.6 to 16.2% in grass and from 17.5 to 29.1% in legumes; while NDF content of grass and legume ranged from 57.2 to 66.2% and from 24.4 to 55.6%, respectively. The DMD, OMD and CPD of grass ranged from 49.1 to 62.2%, 51.9 to 64.4% and 50.5 to 60.3%; while in legumes the values ranged from 59.1 to 71.8%, 65.2 to 72.0% and 68.2 to 71.6%, respectively. The ME content of grass varied from 6.4 to 9.3 MJ/kg and from 6.5 to 8.3 MJ/kg for legumes. In general, within species of grass Cynodon plectostachyus contained higher CP but was lower in NDF that resulted in much higher digestibility; a similar result was also found in Leucaena leucocephala for the legumes. The two forages also contained much higher ME than the others. In conclusion, the nutritive value of forages in North Sumatra, Indonesia during the rainy season was relatively high as ruminant feed, with the best quality noted for Cynodon plectostachyus and Leucaena leucocephala.

• Asian-Australasian Journal of Animal Sciences
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• 제32권6호
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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.

• 동물자원연구
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• 제30권3호
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• pp.111-120
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• 2019

생물학적 평가 방법 중 in situ와 in vitro 평가 방법은 비용 및 시간을 절약할 수 있으며 동물 복지 측면에서 제약이 적어 주목되고 있는 방법이다. In vitro 실험에서 사용되는 사료 접종법으로 배양병에 직접 사료를 담는 dispersion 방법은 실제 반추위 발효 상황에 근접한 조건에서 실험할 수 있는 장점이 있지만, 시간과 노동력이 많이 소모된다. 반대로 사료 주머니를 이용한 방법은 실험 수행의 편이성이 있지만 첨가제 실험 수행의 제약이 있으며, 미생물 활성과 섬유소 소화율이 저하되는 단점이 있다. 본 실험은 dispersion 방법과 사료 주머니 방법을 조합하여 보다 편리한 방법으로 dispersion 에 가까운 높은 재현성의 발효 성상과 소화율 평가 가능할 것이라고 생각하여 in vitro 발효 성상과 미생물 조성을 비교하였다. Dispersion 방법과 nylon bag 을 조합한 실험에서 건물 소화율은 세척 과정에서 미생물 및 사료 입자의 유실로 과대평가될 수 있지만 이론적 최대 가스 발생량 V_max, 가스 발생량에 대한 분해 속도 상수 K_g 에서 유의적 차이가 나타나지 않으며, IVNDFD, pH, NH₃-N, total VFA 발생량 그리고 VFA 조성에서도 유의적 차이가 나지 않았다. Real-time PCR 을 이용한 미생물 조성 분석에서는 general bacteria의 절대량에서 유의적 차이가 나지 않았다. 상기의 결과를 종합해 볼 때, in vitro 실험에서 사료 주머니의 사용은 충분히 적용가능하며 일반적인 방법인 dispersion과 비교해 볼 때, 영양소 이용에 있어 반추미생물의 사료 접촉을 방해하지 않을 것으로 생각된다. 하지만 in vitro bach culture 방법은 배양조건에서 제약이 있어, 향후 연속 배양 방법과 in situ의 실험을 통해 본 실험의 실험결과에 대한 추가적인 규명이 필요할 것으로 사료된다.