• Asian-Australasian Journal of Animal Sciences
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• 제18권7호
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• pp.990-996
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• 2005

The effect of diets varying in level and source of nitrogen (N) and fermentable organic matter on dynamic characteristics of microbial populations in rumen liquor and their impact on substrate fermentation in vitro was studied. The diets tested were straw alone, straw+concentrate mixture and straw+urea molasses mineral block (UMMB) lick. The same diets were taken as substrates and tested on each inoculum collected from the diets. Diet had no effect on the amino acid (AA) composition of either bacteria or protozoa. Differences among the diets in intake, source of N and OM affected bacterial and protozoal characteristics in the rumen. Upper asymptote of gas production (Y$\alpha$) had a higher correlation with bacterial pool size and production rate than with protozoal pool size and production rate. Among the parameters of the gas production model, Y$\alpha$ and lag time in total gas has showed significant (p<0.01) correlation with bacterial characteristics. Though the rate constant of gas production significantly differed (p<0.01) between diet and type of straw, it was least influenced by the microbial characteristics. The regression coefficient of diet and type of straw for Y$\alpha$ indicated that the effect of diet on Y$\alpha$ was threefold higher than that of the straw. As microbial characteristics showed higher correlation with Y$\alpha$, and diet had more influence on the microbial characteristics, gas production on a straw diet could be used effectively to understand the microbial characteristics.

• 한국축산시설환경학회지
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• 제19권2호
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• pp.155-162
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• 2013

The objective of this study is to investigate the effects of Cynanchum wilfordii (CW) on cell viability, anti-oxidant activity, volatile fatty acid (VFA) production and methane gas production. Collected rumen fluid incubated with CW powder (1% w/v) for 12 and 24 hours were analyzed for pH, VFAs and methane. Alamar blue assay showed no significant difference on the viability of 3T3-L1 and C2C12 cells treated with CW for 24 hours. TBARS data showed a dose dependent increase on the antioxidant activity of CW. VFAs increased in the CW-treated groups compared to the control group. In addition, propionate increased more than other VFAs by the treatment with CW. There was a significant decrease in methane gas production in batch culture treated with CW in 12hrs. In conclusion, it was suggested that Cynanchum wilfordii could manipulate rumen fermentation considered by increasing VFA production and inhibition of methanogenesis.

• 생명과학회지
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• 제22권9호
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• pp.1187-1193
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• 2012

본 연구는 할로겐 화합물의 첨가가 in vitro 상의 반추위 발효성상과 메탄생성에 미치는 영향에 대한 효과를 규명하고자 실시하였다. Italian rye grass 및 배합사료를 6:4의 비율로 급여한 반추위 cannula가 시술된 홀스타인에서 반추위액을 채취하여 사용하였고, 채취된 반추위액은 분쇄된 timothy (대조구; C)에 bromochloromethane (BCM구), 2-Bromoethanesulfonic acid (BES구), 3-Bromopropanesulfonic acid (BPS구), chloroform (CLF구) 및 Pyromellitic diimide (PMDI구)의 5가지 할로겐 화합물을 각각 1 ppm씩 첨가하여 in vitro 배양하였다. pH는 6.72에서 6.25 정도로 배양시간이 경과함에 따라 낮아지는 경향을 나타내었고, 배양 48시간에는 처리구간 차이가 없었다. 배양 48시간 후의 총 가스 발생량은 처리구간 유의적인(p<0.05) 차이는 없었고, BPS구를 제외한 모든 처리구에서의 메탄 발생량은 대조구에 비해 유의적으로(p<0.05) 감소하였다. 배양 12시간에서의 총 휘발성 지방산 및 propionic acid의 발생량은 처리구가 대조구에 비해 유의적으로(p<0.05) 높았다. 본 실험의 결과, 할로겐 화합물의 첨가는 반추위 내의 pH, 건물 소화율, 미생물 수 및 총 가스 발생량의 발효 성상에는 영향을 주지 않으면서 메탄의 발생량이 감소 되었다. 실험 결과를 종합해 보면, 할로겐 화합물 첨가는 반추위 내 pH, 가스 발생량, 반추위 미생물 성장량 및 propionic acid 모두 증가하였으며, 반추위내 메탄생성을 억제하였다. 앞으로 할로겐화합물과 다른 메탄억제 물질과 혼합하여 반추위 내 메탄생성 억제에 관한 구체적인 연구가 필요한 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권2호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제14권1호
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• pp.48-53
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• 2001

An in vitro study was conducted to examine the effect of sources and the addition levels of carbohydrates on fermentation characteristics, bacterial growth, and hydrogenation of linoleic acid ($C_{18:2}$) by mixed ruminal bacteria. Starch and cellobiose were added to the 200 ml non-selective basal media at the levels of 0.20 and 0.35% (w/v), respectively. Linoleic acid (66.8~79.6 mg) in the absorbed form into the pieces of nylon cloth was also added to the media of 5 treatments including control which was not added with carbohydrate. Three mls of rumen fluid strained through 12 layers of cheese cloth were added to each medium, and were incubated anaerobically in the shaking incubator of $39^{\circ}C$ for 24 hours. During 24 h incubation the pH in incubation media of all treatments was maintained above 6.6 by the addition of sodium bicarbonate. The pH and ammonia concentration of incubation media were not clearly influenced by the sources and addition levels of carbohydrates while additions of carbohydrates increased (p<0.0001) VFA concentration at the 24 h incubation. Molar proportions of acetate were reduced (p<0.0004) while those of propionate were increased (p<0.0006) by the addition of carbohydrates. But the differences in concentration and molar proportions of the VFA were small between the sources or the addition levels. Bacterial growth was faster (p<0.0004) in the starch added treatments than in the cellobiose added ones and control, but no differences were found between addition levels. Increased (p<0.0487) hydrogenation was observed from the starch added treatments compared to the cellobiose added ones, but there was no difference between addition levels.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1083-1095
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• 2019

Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on $NH_3-N$ at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, $NH_3-N$ and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with $10^6CFU/ml$ C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

• 한국환경과학회지
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• 제28권6호
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• pp.527-533
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• 2019

The aim of present study was to investigate the effect of three types of Chestnut Meals (CM) on chemical composition and rumen fermentation characteristics of the fermented diet. The inoculants consisted of Lactobacillus acidophilus, Bacillus subtilis, and Sacaromyces cerevisiae and were applied to three different types of CM; Whole Chestnut (WC), endodermis (EN), and kernel (KE). All types of CMs were ensiled at $39^{\circ}C$ for 0, 1, 2, 4, or 6 days. After ensiling, the fermented CMs were sub-sampled for laboratory assays. On day six of fermentation, counts of the lactic acid-producing Bacillus subtilis, and yeast were higher (P<0.05) in WC than in the other CM types. On day four, KE had higher (P<0.05) crude protein content but lower (P<0.05) neutral detergent fiber and acid detergent fiber contents than the other treatments. In terms of rumen digestibility, KE had the highest (P<0.05) in vitro digestibility of dry matter (IVDMD), neutral detergent fiber digestibility (IVNDFD), total volatile fatty acid (VFA), propionate, butyrate concentrations, and total gas volume, as well as the lowest (P<0.05) acetate concentration. On the other hand, EN had the highest (P<0.05) pH and ammonia-N concentration in the rumen. In the rumen, even though WC application produced the highest microbial count and fermentation characteristics, it did not have a beneficial effect on rumen digestibility. Therefore, this study concluded that application of KE could be recommended due to the observed improvements in IVDMD and IVNDFD.

• Asian-Australasian Journal of Animal Sciences
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• 제28권1호
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• pp.50-57
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• 2015

This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

• Asian-Australasian Journal of Animal Sciences
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• 제3권1호
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• pp.27-31
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• 1990

Influences of plant phenolic acids and their possible metabolites(non-phenolic aromatic acids involved) in the rumen on the cellulolytic activity of mixed rumen populations were examined by a simple in vitro culture technique. Initial concentrations of aromatic acids were 1, 5, 10 and 20 mM/l. All the tested aromatic acids reduced microbial cellulose digestion especially at the higher initial concentration. P-Coumaric acid, ferulic acid and cinnamic acid, those having unhydrogenated propenoic side chain were more inhibitory than were 3-phenylpropinic acid and phloretic acid, those having hydrogenated propanoic side chain. Lag-time for cellulose digestion was prolonged by former three acids by 16 h. Apparent reduction in p-coumaric acid concentration was observed at 24 h when cellulose digestion began. Volatile fatty acid productions from cellulose fermentation were shifted by former three aromatic acids to produce more acetate and less propionate. This suggests that the selection of celluloytic organisms was induced by these aromatic acids.

• 한국초지조사료학회지
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• 제31권2호
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• pp.177-190
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• 2011

본 연구는 허브 부식토를 이용하여 첨가 수준별 in vitro 반추위 발효특성 평가와 젖소를 이용하여 급여시 유생산성에 미치는 영향을 조사하기 위하여 본 연구를 수행하였다. 시험 1에서는 티머시 건초를 기질로 하여 허브부식토(herbaceous peat)를 0,1 및 5%를 3반복으로 각각 첨가하여 in vitro 반추위내 pH, 가스발생량, VFA (volatile fatty acid), ammonia-N 및 건물분해율을 조사하여 반추위내 발효성상의 변화를 평가하였다. pH 변화는 0, 3, 12 및 24시간 배양에 있어서 모든 처리구에서 유의한 차이를 보이지 않았지만 6시간. 대조구에서 1 및 5% 첨가구와 비교하여 유의하게 낮은 pH를 나타냈다(p<0.05).가스 발생량은 배양 12시간까지 처리구에서 대조구와 비교하여 유의하게 증가 하였으며(p<0.05), 반추위액내 암모니아 농도는 모든 처리구에서 24시간까지 증가하는 경향을 나타냈으며, 처리간 유의한 차이는 없었다. 건물분해율은 모든 시간에서 5%구가 대조구 및 1%구와 비교하여 건물분해율이 높았다(p< 0.05) 따라서 허브부식토를 0, 1 및 5% 수준으로 각각 첨가하여 반추위내 in vitro 배양시간 별 반추위 발효특성은 대부분 조사항목에서 처리구가 대조구에 비해 개선효과를 나타냈으며, 시험 2에서는 젖소를 이용하여 사양시험을 통해 유생산성을 평가하였으며, 우유생산량 변화와 유성분 및 체세포 수 변화를 조사하기 위해 홀스타인 착유우 16두를 공시하여 4주간 사양 시험을 실시하였다. 산유량은 T3구가 25.0 kg으로 대조구(23.2 kg), 부식토 처리구(23.1 kg), 비타민 C 처리구(23.4 kg)와 비슷하여 수치적으로 증가하였다. 대조구와 처리구별 시험 시작 전 유성분(유지방, 유단백, 유당, MUN 및 SNF) 및 체세포 수의 변화는 일부 유의성이 인정되었다(p<0.05). 4주간의 시험 기간 중 전반기(1~2주)와 후반기(3~4주)의 유성분 변화를 보면 부식토 첨가구에 있어서 유단백질과 SNF가 유의적으로 증가하는 경향을 나타내었다. 대조구와 처리구의 시험 시작 전 의 혈액내 영양성분 (총단백질, 콜레스테롤, NEFA, BUN), 간기능 성분(AST, GGT) 및 무기물(Ca, P, Mg)에서 뚜렷한 변화를 나타내지 않았다. 이상의 결과를 종합해 볼 때 착유우에 부식토를 급여하는 것이 유성분, 체세포 수 및 생리적으로 긍정적인 영향을 줄 것으로 판단된다.