• Asian-Australasian Journal of Animal Sciences
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• v.13 no.2
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• pp.176-181
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• 2000

In order to clarify the inhibitory effects of orchardgrass (Dactylis glomerata L.) lipids on ruminal fermentation and digestion, two experiments were carried out in vitro. Experiment 1 was carried out using residues of grass hay from which the lipid fraction was removed by ether extraction. To ground grass samples were added 0, 1.5, 3.0, 4.5 and 6.0% lipids and incubated anaerobically at $39^{\circ}C$ for 24 h, with the mixtures of artificial saliva and rumen fluid. Increasing grass lipid levels remarkably reduced DM and NDF disappearances. Volatile fatty acid concentration was significantly reduced at 3.0, 4.5 and 6.0% lipid levels. Microbial nitrogen proportion to total nitrogen tended to decrease by the addition of the lipids. These results indicated that grass lipids have a marked inhibitory effect on ruminal fermentation and digestion, especially when to the substrate was added 3% or more grass lipids as ether extracts. Experiment 2 was conducted to study the relationship between changes in the free fatty acids and changes in the fermentation traits. Samples were incubated for 3, 6, 9, 12, 15, 18, 21 and 24 h as a sole substrate. The polyunsaturated fatty acids steadily decreased during incubation, whereas the saturated fatty acid ($C_{18:0}$) increased. It was suggested that the hydrogenation was extended during the initial stage of incubation. The unsaturated fatty acids ($C_{18:2}$, $C_{18:3}$) produced at the initial stage of incubation were negatively correlated with the amount of microbial N and DM disappearance, indicating that polyunsaturated fatty acids had the possibility to show an inhibiting effect on ruminal fermentation and digestion.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.7
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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.

• Journal of Animal Environmental Science
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• v.19 no.2
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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.

• Journal of Life Science
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• v.22 no.9
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• pp.1187-1193
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• 2012

This study was conducted to evaluate effects of halogenated compounds on in vitro rumen fermentation characteristics and methane emissions. A fistulated Holstein cow of 650 kg body weight was used as a donor of rumen fluid. Five kinds of halogenated compounds (bromochloromethane (BCM), 2-bromoethane sulfonic acid (BES), 3-bromopropanesulfonic acid (BPS), chloroform (CLF), and pyromellitic diimide (PMDI) known to inhibit methyl-coenzyme M reductase activity were added to an in vitro fermentation incubated with rumen fluid. The microbial population including bacteria, protozoa, and fungi were enumerated, and gas production including methane and fermentation characteristics were observed in vitro. The pH values ranged from 6.25 to 6.72 in all the treatments, and these showed a similar level at 48 hr. The total gas production in the treatments showed a similar pattern with C at 48 hr, whereas methane production in the treatments was lower (p<0.05) than C. Concentrations of total volatile fatty acids (VFAs) and propionic acid were higher (p<0.05) in the treatments than in C at 12 hr. Therefore, halogenated compounds (BCM, BES, BPS, CLF, and PMDI) inhibited in vitro methane emissions by inhibiting methanogens in the rumen. Further studies on safety are needed.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.1
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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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• v.29 no.7
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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.

• Journal of Environmental Science International
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• v.28 no.6
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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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• v.28 no.1
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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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• v.3 no.1
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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.