• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.987-996
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• 2003

This study was conducted to determine the effects of the artificial culture medium of wild-ginsengs on in vitro fermentation characteristics. NH$_3$-N concentration was showed the highest in 3% WGM treatment among all treatments and control. In addition, microbial protein synthesis was significantly different in all treatments throughout the incubation time, and WGM 3% treatment was the highest at the 9 h incubation(P〈0.05). Protozoa numbers within rumen were decreased in all WGM treatments at 9 h incubation time, whereas WGM 3% treatment was always decreased throughout the incubation(P〈0.05). NDF and ADF digestibility were proportionally increased as the incubation time in both control and treatments. NDF digestibility showed no significantly difference between control and the 3% treatment, and ADF digestibility was similar in all. Total volatile fatty acid(VFA) concentrations of WGM treatments without 5% were significantly higher than control (P〈0.05). No differences were observed in total VFA, acetate, propionate and butyrate concentration among the WGM treatments. Acetate/Propionate ratio of WGM treatments was higher than control after 12 h incubation(P〈0.05). As a result of the artificial culture medium of wild-ginseng on rumen fermentation characteristics in vitro, microbial protein synthesis of WGM treatment was higher than control, and WGM 3% was the highest in all treatments(P〈0.05). The effect of saponin in artificial culture medium of wild-ginseng tended to decrease NH$_3$-N concentration, while it increases the microbial synthesis in early incubation. Therefore, artificial cultures medium of wild-ginseng can increase utilization of feed by microbial and anti-protozoal effects of saponin, which may enhance microbial synthesis capacity in early fermentation period in rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.283-294
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• 2007

Conventional culture-based methods of enumerating rumen microorganisms (bacteria, archaea, protozoa, and fungi) are being rapidly replaced by nucleic acid-based techniques which can be used to characterise complex microbial communities without incubation. The foundation of these techniques is 16S/18S rDNA sequence analysis which has provided a phylogenetically based classification scheme for enumeration and identification of microbial community members. While these analyses are very informative for determining the composition of the microbial community and monitoring changes in population size, they can only infer function based on these observations. The next step in functional analysis of the ecosystem is to measure how specific and, or, predominant members of the ecosystem are operating and interacting with other groups. It is also apparent that techniques which optimise the analysis of complex microbial communities rather than the detection of single organisms will need to address the issues of high throughput analysis using many primers/probes in a single sample. Nearly all the molecular ecological techniques are dependant upon the efficient extraction of high quality DNA/RNA representing the diversity of ruminal microbial communities. Recent reviews and technical manuals written on the subject of molecular microbial ecology of animals provide a broad perspective of the variety of techniques available and their potential application in the field of animal science which is beyond the scope of this treatise. This paper will focus on nucleic acid based molecular methods which have recently been developed for studying major functional groups (cellulolytic bacteria, protozoa, fungi and methanogens) of microorganisms that are important in nutritional studies, as well as, novel methods for studying microbial diversity and function from a genomics perspective.

• 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.

• Korean Journal of Organic Agriculture
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• v.20 no.3
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

• Journal of agriculture & life science
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• v.46 no.3
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• pp.79-85
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• 2012

This study was conducted to examine the effect of dietary n-3/n-6 fatty acid (FA) ratio on in vitro dry matter digestibility (IVDMD), fermentation indices and FA profile. Rice bran was mixed with oil sources (cotton seed oil and linseed oil) to make the diets at 0.02, 0.29 and 0.61 of dietary n-3/n-6 FA ratio. These diets (0.5g) were placed into the incubation bottles with 40 ml of anaerobic culture medium, which contained rumen fluid and Van Soest medium at 1:2 ratio. Five replicates of each diet and two blanks were incubated at $39^{\circ}C$ for 48 hours. After incubation, the incubated contents were centrifuged. The residues were freeze-dried for DMD and FA analyses. The supernatant was used for pH, $NH_3-N$ and volatile fatty acid analyses. The concentrations of lactate (p<0.001) and iso-valerate (p<0.001) decreased linearly with increasing dietary n-3/n-6 FA ratio, but acetate concentration (p=0.056) and the ratio of acetate to propionate (p=0.005) was increased linearly. The concentrations of n-3, n-6 FA and the ratio of n-3/n-6 FA in residues increased (p<0.001) linearly with increasing dietary n-3/n-6 FA ratio, but C18:1n-9 FA concentration was decreased (p<0.001) linearly. With these results, it could affect fermentation characteristics and FA profile of rumen content by dietary n-3/n-6 FA ratio.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.2
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• pp.234-239
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• 2012

The objective of this study was to evaluate the effects of forage level and oil supplement on selected strains of rumen bacteria believed to be involved in biohydrogenation (BH). A continuous culture system consisting of four fermenters was used in a $4{\times}4$ Latin square design with a factorial arrangement of treatments, with four 10 d consecutive periods. Treatment diets were: i) high forage diet (70:30 forage to concentrate (dry matter basis); HFC), ii) high forage plus oil supplement (HFO), iii) low forage diet (30:70 forage to concentrate; LFC), and iv) low forage plus oil supplement (LFO). The oil supplement was a blend of fish oil and soybean oil added at 1 and 2 g/100 g dry matter, respectively. Treatment diets were fed for 10 days and samples were collected from each fermenter on the last day of each period 3 h post morning feeding. The concentrations of vaccenic acid (t11C18:1; VA) and c9t11 conjugated linoleic acid (CLA) were greater with the high forage diet while the concentrations of t10 C18:1 and t10c12 CLA were greater with the low forage diet and addition of oil supplement increased their concentrations at both forage levels. The DNA abundance of Anaerovibrio lipolytica, and Butyrivibrio fibrisolvens vaccenic acid subgroup (Butyrivibrio VA) were lower with the low forage diets but not affected by oil supplement. The DNA abundance of Butyrivibrio fibrisolvens stearic acid producer subgroup (Butyrivibrio SA) was not affected by forage level or oil supplement. In conclusion, oil supplement had no effects on the tested rumen bacteria and forage level affected Anaerovibrio lipolytica and Butyrivibrio VA.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.3
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• pp.197-205
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• 2013

An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.

• Animal Bioscience
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• v.34 no.4
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• pp.584-593
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• 2021

Objective: The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. Methods: Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. Results: Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p<0.05) in vitro DM and neutral detergent fibre disappearance, but this difference was not observed between the mid-bloom Alf and 15-wk FG. Regardless plant maturity, in vitro ruminal fermentation of FG produced less (p<0.001) CH4 either on DM incubated or on DM disappeared basis than that of Alf during 48-h incubation. In vitro ruminal fermentation of FG yielded similar amount of total volatile fatty acids with higher (p<0.05) propionate percentage as compared to fermentation of Alf irrespective of plant maturity. Microbial protein synthesis was greater (p<0.001) with 11-wk FG than early bloom Alf as substrate and 16S RNA gene copies of total bacteria was higher (p<0.01) with 15-wk FG than mid-bloom Alf as substrate. Compared to mid-bloom Alf, 15-wk FG had lower (p<0.05 to 0.001) amount of 16S RNA methanogen gene copies in the whole culture during 48-h incubation. Conclusion: In comparison to Alf, FG emerges as a high quality forage that can not only improve rumen fermentation in vitro, but can also remarkably mitigate CH4 emissions likely due to being rich in saponins.

• Journal of Animal Science and Technology
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• v.52 no.5
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• pp.413-426
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• 2010

This study was conducted to investigate the effects on fermentation characteristics of rumen microorganism by different types and levels of lignosulfonate treated soybean meal (LSBM) in in vitro test and rumen simulation continuous culture (RSCC) system in dairy cows. The experiment I was control and 12 treatments (each with 3 replications) in vitro test to demonstrate composition of different types of treatments with lignosulfonate (Desulfonate, Na, Ca and solution) and levels (2, 4 and 8%) of soybean meal in the dairy cow diet. LSBM source treatments in the dairy cow diet showed pH value, $NH_3$-N concentration and total VFA concentration lower than control at all levels and incubation times (p<0.05). Dry matter digestibility of LSBM source treatments showed lower than control (p<0.05). Gas production and rumen microbial synthesis was decreased by rumen microbial fermentation for incubation times. Undegradable protein (UDP) concentration of all LSBM treatments was decreased for incubation times, and significantly higher than control (p<0.05). In the experiment II compared diets of the control, LSBM Na 2%, LSBM Sol 2%, which are high performance to undegradable protein (UDP) concentration experiment I in vitro test, and heated treatment lignosulfonate (LSBM Heat) 2% in the dairy cow diet from four station RSCC system ($4{\times}4$ Latin square). A rumen microbial fermentation characteristic was stability during 12~15 days of experimental period in all treatments. The pH value of LSBM treatments was higher than control treatment (p<0.05). The $NH_3$-N concentration, VFA concentration and rumen microbial synthesis of LSBM treatments were lower than control (p<0.05). The undegradable protein (UDP) showed LSBM Na 2% (45.28%), LSBM Sol 2% (43.52%) and LSBM Heat 2% (43.49%) higher than control (41.55%), respectively (p<0.05). Those experiments were designed to improve by-pass protein of diet and milk protein in the dairy cows. We will conduct those experiments the in vivo test by LSBM treatments in dairy cows diet.

• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.1019-1030
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• 2003

The study was conducted to isolate and identify highly fibrolytic anaerobic fungi from the guts of a Hanwoo steer and a Korean native goat, and then investigate the characterization of cellulolytic activity of an anaerobic fungus. Twenty-one anaerobic fungal colonies were isolated in the study, in which 16 colonies were isolated from the rumen contents of the Hanwoo steer and 5 colonies from the duodenal fluids of the Korean native goat. Four anaerobic fungi were selected based on higher cellulolytic enzyme activities to identify under a optical microscope. NLRI-M003 and -T004 belong to Neocallimastix genus and NLRI-M014 belongs to Piromyces genus based on the morphology of their thallus, sporangia, rhizoid and the number of flagella. NLRI-M001 appeared to be an unknown strain of anaerobic fungi due to its different morphology from existing types of anaerobic fungi, though the morpholgoy is similar to Orpinomyces sp. Supplementation of 2% anaerobic fungal culture(NLRI-M003) in rumen-mixed microorganisms increased in vitro DM degradability of rice straw and filter paper up to 4 and 11%, respectively, compared with non-supplementation(control). CMCase and xylanase activities in in vitro culture were also higher in 2% fungal supplementation than controls in both rice straw and filter paper substrates.