• Korean Journal of Agricultural Science
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• v.47 no.1
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• pp.131-138
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• 2020

The aim of this study was to evaluate the effect of aflatoxin B₁ (AFB₁) on in vitro rumen fermentation at various dose levels of 0 (T₁), 100 (T₂), 200 (T₃), and 300 (T₄) ppb in a wheat straw-based buffalo diet. The results show that the truly degradable dry matter, truly degradable organic matter, gas production, microbial biomass production and partitioning factor values in the control group (T₁) were higher (p < 0.05) than those of the T₂, T₃, and T₄ groups. The total volatile fatty acids, acetate, propionate, and butyrate values in the control group (T₁) were higher (p < 0.05) than those of the T₂, T₃, and T₄ groups. The partitioning factor value in the control group (T₁) was higher (p < 0.05) than those of the T₂, T₃, and T₄ groups. The partitioning factor values of the T₂ and T₃ groups were higher (p < 0.05) than that of the T₄ group. There was no significant variation in the partitioning factor value between the T₂ and T₃ group. The acetate : propionate (A : P) ratio in the control group (T₁) was lower (p < 0.05) than those of the T₂, T₃, and T₄ groups. The A : P ratio in the T₂ group was lower (p < 0.05) than those of the T₃ and T₄ groups. It was concluded that different levels of AFB₁ contamination in feed significantly affect the in vitro rumen fermentation characteristics. Thus, these findings could help to determine the influences of AFB₁ in a wheat straw-based buffalo diet. Additionally, it is necessary to manage AFB₁ contamination in ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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• pp.1280-1286
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• 2016

This study was aimed to evaluate the in vitro effects of medicinal herb extracts (MHEs) on ruminal fermentation characteristics and the inhibition of protozoa to reduce methane production in the rumen. A fistulated Hanwoo was used as a donor of rumen fluid. The MHEs (T1, Veratrum patulum; T2, Iris ensata var. spontanea; T3, Arisaema ringens; T4, Carduus crispus; T5, Pueraria thunbergiana) were added to the in vitro fermentation bottles containing the rumen fluid and medium. Total volatile fatty acid (tVFA), total gas production, gas profiles, and the ruminal microbe communities were measured. The tVFA concentration was increased or decreased as compared to the control, and there was a significant (p<0.05) difference after 24 h incubation. pH and ruminal disappearance of dry matter did not show significant difference. As the in vitro ruminal fermentation progressed, total gas production in added MHEs was increased, while the methane production was decreased compared to the control. In particular, Arisaema ringens extract led to decrease methane production by more than 43%. In addition, the result of real-time polymerase chain reaction indicted that the protozoa population in all added MHEs decreased more than that of the control. In conclusion, the results of this study indicated that MHEs could have properties that decrease ruminal methanogenesis by inhibiting protozoa species and might be promising feed additives for ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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• pp.517-522
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• 2013

The effect on methanogens attached to the surface of rumen ciliate protozoa by the addition of plant extracts (pine needles and ginkgo leaves) was studied with particular reference to their effectiveness for decreasing methane emission. The plant extracts (pine needles and ginkgo leaves) were added to an in vitro fermentation incubated with rumen fluid. The microbial population including bacteria, ciliated-associated methanogen, four different groups of methanogens and Fibrobacter succinogenes were quantified by using the real-time PCR. Gas profiles including methane, carbon dioxide and hydrogen, and runinal fermentation characteristics were observed in vitro. The methane emission from samples with an addition of individual juices from pine needles, ginkgo leaves and 70% ethanol extract from ginko leaves was significantly lower (p<0.05, 27.1, 28.1 and 28.1 vs 34.0 ml/g DM) than that of the control, respectively. Total VFAs in samples with an addition of any of the plant extracts were significantly lower than that of the control (p<0.05) as well. The order Methanococcales and the order Methanosarcinales were not detected by using PCR in any incubated mixtures. The ciliate-associated methanogens population decreased from 25% to 49% in the plant extacts as compared to control. We speculate that the supplementation of juice from pine needles and ginkgo leaves extract (70% ethanol extract) decreased the protozoa population resulting in a reduction of methane emission in the rumen and thus inhibiting methanogenesis. The order Methanobacteriales community was affected by addition of all plant extracts and decreased to less than the control, while the order Methanomicrobiales population showed an increase to more than that of the control. The F. succinogenes, the major fibrolytic microorganism, population in all added plant extracts was increased to greater than that of the control. In conclusion, pine needles and ginkgo leaves extracts appear to have properties that decrease methanogenesis by inhibiting protozoa species and may have a potential for use as additives for ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.5
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• pp.639-646
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• 2015

The effects of Saccharomyces cerevisiae supplementation ($6.6{\times}10^8cfu$) and anhydrous ammonia treatment (3%) of wheat straw (WS) were investigated on in-situ dry matter (DM) degradability, and on rumen fermentation and growth performance of lambs. Rumen-fistulated Menemen sheep fed a diet with and without live yeast were used to assess the DM degradability characteristics of WS and ammonia-treated wheat straw ($WS_{NH3}$). Twenty-six yearling Menemen male lambs were fed in four groups. Lambs of control group (WS) received untreated WS without supplemental yeast, whereas other three groups were fed WS treated with anhydrous ammonia ($WS_{NH3}$ group), untreated WS and yeast (WS+YEAST group) or WS treated with anhydrous ammonia and yeast ($WS_{NH3}$+YEAST group). Supplemented live yeast (4 g/d) was added in the diet. Lambs were offered untreated or ammonia treated WS ad-libitum and concentrate was fed at 1% of live body weight. The degradability of the water-insoluble (fraction B) was significantly increased by all of the treatment groups. Potential degradability (A+B), effective DM degradability's (pe2, pe5, and pe8) and average daily weight gain increased only in $WS_{NH3}$+YEAST group (p<0.05). Voluntary DM intake was not increased by the treatments (p>0.05), but voluntary metabolizable energy and crude protein intake were increased by $WS_{NH3}$ and by $WS_{NH3}$+YEAST (p<0.05). Average daily rumen pH was not affected by any of the treatments, but average daily $NH_3$-N was significantly higher in the $WS_{NH3}$ and $WS_{NH3}$+YEAST groups, and total volatile fatty acids were significantly higher in the WS+YEAST and $WS_{NH3}$+YEAST groups. In conclusion, the improvement of feed value of WS was better by the combination of ammonia-treatment and yeast supplementation compared to either treatment alone.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.607-615
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• 2018

Objective: This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results: The maximum activity of recombinant Cel5A (rCel5A) was observed at $50^{\circ}C$ and pH 4.0. The enzyme was constant at the temperature range of $20^{\circ}C$ to $40^{\circ}C$ but also, at the pH range of 3 to 9. The metal ions including $Ca^{2+}$, $K^+$, $Ni^{2+}$,$Mg^{2+}$, and $Fe^{2+}$ increased the endoglucanase activity but the addition of $Mn^{2+}$, $Cu^{2+}$, and $Zn^{2+}$ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and $45.66{\mu}mol/min/mg$. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was $96.69(s^{-1})$ and 6.88 (mL/mg/s), respectively. Conclusion: Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

• Journal of Animal Science and Technology
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• v.48 no.3
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• pp.393-400
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• 2006

The present study was conducted to evaluate feeding value of brown rice in Korean native beef steers (Hanwoo) fed concentrates and orchardgrass hay (50:50 on a wet basis) as a basal diet. Either 50% or 100% of corn in concentrates was substituted for brown rice, and ruminal fermentation characteristics, in situ degradability of feeds and in vivo digestibility of concentrate were investigated. The digestibility and nutritive values of corn and brown rice feeds were similar, whereas rumen degradability of brown rice was higher than that of corn. Compared with corn-based diets, the brown rice substitution decreased (P<0.05) ruminal NH3-N concentration, but did not affect rumen pH. Total volatile fatty acid content in ruminal digesta for the corn based diet was higher than those for the brown rice-contained diets. The brown rice substitution increased (P<0.05) the ratio of ruminal acetic acid to propionic acid (A/P) compared with corn-contained diets.Present results indicate that corn might nutritionally be substituted for brown rice as cattle feeds. In addition, we expect that relatively high A/P with brown rice may produce affirmative effects in rumen physiological aspect, particularly under the grain based feeding system for beef cattle in Korea.

• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.2
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• pp.165-174
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• 2012

This study was conducted to develop Miscanthus as a new roughage resource for ruminant animals. Miscanthus sacchariflorus var. No 1, a newly developed germtype in Korea, was harvested at late maturity stage and its effect on rumen pH, ammonia nitrogen, gas production, volatile fatty acid (VFA) production and digestibility were evaluated using in vitro rumen fermentation. The effects of Miscanthus were compared with rice straw. Miscanthus showed significantly higher pH compared to rice straw (p<0.01). As for ammonia nitrogen, there was no significant difference after 12 h of incubation (p>0.05). Gas production in Miscanthus was significantly lower than that of rice straw in overall incubation time (p<0.05) after 6 h of incubation. In VFA production, acetate, propionate, butyrate, valerate and total VFA production in Miscanthus were lower than those in rice straw. However, production of iso-butyrate and iso-valerate were not different in between two forage materials. Dry matter digestibility of Miscanthus was significantly lower than rice straw (p<0.05) during 12~24 h of incubation. As a result, the availability of Miscanthus as roughage source showed approximately 80% that of rice straw.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.7
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• pp.937-945
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• 2014

Green and black tea by-products, obtained from ready-made tea industry, were ensiled at $10^{\circ}C$, $20^{\circ}C$, and $30^{\circ}C$. Green tea by-product silage (GTS) and black tea by-product silage (BTS) were opened at 5, 10, 45 days after ensiling. Fermentation characteristics and nutrient composition, including tannins, were monitored and the silages on day 45 were subjected to in vitro ruminal fermentation to assess anti-nutritive effects of tannins using polyethylene glycol (PEG) as a tannin-binding agent. Results showed that the GTS and BTS silages were stable and fermented slightly when ensiled at $10^{\circ}C$. The GTS stored at $20^{\circ}C$ and $30^{\circ}C$ showed rapid pH decline and high acetic acid concentration. The BTS was fermented gradually with moderate change of pH and acid concentration. Acetic acid was the main acid product of fermentation in both GTS and BTS. The contents of total extractable phenolics and total extractable tannins in both silages were unaffected by storage temperatures, but condensed tannins in GTS were less when stored at high temperature. The GTS showed no PEG response on in vitro gas production, and revealed only a small increase by PEG on $NH_3$-N concentration. Storage temperature of GTS did not affect the extent of PEG response to both gas production and $NH_3$-N concentration. On the other hand, addition of PEG on BTS markedly increased both the gas production and $NH_3$-N concentration at any ensiled temperature. It can be concluded that tannins in both GTS and BTS suppressed rumen fermentation, and tannins in GTS did more weakly than that in BTS. Ensiling temperature for both tea by-products did not affect the tannin's activity in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.12
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• pp.1721-1725
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• 2014

The objective of this study was to evaluate the in vitro effects of coconut materials on ruminal methanogenesis and fermentation characteristics, in particular their effectiveness for mitigating ruminal methanogenesis. Fistulated Holstein cows were used as the donor of rumen fluid. Coconut materials were added to an in vitro fermentation incubated with rumen fluid-buffer mixture and timothy substrate for 24 h incubation. Total gas production, gas profiles, total volatile fatty acids (tVFAs) and the ruminal methanogens diversity were measured. Although gas profiles in added coconut oil and coconut powder were not significantly different, in vitro ruminal methane production was decreased with the level of reduction between 15% and 19% as compared to control, respectively. Coconut oil and coconut powder also inhibited gas production. The tVFAs concentration was increased by coconut materials, but was not affected significantly as compared to control. Acetate concentration was significantly lower (p<0.05), while propionate was significantly higher (p<0.05) by addition of the coconut materials than that of the control. The acetate:propionate ratio was significantly lowered with addition of coconut oil and coconut powder (p<0.05). The methanogens and ciliate-associated methanogens in all added coconut materials were shown to decrease as compared with control. This study showed that ciliate-associated methanogens diversity was reduced by more than 50% in both coconut oil and coconut powder treatments. In conclusion, these results indicate that coconut powder is a potential agent for decreasing in vitro ruminal methane production and as effective as coconut oil.

• 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.