• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.30 no.3
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• pp.111-120
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• 2019

The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH₃-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH₃-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

• Journal of The Korean Society of Grassland and Forage Science
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• v.41 no.2
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• pp.119-127
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• 2021

The objective of this study was to evaluate effects of feeding methods on in vitro ruminal fermentation, total gas and methane production in Hanwoo steers. Six Hanwoo steers fitted with rumen cannula (430 ± 21 kg of body weight) were randomly assigned to one of three feeding systems: 1) feeding forage 1 hour after concentrate, 2) feeding concentrate 1 hour after forage, 3) feeding mixed ration. Rumen fluid sampled from each animals was incubated 24 hours with maize or timothy substrates in in vitro. Ruminal pH was increased in feeding method 2 or maize substrate than that of other methods or timothy substrate (P < 0.001). The production of total volatile fatty acid, acetate, propionate, butyrate, and valerate were increased when steers fed diets using feeding method 1 or rumen fluid was incubated with maize substrate (P < 0.001). Increased production of total gas and methane was observed in feeding method 1 and maize substrate compared to those of other methods or timothy substrate (P < 0.001). Due to the inconsistent results between ruminal fermentation and gas production in this study, further research is required to estimate effects of feeding method on enteric fermentation and gas production in in vivo.

• Journal of Animal Science and Technology
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• v.60 no.11
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• pp.26.1-26.9
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• 2018

Background: This experiment aimed at assessing polyphenol-rich plant biomass to use in complete feed making for the feeding of ruminants. Methods: An in vitro ruminal evaluation of complete blocks (CFB) with (Acacia nilotica, Ziziphus nummularia leaves) and without (Vigna sinensis hay) polyphenol rich plant leaves was conducted by applying Menke's in vitro gas production (IVGP) technique. A total of six substrates, viz. three forages and three CFBs were subjected to in vitro ruminal fermentation in glass syringes to assess gas and methane production, substrate degradability, and rumen fermentation metabolites. Results: Total polyphenol content (g/Kg) was 163 in A. nilotica compared to 52.5 in Z. nummularia with a contrasting difference in tannin fractions, higher hydrolysable tannins (HT) in the former (140.1 vs 2.8) and higher condensed (CT) tannins in the later (28.3 vs 7.9). The potential gas production was lower with a higher lag phase (L) in CT containing Z. nummularia and the component feed block. A. nilotica alone and as a constituent of CFB produced higher total gas but with lower methane while the partitioning factor (PF) was higher in Z. nummularia and its CFB. Substrate digestibility (both DM and OM) was lower (P < 0.001) in Z. nummularia compared to other forages and CFBs. The fermentation metabolites showed a different pattern for forages and their CFBs. The forages showed higher TCA precipitable N and lower acetate: propionate ratio in Z. nummularia while the related trend was found in CFB with V. sinensis. Total volatile fatty acid concentration was higher (P < 0.001) in A. nilotica leaves than V. sinensis hay and Z. nummularia leaves. It has implication on widening the forage resources and providing opportunity to use forage biomass rich in polyphenolic constituents in judicious proportion for reducing methane and enhancing green livestock production. Conclusion: Above all, higher substrate degradability, propionate production, lower methanogenesis in CFB with A. nilotica leaves may be considered useful. Nevertheless, CFB with Z. nummularia also proved its usefulness with higher TCA precipitable N and PF. It has implication on widening the forage resources and providing opportunity to use polyphenol-rich forage biomass for reducing methane and enhancing green livestock production.

• Korean Journal of Organic Agriculture
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• v.23 no.2
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• pp.373-390
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• 2015

This study investigated the effect of coated feed using mixed oil on rumen fermentation characteristics. Two experiments were conducted based on materials that were mixed. First, cashew nut and soybean oils were mixed with white mineral oil. And second, different plant extracts were mixed with white mineral oil. At first experiment, inclusion levels of mixed oil on diet (0.03%, 0.1%, 0.5%) were applied as variables. A coated diet was fermented with rumen inoculum according to in vitro rumen fermentation and its parameters were investigated. In the result of first experiment, no negative effects on rumen pH were found. Significantly decreased dry matter digestibility was detected at 0.5% treatment (P<0.05). Total gas productions in control and 0.03% were significantly greater than those of others (P<0.05). Significantly reduced methane productions were found in all treatments compared to the control (P<0.05). Inclusion of mixed oil did not affect on ammonia production. Total volatile fatty acid production was also not influenced by coating with mixed oils. Rumen fermentation parameters were greatly changed according to introduced plant extracts at second experiment. The significantly lowest and greatest ammonia productions were found at treatments with Ixeris dentata and Plantago asiatica, respectively (P<0.05). The significantly greatest acetate and propionate productions were detected at treatments with Crucuma longa and Zizyphus vulgaris 1, respectively (P<0.05). All treatments, except Chrysanthemum idicum, Euyale ferox seed, Moringa leaf and fruit and Zizyphus vulgaris 1, showed significantly increased total volatile fatty acid production compared to the control (P<0.05). Only Paeonia lactiflora showed significantly lesser gas production than the control (P<0.05). In methane production, Ceramium, Zizyphus, Paeonia, Agrimonia, Torilis, Mugwort, Foeniculum, Euphorbia, Taraxacum, Artemisia, Momordica, Curcuma and Moringa reduced methane significantly compared to the control (P<0.05).

• Journal of Environmental Science International
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• v.26 no.5
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• pp.651-659
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• 2017

This study aimed to estimate the effects of replacing Mushroom By-Product (MBP) with Tofu By-Product (TBP) on the chemical composition, microbes, and rumen fermentation indices of Fermented Diets (FDs). The basal diet was formulated using MBP, TBP, rice bran, molasses, and inoculants. The MBP in the basal diet was replaced with TBP at 0, 5, and 10% on Dry Matter (DM) basis for the experimental diets. The experimental diets were fermented at $39^{\circ}C$ for 144 h. Chemical composition, pH, microbes, and rumen fermentation indices of the FDs were analyzed. With increasing TBP replacement, crude protein content of FDs increased (L, P < 0.001), whereas crude ash content decreased (L, P = 0.002). Lactic acid bacteria and Bacillus subtilis contents in the TBP-replaced FDs were higher than those in the control (P < 0.05), whereas pH level and mold count were lower (P < 0.05). With increasing TBP replacement, in vitro rumen digestibility of DM (L, P = 0.053) and neutral detergent fiber (L, P = 0.024) increased, wheres rumen pH changed (P = 0.026) quadratically. Rumen total volatile fatty acid (L, P = 0.001) and iso-butyrate contents (Q, P = 0.003) increased with increasing TBP replacement. In conclusion, this study indicates that the replacement of MBP with TBP could improve the quality of FD.

• Journal of Life Science
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• v.17 no.11
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• pp.1555-1561
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• 2007

In order to know the effects of scoria, germanium, charcoal, ginger, stevia, and CLA(Conjugated Linoleic Acid) as biologically active materials on pathogenic microbes and rumen anaerobic microbes, the growth rate of pathogens (including Escherichia coli O157, Salmonella paratyphi, Listeria monocytogenes and Staphylococcus aureus) and in vitro lumen microbial growth, gas production, ammonia concentration, carboxymethyl-cellulase (CMCase) activity, and microbial populations were investigated. The growth of pathogenic microbes was inhibited by the supplement of 0.10% ginger. Ginger had powerful antimicrobial properties on all the pathogens used in this experiments. Additionally in the antibacterial assay by paper disc method, we could observe the clear zone of similar area with the positive control(antibiotics) for E. coli as applied with the 10% stevia or the 10% CLA only. The supplements of ginger, stevia and CLA in vitro rumen fermentation inhibited populations of rumen bacteria and protozoa. Particularly supplement of ginger resulted in remarkable reduction of the protozoa population, which means it might serve as a source inhibiting material of methane creation in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.10
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• pp.1405-1408
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• 2001

Ca salt of soybean oil (PSO) and that of mustard oil plus mahua oil (PMOMO) (50:50) were prepared using double decomposition method, and further tested for their fatty acid composition and degree of saponification. Furthermore, the different levels of protected fat of PSO and PMOMO were evaluated in wheat straw based total mixed ration (TMR) in vitro. Results indicated that capric, lauric, myristic, palmitic, steric, oleic, linoleic, leinolenic acids were traces, traces, traces, 10.00, 2.00, 25.00, 58.50, 5.0% in PSO while the corresponding values in PMOMO were 1.08, 0.28, 0.45, 16.9, 12.95, 44.38, 17.46 and 6.50%, respectively. The degree of saponification of both protected fat supplements was more than 80%. Six treatment combinations were tested I.e., blank without feed and fat supplement (T1); control diet with out fat supplement (T2); control diet plus bypass fat supplement (PSO) so that diet contain 5% fat (T3); control diet plus bypass fat supplement (PSO) so that diet contain 7.5% fat (T4); two more diets viz. T5 and T6 were formulated using bypass fat supplement from PMOMO containing 5 and 7.5% fat respectively. TMR was prepared using 50% concentrate mixture and 50% wheat straw. Result indicated that TVFA, $NH_3-N$,TCA-N, total-N and total gas production were increased in treatment diets at 7.5% level of supplementation, however, fermentation pattern remain similar at 5.0% level of supplementation with respect to control diet. Nevertheless, IVDMD and IVOMD values remained unchanged, rather non-significant at both fat levels and with the both fat sources. On the basis of results it was concluded that Ca-salt of Soybean oil or Mustard plus Mahua oil did not show any negative effect either on digestibility or on microbial protein synthesis in rumen, hence the dietary fat upto 7.5% level in total mixed ration based on wheat straw, could be safely used without any adverse effect on rumen fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.40-46
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• 2018

Objective: To examine the effects of Rhodobacter sphaeroides (R. sphaeroides) supplementation as a direct-fed microbial (DFM) on rumen fermentation in dairy cows and on coenzyme Q10 (CoQ10) transition into milk, an in vitro rumen simulation batch culture and an in vivo dairy cow experiment were conducted. Methods: The characteristics of in vitro ruminal fermentation were investigated using rumen fluids from six cannulated Holstein dairy cows at 2 h post-afternoon feeding. A control treatment was included in the experiments based on a typified total mixed ration (TMR) for lactating dairy cows, which was identical to the one used in the in vivo study, plus R. sphaeroides at 0.1%, 0.3%, and 0.5% TMR dry matter. The in vivo study employed six ruminally cannulated lactating Holstein cows randomly allotted to either the control TMR (C-TMR) treatment or to a diet supplemented with a 0.5% R. sphaeroides culture (S-TMR, dry matter basis) ad libitum. The presence of R. sphaeroides was verified using denaturing gradient gel electrophoresis (DGGE) applied to the bacterial samples obtained from the in vivo study. The concentration of CoQ10 in milk and in the supernatant from the in vitro study was determined using high performance liquid chromatography. Results: The results of the in vitro batch culture and DGGE showed that the concentration of CoQ10 significantly increased after 2 h of R. sphaeroides supplementation above 0.1%. When supplemented to the diet of lactating cows at the level of 0.5%, R. sphaeroides did not present any adverse effect on dry matter intake and milk yield. However, the concentration of CoQ10 in milk dramatically increased, with treated cows producing 70.9% more CoQ10 than control cows. Conclusion: The CoQ10 concentration in milk increased via the use of a novel DFM, and R. sphaeroides might be used for producing value-added milk and dairy products in the future.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.12
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• pp.1708-1716
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• 2013

The objective of the trial was to study the relationship between the methane ($CH_4$) production and the Cornell Net Carbohydrate and Protein System (CNCPS) carbohydrate fractions of feeds for cattle and the suitability of CNCPS carbohydrate fractions as the dietary variables in modeling the $CH_4$ production in rumen fermentation. Forty-five rations for cattle with the concentrate/roughage ratios of 10:90, 20:80, 30:70, 40:60, and 50:50 were formulated as feed samples. The Menke and Steingass's gas test was used for the measurement of $CH_4$ production. The feed samples were incubated for 48 h and the $CH_4$ production was analyzed using gas chromatography. Statistical analysis indicated that the $CH_4$ production (mL) was closely correlated with the CNCPS carbohydrate fractions (g), i.e. CA (sugars); $CB_1$ (starch and pectin); $CB_2$ (available cell wall) in a multiple linear pattern: $CH_4=(89.16{\pm}14.93)$ $CA+(124.10{\pm}13.90)$ $CB_1+(30.58{\pm}11.72)$ $CB_2+(3.28{\pm}7.19)$, $R^2=0.81$, p<0.0001, n = 45. Validation of the model using 10 rations indicated that the $CH_4$ production of the rations for cattle could accurately be predicted based on the CNCPS carbohydrate fractions. The trial indicated that the CNCPS carbohydrate fractions CA, $CB_1$ and $CB_2$ were suitable dietary variables for predicting the $CH_4$ production in rumen fermentation in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.61-68
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• 2020

Objective: The present study explored the effects of grape seed procyanidin extract (GSPE) on rumen fermentation, methane production and archaeal communities in vitro. Methods: A completely randomized experiment was conducted with in vitro incubation in a control group (CON, no GSPE addition; n = 9) and the treatment group (GSPE, 1 mg/bottle GSPE, 2 g/kg dry matter; n = 9). The methane and volatile fatty acid concentrations were determined using gas chromatography. To explore methane inhibition after fermentation and the response of the ruminal microbiota to GSPE, archaeal 16S rRNA genes were sequenced by MiSeq high-throughput sequencing. Results: The results showed that supplementation with GSPE could significantly inhibit gas production and methane production. In addition, GSPE treatment significantly increased the proportion of propionate, while the acetate/propionate ratio was significantly decreased. At the genus level, the relative abundance of Methanomassiliicoccus was significantly increased, while the relative abundance of Methanobrevibacter decreased significantly in the GSPE group. Conclusion: In conclusion, GSPE is a plant extract that can reduce methane production by affecting the structures of archaeal communities, which was achieved by a substitution of Methanobrevibacter with Methanomassiliicoccus.