• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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• pp.523-528
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• 2013

This study investigates the effects of three branched-chain amino acids (BCAA; valine, leucine, and isoleucine) on the in vitro ruminal fermentation of wheat straw using batch cultures of mixed ruminal microorganisms. BCAA were added to the buffered ruminal fluid at a concentration of 0, 2, 4, 7, or 10 mmol/L. After 72 h of anaerobic incubation, pH, volatile fatty acids (VFA), and ammonia nitrogen ($NH_3$-N) in the ruminal fluid were determined. Dry matter (DM) and neutral detergent fiber (NDF) degradability were calculated after determining the DM and NDF in the original material and in the residue after incubation. The addition of valine, leucine, or isoleucine increased the total VFA yields ($p{\leq}0.001$). However, the total VFA yields did not increase with the increase of BCAA supplement level. Total branched-chain VFA yields linearly increased as the supplemental amount of BCAA increased (p<0.001). The molar proportions of acetate and propionate decreased, whereas that of butyrate increased with the addition of valine and isoleucine (p<0.05). Moreover, the proportions of propionate and butyrate decreased (p<0.01) with the addition of leucine. Meanwhile, the molar proportions of isobutyrate were increased and linearly decreased (p<0.001) by valine and leucine, respectively. The addition of leucine or isoleucine resulted in a linear (p<0.001) increase in the molar proportions of isovalerate. The degradability of NDF achieved the maximum when valine or isoleucine was added at 2 mmol/L. The results suggest that low concentrations of BCAA (2 mmol/L) allow more efficient regulation of ruminal fermentation in vitro, as indicated by higher VFA yield and NDF degradability. Therefore, the optimum initial dose of BCAA for in vitro ruminal fermentation is 2 mmol/L.

• Korean Journal of Agricultural Science
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• v.45 no.3
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• pp.419-427
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• 2018

The present study was conducted to investigate the effect of different levels of Korean corn grain on in vitro ruminal fermentation with total mixed ration (TMR) as a basal feed. Three ruminal cannulated Holstein steers (Body Weight $479{\pm}33.0kg$) were used as rumen fluid donors. Treatments for in vitro fermentation were TMR only (control, 3.0 g), TMR substituted partially with high level (HC, TMR 1.5 and corn 1.5 g), and with low level of Korean corn grain (LC, TMR 2.25 and corn 0.75 g), respectively. To measure in vitro ruminal pH, gas production, ammonia N and volatile fatty acids (VFA), the in vitro fermentation incubation was triplicated at $39^{\circ}C$, 120 rpm for 0, 1, 3, 6, 12, 24 and 48 h, respectively. Mean ruminal pH was significantly lower (p < 0.05) for HC than control. Changes in rumen pH was rather similar between the groups till 6 h after incubation, but the lowest pH for HC (pH 5.10) appeared at 48 h compared with control and LC. Total gas production was tended (p < 0.09) to be higher and ammonia N was significantly lower (p < 0.05) for HC than control and LC. Total VFA was higher (p < 0.05) for HC and LC than control but no differences appeared between HC and LC. Overall, the present data indicate that feeding different levels of Korean domestic corn grain may lead to high and sustainable starch degradation in the rumen.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.6
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• pp.776-782
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• 2019

Objective: Fasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods. Methods: Three ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40. Results: Fasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility. Conclusion: When in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.

• Journal of The Korean Society of Grassland and Forage Science
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• v.39 no.3
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• pp.132-140
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• 2019

Fermented total mixed ration (TMR) is a novel feed for ruminants in South Korea. The purpose of this study was to evaluate the effects of lactic acid bacteria (LAB) on the quality of TMR and in vitro ruminal fermentation. Strains of three LAB spp. (Lactobacillus plantarum, L. brevis, L. mucosae) were used in fermentation of TMR. Inoculations with the three LAB spp. lowered pH and increased concentrations of lactic acid, acetic acid, and total organic acid compared to non-LAB inoculated control (only addition of an equivalent amount of water) (p<0.05). Bacterial composition indicated that aerobic bacteria and LAB were higher. However, E. coli were lower in the fermented TMR than those in the control treatment (p<0.05). Among the treatments, L. brevis treatment had the highest concentration of total organic acid without fungus detection. Gas production, pH, and ammonia-nitrogen during ruminal in vitro incubation did not differ throughout incubation. However, ruminal total VFA concentration was higher (p<0.05) in the LAB spp. treatments than the control treatment at 48 hours. Overall, the use of L. brevis as an inoculant for fermentation of high moisture. TMR could inhibit fungi growth and promote lactic fermentation, and enhance digestion 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.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.4
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• pp.530-537
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• 2015

The objective of this study was to evaluate the in vitro effects of flavonoid-rich plant extracts (PE) on ruminal fermentation characteristics and methane emission by studying their effectiveness for methanogenesis in the rumen. A fistulated Holstein cow was used as a donor of rumen fluid. The PE (Punica granatum, Betula schmidtii, Ginkgo biloba, Camellia japonica, and Cudrania tricuspidata) known to have high concentrations of flavonoid were added to an in vitro fermentation incubated with rumen fluid. Total gas production and microbial growth with all PE was higher than that of the control at 24 h incubation, while the methane emission was significantly lower (p<0.05) than that of the control. The decrease in methane accumulation relative to the control was 47.6%, 39.6%, 46.7%, 47.9%, and 48.8% for Punica, Betula, Ginkgo, Camellia, and Cudrania treatments, respectively. Ciliate populations were reduced by more than 60% in flavonoid-rich PE treatments. The Fibrobacter succinogenes diversity in all added flavonoid-rich PE was shown to increase, while the Ruminoccocus albus and R. flavefaciens populations in all PE decreased as compared with the control. In particular, the F. succinogenes community with the addition of Birch extract increased to a greater extent than that of others. In conclusion, the results of this study showed that flavonoid-rich PE decreased ruminal methane emission without adversely affecting ruminal fermentation characteristics in vitro in 24 h incubation time, suggesting that the flavonoid-rich PE have potential possibility as bio-active regulator for ruminants.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.4
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• pp.544-547
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• 1999

The object of this study was to determine the influence of monensin and virginiamycin (VM) on in vitro ruminal fermentation of rice straw or ammoniated rice straw. Rumen fluid was collected from 4 wethers fed 200 g of concentrate supplement with 400 g of untreated (U) or ammoniated (A) rice straw once daily for 28 days. Mixed ruminal microorganisms were incubated in anaerobic media that contained 20% (vol/vol) ruminal fluid and 0.3 g of either U or A rice straw. Monensin and/or VM, dissolved in ethanol, were added in centrifuge tubes at final concentrations of 0, 15, 30, 15+15 and 30+30 ppm of culture fluid. The addition of monensin and VM combination to A rice straw fermentation decreased (p<0.05) the acetate to propionate ratio, total VFA and lactate production, but increased (p<0.05) pH. Total gas production tended to be decreased by the addition of monensin plus VM. Antimicrobial agents decreased $NH_3$ N concentration and dry matter digestibility.

• Korean Journal of Agricultural Science
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• v.44 no.4
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• pp.541-548
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• 2017

This study was conducted to compare in vitro rumen fermentation characteristics among corn grains imported from America, Brazil, Argentina and Ukraine A and Ukraine B. Two Holstein steers, each surgically fitted with a ruminal cannula, consuming total mixed ration were used as rumen fluid donors. In vitro rumen fermentation experiments were performed in a completely random design which included a control (no corn) and treatments with 3.0 g of corn from different geographical origins, i.e., America, Brazil, Argentina, and Ukraine A and Ukraine B, respectively. Ruminal pH, ammonia-N, volatile fatty acid (VFA) and total gas production were measured at 0, 1, 3, 6, 12, 24 and 48 h post-incubation, respectively. No differences (p > 0.05) in mean ruminal pH appeared among the treated groups, however, ruminal pH patterns differed; i.e. corn treated groups had dramatically lower pH compared with control during the entire incubation period. Similarly, no different patterns between the groups in ammonia-N (p > 0.05) appeared until 6 h post-incubation. Unexpectedly, higher ammonia-N concentration for control than that for the corn treated groups appeared after 12 h post-incubation despite that for all groups increased. Total VFA was similar between the groups until 6 h post-incubation, but VFA after 12 h post-incubation was different (p < 0.05), i.e. VFA for corn from Argentina, Ukraine A, Ukraine B, and Brazil were comparatively higher than for America. Overall, data in this study showed that the corns of different origins may have different feed values to ruminants despite having similar chemical compositions.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.12
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• pp.1726-1735
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• 2014

This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (p<0.05) the total gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (p<0.05) the ratio of acetate to propionate, the concentrations of isovalerate, valerate and ammonia-nitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen.