• 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.

• 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.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.

• Animal Bioscience
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• v.35 no.2
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• pp.184-195
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• 2022

Objective: In this study we aimed to evaluate the effect of dietary live yeast supplementation on ruminal pH pattern, fermentation characteristics and associated bacteria in beef cattle. Methods: This work comprised of in vitro and in vivo experiments. In vitro fermentation was conducted by incubating 0%, 0.05%, 0.075%, 0.1%, 0.125%, and 0.15% active dried yeast (Saccharomyces cerevisiae, ADY) with total mixed ration substrate to determine its dose effect. According to in vitro results, 0.1% ADY inclusion level was assigned in in vivo study for continuously monitoring ruminal fermentation characteristics and microbes. Six ruminally cannulated steers were randomly assigned to 2 treatments (Control and ADY supplementation) as two-period crossover design (30-day). Blood samples were harvested before-feeding and rumen fluid was sampled at 0, 3, 6, 9, and 12 h post-feeding on 30 d. Results: After 24 h in vitro fermentation, pH and gas production were increased at 0.1% ADY where ammonia nitrogen and microbial crude protein also displayed lowest and peak values, respectively. Acetate, butyrate and total volatile fatty acids concentrations heightened with increasing ADY doses and plateaued at high levels, while acetate to propionate ratio was decreased accordingly. In in vivo study, ruminal pH was increased with ADY supplementation that also elevated acetate and propionate. Conversely, ADY reduced lactate level by dampening Streptococcus bovis and inducing greater Selenomonas ruminantium and Megasphaera elsdenii populations involved in lactate utilization. The serum urea nitrogen decreased, whereas glucose, albumin and total protein concentrations were increased with ADY supplementation. Conclusion: The results demonstrated dietary ADY improved ruminal fermentation dose-dependently. The ruminal lactate reduction through modification of lactate metabolic bacteria could be an important reason for rumen pH stabilization induced by ADY. ADY supplementation offered a complementary probiotics strategy in improving gluconeogenesis and nitrogen metabolism of beef cattle, potentially resulted from optimized rumen pH and fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.80-85
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• 2018

Objective: This study was conducted to investigate the effect of corn grain particle size on ruminant fermentation and blood metabolites in Holstein steers fed total mixed ration (TMR) as a basal diet to explain fundamental data of corn grain for cattle in Korea. Methods: Four ruminally cannulated Holstein steers (body weight $592{\pm}29.9kg$) fed TMR as a basal diet were housed individually in an auto temperature and humidity modulated chamber ($24^{\circ}C$ and 60% for 22 h/d). Treatments in a $4{\times}4$ Latin square design were TMR only (control), TMR with whole corn grain (WC), coarsely ground corn grain (CC), and finely ground corn grain (FC), respectively. The corn feeds substituted for 20% energy intake of TMR intake. To measure the ruminal pH, ammonia N, and volatile fatty acids (VFA), ruminal digesta was sampled through ruminal cannula at 1 h intervals after the morning feeding to determine ruminal fermentation characteristics. Blood was sampled via the jugular vein after the ruminal digesta sampling. Results: There was no difference in dry matter (DM) intake between different corn particle size because the DM intake was restricted to 1.66% of body weight. Different corn particle size did not change mean ammonia N and total VFA concentrations whereas lower (p<0.05) ruminal pH and a ratio of acetate to propionate, and higher (p<0.05) propionate concentration were noted when the steers consumed CC compared with WC and FC. Concentration of blood metabolites were not affected by different particle size of corn grain except for blood triglyceride concentration, which was significantly (p<0.05) increased by FC. Conclusion: Results indicate that feeding CC may increase feed digestion in the rumen, whereas the FC group seemed to obtain inadequate corn retention time for microbial degradation in the rumen.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.419-428
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• 2005

This study was carried out to investigate the effect of replacing dried mugwort (Artemisia montana Pampan) in concentrate a time of preparation pellet. The treatments, arranged in a 4${\times}$4 Latin square design, were four additional levels of mugwort at 0, 3, 5 and 10% in concentrate. Four crossbred (CorriedalexPolwarth, t) sheep with a mean body weight of 41.3 kg were used to evaluate nutrients digestibility, palatability, fermentation characteristics and microbial protein synthesis in the rumen. The digestibility of crude protein was improved (p < 0.05) to 6.1 % - 8.6 % in sheep fed 3, 5 and 10% mugwort pellet treatments compared with that of control. That of crude fat and NDF was improved (p < 0.05) to 5.8 % - 7.3 % in sheep fed 3 % compared to other treatments. The ruminal pH was significantly (p < 0.05) decreased in sheep fed 3 % mugwort pellet compared to other treatments when observed at 0.5 hour after feeding. The ammonia nitrogen concentrations were the highest in sheep fed all treatments at 1 hour after feeding. The ruminal concentrations of acetic acid and propionic acids were an improvement (p < 0.05) at the 3% and 5% treatments. Retained nitrogen of 3, 5 and 10% treatment with the value of 2.24 - 2.82 g was higher (p < 0.05) than that of the control with 0.78 g and microbial protein production of 10% treatment was higher (p < 0.05) than that of control. This study suggested that the replacing with 3% dry mugwort (Artemisia montana Pampan) in concentrate a time of preparation pellet will improve nutrient digestibility, palatability, ruminal fermentation characteristics and feed value.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.10
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• pp.1374-1382
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• 2001

Fifteen Inner Mongolian wethers with permanent ruminal and duodenal cannulas were used to study the effects of dietary rumen-undegradable protein (RUP) to rumen-degradable protein (RDP) ratios or protein sources on fiber digestion in the gastrointestinal tract and ruminal fluid characteristics. Fiber digestion and ruminal fermentation were not affected (p>0.05) by dietary RUP to RDP ratios (from 1.54 to 0.72). Soybean meal supplementation improved ruminal digestion. Fish meal supplementation increased (p<0.05) the ruminal degradability of fiber. The different RUP to RDP ratios (from 1.54 to 0.72) did not influence (p>0.05) ruminal fluid pH, but there were differences (p<0.05) in ruminal fluid $NH_3-N$ concentration because of urea replacement. Soybean meal as a dietary protein source decreased (p<0.05) ruminal fluid pH and increased (p<0.05 or p<0.01) $NH_3-N$, acetate, propionate and butyrate concentrations in the rumen. Fish meal as a dietary protein source decreased (p<0.05 or p<0.01) ruminal $NH_3-N$ and acetate concentrations and increased (p<0.05) ruminal propionate concentration. It can be concluded that dietary protein sources have more significant effect on fiber digestion and ruminal fermentation than different dietary RUP to RDP ratios, when the dietary crude protein requirements of growing sheep are satisfied.

• 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.30 no.3
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• pp.355-362
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• 2017

Objective: The use of locally produced forage (LPF) in cattle production has economic and environmental advantages over imported forage. The objective of this study was to characterize the nutritional value of LPF commonly used in Korea. Differences in ruminal fermentation characteristics were also examined for the LPF species commonly produced from two major production regions: Chungcheong and Jeolla. Methods: Ten LPF (five from each of the two regions) and six of the most widely used imported forages originating from North America were obtained at least three times throughout a year. Each forage species was pooled and analyzed for nutrient content using detailed chemical analysis. Ruminal fermentation characteristics were also determined by in vitro anaerobic incubations using strained rumen fluid for 0, 3, 6, 12, 24, and 48 h. At each incubation time, total gas, pH, ammonia, volatile fatty acid (VFA) concentrations, and neutral detergent fiber digestibility were measured. By fitting an exponential model, gas production kinetics were obtained. Results: Significant differences were found in the non-fiber carbohydrate (NFC) content among the forage species and the regions (p<0.01). No nutrient, other than NFC, showed significant differences among the regions. Crude protein, NFC, and acid detergent lignin significantly differed by forage species. The amount of acid detergent insoluble protein tended to differ among the forages. The forages produced in Chungcheong had a higher amount of NFC than that in Jeolla (p<0.05). There were differences in ruminal fermentation of LPF between the two regions and interactions between regions and forage species were also significant (p<0.05). The pH following a 48-h ruminal fermentation was lower in the forages from Chungcheong than from Jeolla (p<0.01), and total VFA concentration was higher in Chungcheong than in Jeolla (p = 0.05). This implies that fermentation was more active with the forages from Chungcheong than from Jeolla. Analysis of gas production profiles showed the rate of fermentation differed among forage species (p<0.05). Conclusion: The results of the present study showed that the nutritional values of some LPF (i.e., corn silage and Italian ryegrass) are comparable to those of imported forages widely used in Korea. This study also indicated that the nutritional value of LPF differs by origin, as well as by forage species. Detailed analyses of nutrient composition and digestion kinetics of LPF should be routinely employed to evaluate the correct nutritional value of LPF and to increase their use in the field.

• Animal Bioscience
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• v.37 no.2_spc
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• pp.360-369
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• 2024

Ruminal methane production functions as the main sink for metabolic hydrogen generated through rumen fermentation and is recognized as a considerable source of greenhouse gas emissions. Methane production is a complex trait affected by dry matter intake, feed composition, rumen microbiota and their fermentation, lactation stage, host genetics, and environmental factors. Various mitigation approaches have been proposed. Because individual ruminants exhibit different methane conversion efficiencies, the microbial characteristics of low-methane-emitting animals can be essential for successful rumen manipulation and environment-friendly methane mitigation. Several bacterial species, including Sharpea, uncharacterized Succinivibrionaceae, and certain Prevotella phylotypes have been listed as key players in low-methane-emitting sheep and cows. The functional characteristics of the unclassified bacteria remain unclear, as they are yet to be cultured. Here, we review ruminal methane production and mitigation strategies, focusing on rumen fermentation and the functional role of rumen microbiota, and describe the phylogenetic and physiological characteristics of a novel Prevotella species recently isolated from low methane-emitting and high propionate-producing cows. This review may help to provide a better understanding of the ruminal digestion process and rumen function to identify holistic and environmentally friendly methane mitigation approaches for sustainable ruminant production.