• Asian-Australasian Journal of Animal Sciences
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• v.11 no.4
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• pp.351-362
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• 1998

Microbial growth efficiency in the rumen was studied in sheep given hourly, 31.25 g oaten chaff with either 0.31 and 0.88 g urea or 1.88 and 5.63 g casein (exp. 1) and 33.33 g oaten chaff with 1.04 casein or 0.3, 0.6 and 0.9 g urea or the mixture of the casein and urea (exp. 2). Concentrations of ruminal fluid ammonia increased with increasing nitrogenous supplements. Organic matter digestibility in sacco in the rumen was not different irrespective of N sources. Isoacids and valeric acid increased with increasing ingested casein but decreased with increasing urea intake. Peptide and amino acid pools in ruminal fluid increased with increasing ammonia concentrations (exp. 2) suggesting that proteolytic activity and transportation of peptides and amino acids across microbial membrane of rumen microbes may be regulated by the metabolite mechanism (intracellular amino acids and $NH_4{^+}$, respectively). Densities of total viable and cellulolytic bacteria in ruminal fluid increased with increasing ammonia levels but that of small Entodinia decreased. The density of fungal sporangia growth on oat leaf blades decreased with increasing ammonia concentrations but appeared to remain constant in the presence of casein. Efficiency of net microbial cell synthesis was 15-28% higher when ammonia concentrations increased from 100 to above 200 mg N/l regardless of N sources. In conclusion, supplementation of preformed protein had no effect on rumen digestion and microbial growth efficiency. This could not be accounted for its effect on ruminal fluid ammonia. Increased microbial growth efficiency with increasing ammonia levels may be due to a reduction in the turnover of microbial cells within the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.3
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• pp.370-378
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• 2017

Objective: An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods: Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results: Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion:The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient digestibility, ruminal fermentation and protozoa and HAP bacteria count, blood cells and metabolites.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.5
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• pp.557-562
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• 1996

The effects of diets differing in protein content through soybean meal supplementation on ruminal fractionation of selenium (Se) were studied. A $3{\times}3$ Latin square design was used with three Japanese Corriedale wethers (45 kg average body weight), three periods, and three dietary treatment. The three dietary treatments were : Diet 1, without soybean meal supplementation (14% crude protein, CP); Diet 2, with 10% soybean meal supplementation (16.5% CP); and Diet 3, with 20% soybean meal supplementation (19% CP). All the diets had a Se supplementation in the form of sodium selenite at 0.2 mg Se/kg dietary DM. The Se supplement and the concentrate mixture were fed only in the morning before the hay was given. Daily feeding schedule for gay was set at 09:00 and 17:00 h. On the final day of collection period, ruminal fluid samples were obtained at 0.5, 2, 6, 12 and 24 h post-feeding starting at 09:00 h. Total ruminal fluid Se was markedly higher (p<0.05) in Diet 3 than those in Diets 1 and 2 at almost all sampling time except at 24 h. The proportion of Se in soluble protein to the total ruminal Se was higher (p< 0.05) in Diet 3 (40%) followed by Diet 2 (28%) and Diet 1 (21%). The proportion of free inorganic Se to the total ruminal Se was the reverse, especially after two hours where Diet 1 (p<0.05) was higher than the other diets. Bacterial Se was lower (p < 0.05) in Diet 1 than those in Diets 2 and 3 at any sampling time. The highest was observed at 2 h postprandially in all diets with a value of 421, 556, $655{\mu}g/kg$ bacterial DM for Diet 1, 2 and 3, respectively. No differences (p>0.05) were observed on ruminal pH, ammonia and total nolatile fatty acids although increasing protein supplementation tended to decline the ruminal pH and increase ruminal ammonia. This study concludes that increasing dietary protein content by soybean meal supplementation can affect the ruminal Se metabolism.

• 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.

• 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.

• 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.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.4
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• pp.619-622
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• 1993

Rumen bacterial amino acids in sheep on urea diet were monitored to assess a possible change in amino acid synthesis as a long term response to high rumen ammonia environment. A sheep was fed a semipurified diet with soybean meal, followed by a diet with urea as a main nitrogen source. Mixed rumen bacteria were harvested from ruminal fluid taken 3 h after feeding (twice in soybean meal feeding and 6 times in urea feeding) and fractionated as cell wall, proteins and protein-free cell supernatant of monitor amino acids in each fraction. Ruminal ammonia concentration at the sampling ranged from 5.7 to 39.5 mgN/dl. Cell wall and protein fractions of mixed rumen bacteria were stable in their amino acid composition regardless of nitrogen sources of diet and the feeding duration. However, protein-free cell supernatant fraction showed a higher alanine proportion with urea feeding (18.6 and 28.2 molar % of alanine for samples from sheep fed soybean meal and urea, respectively) and its duration (20.6 and 32.9 molar % for samples from sheep on urea diet for 1 and 65 days, respectively). Total free amino acid level of bacteria was depressed in the initial period of urea feeding but restored on 65th day of the feeding. These results suggest that an alanine synthesizing system may develop in rumen bacteria as urea feeding becomes longer.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.1
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• pp.67-72
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• 1993

Three sheep fitted with cannulas in the rumen and the caecum were used in a $3{\times}3$ Latin square design to study the changes in ruminal and caecal microbial populations and their metabolite composition with ammoniated rice straw feeding. The 3 diets contained either 80% untreated rice straw (UTS) or ammoniated rice straw (ATS) and 20% formula feed. These were a control diet (C), a urea supplemented diet (U) containing urea at 1.1% and an ammoniated rice straw diet (AT). Data were analyzed by analysis of variance and means separated by the Student Neumann Kuel's multiple comparison. AT feeding increased ruminal bacterial counts, in particular cellulolytic bacterial counts (p < 0.05) which were 1.8, 2.4 and 7.0 (${\times}10^6/ml$ ruminal fluid) for C, U and AT, respectively. There was an increasing tendency (p < 0.10) in ruminal fungal population with U; values were 2.0, 5.2, 3.1 (${\times}10^3/ml$ ruminal fluid) for C, U and AT, respectively. Ruminal protozoa counts were not significantly (p > 0.05) altered with diets. Caecal total viable bacterial count with AT was about thrice the value with C. Total VFA concentration in the rumen was significantly increased (p < 0.025) (7.7 mmol/dl for C and 8.2 mmol/dl for AT) and correspondingly, pH lowered when AT was fed. Sheep on AT tended to produce less acetate and more butyrate in the rumen without significance (p > 0.05). Similar to the rumen, total VFA concentrations of 4.4, 3.8 and 5.2 mmol/dl were detected, respectively, for C, U and AT. Caecal ammonia-nitrogen concentrations were about six-fold of that in the rumen, though there were no differences (p > 0.05) among treatments.