• Animal Bioscience
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• v.37 no.4
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• pp.655-667
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• 2024

Objective: This study aimed to assess the impact of a hydroethanolic extract of walnut green husks (WGH) on rumen fermentation and the diversity of bacteria, methanogenic archaea, and fungi in sheep fed a high-concentrate diet. Methods: Five healthy small-tailed Han ewes with permanent rumen fistula were selected and housed in individual pens. This study adopted a self-controlled and crossover design with a control period and an experimental period. During the control period, the animals were fed a basal diet (with a ratio of concentrate to roughage of 65:35), while during the treatment period, the animals were fed the basal diet supplemented with 0.5% hydroethanolic extract of WGH. Fermentation parameters, digestive enzyme activities, and microbial diversity in rumen fluid were analyzed. Results: Supplementation of hydroethanolic extract of WGH had no significant effect on feed intake, concentrations of total volatile fatty acids, isovalerate, ammonia nitrogen, and microbial protein (p>0.05). However, the ruminal pH, concentrations of acetate, butyrate and isobutyrate, the ratio of acetate to propionate, protozoa count, and the activities of filter paper cellulase and cellobiase were significantly increased (p<0.05), while concentrations of propionate and valerate were significantly decreased (p<0.05). Moreover, 16S rRNA gene sequencing revealed that the relative abundance of rumen bacteria Christensenellaceae R7 group, Saccharofermentans, and Ruminococcaceae NK4A214 group were significantly increased, while Ruminococcus gauvreauii group, Prevotella 7 were significantly decreased (p<0.05). The relative abundance of the fungus Pseudomonas significantly increased, while Basidiomycota, Fusarium, and Alternaria significantly decreased (p<0.05). However, there was no significant change in the community structure of methanogenic archaea. Conclusion: Supplementation of hydroethanolic extract of WGH to a high-concentrate diet improved the ruminal fermentation, altered the structure of ruminal bacterial and fungal communities, and exhibited beneficial effects in alleviating subacute rumen acidosis of sheep.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.1
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• pp.23-30
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• 2000

The effects of grass lipids and long chain fatty acids (LCFA; palmitic, stearic and oleic acids), at low concentrations (0.001~0.02%), on the growth and enzyme activity of two strains of anaerobic fungi, monocentric strain Piromyces rhizinflata B157 and polycentric strain Orpinomyces joyonii SG4, were investigated. The addition of grass lipids to the medium significantly (p<0.05) decreased filter paper (FP) cellulose digestion, cellulase activity and fungal growth compared to control treatment. However, LCFA did not have any significant inhibitory effects on fungal growth and enzyme activity, which, however, were significantly (p<0.05) stimulated by the addition of oleic acid as have been observed in rumen bacteria and protozoa. This is the first report to our knowledge on the effects of LCFA on the rumen anaerobic fungi. Continued work is needed to identify the mode of action of LCFA in different fungal strains and to verify whether these microorganisms have ability to hydrogenate unsaturated fatty acids to saturated fatty acids.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.8
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• pp.1215-1221
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• 1999

Twelve rumen fistulated cross-bred calves were divided into three groups and fed wheat straw and concentrate mixture according to their maintenance requirement. Animals in group II and III were fed 50 and 100mg rumensin per day, in addition to basal feed. Supplementation of rumensin in the diet decreased the dry matter intake significantly (p<0.05) along with a significant decrease in the straw intake. Digestibility coefficients of all the nutrients were not affected significantly except that of CF digestibility which was lower (p<0.05) in groups II and III as compared to group I. Among N-parameters in the rumen fluid, mean $NH_3-N$ was significantly lower in groups II and III (19.13 and 18.63 mg N/100 ml respectively) than in group I (22.68); total-N and TCA-ppt-N did not differ among the three groups. Total VFA concentration did also not differ among the three groups, however, propionate increased from 24.33 molar % to 32.73 while acetate and butyrate decreased respectively from 65.85 to 58.81% and 9.79 to 8.46%. Total VFA, bacteria and protozoa production rates were not affected significantly due to rumensin in diet. Methane production per kg DDM as well as % of methane in total gas were reduced at both the levels of rumensin on different concentrate ratios with wheat straw as roughage. Similar trend was also observed with rice straw and concentrate mixture as substrate with rumensin addition.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.7
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• pp.990-996
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• 2005

The effect of diets varying in level and source of nitrogen (N) and fermentable organic matter on dynamic characteristics of microbial populations in rumen liquor and their impact on substrate fermentation in vitro was studied. The diets tested were straw alone, straw+concentrate mixture and straw+urea molasses mineral block (UMMB) lick. The same diets were taken as substrates and tested on each inoculum collected from the diets. Diet had no effect on the amino acid (AA) composition of either bacteria or protozoa. Differences among the diets in intake, source of N and OM affected bacterial and protozoal characteristics in the rumen. Upper asymptote of gas production (Y$\alpha$) had a higher correlation with bacterial pool size and production rate than with protozoal pool size and production rate. Among the parameters of the gas production model, Y$\alpha$ and lag time in total gas has showed significant (p<0.01) correlation with bacterial characteristics. Though the rate constant of gas production significantly differed (p<0.01) between diet and type of straw, it was least influenced by the microbial characteristics. The regression coefficient of diet and type of straw for Y$\alpha$ indicated that the effect of diet on Y$\alpha$ was threefold higher than that of the straw. As microbial characteristics showed higher correlation with Y$\alpha$, and diet had more influence on the microbial characteristics, gas production on a straw diet could be used effectively to understand the microbial characteristics.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.9
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• pp.1219-1227
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• 2000

Rumination behavior, in vivo digestibility of cell wall constituents, particle size reduction in the rumen, and retention time in the digestive tract of sheep were examined using rice and oat straw as roughage sources. The in sacco digestibility, rumen fermentation, and microbial population and internal adenosine 5-triphosphate (ATP) content were also determined under feeding conditions of high-roughage and high-concentrate diets. Chewing number and time in rumination behavior were higher with rice straw than with oat straw, while the in sacco and in vivo DMD of rice straw were consistently lower than those of oat straw. Rice straw also showed higher frequency of thinner and longer particles in the rumen contents and lower retention time in the whole digestive tract as compared to those of oat straw. Rice straw was more effective to maintain the ruminal pH than oat straw, being reflected in higher internal ATP content of large-type protozoa on the high- concentrate diet. Changes in the ruminal microflora by shifting from the low- to the high- concentrate diet were also different between rice and oat straw.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.139-147
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• 1999

Peptides are formed in the rumen as the result of microbial proteinase activity. The predominant type of activity is cysteine ptoteinase, but others, such as serine proteinases, are also present. Many species of protozoa, bacteria and fungi are involved in ptoteolysis; large animal-to-animal variability is found when proteinase activities in different animals are compared. The peptides formed from proteolysis are broken down to amino acids by peptidases. Different peptides are broken down at different rates, depending on their chemical composition and particularly their N-terminal structure. Indeed, chemical addition to the N-terminus of small peptides, such as by acetylation, causes the peptides to become stable to breakdown by the rumen microbial population; the microorganisms do not appear to adapt to hydrolyse acetylated peptides even after several weeks exposure to dietary acetylated peptides, and the amino acids present in acetylated peptides are absorbed from the small intestine. The amino acids present in some acetylated peptides remain available in nutritional trials with rats, but the nutritive value of the whole amino acid mixture is decreased by acetylation. The genus Prevotella is responsible for most of the catabolic peptidase activity in the rumen, via its dipeptidyl peptidase activities, which release dipeptides rather than free amino acids from the N-terminus of oligopeptides. Studies with dipeptidyl peptidase mutants of Prevotella suggest that it may be possible to slow the rate of peptide hydrolysis by the mixed rumen microbial population by inhibiting dipeptidyl peptidase activity of Prevotella or the rate of peptide uptake by this genus. Peptides and amino acids also stimulate the growth of rumen microorganisms, and are necessary for optimal growth rates of many species growing on tapidly fermented substrates; in rich medium, most bacteria use pre-formed amino acids for more than 90% of their amino acid requirements. Cellulolytic species are exceptional in this respect, but they still incorporate about half of their cell N from pre-formed amino acids in rich medium. However, the extent to which bacteria use ammonia vs. peptides and amino acids for protein synthesis also depends on the concentrations of each, such that preformed amino acids and peptides are probably used to a much lesser extent in vivo than many in vitro experiments might suggest.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.900-907
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• 2007

Three male Gayal (Bos frontalis) and three male Yunnan Yellow cattle (Bos taurus) were fed pelleted lucerne and measurements made of digestibility, nitrogen utilisation, rumen fermentation and microbial population and key plasma metabolites. Total actual dry matter intake was similar but when expressed in terms of live weight or metabolic live weight feed intakes were significantly higher (p<0.05) for Gayal than cattle. Apparent digestibilities of dry matter, organic matter, fibre and dietary nitrogen were similar for both Gayal and cattle. Rumen ammonia nitrogen and total volatile fatty acids were significantly higher (p<0.05) for Gayal than cattle and total numbers of viable rumen bacteria, cellulolytic and amylolytic bacteria, but not proteolytic bacteria nor protozoa, were significantly greater (p<0.05) for Gayal than cattle. Although Gayal have a different rumen ecology to cattle, similar digestive parameters were exhibited. Further research is required to establish relationship between rumen ecology and digestive parameters.

• 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.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1092-1100
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• 2010

Bacteria, fungi, and protozoa inhabiting the rumen, the largest chamber of the ruminants' stomach, release large quantities of hydrogen during the fermentation of carbohydrates. The hydrogen is used by coexisting methanogens to produce methane in energy-yielding processes. This work shows, for the first time, a fundamental possibility of using a hydrogen-rich fermentation gas produced by selected rumen ciliates to feed a low-temperature hydrogen fuel cell. A biohydrogen fuel cell (BHFC) was constructed consisting of (i) a bioreactor, in which a hydrogen-rich gas was produced from glucose by rumen ciliates, mainly of the Isotrichidae family, deprived of intra- and extracellular bacteria, methanogens, and fungi; and (ii) a chemical fuel cell of the polymer-electrolyte type (PEFC). The fuel cell was used as a tester of the technical applicability of the fermentation gas produced by the rumen ciliates for power generation. The average estimated hydrogen yield was ca. 1.15 mol $H_2$ per mole of fermented glucose. The BHFC performance was equal to the performance of the PEFC running on pure hydrogen. No fuel cell poisoning effects were detected. A maximum power density of $1.66\;kW/m^2$ (PEFC geometric area) was obtained at room temperature. The maximum volumetric power density was $128\;W/m^3$ but the coulombic efficiency was only ca. 3.8%. The configuration of the bioreactor limited the continuous operation time of this BHFC to ca. 14 h.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.11
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• pp.1515-1528
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• 2017

The gastrointestinal (GI) tract, including the rumen and the other intestinal segments of cattle, harbors a diverse, complex, and dynamic microbiome that drives feed digestion and fermentation in cattle, determining feed efficiency and output of pollutants. This microbiome also plays an important role in affecting host health. Research has been conducted for more than a century to understand the microbiome and its relationship to feed efficiency and host health. The traditional cultivation-based research elucidated some of the major metabolism, but studies using molecular biology techniques conducted from late 1980's to the late early 2000's greatly expanded our view of the diversity of the rumen and intestinal microbiome of cattle. Recently, metagenomics has been the primary technology to characterize the GI microbiome and its relationship with host nutrition and health. This review addresses the main methods/techniques in current use, the knowledge gained, and some of the challenges that remain. Most of the primers used in quantitative real-time polymerase chain reaction quantification and diversity analysis using metagenomics of ruminal bacteria, archaea, fungi, and protozoa were also compiled.