• Asian-Australasian Journal of Animal Sciences
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• v.9 no.4
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• pp.375-379
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• 1996

The experiment investigated the possibility of using effluent from RUSITEC (rumen simulation technique) inoculated with rumen liquor or cow faeces as sources of micro-organisms for in vitro digestion of forages. Nine forages ${\times}3$ sources of inoculum were used in a factorial arrangement of treatments. Rumen liquor was collected from fistulated sheep and faeces was collected from cows. The RUSITEC apparatus consisted of 4 vessels, 2 vessels were charged with faecal liquor and 2 with rumen liquor. On the 8th day of the experiment RUSITEC effluent were collected to use in in vitro studies. In vitro OMD (g/kg) values using three sources of inoculum (fresh rumen liquor, RUSITEC effluent from rumen liquor or cow faeces) were statistically significant (p < 0.001). The regression relationships between OMD using fresh rumen liquor and RUSITEC effluent were highly significant ($R^2>0.90$). The results suggest that RUSITEC effluent either from rumen liquor or cow faeces can be used as a source of micro-organisms for in vitro digestion of forages.

• Animal Bioscience
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• v.37 no.2_spc
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• pp.370-384
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• 2024

Rumen microbiota play a central role in the digestive process of ruminants. Their remarkable ability to break down complex plant fibers and proteins, converting them into essential organic compounds that provide animals with energy and nutrition. Research on rumen microbiota not only contributes to improving animal production performance and enhancing feed utilization efficiency but also holds the potential to reduce methane emissions and environmental impact. Nevertheless, studies on rumen microbiota face numerous challenges, including complexity, difficulties in cultivation, and obstacles in functional analysis. This review provides an overview of microbial species involved in the degradation of macromolecules, the fermentation processes, and methane production in the rumen, all based on cultivation methods. Additionally, the review introduces the applications, advantages, and limitations of emerging omics technologies such as metagenomics, meta-transcriptomics, metaproteomics, and metabolomics, in investigating the functionality of rumen microbiota. Finally, the article offers a forward-looking perspective on the new horizons and technologies in the field of rumen microbiota functional research. These emerging technologies, with continuous refinement and mutual complementation, have deepened our understanding of rumen microbiota functionality, thereby enabling effective manipulation of the rumen microbial community.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.1
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• pp.51-54
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• 1998

The present study investigated the use of cow faeces as a source of micro-organisms instead of sheep rumen liquor in in vitro digestibility assays of forages. Initially 40 forage samples comprising ryegrass, wheat, maize and barley were screened to select a wide range of digestibility (327-794 g/kg) of forages. Finally 8 forage samples were assessed using rumen liquor as well as faecal liquor. The absolute organic matter digestibility (OMD) values using faecal liquor were lower than those with rumen liquor. However, the relationship between OMD using rumen liquor and faecal liquor was highly significant (p < 0.001). The $R^2$ value exceed 0.90. The results suggest that micro-organisms from faecal liquor are capable of digesting forage samples.

• Asian-Australasian Journal of Animal Sciences
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• v.2 no.3
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• pp.544-545
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• 1989

• Animal Bioscience
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• v.34 no.11
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• pp.1784-1793
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• 2021

Objective: An experiment was conducted to evaluate the effects of bamboo leaf extract (BLE) on the production performance, rumen fermentation parameters, and rumen bacterial communities of heat-stressed dairy cows. Methods: The experiment comprised a 14-day adaptation period and a 21-day experimental period and was conducted in a high-temperature and humidity environment (daily mean ambient temperature = 33.5℃±1.3℃; daily mean relative humidity = 64.9%±0.8%, daily mean temperature-humidity index = 86.2±0.4). Twelve Holstein dairy cows were randomly allocated into two groups. A total mixed ration supplemented with BLE at 0 (CON) and 1.3 g/kg dry matter (DM) were fed, respectively. Feed intake and milk yield were recorded daily. Milk samples were collected on 1, 11, and 21 d of the experimental period to analyze milk performance. Rumen fluid samples were collected on 21 d of the experimental period to analyze rumen fermentation parameters and rumen bacterial communities. Results: Compared with the control group, supplementation of BLE increased milk yield (p<0.01), milk fat yield (p = 0.04), 4% fat-corrected milk (p<0.01) and milk fat content (p<0.01); reduced somatic cell count (p<0.01). No differences in DM intake and milk protein or lactose content were observed between two groups. Supplementation of BLE also increased the rumen total volatile fatty acid (p<0.01), acetate (p<0.01), butyrate (p<0.01), and valerate (p = 0.05) concentrations. However, no significant effects were observed on rumen pH, ammonia nitrogen, propionate, acetate/propionate ratio, isobutyrate, or isovalerate. Furthermore, BLE increased the rumen bacterial abundance and the diversity of the rumen bacterial community. The BLE reduced the Firmicutes/Bacteroidetes abundance ratio and increased the abundances of Butyrivibrio_2 (p<0.01) and Ruminococcus_2 (p<0.01). Conclusion: The BLE supplementation at 1.3 g/kg DM could improve production performance and rumen fermentation in dairy cows during heat stress.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.79-90
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• 2020

Objective: In the present study, an liquid chromatography/mass spectrometry (LC/MS) metabolomics approach was performed to investigate potential biomarkers of milk production in high- and low-milk-yield dairy cows and to establish correlations among rumen fluid metabolites. Methods: Sixteen lactating dairy cows with similar parity and days in milk were divided into high-yield (HY) and low-yield (LY) groups based on milk yield. On day 21, rumen fluid metabolites were quantified applying LC/MS. Results: The principal component analysis and orthogonal correction partial least squares discriminant analysis showed significantly separated clusters of the ruminal metabolite profiles of HY and LY groups. Compared with HY group, a total of 24 ruminal metabolites were significantly greater in LY group, such as 3-hydroxyanthranilic acid, carboxylic acids, carboxylic acid derivatives (L-isoleucine, L-valine, L-tyrosine, etc.), diazines (uracil, thymine, cytosine), and palmitic acid, while the concentrations of 30 metabolites were dramatically decreased in LY group compared to HY group, included gentisic acid, caprylic acid, and myristic acid. The metabolite enrichment analysis indicated that protein digestion and absorption, ABC transporters and unsaturated fatty acid biosynthesis were significantly different between the two groups. Correlation analysis between the ruminal microbiome and metabolites revealed that certain typical metabolites were exceedingly associated with definite ruminal bacteria; Firmicutes, Actinobacteria, and Synergistetes phyla were highly correlated with most metabolites. Conclusion: These findings revealed that the ruminal metabolite profiles were significantly different between HY and LY groups, and these results may provide novel insights to evaluate biomarkers for a better feed digestion and may reveal the potential mechanism underlying the difference in milk yield in dairy cows.

• Journal of agriculture & life science
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• v.46 no.3
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• pp.109-118
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• 2012

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est2R) was 2,120 bp in length, encoding a protein of 516 amino acid residues with a calculated molecular weight of 57,286 Da. The molecular weight of the enzyme was estimated to be 57,000 Da by SDS-PAGE. Est2R shared 35.6% amino acid identity with esterase (CAH19079) of uncultured prokaryote. The Est2R was most active at $20-40^{\circ}C$, and showed optimum at $30^{\circ}C$ and pH 8.0. The most activity of Est2R for the different chain length of p-nitrophenyl ester group as substrate was p-nitrophenyl acetate. Moreover, the enzyme was found to be most active without organic solvent, followed by 98% active with ethanol, and the enzyme activity was highly affected by the acetonitrile. The enzyme was significantly inhibited by $Zn^{2+}$ but stimulated by $Ca^{2+}$. So, novel esterase gene est2R is likely to obtain from cow rumen metagenome and supposed to use for industrial purpose.

• Journal of Practical Agriculture & Fisheries Research
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• v.19 no.1
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• pp.139-151
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• 2017

This study was conducted to confirm the rumen fermentation characteristics of large-scale CoenzymeQ10(CoQ10) producing bacteria R. spharoides in rumen. We conducted in vitro continuous culture test to investigate the characteristics of rumen fermentation with 5% R. spharoides as a direct fed microorganism. A rumen microbial fermentation characteristic has stability at after 12 days for 15 day of experimental period. pH value, NH₃-N, microbial protein synthesis, ADF digestibility and NDF digestibility were not shown significantly differences between control and treatment. However, UDP was significantly higher in treatment than control (p<0.05). CoQ10 concentration was 336.0mg/l with 5% R. spharoides. On the other hands, CoQ10 was not detected without R. spharoides. Our study was shown that R. spharoides can produce CoQ10 in rumen environment without harmful effects on rumen fermentation parameter. CoQ10 in rumen may transfer into cow milk through cow metabolism. This strategy might be helpful for producing functional dairy cow milk.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1044-1053
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• 2012

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est5S) was 1,026 bp in length, encoding a protein of 366 amino acid residues with a calculated molecular mass of 40,168 Da. The molecular mass of the enzyme was estimated to be 40,000 Da. The Est5S protein contains the Gly-X-Ser-X-Gly motif found in most bacterial and eukaryotic serine hydrolases. However, the Asp or Glu necessary for the catalytic triad [Ser-Asp-(Glu)-His] was not present, indicating Est5S represents a novel member of the GHSQG family of esterolytic enzymes. BlastP in the NCBI database analysis of Est5S revealed homology to hypothetical proteins and it had no homology to previous known lipases and esterases. Est5S was optimally active at pH 7.0 and $40^{\circ}C$. Among the p-nitrophenyl acylesters tested, high enzymatic activities were observed on the short-chain p-nitrophenyl acylesters, such as p-nitrophenyl acetate, etc. The conserved serine residue ($Ser_{190}$) was shown to be important for Est5S activity. The primers that amplified the est5S gene did not show any relative band with 49 species of culturable rumen bacteria. This implies that a new group esterase gene, est5S, may have come from a noncultured cow rumen bacterium.

• Journal of agriculture & life science
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• v.46 no.2
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• pp.129-137
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• 2012

A carboxymethyl cellulase gene, cel5B, was cloned, sequenced, and expressed in Escherichia coli. pRCS20 in E. coli was identified from metagenomic cosmid library of cow rumen for cellulase activity on a carboxymethyl cellulose agar plates. Cosmid clone (RCS20) was partially digested with Sau3AI, ligated into BamHI site of pBluescript II SK+ vector, and transformed into E. coli $DH5{\alpha}$. The insert DNA of 1.3 kb was obtained, designated cel5B, which has the activity of hydrolyzation of CMC. The cel5B gene had an open reading frame (ORF) of 1,059 bp encoding 352 amino acids with a signal peptide of 48 amino acids and the conserved region, VIYEIYNEPL, belongs to the glycosyl hydrolase family 5. The molecular mass of Cel5B protein expressed from E. coli $DH5{\alpha}$ exhibited to be about 34 kDa by CMC-SDS-PAGE. The optimal pH was 8.0, and the optimal temperature was about $50^{\circ}C$ for its enzymatic activity.