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- W957796448 abstract "The objective of these studies were to identify ruminal yeast in varying ratios of roughage to concentrate in TMR diets in order to explore yeast diversity by using molecular technique with similarity of rDNA sequence. The experiment was assigned to four 98.6% of cross bred Holstein Friesian heifers with 2 levels and two replicates of roughage to concentrate ratios as: 10:90 (T1) and 50:50 (T2). The experimental period was 14 days. Rumen fluid sample was collected by stomach tube for total DNA extraction by using silica gel method, and analysis of quantity and quality of DNA by Nanovue and agarose gel electrophoresis. The divergent D1/D2 domain of 26S rDNA was amplified by primers NL-1(5'-GCA TAT CAA TAA GCG GAG GAA AAG-3') and NL4 (5'-GGT CCG TGT TTCAAG ACG G-3') by polymerase chain reaction (PCR). The nucleotide sequences of D1/D2 domain of 26S rDNA were determined using PCR products. Generated sequences were aligned with related species by using the CLUSTAL W. The result showed that an average dry matter intake of T1 was 7.00 kg/d and T2 was 6.99 kg/d. DNA concentrate from T1R1, T1R2, T2R1 and T2R2 were 106, 131.5, 84 and 182.5 ng/μL, respectively. The purity of DNA was 1.57, 1.76, 1.78 and 1.86, respectively. The divergent D1/D2 domain of 26S rDNA of treatment could be amplified for T1R1 and T2R1 but could not for T1R2 and T2R2. The sequences of D1/D2 domain of 26S rDNA were compared with nucleotide database by BLAST programs (http://www.ncbi.nlm.nih.gov/BLAST), the T2R1 yeast-strain was closest to Yarrowia lipolytica. However, yeast strain in T1R1 could not be specifically identified because D1/D2 domain of 26S rDNA seem to represent variable region with number of nucleotide sequence showing 2-3 substitution from known species. The phylogenetic tree based on the sequences of the D1/D2 domain of 26S rDNA showed that T1R1 was related to Pichia and Candida (96%) and T2R1 was related to Yarrowia lypolytica (100%). This study indicated that ruminal yeast strains could be found varying in different ratio of roughage to concentrate." @default.
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- W957796448 date "2011-12-01" @default.
- W957796448 modified "2023-09-27" @default.
- W957796448 title "Screening Yeasts from Ruminal Fluid of Dairy Heifer Fed a Different Ratio Roughage to Concentrate Diets" @default.
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