Determination of assay parameters for RNA analysis in bacterial and duodenal samples by spectrophotometry. Influence of sample treatment and preservation
Year: 1985
Type: article
Abstract: A simple analytical procedure derived from that described by Zinn and Owens (1980), based on the determination of nucleic purine bases (RNA eq), was carried out to measure microbial nitrogen flow in the ruminant duodenum.Several procedures for sample preservation were tested ; the efficiency of each step of the analytical method was also determined using high performance liquid chromatography (HPLC).The proposed method (RNA eq) was compared with two other methods considered as references (2-6 diaminopimelic acid and 35 S incorporation) and microbial nitrogen flow was measured in defaunated sheep.The recovery of purine bases analysed by the Zinn and Owens method was generally good (101 % pure bases ; 90 % when bases were added to bacterial samples ; 96 % when added to yeast RNA).The HPLC measurements allowed us to conclude that this spectrophotometric method is specific for purine bases, all pyrimidine bases being eliminated.Moreover, it was found that the method must be used on freeze-dried samples ; storage at + 4 °C, defatting or freezing gave incorrect results.Using the described assay, we observed that microbial nitrogen flow at the duodenum of defaunated sheep was not significantly different from that obtained using more traditional markers such as sulphur-35 incorporation or diaminopimelic acid. Introduction.The utilization of nucleic acids as a marker for quantifying microbial synthesis in the rumen was first suggested by Gausseres and Fauconneau 119651.A series of studies on the utilization of this marker was completed by McAllan and Smith (1969, 1972, 1973, 1983, 1984) in which these authors reported its advantages and disadvantages.The analytical procedure for determining acids is so long and laborious that it is difficult to apply this method to trials which provide large quantities of samples.Some workers use a simpler analytical procedure (Topps and Elliott, 1965) or determine nucleic acid purine bases (McDonald, 1954).However, McAllan and Smith (1969) pointed out that these Reproduction Nutrition D6veloppement, n° 6-85.-3. 3.
DOI: 10.1051/rnd:19850804
Authors: K. USHIDA, Bernadette LASSALAS, J.-P. JOUANY
Language: English
Subfield: Animal Science and Zoology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W2021202014