Abstract
Biotin is present in nature either free or as biocytin, which is only degraded under the action of a specific enzyme: biotinidase. This enzyme is not included in analytical assays generally used. A method for sample preparation using biotinidase was developed in our laboratory before analysis by ELISA. Three cows equipped with duodenal and ileal cannulae were used to compare the effects of methods of sample preparation on calculations of apparent ruminal synthesis and intestinal absorption of biotin. There was no apparent ruminal synthesis of biotin, no matter whether free or total biotin was measured (p = 0.84). Results also suggested that rumen microbes cannot utilize nor degrade biocytin present in the feed. Estimates of apparent intestinal absorption were influenced by the sample preparation method (p = 0.002). Analysis of free biotin caused an artefact, suggesting intestinal synthesis of this vitamin; whereas determination of total biotin concentrations showed that absorption was taking place in the small intestine.
Acknowledgements
The authors would like to acknowledge the work of Véronique Roy for the tests performed with this assay, as well as for analysis of the samples; Chrystiane Plante for technical support; Steve Méthot for statistical advice; and the staff from the dairy unit of Agriculture and Agri-Food Canada Dairy and Swine Research and Development Centre, Sherbrooke, for animal care.