At wk of age, a blood sample was collected from each animal in disposable evacuated tubes containing EDTA as the anticoagulant, and the plasma was frozen at C for subsequent vitamin B determination.During this last period, pigs were offered a g pellet made of the basal diet mixed with unsweetened apple sauce to induce rapid consumption and g, corresponding to or gkg of diet, before the daily meal.A representative sample of each carcass was then collected and placed in molds of cm and refrozen at C.These samples were further ground to mm in a centrifugal grinder, which was located in a freezer, and liquid nitrogen was used to keep the sample frozen.The same procedure was used for the digestive tract.For vitamin B determination, carcass samples were homogenized in a glass grinder tube with mL of sodium acetate buffer. One mililiter of this homogenate was incubated in a water bath at C for min.The mixture was then autoclaved at C for min and cooled in an icecold water bath.with approximately L of. Two homogenates were prepared for each tissue sample and vitamin B was determined in duplicate for each homogenate in water for liver or undiluted, using a radioassay procedure, for which pig intrinsic factor as the vitamin B binding protein.For each B or B pig, urinary excretion of vitamin B was calculated as the amount excreted during the experimental period of dminus the average daily excretion measured dbefore the initiation of treatments.Similarly, vitamin B retention in each body portion of B and B pigs was calculated as the amount measured in these animals minus the amount present in tissues of the CONT pig within the same block.The total amount of vitamin B absorbed corresponded to the summation of retention and urinary excretion.The bioavailability of dietary cyanocobalamin was expressed in percentage as follows. During the whole experimental period, catheters were rinsed times per week with heparinized saline. The treatments were given as a g pellet as <a href="http://www.targetmol.com/compound/Mirin">buy
Mirin</a> described for trial, providing just before the meal were collected simultaneously from the catheters and min before the meal, every min for the first hpostfeeding, and every hour for the next h.Plasma was collected after centrifugation at, gfor min at C and frozen at C for further analysis.A positive net flux indicates a release of a nutrient from the portaldrained viscera, whereas a negative flux indicates an uptake.Arterial concentrations and net flux of vitamin B across the portaldrained viscera recorded and min before the meal were averaged and used as time. Arterial concentrations and net flux of vitamin B across the portaldrained viscera were analyzed using the MIXED procedure of SAS according to crossover design, and pigs, periods, and treatments were included in the model along with repeated measures in time.Because the time intervals were unequally spaced, the following covariance structures were compared: space, and space. For each variable, the statistical analysis retained was with the best fit statistic value.It seems that the postweaning period, during which the animals were fed the diet unsupplemented with vitamin B, was not sufficient to deplete liver reserves.