Effect of exogenous phytase on degradation of inositol phosphate in dairy cows.

The effect of exogenous phytase on inositol phosphate degradation in the rumen of dairy cows was investigated in a 4 × 4 Latin square design. Four lactating Danish Holstein cows fitted with ruminal, duodenal, and ileal cannulas were offered a total mixed ration (TMR) with a high content of inositol phosphate and supplemented with 1 of 4 concentrations of phytase [none, low, medium, or high, corresponding to 23, 2,023, 3,982, and 6,015 phytase units/kg of dry matter (DM)]. Exogenous phytase lead to a higher rumen pool of phytase. Inositol phosphate content in digesta samples from rumen, duodenum, ileum, and feces was almost entirely composed of myo-inositol hexakisphosphate (InsP(6)), indicating that degradation of this compound is the rate-limiting step in inositol phosphate degradation in the digestive tract. Ruminal and total-tract degradations of InsP(6) were higher when exogenous phytase was added to the TMR. Degradation of InsP(6) occurred mainly before the duodenum. The ruminal degradability of InsP(6) was increased with increasing dietary concentrations of phytase: 86.4, 93.7, 94.5, and 96.3% for none, low, medium, or high, respectively. A comparison of the InsP(6) content in individual feedstuffs and in samples of the TMR revealed that the exogenous phytase started degrading the inositol phosphate when feeds and phytase were mixed, and thus the InsP(6) phosphorus (InsP(6)-P) content in the TMR was found to decrease with higher doses of phytase (1.69, 1.51, 1.39, and 1.25 g/kg of DM for the none, low, medium, and high phytase doses, respectively). It was not possible to distinguish between the degradation of inositol phosphate occurring in the TMR and in the rumen. Exogenous phytase had no effect on total P intake or flow of total P to the duodenum and ileum, whereas exogenous phytase increased flow of microbial P to the duodenum and total fecal P excretion. None of the investigated rumen variables (pH, degradability of neutral detergent fiber, and rumen kinetics for neutral detergent fiber) were affected by treatment. Rumen and total-tract degradations of inositol phosphate were increased when exogenous phytase was added to the TMR, which offers the potential for reducing P excretion through reduced dietary P.

Effect of exogenous phytase on feed inositol phosphate hydrolysis in an in vitro rumen fluid buffer system.

Three in vitro experiments using a rumen fluid buffer system were performed to investigate the effect of addition of 4 experimental phytases (Phy1, Phy2, Phy3, and Phy4) compared with no addition of phytase on feed inositol phosphate hydrolysis in wheat and rapeseed cake to determine which of the 4 phytases was most suitable under rumen-like conditions. The feedstuffs were incubated with a mixture of physiological buffer, ruminal fluid, and exogenous phytase at pH 6.2, after which the samples were incubated for different periods. Incubations were stopped using HCl, and the samples were analyzed for inositol phosphates via high performance ion chromatography. Addition of phytase (Phy1) resulted in enhanced degradation of myo-inositol hexakisphosphate (InsP(6)) in rapeseed cake, whereas addition of exogenous phytase did not improve the degradation of InsP(6) in wheat. Only rapeseed cake was therefore used subsequently. All 4 phytases increased degradation of InsP(6) in rapeseed cake in the in vitro system, and degradability of InsP(6) increased with higher incubation time and higher phytase dosages, independent of phytase. Addition of 2 units of phytase per gram of substrate of the phytases Phy1, Phy2, Phy3, and Phy4 led to an undegraded InsP(6) content of 56, 49, 70, and 18%, respectively, when incubated with rapeseed cake for 6h, indicating that Phy2 and Phy4 were the most effective phytases. However, Phy2 had a higher specific activity than Phy4, as 60% of the original InsP(6) content was remaining after 3h when 5mg of enzyme protein per gram of substrate of Phy2 was added to rapeseed cake, whereas 150 mg of enzyme protein per gram of substrate of Phy4 was necessary to achieve a similar result. Therefore, Phy2 appeared to be most applicable under rumen-like conditions.

Intestinal metabolism of rye lignans in pigs.

To study the intestinal metabolism of lignans, the concentrations of plant and mammalian lignans in intestinal digesta sampled along the intestinal tract of pigs were determined by isotope dilution GC-MS. The pigs were fed rye-bread diets made from either whole rye-grains or rye-grain milling fractions enriched in pericarp-testa, aleurone or endosperm cells. The content and characteristics of dietary fibre varied between diets and had been shown to induce different colon fermentation patterns. As the metabolism of lignans depends on the action of the intestinal flora, we tested whether the rye-bread diets influence the metabolism of lignans. In the ileum, the lignans were mainly present as conjugated plant lignans, which were determined only when the analytical procedure included a hydrolysis step. High recovery of dietary lignans in the ileum may indicate that the lignans enter the enterohepatic circulation. In addition, two to three times the intake of lignans were recovered in the faeces when the diets had a high content of dietary fibre suggesting underestimation of plant lignans in the diet. Most of the plant lignans disappeared from the intestinal tract between the terminal ileum and the caecum. The intestinal concentrations and the disappearance of lignans correlated with the content of lignans in the diet, being highest on the pericarp-testa diet and lowest on the endosperm diet. No effect of fermentation pattern on the intestinal metabolism of lignans was observed. The lignans were liberated from the pericarp-testa diet although the plant cell walls remained largely undegraded.

Intestinal degradation in pigs of rye dietary fibre with different structural characteristics.

In order to investigate the effects of dietary fibre (DF) characteristics on carbohydrate degradation and the metabolism in the large intestine, pigs were fed on four rye-bread diets (based on whole rye, pericarp/testa, aleurone or endosperm) with differences in characteristics and amount of DF. The degradability of DF in the large intestine varied greatly between diets. The pericarp/testa DF was hardly degraded in the large intestine, whereas endosperm DF was extensively and rapidly degraded in the caecum. Caecal degradation of aleurone DF was also limited, leaving more material to be degraded in the colon. The undegradable pericarp/testa DF was characterized by high contents of lignin, cellulose, ferulic acids and highly substituted arabinoxylans (the major DF component in rye). Ingestion of this diet resulted in increased faecal bulk and reduced transit time, but with low colonic pH and the lowest concentrations of shortchain fatty acids (SCFA). The aleurone diet, on the other hand, led to a fermentation pattern which may be considered more optimal, with lower colonic pH and higher concentrations of SCFA, in particular butyric acid. Despite the large differences in carbohydrate fermentation only minor significant effects on the presence of protein degradation products and on histological measurements (height and diameter of colonic crypts and thickness of the muscularis external) were observed.