In vitro versus in situ evaluation of the effect of phytase supplementation on calcium and phosphorus solubility in soya bean and rapeseed meal broiler diets

Abstract

1. In vitro assays provide a rapid and economical tool to evaluate dietary effects, but have limitations. In this study, the effect of phytase supplementation on solubility, and presumed availability, of calcium (Ca) and phosphorus (P) in soya bean meal (SBM) and rapeseed meal (RSM) based diets were evaluated both in situ and by a two-step in vitro digestion assay that simulated the gastric and small intestine (SI) phases of digestion. 2. Comparison of the in vitro findings to in situ findings was used to evaluate the in vitro assay. Ross 308 broilers (n = 192) were fed on one of 6 SBM or RSM diets supplemented with 0, 500 or 5000 FTU/kg phytase from 0 to 28 d post hatch. The 6 diets and raw SBM and RSM were exposed to a two-step in vitro assay. Ca and P solubility and pH in the gizzard and jejunal digesta and in the gastric and SI phase of in vitro digestion were measured. 3. Both in vitro and in situ analyses detected that Ca solubility was lowest when diets were supplemented with 500 FTU/kg phytase, compared to the control diets and diets supplemented with 5000 FTU/kg phytase. Phosphorus solubility increased with increasing phytase level. Both methods also identified that mineral solubility plateaus in the gastric phase. 4. Overall relationship of the two methods was strong for both determination of gastric phase Ca and P solubility (r = 0.96 and 0.92, respectively) and also SI phase Ca and P solubility (r = 0.71 and 0.82, respectively). However, mineral solubility and pH were higher when measured in vitro than in situ, and the in situ assay identified an interaction among the effects of phase, protein source and phytase inclusion level on Ca solubility that the in vitro assay did not detect. 5. This two-step in vitro assay successfully predicted phytase efficacy, but to determine detailed response effects in the animal, in situ data is still required.