Monitoring Phytate Hydrolysis Using Serial Blood Sampling and Feather Myo-Inositol Levels in Broilers.

Phytate forms insoluble precipitates with various cations that are recalcitrant to digestion in poultry. Dietary supplementation with exogenous phytase has been shown to improve phytate solubility and digestibility and, in turn, improve animal growth performance. Although the kinetics of phytate hydrolysis by exogenous phytase are well described in vitro, the progression of the reaction in vivo is still not well defined. The aim of the present study was, therefore, to monitor the kinetic variation of myo-inositol (myo-Ins) levels in both circulation and feather following exogenous phytase supplementation. In experiment 1, 4 week-old male broilers were individually housed with ad libitum access to water and a standard commercial diet. Birds were maintained under environmental temperature of 24°C and 30% RH. Birds were cannulated in the cutaneous ulnar vein on the right wing and remained untouched for 3 days. On the day of the experiment, birds were randomly divided into three body weight-matched groups and fed either the control diet, the control diet-supplemented with myo-Ins or Ronozyme HiPhos (0.06%, DSM Nutritional Products, Switzerland) for 10 h. In the experiment 2, birds were fed only HiPhos for 30 h. Growing feathers and blood were collected at baseline and then every 2 h for 10 h (experiment 1) and 30 h (experiment 2) post-prandially. Plasma and feather myo-Ins levels were determined by UHPLC-MS/MS. The relative expression of inositol polyphosphate-1-phosphatase (INPP1), inositol hexakisphosphate kinase 1-3 (IP6K1-3), inositol-3-phosphate synthase (ISYNA), and multiple inositol-polyphosphate phosphatase 1 (MNPP1) genes in blood and feathers was determined by real-time qPCR using 2-ΔΔCt method. Plasma and feather myo-Ins levels were significantly increased by HiPhos at 6 h to 8 h post-prandial. The mRNA abundances of INPP1, IP6K1, and ISYNA in the circulation were significantly down regulated at all periods compared to the baseline levels. IP6K2, IP6K3, and MINPP1 gene expression, however, was up regulated at 8 h post-prandial and then returned to the baseline levels. In feathers, the expression of INPP1 was induced at 8 h post-prandial and remained higher compared to the baseline. The expression of IP6K2, IP6K3, and MINPP1 was down regulated during the first 10 h and then returned to baseline levels for the rest of the post-prandial period. Taken together, our data show that phytase modulates the expression of genes associated with myo-Ins metabolism and generates release of myo-Ins in both circulation and feather at 6-10 h post-feeding. Feather myo-Ins concentration could be used as a non-invasive method to monitor phytate hydrolysis in practice.

Extra-phosphoric effects of super dosing phytase on growth performance of pigs is not solely due to release of myo-inositol.

Two experiments were conducted to determine the effects of myo-inositol and phytase on growth performance, plasma metabolites, and nutrient digestibility of growing pigs. In experiment 1, 96 growing pigs with average initial body weight (BW) of 26.2 kg were used in a 25-d growth performance study. Pigs were assigned to four dietary treatments with three pigs per pen and eight replicate pens per treatment in a randomized complete block design. The four treatments were control diet (CD); CD + 2 g/kg inositol; CD + 1,000 FYT/kg phytase and CD + 3,000 FYT/kg phytase. Pigs were weighed individually every week. On day 25, blood sample was collected from one pig per pen to measure plasma metabolites concentrations. In experiment 2, 16 barrows (initial BW 34.8 ± 8.2 kg) were surgically fitted with T-cannulas. Pigs were allotted to four blocks based on BW and assigned to a quadruplicate 4 × 2 incomplete Latin square design with same four dietary treatments and two periods. Ileal digesta samples were collected from each pig on days 6 and 7 of each period to determine apparent ileal digestibility (AID) of nutrients. Phytase supplementation increased final BW and average daily gain (ADG) compared with CD (P < 0.05) with no effects on average daily feed intake (ADFI) and gain to feed (G:F) was higher in 3,000 FYT/kg phytase (P < 0.05). Inositol supplementation had no effects on growth performance. Plasma myo-inositol concentration was increased by inositol supplementation, and 3,000 FYT/kg phytase increased myo-inositol in the plasma by 97.2% (P < 0.05). Plasma P concentration was increased by 1,000 or 3,000 FYT/kg phytase with no effects on alkaline phosphatase (ALP), glucose, triglycerides (TAG), calcium (Ca), and urea concentrations. Phytase supplementation reduced (P < 0.05) the phytate-P concentration in the ileal digesta and increased the digestibility of phytate-P and total P with no effects on the AID of dry matter (DM), gross energy (GE), nitrogen (N), and Ca. In conclusion, the beneficial effects of 3,000 FYT/kg phytase on feed efficiency may due to the increased release of both myo-inositol and phosphorus (P), and may not be solely due to myo-inositol release by this level of phytase.

Time-series responses of swine plasma metabolites to ingestion of diets containing myo-inositol or phytase.

The effect of the ingestion of diets containing either myo-inositol or exogenous phytase on plasma metabolites was examined using 29 kg barrows. The diets were: control (maize, soya, rapeseed, rice bran), control plus 2 g/kg myo-inositol, control plus 1000 phytase units (FYT)/kg or 3000 FYT/kg exogenous phytase. Pigs were housed in a PigTurn device and blood was collected, from jugular catheters, via an automated system at -30, (30 min before feeding), 0, 15, 30, 45, 60, 90, 120, 150, 180, 240, 300 and 360 min post-feeding. The addition of 2 g/kg myo-inositol to the basal diet resulted in an increase in plasma myo-inositol concentration that was evident 45-60 min after diet introduction and persisted to 360 min post-feeding. Similarly, supplementation of the basal diet with either 1000 or 3000 FYT/kg exogenous phytase resulted in an increase in plasma myo-inositol concentration that was still rising 360 min post-feeding. Plasma P concentration was increased over time by the addition of 1000 and 3000 FYT/kg phytase, but not by the addition of myo-inositol. Other plasma metabolites examined were not affected by dietary treatment. It can be concluded that oral delivery of myo-inositol results in rapid increase in plasma myo-inositol concentrations that peak approximately 45-60 min after feeding. Use of supplemental phytase achieves similar increases in myo-inositol concentration in plasma but the appearance is more gradual. Furthermore, supplementation of pig diets with exogenous phytase results in rapid appearance of P in plasma that may be sustained over time relative to diets with no added phytase.