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

1. In vitro assays provide a sensitive and economic 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 soybean meal (SBM) and rapeseed meal (RSM) based diets was evaluated both in situ and by a 2-step in vitro digestion assay that simulated the gastric and small intestine (SI) phases of digestion

Contribution of intestinal- and cereal-derived phytase activity on phytate degradation in young broilers

There is little consensus as to the capability of poultry to utilize dietary phytate without supplemental phytase. Therefore, an experiment was conducted to examine the extent to which endogenous phytase of intestinal and cereal origin contributes to phytate degradation in birds aged 0 to 14 d posthatch. Ross 308 broilers (n = 720) were fed one of 4 experimental diets with differing dietary ingredient combinations and approximate total phytate levels of 10 g/kg, dietary phytase activity analyzed at 460 U/kg, dietary calcium (Ca) levels of 11 g/kg, and nonphytate-phosphorus (P) levels of 4 g/kg. Broiler performance, gizzard, duodenum, jejunum and ileum pH, Ca and P digestibility and solubility, amount of dietary phytate hydrolyzed in the gizzard, jejunum, and ileal digesta phytase activity were analyzed at d 4, 6, 8, 10, 12, and 14 posthatch. Intestinal endogenous phytase activity increased significantly (P < 0.001) between d 4 and 6, resulting in increased phytate hydrolysis in the gizzard (P = 0.003), jejunum (P < 0.001), and ileum (P < 0.001). Phytase activity and phytate hydrolysis continued to increase with age, with a greater phytase activity and associated increase in phytate hydrolysis and mineral utilization between d 10 and 12. Gizzard and jejunum Ca and P solubility and ileal Ca and P digestibility increased significantly (P < 0.001), and gastrointestinal pH decreased significantly (P < 0.001) between d 4 and 6. By d 14, phytase activity recovered in the ileum was approximately 45 U/kg. There were strong correlations between phytase activity measured in the ileum and phytate hydrolyzed in the gizzard (r = 0.905, P < 0.001), jejunum (r = 0.901, P = 0.023), and ileum (r = 0.938, P = 0.042). This study shows intestinal- and dietary-derived endogenous phytase activity is responsible for phytate-P hydrolysis in broilers

Effect of phytase on growth performance, phytate degradation and gene expression of myo-inositol transporters in the small intestine, liver and kidney of 21 day old broilers

An experiment was conducted to evaluate phytase on growth performance, phytate degradation and the gene expression of myo-inositol transporters in 21-day old broilers. Ross 308, male broilers (n = 240) were obtained and assigned to one of four diets, 10 with pens/diet and six birds/pen from day one to 21. The diets consisted of a negative control (NC) formulated to meet or exceed Ross 308 nutrient requirements, with the exception of calcium (Ca) and available P (avP), which was reduced by 0.16 and 0.15%, respectively. The NC diet was supplemented with 0, 500, 1,500 or 4,500 FTU/kg of phytase to create four experimental diets. On day 21, all birds per pen were euthanized to obtain digesta and tissue samples for phytate degradation and gene expression. Data were analysed as an analysis of variance using the fit model platform in JMP v 13.0. The model included phytase and significant means were separated using orthogonal linear and quadratic contrasts. Phytase supplementation increased gain (linear, P &lt; 0.05) but had no effect on feed intake or feed conversion ratio. Phytate (IP6; quadratic, P &lt; 0.05), phytate ester (IP5, IP4, IP3; quadratic, P &lt; 0.05) and inositol (linear, P &lt; 0.05) concentration in the gizzard was influenced by phytase supplementation. Phytase supplementation decreased IP6 (linear, P &lt;0.05) and IP5, IP4, IP3 (linear or quadratic, P &lt; 0.05) and increased inositol (quadratic, P &lt; 0.05) concentration in the ileal digesta. The expression of the H+-dependent myo-inositol transporter, HMIT, was decreased (linear, P &lt; 0.05) in the kidney and increased (linear, P &lt; 0.05) in the ileum as phytase dose increased. Expression of the sodium-dependent myo-inositol transporter, SMIT2, increased in the liver (quadratic, P &lt; 0.10) and the jejunum (quadratic, P &lt; 0.05) as phytase dose increased. Intestinal alkaline phosphatase expression increased in the ileum (linear, P &lt; 0.05) as phytase dose increased. The influence of phytase on phytate, phytate esters and inositol may influence intestinal alkaline phosphatase activity and the expression of myo-inositol transporters in the small intestine and kidney

Hydrolysis of phytate to its lower esters can influence the growth performance and nutrient utilization of broilers with regular or super doses of phytase

The aim of the study was to observe the effects of dietary available phosphorus (aP) and calcium (Ca), with regular or super doses of phytase, on phytate hydrolysis and subsequent influences on broiler growth performance and nutrient utilization. In a 2 × 3 factorial design, 384 Ross-308 broilers were allocated to one of 6 dietary treatments with 8 replicates in a randomized complete block design for 21 days. Diets were nutritionally adequate (positive control, PC) or marginally deficient in aP and Ca (negative control, NC), with 0, 500 or 1,500 FTU/kg phytase. Bird and feed weights were recorded on d 0 and 21, excreta were collected on d 19 and 20, and gizzard and ileal contents were collected on d 21. Body weight gain (P &lt; 0.01) increased linearly with phytase in the PC and quadratically in the NC. There was an interactive effect on ileal DM, N, and P utilization, increasing quadratically with phytase supplementation in the NC, but there was no phytase influence in the PC (P &lt; 0.05). Phytase linearly increased copper (P &lt; 0.001) and linearly decreased Ca (P &lt; 0.05) utilization in the ileum. Phytase decreased ileal (IPx, inositol x-phosphate) IP6 and IP5 and increased inositol (quadratic, P &lt; 0.001) but had no effect on IP4 or IP3. The influence of the dietary aP was more apparent on the hydrolysis of phytate and phytate esters after the ileum, with increasing (linear and quadratic, P &lt; 0.05) IP4 and IP3 content in the excreta of birds fed the NC or PC when phytase was added. Phytate hydrolysis improves the growth potential of birds fed NC diets, allowing them to match the growth performance of birds fed PC diets and improve nutrient utilization. These results indicate that dietary Ca and aP concentrations can be reduced when phytase is supplemented. It also may be beneficial to apply the enzyme nutrient matrix to other nutrients in the diet to maintain an optimal balance of nutrients in the digesta

Low-level regulatory T-cell activity is essential for functional type-2 effector immunity to expel gastrointestinal helminths

Helminth infection is frequently associated with the expansion of regulatory T cells (Tregs) and suppression of immune responses to bystander antigens. We show that infection of mice with the chronic gastrointestinal helminth Heligmosomoides polygyrus drives rapid polyclonal expansion of Foxp3(+)Helios(+)CD4(+) thymic (t)Tregs in the lamina propria and mesenteric lymph nodes while Foxp3(+)Helios(-)CD4(+) peripheral (p)Treg expand more slowly. Notably, in partially resistant BALB/c mice parasite survival positively correlates with Foxp3(+)Helios(+)CD4(+) tTreg numbers. Boosting of Foxp3(+)Helios(+)CD4(+) tTreg populations by administration of recombinant interleukin-2 (rIL-2):anti-IL-2 (IL-2C) complex increased worm persistence by diminishing type-2 responsiveness in vivo, including suppression of alternatively activated macrophage and granulomatous responses at the sites of infection. IL-2C also increased innate lymphoid cell (ILC) numbers, indicating that Treg functions dominate over ILC effects in this setting. Surprisingly, complete removal of Tregs in transgenic Foxp3-DTR mice also resulted in increased worm burdens, with "immunological chaos" evident in high levels of the pro-inflammatory cytokines IL-6 and interferon-γ. In contrast, worm clearance could be induced by anti-CD25 antibody-mediated partial depletion of early Treg, alongside increased T helper type 2 responses and without incurring pathology. These findings highlight the overarching importance of the early Treg response to infection and the non-linear association between inflammation and the prevailing Treg frequency

The Toxic Effects of Cigarette Additives. Philip Morris' Project Mix Reconsidered: An Analysis of Documents Released through Litigation

Stanton Glantz and colleagues analyzed previously secret tobacco industry documents and peer-reviewed published results of Philip Morris' Project MIX about research on cigarette additives, and show that this research on the use of cigarette additives cannot be taken at face value