P and Ca digestibility is increased in broiler diets supplemented with the high-phytase HIGHPHY wheat

Around 70% of total seed phosphorus is represented by phytate which must be hydrolysed to be bioavailable in non-ruminant diets. The limited endogenous phytase activity in non-ruminant animals make it common practice to add an exogenous phytase source to most poultry and pig feeds. The mature grain phytase activity (MGPA) of cereal seeds provides a route for the seeds themselves to contribute to phytate digestion, but MGPA varies considerably between species and most varieties in current use make negligible contributions. Currently, all phytases used for feed supplementation and transgenic improvement of MGPA are derived from microbial enzymes belonging to the group of histidine acid phosphatases (HAP). Cereals contain HAP phytases, but the bulk of MGPA can be attributed to phytases belonging to a completely different group of phosphatases, the purple acid phosphatases (PAPhy). In recent years, increased MGPAs were achieved in cisgenic barley holding extra copies of barley PAPhy and in the wheat HIGHPHY mutant, where MGPA was increased to ~6200 FTU/kg. In the present study, the effect of replacing 33%, 66% and 100% of a standard wheat with HIGHPHY wheat was compared with a control diet with and without 500 FTU of supplemental phytase. Diets were compared by evaluating broiler performance, ileal Ca and P digestibility and tibia development, using nine replicate pens of four birds per diet over 3 weeks from hatch. There were no differences between treatments in any tibia or bird performance parameters, indicating the control diet did not contain sufficiently low levels of phosphorus to distinguish effect of phytase addition. However, in a comparison of the two wheats, the ileal Ca and P digestibility coefficients for the 100% HIGHPHY wheat diets are 22.9% and 35.6% higher, respectively, than for the control diet, indicating the wheat PAPhy is functional in the broiler digestive tract. Furthermore, 33% HIGHPHY replacement of conventional wheat, significantly improved Ca and P digestibility over the diet-supplemented exogenous phytase, probably due to the higher phytase activity in the HIGHPHY diet (1804 v. 1150 FTU). Full replacement by HIGHPHY gave 14.6% and 22.8% higher ileal digestibility coefficients for Ca and P, respectively, than for feed supplemented with exogenous HAP phytase at 500 FTU. This indicates that in planta wheat PAPhys has promising potential for improving P and mineral digestibility in animal feed.     

Zinc availability and digestive zinc solubility in piglets and broilers fed diets varying in their phytate contents, phytase activity and supplemented zinc source

The study was conducted to evaluate the effects of dietary zinc addition (0 or 15 mg/kg of Zn as inorganic or organic zinc) to three maize-soybean meal basal diets varying in their native Zn, phytic P contents and phytase activity (expressed in kg of feed: P- with 25 mg Zn and 1.3 g phytic P, P+ with 38 mg Zn and 2.3 g phytic P or P+/ENZ being P+ including 500 units (FTU) of microbial phytase per kg) in two monogastric species (piglets, broilers). Measured parameters were growth performance, zinc status (plasma, and bone zinc) and soluble zinc in digesta (stomach, gizzard and intestine). The nine experimental diets were fed for 20 days either to weaned piglets (six replicates per treatment) or to 1-day-old broilers (10 replicates per treatment). Animal performance was not affected by dietary treatments (P > 0.05) except that all P- diets improved body weight gain and feed conversion ratio in piglets (P < 0.05). Piglets fed P- diets had a better Zn status than those fed P+ diets (P < 0.05). In both species, Zn status was improved with supplemental Zn (P < 0.05), irrespective of Zn source. Phytase supplementation improved piglet Zn status to a higher extent than adding dietary Zn, whereas in broilers, phytase was less efficient than supplemental Zn. Digestive Zn concentrations reflected the quantity of ingested Zn. Soluble Zn (mg/kg dry matter) and Zn solubility (% of total Zn content) were highest in gizzard contents, which also presented lower pH values than stomach or intestines. The intestinal Zn solubility was higher in piglet fed organic Zn than those fed inorganic Zn (P < 0.01). Phytase increased soluble Zn in piglet stomach (P < 0.001) and intestine (P = 0.1), but not in broiler gizzard and intestinal contents. These results demonstrate (i) that dietary zinc was used more efficiently by broilers than by piglets, most probably due to the lower gizzard pH and its related higher zinc solubility; (ii) that zinc supplementation, irrespective of zinc source, was successful in improving animal's zinc status; and (iii) suggest that supplemented Zn availability was independent from the diet formulation. Finally, the present data confirm that phytase was efficient in increasing digestive soluble Zn and improving zinc status in piglets. However, the magnitude of these effects was lower in broilers probably due to the naturally higher Zn availability in poultry than in swine.     

Sparing effect of microbial phytase on zinc supplementation in maize-soya-bean meal diets for chickens

The experiment was conducted to evaluate the sparing effect of microbial phytase on the need for dietary zinc supplementation in chicks. A maize–soya-bean meal basal diet, containing 33 mg of zinc and 16 mg of copper per kg, supplemented with 0, 6, 12, 18, 24, 30 or 60 mg of zinc as sulphate per kg or with 250, 500, 750 or 1000 units (FTU) of microbial phytase (3-phytase from Aspergillus niger, Natuphos®) per kg was given to 1-day-old chicks for 20 days. Sixteen chicks placed in individual cages were assigned to each diet except the unsupplemented basal diet which was assigned to 32 cages. Actual range of phytase supplementation was 280 to 850 FTU per kg diet. Growth performance was not affected by microbial phytase. Chicks given the unsupplemented basal diet and the basal diet supplemented with 60 mg of zinc per kg displayed similar performance. Bone weight, bone ash, liver weight and liver dry matter were independent (P > 0.1) of zinc and phytase supplementations. Plasma, bone and liver zinc concentrations increased linearly (P < 0.001) and quadratically (P < 0.001; P < 0.001 and P < 0.05, respectively) with zinc added. Plasma zinc tended to increase linearly (P = 0.07) and bone zinc increased linearly (P < 0.01) with phytase added but no quadratic response was detected (P > 0.1). Liver zinc was unresponsive to phytase added (P > 0.1). Liver copper decreased linearly (P < 0.001) and quadratically (P < 0.01) with zinc supplementation. Mathematical functions were fitted to the responses of plasma and bone zinc to zinc and phytase added and used to calculate zinc equivalency values of phytase. The models included a linear plateau response to zinc added and a linear response to phytase added. In diets without phytase, plasma and bone zinc concentrations were maximised for a dietary zinc concentration of 55 and 51 mg/kg, respectively. Over the range of 280 to 850 FTU, 100 FTU was equivalent to 1 mg of zinc as sulphate. Consequently, in a maize–soya-bean meal chicken diet formulated to contain 60 mg zinc per kg, zinc ingested, and in turn, zinc excreted may be reduced by around 10% if the diet contains 500 FTU as Natuphos® per kg.     

Dietary phytase and myo-inositol supplementation are associated with distinct plasma metabolome profile in broiler chickens

Phytase enzyme is used as a dietary supplement in broiler nutrition to improve phosphorous bioavailability. Phytase deliberates phosphate groups from phytic acid and produces myo-inositol after total dephosphorylation. Myo-inositol is a bioactive compound having beneficial modulatory effects on metabolism in humans. However, it is not well understood if and how phytic acid degradation products, particularly myo-inositol, can modulate metabolism in broiler chicken. The purpose of this study was to investigate effects of dietary supplements of phytase and myo-inositol on the blood plasma metabolome profile of broiler chickens. Broilers were provided a nutrient-adequate control diet or the same diet supplemented with either 3.5 g myo-inositol or 500, 1500 or 3000 units of phytase, per kilogram of feed (grower diet). Broilers were group-housed in floor pens (eight pens per diet) and provided one of the treatment diets for 22 days. Then, blood was collected from one bird per pen, resulting in eight replicated measurements per diet. A targeted metabolomics approach was applied to the heparin plasma. Body weight of the birds was not significantly affected by the treatments. Plasma myo-inositol concentrations were significantly increased by myo-inositol supplementation and phytase supplementation at 500 and 1500 units/kg. Metabolites generally affected by phytase supplementation belonged to the groups of acyl-carnitines, phosphatidylcholines, sphingomyelins, lysophosphatidylcholine, biogenic amines and amino acids. Compared to the control diet, phytase supplements had significantly higher plasma concentrations of kynurenine and creatinine, but lower concentrations of histamine and cis-4-hydroxyproline. Myo-inositol supplementation significantly increased plasma concentrations of dopamine and serotonine. While some metabolites were similarly affected by myo-inositol and phytase supplementation, others were distinctly differently affected. We conclude that myo-inositol, either as a directly added supplement or indirectly released from phytate upon phytase supplementation, can affect specific metabolic pathways. Additional effects found on phytase supplementation may be related to intermediary phytate degradation products. Results are indicative for innovative hypothesis to be tested in future experiments, for instance, with regard to relationships between phytase or myo-inositol supplements and bird immunity or behaviour.     

Microbial phytase addition resulted in a greater increase in phosphorus digestibility in dry-fed compared with liquid-fed non-heat-treated wheat–barley–maize diets for pigs

The objective was to evaluate the effect of microbial phytase (1250 FTU/kg diet with 88% dry matter (DM)) on apparent total tract digestibility (ATTD) of phosphorus (P) in pigs fed a dry or soaked diet. Twenty-four pigs (65±3 kg) from six litters were used. Pigs were housed in metabolism crates and fed one of four diets for 12 days; 5 days for adaptation and 7 days for total, but separate collection of feces and urine. The basal diet was composed of wheat, barley, maize, soybean meal and no mineral phosphate. Dietary treatments were: basal dry-fed diet (BDD), BDD with microbial phytase (BDD+phy), BDD soaked for 24 h at 20°C before feeding (BDS) and BDS with microbial phytase (BDS+phy). Supplementation of microbial phytase increased ATTD of DM and crude protein (N×6.25) by 2 and 3 percentage units (P<0.0001; P<0.001), respectively. The ATTD of P was affected by the interaction between microbial phytase and soaking (P=0.02). This was due to a greater increase in ATTD of P by soaking of the diet containing solely plant phytase compared with the diet supplemented with microbial phytase: 35%, 65%, 44% and 68% for BDD, BDD+phy, BSD and BSD+phy, respectively. As such, supplementation of microbial phytase increased ATTD of P in the dry-fed diet, but not in the soaked diet. The higher ATTD of P for BDS compared with BDD resulted from the degradation of 54% of the phytate in BDS by wheat and barley phytases during soaking. On the other hand, soaking of BDS+phy did not increase ATTD of P significantly compared with BDD+phy despite that 76% of the phytate in BDS+phy was degraded before feeding. In conclusion, soaking of BDS containing solely plant phytase provided a great potential for increasing ATTD of P. However, this potential was not present when microbial phytase (1250 FTU/kg diet) was supplemented, most likely because soaking of BDS+phy for 24 h at 20°C did not result in a complete degradation of phytate before feeding.     

Improvement in overall performance of Catla catla fingerlings fed phytase included low cost plant by products-based diet

Phytic acid’s presence in low-cost Moringa by-products effect the availability of important nutrients, diminishing the fish quality and blood composition in fish. Phytate having chelating effects with nutrients and minerals, can be reduced by the supplementation of phytase enzyme. Without the use of enzyme, plant meal may cause water pollution and decrease the fish health that results in higher culture cost. Therefore, current study was designed to check improvement in overall performance of Catla catla fingerlings fed Moringa by product-based diets supplemented with phytase (0, 300, 600, 900, 1200 and 1500, FTU/kg). All diets were integrated with non-digestible marker (Cr₂O₃) at the rate of 1%. The fingerlings were fed couple of times a day (4% of live wet weight). Results showed significant (p < 0.05) improvement in nutrient digestibility (i.e. EE, CP and GE), carcass composition and hematological parameters (i.e. RBCs, PLT and Hb) at 900, FTU/kg of phytase in contrast with other treatments. Moreover, phytase addition improves the water quality by reducing the nutrients leaching through feces at low cost. Current results indicated that, using mixture of Moringa seed meal and Moringa leaf meal based diet supplemented with phytase at 900, FTU/kg concentration is the most optimum level to develop a cost-effective as well as eco-friendly fish feed with maximum absorption of important nutrients and minerals in fish body resultantly high higher fish performance.     

Complete hydrolysis of <Emphasis Type="Italic">myo</Emphasis>-inositol hexakisphosphate by a novel phytase from <Emphasis Type="Italic">Debaryomyces castellii</Emphasis> CBS 2923

Debaryomyces castellii phytase was purified to homogeneity in a single step by hydrophobic interaction chromatography. Its molecular mass is 74 kDa with 28.8% glycosylation. Its activity was optimal at 60°C and pH 4.0. The K _m value for sodium phytate was 0.532 mM. The enzyme exhibited a low specificity and hydrolyzed many phosphate esters. The phytase fully hydrolyzed myo-inositol hexakisphosphate (or phytic acid, Ins P₆) to inositol and inorganic phosphate. The sequence of Ins P₆ hydrolysis was determined by combining results from high-performance ionic chromatography and nuclear magnetic resonance. D. castellii phytase is a 3-phytase that sequentially releases phosphate groups through Ins (1,2,4,5,6) P₅, Ins (1,2,5,6) P₄, Ins (1,2,6) P₃, Ins (1,2) P₂, Ins (1 or 2) P₁, and inositol (notation 3/4/5/6/1 or 2).     

Influence of Phytase Transgenic Corn on the Intestinal Microflora and the Fate of Transgenic DNA and Protein in Digesta and Tissues of Broilers

An experiment was conducted to investigate the effect of phytase transgenic corn (PTC) on intestinal microflora, and the fate of transgenic DNA and protein in the digesta and tissues of broilers. A total of 160 1-day-old Arbor Acres commercial male broilers were randomly assigned to 20 cages (8 chicks per cage) with 10 cages (replicates) for each treatment. Birds were fed with a diet containing either PTC (54.0% during 1–21 days and 61.0% during 22–42 days) or non-transgenic isogenic control corn (CC) for a duration of 42 days. There were no significant differences (P>0.05) between birds fed with the PTC diets and those fed with the CC diets in the quantities of aerobic bacteria, anaerobic bacteria, colibacillus and lactobacilli, or microbial diversities in the contents of ileum and cecum. Transgenic phyA2 DNA was not detected, but phyA2 protein was detected in the digesta of duodenum and jejunum of broilers fed with the PTC diets. Both transgenic phyA2 DNA and protein fragments were not found in the digesta of the ileum and rectum, heart, liver, kidney, and breast or thigh muscles of broilers fed with the PTC diets. It was concluded that PTC had no adverse effect on the quantity and diversity of gut microorganisms; Transgenic phyA2 DNA or protein was rapidly degraded in the intestinal tract and was not transferred to the tissues of broilers.     

Effect of microbial phytase supplementation on P digestibility in pigs: a meta-analysis

ABSTRACT

The objectives of this meta-analysis were to determine to which extent phosphorus (P) digestibility and digestible P concentration in pig diets were increased by phytase supplementation and to quantify factors that potentially influence effects of phytase supplementation. A data set with a total of 547 data lines was compiled from 88 experiments published in 74 peer-reviewed papers between 2007 and April 2019. An exponential model was determined as more suitable to describe the response of P digestibility to phytase supplementation than a polynomial model. Phytase supplementation increased P digestibility by 25.6 percentage points (standard error (SE) = 1.54) to a plateau at 64.9% (SE = 1.82). The digestible P concentration was increased by phytase supplementation in the order of 1.01 g/kg (SE = 0.102) to a plateau at 2.62 g/kg (SE = 0.122). Goodness-of-fit criteria were R² = 0.780 and root mean square error = 7.55% for P digestibility, and R² = 0.691 and root mean square error = 0.48 g/kg for digestible P concentration. Consideration of further factors such as mineral P supplementation (yes or no), ad libitum vs. restrictive feeding, mixed diets vs. single feed ingredients, sex and age of pigs did not increase the accuracy of prediction in this data set. Some of these traits exhibited responses, but they likely are artefacts generated through the imbalanced structure of the data set. Effects of dietary total P, phytate (InsP₆), InsP₆-P to total P ratio, and Ca on the effect of supplemented phytase were not quantifiable. The present meta-analysis showed that responses to phytase supplementation can be well predicted although variation in P digestibility and digestible P concentration in the data set was high. Overall, predicted effects of phytase on P digestibility well corresponded to predictions made 25 years ago.     

Effect of inorganic phosphate supplementation on egg production in Hy-Line Brown layers fed 2000 FTU/kg phytase

Phytase has long been used to decrease the inorganic phosphorus (Pi) input in poultry diet. The current study was conducted to investigate the effects of Pi supplementation on laying performance, egg quality and phosphate–calcium metabolism in Hy-Line Brown laying hens fed phytase. Layers (n = 504, 29 weeks old) were randomly assigned to seven treatments with six replicates of 12 birds. The corn–soybean meal-based diet contained 0.12% non-phytate phosphorus (nPP), 3.8% calcium, 2415 IU/kg vitamin D₃ and 2000 FTU/kg phytase. Inorganic phosphorus (in the form of mono-dicalcium phosphate) was added into the basal diet to construct seven experimental diets; the final dietary nPP levels were 0.12%, 0.17%, 0.22%, 0.27%, 0.32%, 0.37% and 0.42%. The feeding trial lasted 12 weeks (hens from 29 to 40 weeks of age). Laying performance (housed laying rate, egg weight, egg mass, daily feed intake and feed conversion ratio) was weekly calculated. Egg quality (egg shape index, shell strength, shell thickness, albumen height, yolk colour and Haugh units), serum parameters (calcium, phosphorus, parathyroid hormone, calcitonin and 1,25-dihydroxyvitamin D), tibia quality (breaking strength, and calcium, phosphorus and ash contents), intestinal gene expression (type IIb sodium-dependent phosphate cotransporter, NaPi-IIb) and phosphorus excretion were determined at the end of the trial. No differences were observed on laying performance, egg quality, serum parameters and tibia quality. Hens fed 0.17% nPP had increased (P < 0.01) duodenum NaPi-IIb expression compared to all other treatments. Phosphorus excretion linearly increased with an increase in dietary nPP (phosphorus excretion = 1.7916 × nPP + 0.2157; R² = 0.9609, P = 0.001). In conclusion, corn–soybean meal-based diets containing 0.12% nPP, 3.8% calcium, 2415 IU/kg vitamin D₃ and 2000 FTU/kg phytase would meet the requirements for egg production in Hy-Line Brown laying hens (29 to 40 weeks of age).     

Effect of crude protein and phosphorus level on growth performance, bone mineralisation and phosphorus, calcium and nitrogen utilisation in grower-finisher pigs

Two experiments in a 2 x 2 factorial arrangement were conducted to evaluate the effect of crude protein (CP) (130 vs. 200 g/kg) and phosphorus (P) (4.0 vs. 6.0 g total P/kg) level in a phytase supplemented diet (500 FTU [phytase units]/kg) in grower-finisher pigs. Owing to the design of the experiment, as dietary P level increased, there was also an increase in dietary calcium (Ca) level in order to maintain a dietary Ca to P ratio of 1.6:1. In Experiment 1, four diets were fed to 56 pigs (n = 14, initial body weight [BW] 36.7 +/- 4.2 kg) to investigate the interaction between CP and P on growth performance, bone mineralisation and digesta pH. Experiment 2 consisted of 16 entire male pigs (n = 4; offered identical diets to that offered in Experiment 1) for the determination of total tract apparent digestibility and nitrogen (N), P and Ca utilisation. There was an interaction between CP and P level on bone ash, bone P and bone Ca concentrations (p < 0.05). Pigs offered low CP-low P diets had a higher bone ash, P and Ca concentrations than pigs offered high CP-low P diets. However, there was no effect of CP level at high P levels on bone ash, P and Ca concentrations. Pigs offered low P diets had a lower ileal pH compared with pigs offered high P diets (p < 0.05). In conclusion, offering pigs a high CP-low P, phytase-supplemented diet resulted in a decrease in bone mineralisation.     

Effect of crude protein and phosphorus level on growth performance,bone mineralisation and phosphorus,calcium and nitrogen utilisation in grower-finisher pigs

Two experiments in a 2?×?2 factorial arrangement were conducted to evaluate the effect of crude protein (CP) (130 vs. 200 g/kg) and phosphorus (P) (4.0 vs. 6.0 g total P/kg) level in a phytase supplemented diet (500 FTU [phytase units]/kg) in grower-finisher pigs. Owing to the design of the experiment, as dietary P level increased, there was also an increase in dietary calcium (Ca) level in order to maintain a dietary Ca to P ratio of 1.6:1. In Experiment 1, four diets were fed to 56 pigs (n?=?14, initial body weight [BW] 36.7?±?4.2 kg) to investigate the interaction between CP and P on growth performance, bone mineralisation and digesta pH. Experiment 2 consisted of 16 entire male pigs (n?=?4; offered identical diets to that offered in Experiment 1) for the determination of total tract apparent digestibility and nitrogen (N), P and Ca utilisation. There was an interaction between CP and P level on bone ash, bone P and bone Ca concentrations (p?<?0.05). Pigs offered low CP–low P diets had a higher bone ash, P and Ca concentrations than pigs offered high CP–low P diets. However, there was no effect of CP level at high P levels on bone ash, P and Ca concentrations. Pigs offered low P diets had a lower ileal pH compared with pigs offered high P diets (p?<?0.05). In conclusion, offering pigs a high CP–low P, phytase-supplemented diet resulted in a decrease in bone mineralisation.     

Role of inositol polyphosphates in programmed cell death

The role of inositol polyphosphates (InsPs) in the mediation of cellular apoptosis was investigated in mouse MC3T3 osteoblastic cell line. Extracellular administration of InsP₄, InsP₅, and InsP₆ increased apoptosis in a dose-dependent manner. InsP₆ was more potent than InsP₅ and InsP₄ in promoting apoptosis. Inositol hexasulfate (InsS₆), a structural analog of InsP₆, was used to determine specificity of InsP₆-induced apoptosis as measured by acridine orange/ethidium bromide, flow cytometry, and DNA degradation. In order to study the effects of endogenous InsPs on apoptosis, we used NaF and antimycin A as treatment agents to manipulate intracellular levels of InsPs. NaF is known to increase levels of higher InsPs by inhibiting InsPs phosphatases, a process that is reversed by antimycin A because InsPs kinases are inhibited as a result of depletion of cellular ATP pools. Apoptosis was induced in MC3T3 cells in a NaF dose- and time-dependent manner. Approximately 50% apoptosis was observed at 1 mM NaF in 8 h. Prior treatment with 10 μM antimycin A for 30 min significantly reduced the NaF-induced apoptosis as compared with its control. Additionally, we measured changes in AKT phosphorylation, cleavage of caspase-3 and caspase-9, and release of cytochrome C from mitochondria into cytosol. These changes coincided with total cellular InsPs under similar conditions. The data indicated that NaF-induced changes in apoptotic markers could be due to an increased endogenous InsPs that were partially reversed by antimycin A treatment.     

Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

The aim was to evaluate the effect of heat-treatment, microbial phytase addition and feeding strategy (dry feeding v. fermented liquid feeding) on degradation of phytate (myo-inositol hexakisphosphate, InsP6) and formation and further degradation of lower inositol phosphates (myo-inositol pentakisphosphate-myo-inositol bisphosphate, InsP5-InsP2) at the distal ileum of pigs. Furthermore, the apparent ileal digestibility/degradability (AID) of phosphorus (P), InsP6-P and calcium (Ca) and the apparent total tract digestibility (ATTD) of P and Ca were studied. Pigs were fitted with a T-shaped ileal cannula for total collection of digesta at 2 h intervals during an 8 h sampling period after feeding the morning meal. Each period lasted for 2 weeks: 8 days of adaptation followed by 3 days of total collection of faeces and 3 days of total collection of ileal digesta. The experiment was designed as a 4 × 4 Latin square with four pigs fed four diets. A basal wheat/barley-based diet was fed either as non-heat-treated or heat-treated (steam-pelleted at 90°C). The heat-treatment resulted in an inactivation of plant phytase below detectable level. Diet 1 (non-heat-treated basal diet fed dry); diet 2 (heat-treated basal diet fed dry); diet 3 (as diet 2 but with microbial phytase (750 FTU/kg as fed) fed dry); diet 4 (as diet 3 fed liquid (fermented for 17.5 h nighttime and 6.5 h daytime at 20°C with 50% residue in the tank)). Chromic oxide (Cr2O3) was included as marker and ATTD was determined both by total collection of faeces (ATTDTotal) and Cr2O3 (ATTDCr). InsP6 was completely degraded in diet 4 before feeding resulting in no InsP6-P being present in ileal digesta. InsP6-P concentration in ileal digesta decreased with increasing dietary levels of plant or microbial phytase in pigs fed the dry diets. Consequently, AID and ATTD of P and Ca were greatest for pigs fed diet 4 followed by diets 3, 1 and 2. The ATTD of P depended on the used method as ATTDTotal of P was 72%, 61%, 44% and 34%, whereas ATTDCr of P was 65%, 52%, 38% and 23% for diets 4, 3, 1 and 2, respectively. In all pigs the ileal concentration of InsP5-InsP2-P was extremely small, and thus unimportant for maximisation of ATTD of plant P. In conclusion, fermented liquid feeding with microbial phytase seems to be an efficient approach to improve ATTD of plant P compared with dry feeding. This opens up for further reductions in P excretion.     

Phytase Modulates Ileal Microbiota and Enhances Growth Performance of the Broiler Chickens

Phytase is well studied and explored, however, little is known about its effects on the microbial ecology of the gastrointestinal tract. In total, 400 one-day-old female Ross 308 chicks were randomly distributed to four experimental groups. The dietary treatments were arranged as a 2 × 2 complete factorial design, with the factors being adequate (PC) or insufficient calcium (Ca) and digestible phosphor (dP)(NC) and with or without 5000 phytase units (FTU)/kg of Escherichia coli 6-phytase. The gastrointestinal tract pH values, ileal microbial communities and short-chain fatty acid concentrations in the digesta were determined. The reduction in Ca and dP concentration significantly affected pH in the crop and caeca, and addition of phytase to the NC resulted in a pH increase in the ileum. The reduction in Ca and dP concentration significantly lowered, while phytase supplementation increased ileal total bacterial counts. Additionally, the deficient diet reduced butyrate- but increased lactate-producing bacteria. The addition of phytase increased Lactobacillus sp./Enterococcus sp. whereas in case of Clostridium leptum subgroup, Clostridium coccoides - Eubacterium rectale cluster, Bifidobacterium sp. and Streptococcus/Lactococcus counts, a significant Ca and dP level x phytase interaction was found. However, the recorded interactions indicated that the effects of phytase and Ca and dP levels were not consistent. Furthermore, the reduction of Ca and dP level lowered Clostridium perfringens and Enterobacteriaceae counts. The analysis of fermentation products showed that reducing the Ca and dP content in the diet reduced total SCFA, DL-lactate, and acetic acid in the ileum whereas phytase increased concentrations of these acids in the NC group. This suggests that P is a factor which limits fermentation in the ileum. It may be concluded that phytase plays a role in modulating the gut microbiota of chicken, however, this is clearly linked with the levels of P and Ca in a diet.     

Partial replacement of monocalcium phosphate with neutral phytase in diets for grass carp,Ctenopharyngodon idellus

A 9‐week experiment was designed to study the effects of partial replacement of monocalcium phosphate (MCP) with neutral phytase on growth, body compositions, serum biochemical statuses and intestinal digestive enzyme activities of grass carp, Ctenopharyngodon idellus. The control diet (designated as P2.0) was prepared with 2.0% MCP but without phytase. The three other diets (designated as PP1.5, PP1.0 and PP0.5, respectively) were supplemented with 1.5, 1.0 and 0.5% MCP, respectively, along with 500 FTU of neutral phytase kg^?1 diet in each. After a 9‐week feeding trial, fish (initial body weight: 43.44 ± 2.37 g) fed with PP1.5 and PP1.0 had no significant change in weight gain (WG), specific growth rate (SGR), protein efficiency rate (PER) or feed conversion ratio (FCR) compared with the control (P > 0.05) whereas fish fed with PP0.5 showed significantly lower growth performance in the above parameters. The crude lipid content in whole body or muscle of the fish fed with PP1.5 was significantly lower than the control while significantly higher in fish fed with PP0.5 (P < 0.05), whereas no obvious change was observed in the fish fed with PP1.0. For serum indices, higher serum alkaline phosphatase (Alkp), phosphorus (P) and calcium (Ca) contents were observed in fish fed with phytase‐supplemented diets in comparison with the control. In addition, dietary phytase supplementation increased amylase activity and decreased lipase activity in both foregut and hindgut. The present study suggests that dietary MCP can be reduced when neutral phytase is added to the grass carp diet, and that the maximum MCP reduction level can be up to 1% when neutral phytase is supplemented at 500 FTU kg^?1 diet.     

Effects of ginsenosides on carbachol-stimulated formation of inositol phosphates in rat cortical cell cultures

We examined the effect of ginseng total saponins (GTS) on phosphoinositide metabolism stimulated by activation of muscarinic receptor using rat cortical cultures. Carbachol stimulated formation of [³H]inositol phosphates ([³H]InsPs) by 3.3-fold over basal level in [³H]inositol-prelabeled cells. Pretreatment of GTS inhibited formation of [³H]InsPs evoked by carbachol by 70%–90%. Addition of GTS alone had no effect on the basal formation of [³H]InsPs. The inhibitory effect of the GTS on carbachol-stimulated formation of [³H]InsPs was dose- and time-dependent. IC₅₀ was 6.0 ± 2.8 g/ml. We also examined the effect of GTS on [³H]InsP₁, [³H]InsP₂, or [³H]InsP₃ formation evoked by carbachol. Although GTS had no effect on the basal [³H]InsP₁, [³H]InsP₂, or [³H]InsP₃ formation, pretreatment of GTS inhibited [³H]InsP₁, [³H]InsP₂, or [³H]InsP₃ formation evoked by carbachol, respectively. Addition of individual ginsenosides such as ginsenoside Rb₁, Rc, Rd, Re, or Rg₂ had no effect on the basal formation of [³H]InsPs, whereas pretreatment of ginsenoside Rb₂, Rc, Rd, Re, Rf, Rg₁ or Rg₂ inhibited formation of [³H]InsPs evoked by carbachol by 79%–89%. The results suggest that the inhibitory effect of GTS and its individual ginsenosides on carbachol-stimulated formation of [³H]InsPs in cortical neurons could be one pharmacological action of Panax ginseng.     

Digestibility and energy value of cereal-based diets in relation to digesta viscosity and retention time in turkeys and chickens at different ages estimated with different markers

Digesta viscosity, ileal transit time of digesta, apparent ileal protein digestibility and apparent metabolisable energy (AME_N) of diets based on wheat and dehulled barley (WB), oats (O) or a mixture of these (WBO) fed as such or with enzyme supplementation in three- and six-week old turkeys and broilers were investigated. In addition, differences between ileal digestibility and AME_N calculated by using titanium dioxide (TiO₂), chromic oxide (Cr₂O₃) or acid insoluble ash (AIA) as indigestible markers were compared. Digesta viscosities were generally moderate reaching from 2.5 mPa · s to 7.3 mPa · s. The highest viscosities were observed in WBO diets. Viscosities were reduced with age in broilers, and were generally higher in turkeys than in broilers, especially at six weeks of age. Digesta retention time in ileum was elongated with age of the birds, pronouncedly in broilers. Oat inclusion to the diets decreased retention time especially in broilers at six weeks of age. Apparent ileal digestibility of protein ranged from 0.64–0.83, was lower at six weeks of age than at three weeks of age and generally lowest in O diets, especially in turkeys. AME_N of the diets ranged from 11.2–13.4 MJ/kg being higher at six weeks of age than at three weeks of age. AME_N of Diets O was the lowest but AME_N of WBO diets was higher than that of WB diets indicating a synergistic interaction of cereals, this trend being more pronounced in broilers. Enzyme supplementation decreased viscosity and improved AME_N in most diets, but did not affect ileal protein digestibility. Differences between ileal digestibility estimates obtained with TiO₂ or Cr₂O₃ were small and mainly not significantly different from 0. AME_N estimates were generally higher when calculated with Cr₂O₃ than with TiO₂. AIA gave remarkably lower AME_N values than TiO₂ (the significant differences ranging from 0.24–0.94 MJ/kg). In addition, effects of markers on AME_N estimates interacted with age of the birds and dietary treatments.     

Different Patterns of Agonist-Stimulated Increases of 3H-Inositol Phosphate Isomers and Cytosolic Ca2+ in Bovine Adrenal Chromaffin Cells: Comparison of the Effects of Histamine and Angiotensin II

Abstract: Bovine adrenal chromaffin cells (BCC) were used to compare histamine- and angiotensin II-induced changes of inositol mono-, bis-, and trisphosphate (InsP₁, InsP₂, and InsP₃, respectively) isomers, intracellular free Ca²⁺ ([Ca²⁺]_i), and the pathways of inositol phosphate metabolism. Both agonists elevated [Ca²⁺]_i by 200 nM 3–4 s after addition, but afterwards the histamine response was much more prolonged. Histamine and angiotensin II also produced similar four- to fivefold increases of Ins(1,4,5)P₃ that peaked within 5 s. Over the first minute of stimulation, however, Ins(1,4,5)P₃ formation was monophasic after angiotensin II, but biphasic after histamine, evidence supporting differential regulation of angiotensin II- and histamine-stimulated signal transduction. The metabolism of Ins(1,4,5)P₃ by BCC homogenates was found to proceed via (a) sequential dephosphorylation to Ins(1,4)P₂ and Ins(4)P, and (b) phosphorylation to inositol 1,3,4,5-tetrakisphosphate, followed by dephosphorylation to Ins(1,3,4)P₃, Ins(1,3)P₂, and Ins(3,4)P₂, and finally to Ins(1 or 3)P. In whole cells, Ins(1 or 3)P only increased after histamine treatment. Additionally, Ins(1,3)P₂ was the only other InsP₂ besides Ins(1,4)P₂ to accumulate within 1 min of agonist treatment [Ins(3,4)P₂ did not increase]. These results support a correlation between the time course of Ins(1,4,5)P₃ formation and the time course of [Ca²⁺]_i transients and illustrate that Ca²⁺-mobilizing agonists can produce distinguishable patterns of inositol phosphate formation and [Ca²⁺], changes in BCC. Different patterns of second-messenger formation are likely to be important in signal recognition and may encode agonist-specific information.     

Effects of mineral and rapeseed phosphorus supplementation on phytate degradation in dairy cows

The objective of this study was to evaluate the effects of diet composition on phytate (InsP₆) degradation in dairy cows. In Experiment 1, four diets that differed in the amount and source of phosphorus (P) were fed to 24 lactating cows in a 4 × 4 Latin Square design. The control diet (Diet C) contained 4.18 g P/kg dry matter (DM). Diet MP contained additional mineral P (5.11 g P/kg DM), Diet RS contained rapeseed and rapeseed meal as organic P sources (5.26 g P/kg DM) and Diet RSM contained rapeseed meal and rapeseed oil (5.04 g P/kg DM). Total P (tP) and InsP₆ excretion in faeces were measured. In Experiment 2, we used a rumen simulation technique (Rusitec) to estimate ruminal disappearance of tP and InsP₆ from Diets C, MP and RSM. In Experiment 1, tP concentration in faeces increased with tP intake and was highest for Diets RS and RSM. The source of supplemented P had no influence on tP digestibility, but tP digestibility was reduced for Diets MP, RS and RSM in comparison to that for Diet C. InsP₆ disappearance decreased in Diet MP (85.0%) and increased in Diets RS (92.7%) and RSM (94.0%) compared to that in Diet C (90.0%). In Experiment 2, P source influenced ruminal tP disappearance (Diet MP, 78.6%; Diet RSM, 75.3%). InsP₆ disappearance for Diet C (98.1%) was higher than that for Diets MP (95.6%) and RSM (94.9%). The results confirmed the high potential of ruminants to degrade InsP₆, but differences in diet composition influenced InsP₆ disappearance. Further studies of the site of InsP₆ degradation are required to understand the relevance of InsP₆ degradation for the absorption of P.