Safety assessment of phytase transgenic maize 11TPY050 in Sprague-Dawley rats by 90-day feeding study.

The present study aimed to evaluate the subchronic toxicity of feeding with phytase-transgenic maize line 11TPY050 in Sprague-Dawley (SD) rats. Rats (n = 10/sex/group) were fed with 12.5%, 25% or 50% (w/w) transgenic maize diet, 12.5%, 25% or 50% (w/w) non-transgenic isoline OSL940 maize diet, or 50% (w/w) commercially available Zhengdan958 maize diet for 90 days. Daily clinical observations and weekly measurements of body weights and food consumption were conducted. Blood samples were collected on day 46 and day 91 for hematology and clinical chemistry evaluations. At the end of the study, macroscopic and microscopic examinations were performed. No effects on body weight and food consumption were observed. The results of hematology, clinical chemistry, and absolute and relative organ weights in the transgenic maize group were comparable to those in the parental maize group. Several statistical differences were not dose-related and were not considered to be biologically significant. Furthermore, the terminal necropsy and histopathological examination showed no treatment-related changes among the groups. The results from the present 90-day feeding study of phytase-transgenic maize 11TPY050 indicated no unexpected adverse effects in SD rats. The phytase transgenic maize 11TPY050 has substantial equivalence with non-transgenic maize.

Combined effects of enzymes and probiotics on hemato-biochemical parameters and immunological responses of juvenile Siberian sturgeon (Acipenser baerii).

A 10-week feeding trial was run to investigate the separate and simultaneous effects of exogenous enzymes (Enz), probiotics (Pro), and Pro-Enz mixtures on the hematology indices, serum biochemical parameters, and innate-immunity status of juvenile Siberian sturgeon. The fish (138.06 ± 3.64 g) were randomly dispersed into 12 tanks (20 individuals per tank) and fed with Enz (Phytase, protease, and xylanase), Pro (Pediococcus pentosaceus and Lactococcus lactis), and Pro-Enz cocktail. At the end of the feeding bioassay, the highest values of red blood cell count, hemoglobin concentration, hematocrit level, and lymphocyte percentage followed by the lowest neutrophil percentage were obtained in Pro-Enz treatment (P < 0.05). Despite a significantly lower level of alkaline phosphatase in the fish fed with Pro supplemented diet (P < 0.05), no significant difference was found in the serum level of alanine aminotransferase and aspartate aminotransferase among the experimental groups (P > 0.05). Total protein content was significantly upregulated in serum and skin mucus samples from those fed with supplemented diets compared to the control group (P < 0.05). In both serum and skin mucus samples, higher immune responses in terms of lysozyme activity, immunoglobulin M, total protein was seen in Pro-Enz treatment compared to the control group followed by the serum complement components (P < 0.05). The results indicate that the combinational supplementation of Siberian sturgeon diet with the exogenous enzymes and probiotics modulates the physiometabolic responses and innate immune system to a higher grade than their individual supplementation.

Dietary Phytase- and Lactic Acid-Treated Cereals Caused Greater Taxonomic Adaptations than Functional Adaptations in the Cecal Metagenome of Growing Pigs.

Phosphorus (P) is an essential nutrient for the gut bacteria and the host. Nevertheless, little information exists that indicates to what extent an improved level of P availability in the small intestine leads to functional adaptations in bacterial metabolic pathways in the large intestine. Therefore, we investigated the changes in the taxonomic and functional bacterial metagenome in cecal digesta of growing pigs fed diets containing phytase and/or cereals treated with 2.5% lactic acid (LA) for 19 days (n = 8/diet) using shotgun metagenome sequencing. The phytase supplementation resulted in strikingly distinct bacterial communities, affecting almost all major bacterial families, whereas functional changes were less dramatic among the feeding groups. While phytase treatment decreased predominant Prevotellaceae levels, it seemed that Clostridiaceae, Ruminococcaceae, and Lachnospiraceae filled the opening metabolic niches (P < 0.05). The LA-treated cereals mediated reduced levels of Bacteroidaceae and increased levels of Veillonellaceae, but those results were mainly seen when the cereals were fed as a single treatment (P < 0.05). In association with the taxonomic alterations, phytase caused changes within the major functional pathways corresponding to amino acid metabolism; translation; membrane transport; folding, sorting, and degradation; and energy metabolism, whereas the LA treatment of cereals resulted in decreased enzymatic capacities within the carbohydrate metabolism and energy metabolism pathways (P < 0.05). Metabolic dependencies corresponding to the starch and sucrose metabolism, glycolysis/gluconeogenesis, and citrate cycle pathways were indicated by diet-associated changes in enzymatic capacities related to short-chain fatty acid, methane, vitamin, and bacterial antigen synthesis. Accordingly, the present results support the idea of the importance of the availability of intestinal P for bacterial metabolism. However, the functional profiles were less different than the taxonomic profiles among the dietary treatment results, indicating a certain degree of metabolic plasticity within the cecal metagenome.IMPORTANCE Dietary strategies (e.g., phytase supplementation and lactic acid [LA] treatment of cereals) used to improve the availability of phytate-phosphorus (P) from pig feed reduce the amount of P flowing into the large intestine, whereas LA treatment-induced changes in nutrient fractions alter the substrate being available to the microbiota. In ruminants, lower intestinal P availability compromises the fibrolytic activity of the microbiome. Here, we report that the functional capacities were less dramatically affected than the taxonomic composition by phytase-supplemented and LA-treated cereals. The bacterial community appeared to be partly capable of functionally compensating for the altered flow of P by replacing taxa with higher P needs by those with lower P needs. Therefore, by acting as mucosal immune stimulants, alterations in microbiota-associated molecular patterns (MAMPs) due to the taxonomic shifts may play a greater role for host physiology and health than functional differences caused by differing intestinal P availabilities, which merits further research.

Alterations of the Viable Ileal Microbiota of the Gut Mucosa-Lymph Node Axis in Pigs Fed Phytase and Lactic Acid-Treated Cereals.

The gut-lymph node axis is a critical player in the symbiotic relationship between gut microbiota and the host. However, little is known about the impact of diet-related bacterial shifts in the gut lumen on bacterial translocation into lymph nodes. Here, we (i) characterized changes in the viable microbiota composition along the ileal digesta-mucosa-lymph node axis and (ii) examined the effect of dietary phytase supplementation and lactic acid (LA) soaking of cereals on the bacterial taxonomy along this axis, together with their effect on the mucosal expression of innate immune and barrier function genes in pigs (n = 8/diet). After 18 days on diets, ileal digesta, mucosa, and ileocecal lymph nodes (ICLNs) were collected for RNA isolation and 16S rRNA-based high-resolution community profiling. Bacterial communities were dominated by Lactobacillaceae and Clostridiaceae, with clearly distinguishable profiles at the three sampling sites. Specific bacterial subsampling was indicated by enrichment of the ICLNs with Lactobacillaceae, Lachnospiraceae, Veillonellaceae, and Methanobacteriaceae and less Clostridiaceae, Pasteurellaceae, Helicobacteraceae, and Enterobacteriaceae compared to that of the mucosa. LA treatment of cereals reduced proteolytic taxa in the lumen, including pathobionts like Helicobacteraceae, Campylobacteraceae, and Fusobacteriaceae When combined, phytase- and LA-treated cereals largely increased species richness, while the single treatments reduced Actinobacteria and Bacteroidetes in ICLNs and increased mucosal MUC2 expression. In contrast, phytase reduced mucosal CDH1 expression, indicating altered barrier function with potential effects on bacterial translocation. Overall, both treatments, although often differently, changed the viable microbiome along the digesta-mucosa-lymph node axis in the ileum, probably due to altered substrate availability and microbial-host interactions.IMPORTANCE A host's diet largely determines the gut microbial composition and therefore may influence bacterial translocation into ICLNs. Due to its importance for cell metabolism, the intestinal phosphorus availability, which was modified here by phytase and LA treatment of cereals, affects the intestinal microbiota. Previous studies mainly focused on bacteria in the lumen. The novelty of this work resides mainly in that we report diet-microbe effects along the digesta-mucosa-ICLN axis and linked those effects to mucosal expression of barrier function genes as crucial components for host health. Lymph nodes can serve as reservoir of pathobionts; therefore, present diet-microbiome-host interactions have implications for food safety.

Thermoregulatory, behavioral and productive responses of laying hens supplemented with different types and dosages of phytases raised in a hot environment: An integrative approach.

This study had the following objectives: (i) to evaluate the thermoregulatory and behavioral responses of light laying hens supplemented with different types and dosages of phytases in the two day shifts; and (ii) to integrate the thermoregulatory and behavioral responses with performance of these birds raised in a hot environment. 270 light laying hens of the Hy-Line White lineage, with a body weight of 1.60 ± 0.092 kg were distributed in a completely randomized design in a 2 × 2 + 1 factorial model with two types of phytases (bacterial and fungal) and two dosages (450 and 900 FTU), and a control diet. The day shift (morning and afternoon) was considered as a fixed effect in the factorial arrangement. Principal component analysis (PCA), correspondence analysis (CA) and canonical discriminant analysis (CDA) were used. There was no interaction (P > 0.05) between phytases and dosages for thermoregulatory responses. Respiratory rate (RR), cloacal temperature (CT), and surface temperature with feathers (STWF) and featherless (STF) were higher (P < 0.001) in the afternoon. Birds show different thermoregulatory and behavioral responses in the two shifts of the day. We also observed that birds supplemented with bacterial and fungal phytase showed similar thermoregulatory and behavioral responses to the control group in both day shifts. Expression of the "eating" activity was greater in the morning, while the birds remained sitting longer in the afternoon. Egg production was higher (P < 0.001) in birds supplemented with bacterial phytase. The phytase dosages had no effect on thermoregulatory, behavioral or performance responses. Egg production, feed conversion per dozen eggs corresponded to 81.1% of the differences between bacterial and fungal phytase supplementation and group control. Thus, we conclude that: (i) phytase dietary supplementation has no effect on the thermoregulatory responses of laying hens reared in a hot environment; (ii) birds supplemented with bacterial phytase showed higher egg production; and (iii) phytases (450 and 900 FTU) do not interfere with productive, behavioral and thermoregulatory responses.

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

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.

Feeding value of rice distiller's dried grains with solubles as protein supplement in diet of laying hens.

A feeding trial of 10 weeks duration was undertaken on laying hens (n = 240) to evaluate feeding value of rice distiller's dried grains with soluble (rDDGS) with or without enzyme supplementation (α-amylase, ß-glucanase, xylanase, carboxymethylcellulase, pectinase, proteinase, α-galactosidase, ß-galactosidase, lipase, and phytase), following 4 × 2 factorial design, on egg production, nutrient utilization, and cost economics of egg production. The birds were randomly assigned to eight dietary treatments with 30 birds/treatment. The birds were housed individually in layer cages and each bird was taken as an experimental unit. Eight experimental diets were prepared by incorporating four levels (0, 50, 75, and 100 g/kg) of rDDGS with and without enzyme supplementation. The results revealed a significant (P < 0.01) increase of egg mass, feed intake, egg production, and body weight gain in dietary treatments with up to 75 g rDDGS though the values were statistically similar to the hens fed 100 g rDDGS. Enzyme supplementation resulted in significant (P < 0.01) improvement of egg mass, egg production, feed conversion ratio (FCR) per dozen eggs, FCR per kilogramme egg mass, and net FCR. The significantly (P < 0.01) higher yolk index was observed at 100 g rDDGS level, while shell thickness improved significantly (P < 0.01) up to 75 g rDDGS level. No significant effect of rDDGS inclusion was observed on shape index, albumin index, and Haugh unit. Enzyme supplementation significantly improved the shell thickness and yolk colour of eggs. Nitrogen, calcium, and phosphorus retention and dry matter metabolizability did not show any significant treatment effects. There was significant (P < 0.01) reduction in feed-cost per kilogramme egg mass or per dozen eggs with the increased DDGS levels and dietary enzyme supplementation. It was concluded that rDDGS can be used up to 100 g/kg diet of laying hens along with enzyme supplementation for better productivity of layer hens.

The ranked importance of dietary factors influencing the performance of broiler chickens offered phytase-supplemented diets by the Plackett-Burman screening design.

1.The objective of the present study was to rank the importance of the following dietary factors; canola meal, wheat, whole barley, digestible lysine, phytate-P, calcium, available P, sodium and three NSP-degrading feed enzymes. Their influence on growth performance, gastro-intestinal tract parameters, energy utilisation, ileal N digestibility and disappearance rates were determined via the Plackett-Burman design in broiler chickens offered phytase-supplemented diets. 2. The eleven dietary factors were assigned two levels in the Plackett-Burman design matrix. The resulting twelve dietary treatments were offered to six replicates per treatment (six birds per cage) with a total of 468 male Ross 308 broiler chicks from 7 to 28 d post-hatch. 3. Increasing digestible lysine levels improved weight gain by 15.6% (P < 0.001) and gain:feed by 9.36% (P < 0.001). Increasing calcium levels reduced weight gain by 6.36% (P < 0.001) and gain:feed by 2.60% (P < 0.001). The high calcium level increased gizzard pH from 2.78 to 3.01 (P < 0.005). Whole barley significantly increased relative gizzard weights and contents, pancreas weights and both ileal N digestibility coefficients (0.774 versus 0.803; P < 0.001) and ileal N disappearance rates (23. 3 versus 24.5 g/bird/day; P < 0.001). 4. Overall, digestible lysine level and calcium level were identified as the most influential dietary factors to influence growth performance of broilers offered phytase-supplemented diets, which hold implications for practical diet formulations.

The effect of exogenous phytase supplementation on nutrient digestibility, ruminal fermentation and phosphorous bioavailability in Rambouillet sheep.

BACKGROUND: The effect of phytase supplementation with respect to a high sorghum grain diet on sheep voluntary feed intake, apparent nutrient digestibility, ruminal fermentation, phosphorus (P) excretion and blood serum P concentration was evaluated. RESULTS: Phytase supplementation significantly decreased fecal P excretion (P = 0.003), resulting in a 26% decrease in relation to the phytase free diet. Dry matter intake, nutrient digestibility, ruminal butyrate and serum P were not (P > 0.10) affected by the phytase level. Neutral detergent digestibility showed a tendency to increase linearly (P = 0.10) with increasing phytase levels. Ruminal pH was lower for phytase supplemented sheep, with a significant decrease (P = 0.007) at 9 h post feeding, whereas ruminal ammonia-N at 3 h post feeding was lower (P = 0.004) for the phytase treatment groups, resulting in a decreasing linear response (P = 0.001) with an increasing phytase dose. Duodenal pH was significantly reduced at 6 h post feeding. Propionate tended (P = 0.051) to be increased linearly as the phytase supplementation level increased. CONCLUSION: Exogenous phytase supplementation of high sorghum grain diets significantly decreased fecal P excretion in Rambouillet rams. Phytase supplementation appears to affect neutral detergent fiber digestibility, duodenal and ruminal pH, ammonia and propionate. © 2018 Society of Chemical Industry.

Superdosing phytase reduces real-time gastric pH in broilers and weaned piglets.

1. The current study was conducted to investigate the effect of high phytase doses on growth performance and real-time gastric pH measurements in broiler chickens and pigs. 2. In the first experiment, 576 male Ross 308 broilers were fed in two phases (0-21 and 21-42 d) with 4 treatment groups, with diets meeting nutrient requirements containing 0, 500, 1500 or 2500 FTU/kg phytase. In the second, 64 Landrace weaners were fed on diets meeting nutrient requirements with or without phytase (0 or 2500 FTU/kg) in two phases (0-21 and 21-42 d). Heidelberg pH capsules were administered to 7 broilers and approximately 13 pigs per treatment group, pre- and post-phase change, with readings monitored over several hours. 3. Addition of phytase into an adequate Ca and P diet had no significant effect on broiler performance although phytase tended (P < 0.07) to improve feed conversion in pigs over the entire experimental period. Real-time pH capsule readings in broilers demonstrated an increase (P < 0.05) in gizzard pH when phytase was dosed at 500 or 1500 FTU/kg, while higher doses of 2500 FTU/kg phytase lowered pH to a level comparable to control birds. Gastric pH increased (P < 0.01) when animals were exposed to dietary phase change, signifying a potential challenge period for nutrient digestibility. However, pigs fed 2500 FTU/kg were able to maintain gastric pH levels through diet phase change. In contrast, spear-tip probe measurements showed no treatment effect on gastric pH. 4. These findings demonstrate dietary manipulation of gastric pH and the value of real-time pH capsule technology as a means of determining phytase dose response.

Supplementation of broiler diets with high levels of microbial protease and phytase enables partial replacement of commercial soybean meal with raw, full-fat soybean.

A 3 × 3 + 1 factorial, involving three levels of protease (0, 15,000 or 30,000 PROT/kg) and three levels of phytase (1,000, 2,000 or 3,000 FYT/kg), was used to evaluate the effect of replacing commercial soybean meal (SBM) with raw, full-fat soybean (RFSB) at 75 g/kg of diet for broilers. A control diet was used for comparison. Each treatment was replicated six times, with nine birds per replicate. The concentration of trypsin inhibitors (TIs) in the test diets was approximately 10,193.4 TIU/kg. Regardless of enzyme supplementation, feed intake (FI) and body weight gain (BWG) of birds in the control group were superior to those on the test diets. Birds that received the protease-free test diets had reduced FI and BWG, but when supplemented with protease, were similar to the control diet in BWG, FI (except 0-35 days) and feed conversion ratio (FCR). When the test diet was supplemented with elevated levels (extradose) of protease and phytase, the BWG was improved during 0-10 days (p = .05) and 0-24 days (p < .01). Regardless of protease supplementation, the weight of thighs was lower for birds fed the test diets. Birds that received the control diet had smaller weight of pancreas. Increasing the level of phytase supplementation reduced (p < .05) the weight of the pancreas. The apparent ileal digestibility (AID) of CP and AA was higher in birds on the control diets, but this was also improved in test diets by protease supplementation. The activities of trypsin (7%), general proteolytic (11%) and lipase (12%) were slightly increased because of protease supplementation. Mucosal depth and apparent villus surface areas were increased by about 2.9% and 20%, respectively, due to supplementation of elevated level of phytase. It can be concluded that RFSB could partially replace SBM in broiler diets, provided the diets are supplemented with elevated levels of protease and 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.

Increased iron level in phytase-supplemented diets reduces performance and nutrient utilisation in broiler chickens.

1. The effect of different levels of dietary iron on phytase activity and its subsequent effect on broiler performance were investigated in a 3 × 2 factorial arrangement. A total of 360 day-old Ross 308 male broiler chicks were distributed to 6 experimental diets, formulated with three levels of Fe (60, 80 and 100 mg/kg) and two levels of phytase (0 and 500 FTU/kg). 2. Phytase supplemented to mid-Fe diets increased feed consumption more than the non-supplemented diet at d 24. From hatch to d 35, Fe × phytase interaction significantly influenced the feed intake (FI), body weight gain (BWG) and feed conversion ratio (FCR). The high-Fe diet supplemented with phytase significantly reduced FI and BWG of broilers than those supplemented with low- or mid-Fe diets. The overall FCR was significantly better in birds fed on the mid-Fe diets with phytase supplementation. 3. A significant improvement in ileal digestibility of N, P, Mg and Fe was observed in birds feed diets containing 60 mg Fe/kg, with significant interaction between Fe and phytase. 4. Phytase improved the bone breaking strength when supplemented to low- or mid-Fe diets, compared to the non-supplemented diets. There was a significant Fe × phytase interaction effect. Tibia Fe content was higher in birds fed on phytase-free diets with high Fe but the reverse was the case when phytase was added and their interaction was significant. High dietary Fe significantly increased the accumulation of Fe in liver. 5. Phytase improved Ca-Mg-ATPase, Ca-ATPase and Mg-ATPase activities in jejunum when supplemented to the diet containing 80 mg Fe/kg. 6. This study indicates that high (100 mg/kg) dietary Fe inhibited phytase efficacy and subsequently reduced the overall performance and nutrient utilisation of broilers.

Effect of diet phase change, dietary Ca and P level and phytase on bird performance and real-time gizzard pH measurements.

In this study, a novel capsule technique was used to capture real-time pH readings from the gizzard over several hours, in response to different dietary treatments. 1. The first experiment was a preliminary study into capsule administration and pH recordings using 9 male Ross 308 broilers from 20 d. In the second experiment, broilers (576) were fed in two phases (0-21 and 21-42 d) with 4 treatment groups; low and adequate Ca and AvP diets with and without Quantum Blue phytase (1500 FTU/kg). Capsules were administered to 8 birds from each treatment group, pre and post diet phase change, with readings captured over a 2.5 h period. 2. Phytase addition improved body weight gain (BWG) and feed conversion ratio (FCR) of birds fed low dietary Ca, while having no significant effect on birds fed adequate Ca diets. Unexpectantly, diets with higher Ca levels gave a lower average gizzard pH compared to the low Ca diet. Phytase addition, irrespective of Ca level, increased average gizzard pH. Fluctuations in gizzard pH (0.6-3.8) were observed across all treatment groups. Higher frequencies of pH readings below pH 1.0 were seen in birds fed an adequate Ca diet and with phytase supplementation of a low Ca diet. 3. These results signify the potential use of capsule techniques to monitor real-time pH changes. The implication on gastric protein and fibre hydrolysis as a result of large fluctuations in pH should be considered.

Exploratory transcriptomic analysis in muscle tissue of broilers fed a phytase-supplemented diet.

The effect of phytase on phosphorus retention, broiler (Gallus gallus) performance and bone mineralization in diets with reduced inorganic phosphate concentration is well documented. Furthermore, so-called 'extra-phosphoric' effects of phytase have been described in the literature that may be associated with changes in mineral and amino acid partitioning and requirements per se. In particular, the role of myo-inositol in phytase responses is implied but not well elucidated. It was the purpose of the experiment reported herein to explore the effect of phytase on broiler growth, nutrient digestibility, blood biochemistry and gene expression. A 5-week broiler floor pen trial was conducted to evaluate the effect of supplementation of a moderately phosphorus-deficient diet with 1000 U/kg of a 6-microbial phytase. Parameters measured were growth performance, phosphorus (P), calcium (Ca) and myo-inositol plasma concentrations, apparent ileal P digestibility, bone mineralization, breast meat weight and Pectoralis major muscle transcriptome. Supplementation of the diet with phytase improved weight gain during the starter period (18%) and the whole period (24%) compared with animals that received the control diet (p < 0.05). Improved feed conversion ratio, increased myo-inositol plasma concentration, tibia ash contents and breast meat weight were also observed in animals fed phytase. The transcriptomic analysis revealed that some differentially expressed genes (DEG) in broilers, receiving phytase in comparison with animals fed reduced phosphorus diet without phytase, were part of pathways involved in muscle development, via calmodulin/calcineurin and insulin-like growth factor. Microarray data confirmation was performed on six genes by quantitative PCR (qPCR): PI3K regulatory and catalytic subunit, Phospholipase C beta, Myocyte Enhancer Factors 2A and 2C, and calcineurin A. The results suggested that dietary supplementation with this phytase could generate low molecular weight phytate esters and indirectly myo-inositol, and could help us to understand how muscle metabolism may be affected at a gene level.

Femur morphometry, densitometry, geometry and mechanical properties in young pigs fed a diet free of inorganic phosphorus and supplemented with phytase.

The study investigated in piglets the effect of replacing dietary inorganic P by addition of microbial phytase and its impact on performance, nutrient digestibility and on the geometrical characteristics and mineralisation of the femur. Sixteen pigs on day 58 of age were divided into two groups and fed either a diet free of additional inorganic phosphorus (P) and supplemented with phytase (Diet LP, 4.23 g total P/kg diet) or a diet with a mineral source of P and not supplemented with phytase (Diet SP, 5.38 g total P/kg diet). Performance data and the apparent total tract digestibility of nutrients were estimated between days 58 and 114, and 72 and 86 of age, respectively. On day 114 of age, the pigs were slaughtered, the femur was dissected and the mineral content and mineral density, maximum strength and maximum elastic strength, cortical wall thickness, cross-sectional area and cortical index were analysed. The growth performance and digestibility of nutrient fractions (with exception of P) did not differ between treatment groups. The P-digestibility was significantly higher in Group LP. The femur of pigs in Group LP had significantly greater cortical wall thickness, cortical index, bone mineral content, bone mineral density, maximum strength and maximum elastic strength than Group SP. Femur maximum strength and maximum elastic strength were correlated with cortical wall thickness and cortical index. Resulting from the different supply of digestible P, the femur geometrical, densitometric and mechanical properties of Group LP were better than those of Group SP. The mechanical properties of the femur of pigs depended more on its geometrical characteristics than on the degree of its mineralisation.

Response of broiler chickens to different levels of calcium, non-phytate phosphorus and phytase.

1. Five hundred and seventy six-d old Ross 308 broiler chicks (6 cages per diet, 8 birds per cage in 3 × 2 × 2 factorial arrangement) were fed on maize-soybean meal-based diets containing three concentrations of Ca (6, 8 or 10 g/kg), two concentrations of non-phytate phosphorus (NPP) (3 or 4 g/kg) and two levels of exogenous microbial phytase (0 or 500 FTU/kg) from d 0 to 35. 2. Body weight (BW), feed intake (FI) and mortality records were collected. Two birds per replicate were killed at 24 d of age to obtain tibia samples. 3. Increasing Ca level significantly reduced the FI and body weight gain (BWG) between hatch and 10 and 24 d, especially with the phytase-supplemented diets. However, phytase supplementation of the diet containing 4 g NPP/kg improved the FI and BWG at d 10 and 24. At d 24, phytase supplementation improved feed conversion ratio (FCR) of birds that consumed diets containing high NPP. The overall FCR was better in birds offered the phytase-supplemented, medium-Ca diet. 4. There was a significant reduction in length, width and breaking strength of the tibia bone in birds fed on a diet with high Ca and low NPP. Phytase supplementation improved the tibia ash content and bone breaking strength of chicks fed on the diet containing 8 and 4 g/kg Ca and NPP, respectively. The Ca content of the tibia bone was low in birds fed on diets with 6 and 4 g/kg Ca and NPP, respectively, but this was counteracted by phytase supplementation. 5. Birds fed on diets with 4 g/kg NPP had the best carcass percentage and parts yield. Phytase supplementation to high-Ca diets significantly reduced the carcass yield of birds. 6. These results confirmed the detrimental effect of high dietary Ca on phytase activity and subsequent growth and bone development of birds, especially when NPP is in short supply.

The effects of transportation stress on Japanese quail (Coturnix Coturnix japonica) fed corn-based diet in comparison with wheat-based diet supplemented with xylanase and phytase.

Harvesting, handling and transporting quails to the slaughterhouses, other farms and laboratories might covertly reduce their welfare. The aim of this study was to evaluate the effects of two major sources of energy in poultry nutrition on reducing transportation stress in Japanese quail (Coturnix Coturnix japonica). Male quails (n = 60) were divided into two groups. The first group was fed corn-based diet, and the second was fed wheat-based diet supplemented with xylanase and phytase. At the end of the experiment (day 35), quails were subjected to 80 km of transportation. Immediately on arrival and after 24 h, heterophil counts, lymphocyte counts and H:L ratios were measured. On arrival, H counts were lower, L counts were higher, and H:L ratios were lower for corn-fed group. After 24 h, wheat-fed group showed lower increment of H counts, greater increment of L counts and also decrement of H:L ratios rather than corn-fed group which showed increment of H:L ratios. However, these ratios were still lower in corn-fed group. Results indicate that corn-based diets can help Japanese quail to better resist transportation stress, although it seems that feeding wheat-based diets supplemented with xylanase and phytase could have positive effects for coping better with stress after journeys.

Influence of partial replacement of ground wheat with whole wheat and exogenous enzyme supplementation on growth performance, nutrient digestibility and energy utilization in young broilers.

This study investigated the influence of pre-pelleting inclusion of whole wheat (WW) and exogenous enzyme supplementation on growth performance, coefficient of apparent ileal nutrient digestibility (CAID) and apparent metabolizable energy (AME) in broilers fed wheat-based pelleted diets. A 2 × 3 factorial arrangement of treatments was used with two methods of wheat inclusion [622 g/kg ground wheat (GW) and 250 g/kg WW replaced GW (wt/wt) pre-pelleting (PWW)] and three enzymes (xylanase, phytase and xylanase plus phytase). A total of 288, one-day-old male broilers (Ross 308) were individually weighed and allocated to 36 cages (8 broilers/cage), and the cages were randomly assigned to the six dietary treatments. Birds fed PWW diets gained more (p < 0.05) weight than those fed GW diets. There was no effect (p > 0.05) of WW inclusion on feed intake (FI). Phytase alone increased (p < 0.05) FI compared to xylanase or the combination. Whole wheat inclusion increased (p < 0.05) the gain-to-feed ratio (G:F). Feeding xylanase plus phytase and phytase-added diets resulted in the greatest and lowest G:F, respectively, with xylanase supplemented diets being intermediate. Birds fed PWW diets had greater (p < 0.05) relative gizzard weights than those fed GW diets. There was no effect (p > 0.05) of WW inclusion on the CAID of nitrogen (N), starch and fat. Combination of xylanase and phytase resulted in greater (p < 0.05) digestibility of N, starch and fat than that of individual additions. Feeding PWW diets resulted in greater (p < 0.05) AME values than GW diets. Combination of xylanase and phytase increased (p < 0.05) the AME compared to the diets with individual additions of xylanase or phytase. The current results suggest that the influence of pre-pelleting WW inclusion and exogenous enzymes on nutrient digestibility and broiler performance is not additive.

Effect of super dosing of phytase on growth performance, ileal digestibility and bone characteristics in broilers fed corn-soya-based diets.

A feeding trial was designed to assess the effect of super dosing of phytase in corn-soya-based diets of broiler chicken. One hundred and sixty-eight day-old broilers were selected and randomly allocated to four dietary treatment groups, with 6 replicates having 7 chicks per treatment group. Two-phased diets were used. The starter and finisher diet was fed from 0 to 3 weeks and 4 to 5 weeks of age respectively. The dietary treatments were consisted of normal phosphorus (NP) group without any phytase enzyme (4.5 g/kg available/non-phytin phosphorus (P) during starter and 4.0 g/kg during finisher phase), three low-phosphorus (LP) groups (3.2 g/kg available/non-phytin P during starter and 2.8 g/kg during finisher phase) supplemented with phytase at 500, 2500, 5000 FTU/kg diet, respectively, to full fill their phosphorus requirements. The results showed that super doses of phytase (at 2500 FTU and 5000 FTU/kg) on low-phosphorus diet improved feed intake, body weight gain, ileal digestibility (serine, aspartic acid, calcium, phosphorus), blood P levels and bone minerals such as calcium (Ca), P, magnesium (Mg) and zinc (Zn) content. It could be concluded that super doses of phytase in low-phosphorus diet were beneficial than the normal standard dose (at 500 FTU/kg) of phytase in diet of broiler chicken.