Phytase Inclusion Research Articles

Accentuating the positive and eliminating the negative: Efficacy of TiO2 as digestibility index marker for poultry nutrition studies.

Inert digestibility index markers such as titanium dioxide are universally accepted to provide simple measurement of digestive tract retention and relative digestibility in poultry feeding trials. Their use underpins industry practice: specifically dosing regimens for adjunct enzymes added to animal feed. Among these, phytases, enzymes that degrade dietary phytate, inositol hexakisphosphate, represent a billion-dollar sector in an industry that raises ca. 70 billion chickens/annum. Unbeknown to the feed enzyme sector, is the growth in cell biology of use of titanium dioxide for enrichment of inositol phosphates from extracts of cells and tissues. The adoption of titanium dioxide in cell biology arises from its affinity under acid conditions for phosphates, suggesting that in feeding trial contexts that target phytate degradation this marker may not be as inert as assumed. We show that feed grade titanium dioxide enriches a mixed population of higher and lower inositol phosphates from acid solutions. Additionally, we compared the extractable inositol phosphates in gizzard and ileal digesta of 21day old male Ross 308 broilers fed three phytase doses (0, 500 and 6000 FTU/kg feed) and one inositol dose (2g/kg feed). This experiment was performed with or without titanium dioxide added as a digestibility index marker at a level of 0.5%, with all diets fed for 21 days. Analysis yielded no significant difference in effect of phytase inclusion in the presence or absence of titanium dioxide. Thus, despite the utility of titanium dioxide for recovery of inositol phosphates from biological samples, it seems that its use as an inert marker in digestibility trials is justified-as its inclusion in mash diets does not interfere with the recovery of inositol phosphates from digesta samples.

Evaluation of phytase dose effect on performance, bone mineralization, and prececal phosphorus digestibility in broilers fed diets with varying metabolizable energy, digestible amino acids, and available phosphorus concentration

The nutritional composition of diets and the provision of exogenous phytases play important roles in animal performance. Therefore, we evaluated the individual and combined impact of metabolizable energy (ME), digestible lysine (dLys), available phosphorus (avP) and calcium (Ca), and phytase dose (1,000 or 2,000 FTU/kg) on the growth performance, feed efficiency, phosphorus digestibility, and bone ash content of broiler chickens from 10 to 42 d of age. Experimental diets were formulated in a Box-Behnken design to contain various levels of ME (11.9, 12.2, 12.54, or 13.1MJ/kg), dLys (0.91, 0.93, 0.96, or 1.00%) and avP/Ca (0.12/0.47, 0.21/0.58, or 0.33/0.68%). The effect of phytase was expressed in terms of the extra nutrients released. The diets were formulated to have consistent phytate substrate contents (0.28% in average). Body weight gain (BWG) and feed conversion ratio (FCR) were described via polynomial equations (R2=0.88 and 0.52, respectively), with interconnections between variables (ME, dLys, and avP/Ca). No interaction was observed among variables (P > 0.05). Metabolizable energy was the most important factor affecting BWG and FCR (linearly; P < 0.001). Reducing ME content from 13.1 to 11.9 MJ/kg in control diet resulted in a 6.8% decrease in BWG and a 3.1% increase in FCR (P < 0.001). The dLys contents also affected performance linearly (P < 0.001), but to a lesser degree; BWG decreased by 160g when the dLys was reduced by 0.09% units, while the same reduction in dLys increased the FCR by 0.108 points. The inclusion of phytase alleviated the negative effects on feed intake (FI), BWG, and FCR. Phytase improved phosphorus digestibility and bone ash content according to a quadratic relationship. When phytase was added, ME negatively affected FI (r=-0.82,P<0.001), whereas the dLys content was correlated with FCR (r=-0.80,P<0.001). Supplementing phytase allowed the reduction of ME, dLys, and avP-Ca in the diet without affecting performance. The addition of phytase increased of ME, dLys, and avP by 0.20 MJ/kg, 0.04 and 0.18% units for 1,000 FTU/kg and 0.4 MJ/kg, 0.06 and 0.20% units for 2,000 FTU/kg.

Effects of Low-Phosphorus Diets Supplemented with Phytase on the Production Performance, Phosphorus-Calcium Metabolism, and Bone Metabolism of Aged Hy-Line Brown Laying Hens

This study was conducted to evaluate the effects of phytase supplementation in low-phosphorus diets on the production performance, phosphorus-calcium metabolism, and bone metabolism in laying hens from 69 to 78 weeks of age. Hy-Line Brown laying hens (n = 1350) were assigned randomly to six treatments with five replicates of 45 birds. A corn-soybean meal-based diet with no inorganic phosphates was formulated to contain 0.12% non-phytate phosphorus (NPP) and 1470 FTU/kg phytase (Released phytate phosphorus content ≥ 0.1%). Inorganic phosphorus (dicalcium phosphate) was supplemented into the basal diet to construct five test diets (level of NPP supplementation = 0.10%, 0.15%, 0.20%, 0.25%, and 0.30%). The level of calcium carbonate was adjusted to ensure that all six experimental diets contained the same calcium percentage (3.81%). The feeding trial lasted 10 weeks (hens from 69 to 78 weeks of age). Upon supplementation with phytase (1470 FTU/kg), supplemental inorganic phosphates (dicalcium phosphate) had no significant effects (p > 0.05) on the production performance or egg quality. Significant differences in serum levels of calcium, phosphorus, copper, iron, zinc, or manganese were not detected across treatments (p > 0.05). Hens fed NPP (0.15%, 0.20%, 0.25%, and 0.30%) had higher levels (p < 0.0001) of tibial ash, calcium, and phosphorus than those not fed inorganic phosphates. The tibial breaking strength of the group without inorganic phosphates was significantly lower than that of the other groups (p < 0.01). Dietary supplementation with inorganic phosphates had no effect (p > 0.05) on serum levels of calcitonin (CT) and 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3). Hens that did not receive supplementation with inorganic phosphates had higher serum levels of parathyroid hormone (PTH), osteoprotegerin (OPG), type-I collagen c-telopeptide (CTX-I), and tartrate-resistant acid phosphatase 5b (TRACP-5b) compared with those in the other groups (p < 0.01). Serum levels of CTX-I and TRACP-5b were significantly lower in the NPP-supplementation groups of 0.25% and 0.30% than in the 0.10% NPP-supplementation group (p < 0.01). Dietary supplementation with inorganic phosphates had no effect (p > 0.05) on serum levels of bone-alkaline phosphatase (BAP), osteocalcin (OCN), or osteopontin (OPN). Hens not fed inorganic phosphate had the highest renal expression of phosphorus transporter type IIa Na/Pi cotransporter (NaPi-Ⅱa). Renal expression of NaPi-Ⅱa was increased significantly in NPP-supplementation groups of 0.10-0.20% compared with that in NPP-supplementation groups of 0.25% and 0.30% (p < 0.0001). The results indicated that a reduction in NPP supplementation to 0.15% (dietary NPP level = 0.27%) with phytase inclusion did not have an adverse effect on the production performance or bone health of laying hens from 69 to 78 weeks of age, which might be attributed to renal phosphorus reabsorption and bone resorption. These findings could support the application of low-phosphorus diets in the poultry industry.

Influence of Dietary Phytase Inclusion Rates on Yolk Inositol Concentration, Hatchability, Chick Quality, and Early Growth Performance

The aim of this study was to determine the influence of dietary phytase in breeder hens on yolk nutrients, hatchability, chick quality, and growth rate of their progeny, and their subsequent performance to 42 d post-hatch when fed diets with the same phytase concentrations. Breeder hens (n = 216) were divided into 3 groups receiving nutrient-adequate diets with reduced calcium (Ca) and phosphorus (P) (by 0.16% and 0.15%, respectively), supplemented with either 500, 1500, or 4500 FTU/kg phytase from 27 to 50 weeks of age. Eggs were collected at 38 weeks of age and incubated. On the day of hatch, the chick quality and hatchability were determined, and 18 chicks/group were euthanized for yolk sac collection and the determination of inositol and glycerol concentrations. The remaining chicks were divided into three groups, receiving different diets with reduced Ca and P (by 0.16% and 0.15%, respectively), supplemented with 0, 500, or 1500 FTU/kg phytase to 42 d post-hatch. Increasing the phytase concentration in the breeder hen diet linearly (p < 0.05) increased the number of early embryo deaths and decreased the number of late deaths and pips. The inositol concentration in the yolk sac at day of hatch increased (quadratic; p < 0.05) as the phytase dose increased in the breeder hen diet. The breeder hen diet (p < 0.05) influenced the body weight (BW), feed intake (FI), and feed conversion ratio (FCR) up to 21 days of age. The supplementation of breeder hen diets with 1500 FTU/kg phytase increased the concentration of sodium (Na), magnesium (Mg), potassium (K), manganese (Mn), and zinc (Zn) in the yolk sac. The inclusion of phytase doses up to 4500 FTU/kg appeared to influence embryo mortality, chick feed intake, and BW gain to 21 days and the FCR throughout the entire production phase.

Also in Crayfish: How Phytase Inclusion Avoids Phytic Acid Effects on Hepatopancreas Enzymes of Redclaw Cherax quadricarinatus

This work aimed to evaluate the effects of dietary phytic acid on the enzymatic activities of the hepatopancreas of the redclaw crayfish, Cherax quadricarinatus. For this purpose, a completely randomized in vitro trial was conducted with three phytic acid levels (0.56, 1.68, and 2.80%) and three phytase doses (0, 250, and 500 PU/kg DM). Solubilized protein, reducing sugars, and soluble phosphorus showed significant responses to the interaction between phytic acid and phytase p &lt; 0.001 . Only the main effects were detected on the released amino acids, in keeping with the main effects of alkaline protease activity, which are negatively affected by phytic acid p &lt; 0.001 and improved by phytase inclusion p &lt; 0.001 . Differences in released reducing sugars were attributed to a reduction in amylase activity by increased levels of phytic acid and not to cellulase activity, where only a negative trend of phytic acid was found p = 0.068 . Phytic acid depresses calcium availability, which would explain the decrease in amylase activity. A 500 PU/kg DM dose improved amino acid, reduced sugars, and phosphorus release. These in vitro results might have in vivo implications for the digestibility of proteins, minerals, and energy. Further investigations are required to determine the chelated calcium effect on redclaw amylase activity, molting, and survival.

Impact of Exogenous Xylanase and Phytase, Individually or in Combination, on Performance, Digesta Viscosity and Carcass Characteristics in Broiler Birds Fed Wheat-Based Diets.

The current study was conducted to evaluate the effects of stored wheat-based diet (1.5 and 2.5 years stored wheat) with and without the supplementation of xylanase and phytase enzymes in combination or individually on performance parameters, digestibility, digesta viscosity and carcass characteristics of broilers. For this purpose, a total of 640-day-old male broilers were randomly distributed to the 64 pens, and each pen had 10 birds. Two basal isocaloric and isonitrogenous diets contained 1.5 and 2.5 years stored wheat were formulated in this experiment. In the current study, experimental feeds were prepared by supplementing exogenous enzymes in both basal diets with xylanase (500 XU), phytase (500 FTU) alone or in a combination of phytase and xylanase. Performance parameters data represents that both in starter phase and finisher phase, inclusion of exogenous enzymes xylanase and phytase in both basal diets alone or in combination enhance the feed intake, and body weight gain (p &lt; 0.05) and improve the feed conversion ratio in overall phase (p &lt; 0.05). Similarly, supplementation of exogenous xylanase and phytase alone or in combination enhance the nutrient digestibility and reduce the digesta viscosity (p &lt; 0.05). Based on the results of this experiment, it is concluded that supplementation of exogenous xylanase and phytase enzymes alone or in combination in wheat-based diets (stored wheat 1.5 and 2.5 years) enhance and improves the performance of birds.

Phytate-phosphorus and phytase on performance, bone characteristics, tissue and serum mineral concentration on broilers.

Male broiler chickens (384), Cobb 500, were housed in metabolic cages to assess the efficacy of phytase in diets with low and high phytate-phosphorus on the performance, bone physical characteristics, tissue and serum mineral deposits. Birds were distributed in four treatments with a 2 x 2 factorial arrangement in a completely randomized block design. Experimental diets based on maize-soybean meal were T1 - diet low phytate-phosphorus; T2 - diet low phytate-phosphorus and phytase (500 FTU/kg); T3 - diet high phytate-phosphorus; T4 - diet high phytate-phosphorus and phytase (500 FTU/kg). Feed intake, body weight, weight gain and feed conversion ratio were assessed. Two left tibias per experimental unit were analyzed for physical characteristics and mineral concentration; a section of skinless breast muscle and blood were collected to measure the concentration of calcium, phosphorus and sodium. Results showed interaction between bone stiffness and serum calcium. The inclusion of phytase in diets with low and high phytate-phosphorus did not alter performance, bone resistance and flexibility, mineral deposits in the tibia and breast muscle, but increased bone stiffness after 22 days of age. It also provided a higher serum calcium rate in broilers fed diets with low phytate-phosphorus up to 32 days of age.

Plant-based polymer as a thermoresistant carrier of phytase in Nile tilapia (Oreochromis niloticus) diets

A biodegradable soybean oil-based polymer produced following the Green Chemistry principles, was assessed as a thermoresistant vehicle of phytase aiming to improve the enzyme stability during fish diet processing. Juvenile Nile tilapia (70 ± 4.3 g) were submitted to a digestibility assay designed in a factorial model (3 ×2) to assess the polymer efficiency. The treatments comprised the combination of three phytase inclusion methods with two feed processing methods. The inclusion methods of phytase were: CPP- coating post processing, in which the enzyme was spread on top of pellets after processing, PAE- polymer associated to enzyme, which was adsorbed by the polymer and included in the ingredients mash prior feed processing and NE- no enzyme inclusion, playing a control role as phytase was not included. The feed processing methods were extrusion (120 °C) or pelleting (40 °C), exposing the polymer to different temperatures. Phytase was supplemented at 1500 units kg−1 using the CPP or PAE methods. There was a significant interaction for the apparent digestibility coefficient (ADC) values of phosphorus (P), calcium (Ca) and Ash (P < 0.05). CPP resulted in higher ADC values for P and Ca, compared to PAE and NE in extruded diets (P < 0.05). The PAE method resulted in higher ADC values of P, Ca, and Ash than NE in extruded diets (P < 0.05). In pelleted diets, PAE resulted in higher ADC values for P and Ca, and a similar ADCAsh in comparison to CPP (P < 0.05). As verified in extruded diets data, the polymer partially protected phytase from heat denaturation, resulting in higher ADC values in comparison to NE (P < 0.05). The polymer improved phytase effects in pelleted diets probably by reducing degradation in fish stomach. These results suggest that the polymer may be a potential protective carrier of phytase and other thermolabile additives in extruded and pelleted diets.

Research Note: Evaluation of a precision-fed rooster assay for determination of phytic acid disappearance in feedstuffs

The objective of this study was to evaluate a precision-fed rooster assay that is suitable for determination of phytic acid (myo-inositol 1,2,3,4,5,6-hexakis; InsP6) disappearance in plant-based feed ingredients. A 48-h precision-fed rooster assay was used to measure InsP6 disappearance using conventional White Leghorn roosters. A minimum of 4 individually-caged roosters per treatment were fasted for 26 h prior to crop intubation with 15 to 30 g of sample, and excreta were quantitatively collected for 48 h. Soybean meal, soybean hulls, canola meal, conventional distillers dried grains with solubles (DDGS), palm kernel meal (PKM), and wheat bran were evaluated in Experiment 1, whereas wheat middlings (WM) and rice bran (RB) were evaluated without and with 1,000 and 1,800 U/kg phytase in Experiment 2. Data from Experiment 1 were subjected to a one-way ANOVA for a completely randomized design, while data from Experiment 2 were subjected to two-way ANOVA for a 2×3 factorial arrangement of treatments. In Experiment 1, InsP6 disappearance ranged from 3 to 95% among all ingredients. The InsP6 disappearance for conventional DDGS (95%) was the highest (P < 0.05), wheat bran and soybean hulls were intermediate (47-48%), PKM was low (24%), and soybean meal and canola meal were very low (3-5%). In Experiment 2, there was a significant ingredient × phytase interaction (P < 0.05). Phytase inclusion at both 1,000 and 1,800 U/kg resulted in a significant improvement (P < 0.05) in InsP6 disappearance for RB; however, only the addition of 1,800 U/kg resulted in an increase in InsP6 disappearance for WM. The addition of 1,800 U/kg phytase increased the InsP6 disappearance from 58 to 74% for WM and from 26 to 53% for RB. These results suggest the precision-fed rooster assay can be used to evaluate phytic acid disappearance in plant-based feed ingredients and the assay was able to detect a significant effect of 1,800 U/kg of exogenous phytase on phytic acid disappearance for WM and RB.

Ileal and total tract digestibility of energy and nutrients in pig diets supplemented with a novel consensus bacterial 6-phytase variant.

An experiment was conducted to test the hypothesis that increasing levels of a novel phytase increases the apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients in diets fed to young pigs. A negative control (NC) diet based on corn, soybean meal, and canola meal that contained approximately 0.83% phytate (i.e., 0.23% phytate-bound P) was formulated to be deficient in Ca, P, and standardized ileal digestible amino acids (AA). Five additional diets were formulated by adding 250, 500, 1,000, 2,000, or 4,000 phytase units/kg of the novel phytase to the NC diets. Eighteen ileal-cannulated pigs (17.81 ± 1.71 kg) were allotted to a 6 × 3 incomplete Latin square design with six diets and three 11-day periods. There were three pigs per diet in each period; therefore, there were nine replicate pigs per diet. The initial 5 d of each period was considered an adaptation period to the diet. For each period, fecal samples were collected via anal stimulation on days 6, 7, 8, and 9, whereas ileal digesta were collected on days 10 and 11 using standard procedures. Results indicated that the AID of crude protein, indispensable AA, and dispensable AA was increased (quadratic, P < 0.05) as the concentration of microbial phytase increased in the diets. Dietary inclusion of the novel phytase at 1,000 or 2,000 FTU/kg increased the AID of total AA from 73.7% to 79.8%. Increasing levels of microbial phytase increased (quadratic, P < 0.05) the AID of dry matter and minerals (i.e., Ca, P, K, Mg, Cu) in the diets. Likewise, a linear increase (P < 0.05) in the AID of ash and Na was observed as the inclusion level of phytase increased in the diets. Increasing levels of microbial phytase increased (linear, P < 0.01) the AID of gross energy (GE) and starch in the diets. A quadratic (P < 0.05) increase in the ATTD of ash, Ca, P, K, and Cu in experimental diets was observed as the concentration of microbial phytase increased in the diets. The ATTD of Mg and GE also increased (linear; P < 0.05) as concentration of dietary phytase increased. In conclusion, the novel microbial phytase used in this experiment was effective in increasing the AID of dry matter, GE, starch, minerals, and AA, as well as the ATTD of gross energy and minerals in diets formulated to be deficient in Ca, P, and AA.

Effects of microbial phytase on standardized total tract digestibility of phosphorus in feed phosphates fed to growing pigs.

An experiment was conducted to test the hypothesis that the apparent total tract digestibility (ATTD) and the standardized total tract digestibility (STTD) of P in feed phosphates are increased by microbial phytase when fed to growing pigs. Monocalcium phosphate (MCP), monosodium phosphate (MSP), and magnesium phosphate (MgP) from volcanic deposits were used in the experiment. Three corn-soybean meal based diets that contained 0, 500, or 4,000 units of microbial phytase (FTU), but no feed phosphates, were formulated. Nine additional diets were formulated by adding each of the three feed phosphates to the three basal diets. A P-free diet was also formulated to estimate the basal endogenous loss of P, and therefore, 13 diets were used in the experiment. A total of 117 growing barrows (initial body weight: 15.56 ± 1.68 kg) were allotted to the 13 diets with 9 pigs per diet. Pigs were housed individually in metabolism crates equipped with a feeder and a nipple drinker. Installation of a screen floor under the slatted floor allowed for collection of feces. Diets were fed for 10 d, with the initial 5 d being a period of adaptation to the diet followed by a collection period of 4 d. During the experiment, pigs were fed equal amounts of feed twice daily at 0800 and 1600 h. Results indicated that the ATTD and STTD of P in all diets increased with the inclusion of 500 or 4,000 FTU, but the ATTD and STTD of P in the feed phosphates were not affected by the inclusion of phytase. This indicates that the increases in ATTD and STTD of P that were observed in the mixed diets when phytase was used were due to the release of P from phytate in corn and soybean meal and not from an increase in digestibility of P in feed phosphates. However, MgP had a lower (P < 0.05) ATTD and STTD of P than MCP and MSP. In conclusion, microbial phytase does not increase the digestibility of P in MCP, MSP, or MGP, but the digestibility of P in MgP is less than in MCP and MSP.

Phytase dose-dependent response of kidney inositol phosphate levels in poultry.

Phytases, enzymes that degrade phytate present in feedstuffs, are widely added to the diets of monogastric animals. Many studies have correlated phytase addition with improved animal productivity and a subset of these have sought to correlate animal performance with phytase-mediated generation of inositol phosphates in different parts of the gastro-intestinal tract or with release of inositol or of phosphate, the absorbable products of phytate degradation. Remarkably, the effect of dietary phytase on tissue inositol phosphates has not been studied. The objective of this study was to determine effect of phytase supplementation on liver and kidney myo-inositol and myo-inositol phosphates in broiler chickens. For this, methods were developed to measure inositol phosphates in chicken tissues. The study comprised wheat/soy-based diets containing one of three levels of phytase (0, 500 and 6,000 FTU/kg of modified E. coli 6-phytase). Diets were provided to broilers for 21 D and on day 21 digesta were collected from the gizzard and ileum. Liver and kidney tissue were harvested. Myo-inositol and inositol phosphates were measured in diet, digesta, liver and kidney. Gizzard and ileal content inositol was increased progressively, and total inositol phosphates reduced progressively, by phytase supplementation. The predominant higher inositol phosphates detected in tissues, D-and/or L-Ins(3,4,5,6)P4 and Ins(1,3,4,5,6)P5, differed from those (D-and/or L-Ins(1,2,3,4)P4, D-and/or L-Ins(1,2,5,6)P4, Ins(1,2,3,4,6)P5, D-and/or L-Ins(1,2,3,4,5)P5 and D-and/or L-Ins(1,2,4,5,6)P5) generated from phytate (InsP6) degradation by E. coli 6-phytase or endogenous feed phytase, suggesting tissue inositol phosphates are not the result of direct absorption. Kidney inositol phosphates were reduced progressively by phytase supplementation. These data suggest that tissue inositol phosphate concentrations can be influenced by dietary phytase inclusion rate and that such effects are tissue specific, though the consequences for physiology of such changes have yet to be elucidated.

Apparent digestibility of energy and nutrients and efficiency of microbial phytase is influenced by body weight of pigs.

An experiment was conducted to test the hypothesis that regardless of pig body weight (BW), increasing dietary phytase results in increased phytate degradation and improved digestibility of minerals, amino acids (AA), and gross energy (GE). Eighteen pigs were equipped with a T-cannula in the distal ileum and allotted to a triplicated 6 × 3 Youden square design with six diets and three collection periods of 7 d, for a total of nine replicate pigs per diet. This design was repeated four times to simulate four production phases, and there was a 7-d resting period before each collection phase started (BW at start of collections: 29.3, 53.6, 85.1, and 114.4 kg for phases 1, 2, 3, and 4, respectively). Six corn-soybean meal diets were formulated by including 0, 250, 500, 1,000, 2,000, or 4,000 phytase units/kg feed (FTU). The six diets were used throughout the experiment. Samples of feces and ileal digesta were collected in each period. Results indicated that regardless of pig BW, increasing inclusion of phytase increased (quadratic; P < 0.05) apparent ileal digestibility (AID) of crude protein (CP) and most AA, increased apparent total tract digestibility (ATTD) of Ca, P, K, Mg (linear and quadratic; P < 0.05), and Na (linear; P < 0.05), but decreased (linear and quadratic; P < 0.05) AID and ATTD of GE. In all phases, ileal concentrations of inositol phosphate (IP) 6, IP5, IP4, and IP3 decreased (linear and quadratic; P < 0.05), whereas ileal inositol increased (linear and quadratic; P < 0.05) with increasing dietary phytase. However, as pig BW increased, AID of GE, CP, and AA increased (linear, P < 0.05), and the AID of a few AA (Met, Phe, Thr, Trp, Ala, Asp, Gly, and Ser) also increased quadratically (P < 0.05). The ATTD of GE, K, and Mg increased (linear and quadratic; P < 0.05), but ATTD of Ca and Na (linear; P < 0.05) and of P (linear and quadratic; P < 0.05) decreased as pig BW increased. Ileal IP6 and IP3 (linear and quadratic; P < 0.05) and ileal IP5 and IP4 (linear; P < 0.05) increased, whereas ileal inositol decreased (linear; P < 0.05) as pig BW increased. In conclusion, regardless of pig BW, increasing dietary phytase increased phytate degradation and inositol release in the small intestine, and consequently increased mineral and AA digestibility. Older pigs have reduced Ca, P, and Na digestibility, but increased K, Mg, AA, and GE digestibility compared with younger pigs. The efficiency of dietary phytase to degrade phytate appears to decrease as pigs get older.

High dietary Ca and microbial phytase reduce expression of Ca transporters while enhancing claudins involved in paracellular Ca absorption in the porcine jejunum and colon.

Expression levels of genes (RT-qPCR) related to Ca and P homeostasis (transporters and claudins (CLDN)) were determined in porcine jejunal and colonic mucosa. Forty growing pigs (BW 30.4±1.3 kg) received a low and high Ca content (2.0 and 9.6 g/kg, respectively) diet with or without microbial phytase (500 FTU/kg) for 21 days. Dietary Ca intake enhanced serum Ca and alkaline phosphatase concentration and reduced P, 1,25(OH)2D3, and parathyroid hormone concentration. Jejunal TRPV5 mRNA expression was decreased (32%) with phytase inclusion only, while colonic transient receptor potential vanilloid 5 (TRPV5) mRNA was reduced by dietary Ca (34%) and phytase (44%). Both jejunal and colonic TRPV6 mRNA expression was reduced (30%) with microbial phytase. Calbindin-D9k mRNA expression was lower in colonic but not jejunal mucosa with high dietary Ca (59%) and microbial phytase (37%). None of the mRNAs encoding the Na-P cotransporters (NaPi-IIc, PiT-1, PiT-2) were affected. Jejunal, but not colonic expression of the phosphate transporter XPR1, was slightly downregulated with dietary Ca. Dietary Ca downregulated colonic CLDN-4 (20%) and -10 (40%) expression while CLDN-7 was reduced by phytase inclusion in pigs fed low dietary Ca. Expression of colonic CLDN-12 tended to be increased by phytase. In jejunal mucosa, dietary Ca increased CLDN-2 expression (48%) and decreased CLDN-10 (49%) expression, while phytase slightly upregulated CLDN-12 expression. In conclusion, compared to a Ca deficient phytase-free diet, high dietary Ca and phytase intake in pigs downregulate jejunal and colonic genes related to transcellular Ca absorption and upregulate Ca pore-forming claudins.

Effect of Phytase Level and Form on Broiler Performance, Tibia Characteristics, and Residual Fecal Phytate Phosphorus in Broilers from 1 to 21 Days of Age.

Simple SummaryThe demand for poultry products is increasing at the same time as the prices of raw materials and other ingredients used in feed manufacturing, leading to the implementation of strategies to improve nutrient utilization in broiler diets and reduce feeding costs. One strategy used in broiler production is the dietary supplementation of phytase, an enzyme that assists in phytate degradation. Phytate is the primary storage form of phosphorus (P) in seeds and it accounts for two-thirds of the P in vegetable feedstuffs. Dietary phytase supplementation has become a common practice because the activity of endogenous phytase in broilers does not allow for the adequate utilization of phytate-bound P. Consequently, the effects of phytase have been widely studied and the beneficial effects on broiler growth performance and reduced P excretion into the environment have been previously described. In the present study, broilers were fed a diet with a combination of two phytase forms (coated and uncoated) to assess the effects on broiler performance, tibia characteristics, and residual fecal phytate P. The results indicated no distinct advantage of combining phytase forms, but this information may be useful for the direction of future research.The present study evaluated the individual and combined effects of coated and uncoated phytase on broiler performance, tibia characteristics, and residual phytate phosphorus (P) in manure. Two repeated studies were conducted using 240-day-old Cobb 500 by-product male broilers per trial. For each trial, birds were assigned to four treatments with four replicate battery cages per treatment (60 birds/trt) and grown for 21 days. Treatments included: (1) negative control (NC), (2) NC + 1000 phytase units (FTU) coated phytase (C), (3) NC + 1000 FTU uncoated phytase (U), and (4) NC + 500 FTU coated + 500 FTU uncoated phytase (CU). Data were analyzed with a one-way ANOVA and means were separated using Tukey’s HSD. In the pooled data for both trials, all treatments with dietary phytase had a higher body weight (BW) and feed consumption (FC) than the NC on day 21 (p < 0.05). Similarly, a six-point reduction was observed for day 1 to 21 feed conversion (FCR) for U and CU (p < 0.05). All treatments with phytase inclusion differed from the NC in every evaluated parameter for bone mineralization (p < 0.05) and had significantly lower fecal phytate P concentrations compared to the NC (p < 0.05). Overall, bird performance was essentially unaffected by phytase form, indicating that combining phytase forms does not appear to offer any advantage to the evaluated parameters from day 1 to 21.

123 Superdosing Phytase Fed During Transition Phase in Sows Increased Piglet Weight Gain

Abstract Farrowing and lactation periods require a high demand for nutrients. Superdose (≥1500 FTU/kg) of phytase may increase the nutritive value of diets and improve litter performance. The objective of this study was to determine whether the superdosing of phytase (Quantum Blue; AB Vista) in the peripartum diet affects sow and piglet productivity and serum biomarkers (urea, cortisol, and calcium). A total of 54 sows, Topigs TN70 (27 control group, 27 superdosing group) were blocked by parity and randomly assigned to the treatments. The control lactation diet was a mixed-cereal and soybean meal-based, formulated to meet sow’s requirement with 500 FTU/kg phytase inclusion. The superdosing diet, was supplemented with a further 2,000 FTU/kg phytase to give a total of 2,500 FTU/kg. Peripartum treatment diets were provided from 10 d before farrowing until 10 d after, after which a common standard lactation was fed. Piglets were weaned at 26 d of age. Data were analyzed with ANOVA evaluating the effects of phytase dose and parity using the fit model platform in JMP15.1. Significant treatment means were separated using Student’s t-tests, with significance accepted at P ≤ 0.05. Birth weights of piglets were significantly affected by parity, with piglets born from parity 1 or 4+ sows being around 180g lighter in weight (P &amp;lt; 0.01) than piglets born from parity 2 or 3. The superdosing treatment increased piglet weight at weaning by 490g/weaned piglet (P &amp;lt; 0.01). Cortisol levels were greater (3.7 vs. 1.3 µg/dl, P &amp;lt; 0.01) at farrowing compared with 10d post-farrowing. Urea concentrations were 37% higher in the superdosing treatment at farrowing compared with control (P = 0.02). It can be concluded that the use of phytase at superdosing levels can be used in peripartum diets to improve piglet weight gain during lactation which is linked with greater utilization of nutrients that has been associated with greater phytate breakdown.

Beneficial effects of phytase and/or protease on growth performance, digestive ability, immune response and muscle amino acid profile in low phosphorus and/or low fish meal gibel carp (Carassius auratus gibelio) diets

A 10-week feeding trial was conducted to investigate the effects of individual and combined inclusion of phytase and protease on the growth performance, digestive ability, immune response and muscle amino acid profile of gibel carp (Carassius auratus gibelio) fed low phosphorus (LP) and/or low fish meal (LFM) diets. Seven experimental diets were designed: 1) Normal diet [PC, 15 g/kg Ca(H2PO4)2 and 80 g/kg fish meal], 2) Low P diet [10 g/kg Ca(H2PO4)2 and 80 g/kg fish meal], 3) LP + 0.4 g/kg phytase diet (LP + Ph), 4) Low fish meal diet [LFM, 15 g/kg Ca(H2PO4)2 and 50 g/kg fish meal], 5) LFM + 0.5 g/kg protease diet (LFM + Pr), 6) Low P and Low fish meal diet [LP-LFM, 10 g/kg Ca(H2PO4)2 and 50 g/kg fish meal] and 7) LP-LFM + 0.4 g/kg phytase +0.5 g/kg protease diet (LP-LFM + Ph + Pr). The results showed that the growth performance, whole body composition, nutrient digestibility, immunological indices, and muscle amino acid contents of gibel carp fed the LP, LFM and LP-LFM diets were dramatically lower than those of carp fed the normal diet (P<0.05). However, compared the LP-LFM, LFM and LP groups, dietary addition of phytase + protease, protease and phytase, respectively, (1) improved the special growth rate (SGR), weight gain rate (WGR), ADC of dry matter, crude protein and P; (2) upregulated the activities of lipase, trypsin, amylase and chymotrypsin in the liver and intestine; (3) increased liver lysozyme (LZM), superoxide dismutase (SOD), glutathione peroxidase (GPx) and total antioxidant capacity (T-AOC) levels, intestinal secreted immunoglobulin A (sIgA), complement 4 (C4) and C3 levels; (4) increased the contents of amino acids, for example, valine, leucine and methionine, in the muscle. Moreover, when both phytase and protease were added simultaneously, they had a better promoting effect than when individual added. It can be concluded that a reduction of dietary P and fish meal for gibel carp has side effects on growth performance, while phytase and protease could mitigate these effects by improving the digestive ability and immune response of gibel carp.

Effect of Dietary Phytase Supplementation on Growth Performance, Organ Weight and Tibia Ash of Broilers

This study was conducted to investigate the effects of dietary inclusion of phytase on the growth performance, organ weight, and tibia ash of broilers. A total of 1008 one-day-old Ross 308 broiler chicks (mixed gender) with body weight 42 ± 0.90 g (mean ± SD) were used in a trial for 32 d. Chicks were assigned to one of the two treatment diets. Each treatment consisted of 28 replicate cages, with 18 birds per cage. The dietary treatments were as follows: CON, basal diet; TRT1, basal diet + phytase 1,500 Fan Terminal Unit (FTU)/kg diet. Dietary inclusion of 1,500 FTU/kg phytase supplementation in broilers showed only slight improvements in daily feed intake (<i>P</i>=0.086) and feed conversion ratio (<i>P</i>=0.065) on day 9 compared with that in the control group. However, organ weights of the breast muscle, liver, spleen, kidney, and bursa of Fabricius were not affected by the dietary treatments. In addition, the dietary inclusion of 1,500 FTU/kg phytase supplementation in the broiler diet had no effect on tibia ash. The addition of 1,500 FTU/kg phytase in the basal diet of broilers did not have any adverse effect on growth performance, organ weight, and tibia ash, suggesting that phytase supplementation in broiler diets will exhibit comparable effects as that of corn-soybean meal-based diet.

The anti-nutritional effect of phytates - the extraphosphorus effect of phytase (review)

An analytical review was conducted on the availability of phosphorus from phytates, which increase the inclusion of phytase in the feed, while not only phytic acid is broken down, but also as a result of a decrease in concentration reduces its anti-nutritional effect.Phosphorus from plant feeds is not fully available to animals, as it is part of phytates, the cleavage of which in the gastrointestinal tract (gastrointestinal tract) of animals is limited. Phytates, getting into the acidic environment of the stomach, ionize and react with positively charged minerals, proteins, amino acids, creating compounds inaccessible for further digestion. The inclusion of phytase in compound feed is accompanied by an extra phosphoric effect, which is expressed in an increase in the availability of amino acids and energy. The decision on the feasibility of including phytase in feed is made on the basis of production tests of the proposed drugs.The studies carried out by the authors of invitro confirm the assumption that phytates enter into chemical interactions with peptides, as well as amino acids formed in the processes of protein digestion, turning them into compounds inaccessible to absorption. The proposed explanation does not affect the results of the balance experiment, but allows us to develop the study of the mechanism in terms of the interaction of digested amino acids with phytates. Based on this, another conclusion can be seen: the destruction of phytates should occur in the body before the digestion and dissolution of protein. In birds, this place is the goiter and stomachs, in pigs - the stomach. The hydrolysis of newly formed phytates (FC-amino acid) requires phytases that are active in the environment of the small intestine at pH 6 -7. As a result, the bound amino acids will be re-released and available for absorption. The development of new phytase preparations, taking into account the proposed explanation, will make it possible to create more effective feed preparations.

Supplemental Effects of Phytase on Modulation of Mucosa-Associated Microbiota in the Jejunum and the Impacts on Nutrient Digestibility, Intestinal Morphology, and Bone Parameters in Broiler Chickens.

Simple SummaryA positive modulation of the gut microbiota is associated with benefits regarding intestinal health and subsequent growth performance. The supplementation of phytase has been studied for improving nutrient digestibility, bone parameters, and growth performance. This study aimed to determine the effects of increasing doses of phytase on the modulation of mucosa-associated microbiota in the jejunum enhancing intestinal health and the impacts on nutrient digestibility, bone parameters, and growth performance of broiler chickens. It was demonstrated that the use of phytase enhanced the intestinal health of the broiler chickens by potentially increasing beneficial and reducing harmful bacteria, enhancing intestinal morphology, and consequently improving nutrient digestibility and bone parameters. Our results suggest that the use of phytase can positively modulate the jejunal mucosa-associated microbiota in the jejunum, enhance intestinal integrity, nutrient digestibility, and bone parameters of broiler chickens.This study aimed to determine supplemental effects of phytase on modulation of the mucosa-associated microbiota in the jejunum, intestinal morphology, nutrient digestibility, bone parameters, and growth performance of broiler chickens. Three hundred and sixty newly hatched broiler chickens (Ross 308) (44 ± 2 g BW) were randomly allotted in 6 treatments with 10 birds per cage based on a completely randomized design and fed for 27 d. The treatments consisted of one negative control (NC), diet formulated meeting the requirements suggested by Ross recommendations (2019), and without phytase supplementation. The other treatments consisted of a positive control diet (PC) formulated with 0.15% deficient Ca and P and split into 5 treatments with different phytase inclusion levels (0, 500, 1000, 2000, 4000 FTU/kg feed). Titanium dioxide (0.4%) was added to feeds as an indigestible marker to measure apparent ileal digestibility (AID) of nutrients. On d 27, 3 birds were randomly selected from each cage and euthanized to collect samples for analyzing the mucosa-associated microbiota in the jejunum, oxidative stress status, AID, and bone parameters. Data were analyzed using the proc Mixed of SAS 9.4. Phytase supplementation tended to have a quadratic effect (p = 0.078) on the overall ADG (maximum: 41 g/d at 2833 FTU/kg of feed). Supplementation of phytase at 2,000 FTU/kg increased (p < 0.05) the relative abundance of Lactobacillus and reduced (p < 0.05) Pelomonas. Moreover, it tended to reduce Helicobacter (p = 0.085), Pseudomonas (p = 0.090) Sphingomonas (p = 0.071). Phytase supplementation increased (p < 0.05) the villus height and the AID of CP; and tended to increase (p = 0.086) the AID of P. Phytase supplementation increased (p < 0.05) breaking strength and P content in the tibia. In conclusion, phytase supplementation showed potential benefits on the modulation of the mucosa-associated microbiota in the jejunum by tending to reduce harmful bacteria (Pelomonas, Helicobacter, and Pseudomonas) and increase beneficial bacteria (Lactobacillus). In addition, it showed positive effects increasing apparent ileal digestibility of CP and P, enhancing intestinal morphology (villus height), and improving the bone parameters (bone breaking strength, ash, and P content). Phytase supplementation at a range of 38 to 59 FTU/d or 600 to 950 FTU/kg of feed provided the most benefits related to nutrient digestibility.