Hydrolysis of phytic acid by intrinsic plant and supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re-entrant cannulas. 2. Phytase activity

Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (A. niger) were investigated with six minipigs (40-50 kg initial BW) fitted with re-entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5, and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2, and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet (43 U phytase/kg DM); (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM) and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 0730 and 1930 per animal 350 g diet mixed with 1050 ml de-ionized water were fed. Digesta were collected continuously and completely during 12 h after feeding. In the duodenal digesta, 70% of the microbial phytase (diet 2) and 45% of the wheat phytase (diet 3) were recovered within 12 h after ingestion of the phytases, whereas only negligible amounts were detected in the digesta of pigs fed the phytase-poor corn-based diet 1. Most phytase activity passed through the stomach within the first hour after feeding. Microbial phytase activity at pH 2.8 was less sensitive to acidic pHs, such as those found in the stomach, than phytase activity at pH 5.3. Phytase activities in the digesta of the distal ileum did not depend either on source or amount of dietary phytase activity.     

Hydrolysis of phytic acid by intrinsic plant or supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re-entrant cannulas

Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (A. niger) were investigated with six minipigs (40-50 kg initial BW) fitted with re-entrant-cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5, and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2, and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet (43 U Phytase/kg DM); (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM) and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 0730 and 1930 per animal 350 g diet mixed with 1050 ml de-ionized water were fed. Digesta were collected continuously and completely during 12 h after feeding. Duodenal recovery of dry matter and total phosphorus were 100% in the period between two feedings, irrespective of dietary treatment. In animals fed the wheat-based diet, dry matter left the stomach faster (p < 0.05) during the first hour after feeding than in animals fed the corn-based diets (41.3 vs. 31.0 and 25.8% of intake, respectively). Supplemented microbial phytase did not affect ileal dry matter digestibility of the corn-based diet. In the first hour after feeding, phosphorus concentration of the duodenal digesta of animals fed corn-based diets with or without supplemented microbial phytase (5.86, 6.19 mg total P/g DM) exceeded the dietary level considerably (4.30 and 4.21 mg total P/g DM) indicating a higher solubility of corn than wheat phosphorus in the stomach. Apparent ileal P absorption was higher (p < 0.05) in the wheat-based diet (37.6%) and corn-based diet supplemented with microbial phytase (34.3%) than in the unsupplemented corn-based diet (17.6%).     

The effect of supplementary Aspergillus niger phytase in diets for pigs on concentration and apparent digestibility of dry matter, total phosphorus, and phytic acid in different sections of the alimentary tract.

Six barrows of approximately 37 kg BW, fitted with two simple T-cannulas in the duodenum (25 cm posterior to the pylorus) and terminal ileum (12 to 15 cm anterior to the ileocecal junction), were fed two diets containing 2.1 g of P/kg in the form of phytic acid and a low intrinsic phytase activity (corn-soybean meal based diet [Diet A] or a typical Dutch diet [Diet B]) without or with supplementary microbial phytase from Aspergillus niger (var. ficuum) equal to 1,500 phytase units per kilogram of diet, in a crossover design. The apparent duodenal, ileal, and total tract (overall) digestibilities of DM, total P, and phytate P (phytic acid x .282) were calculated using both Cr-NDR (neutral detergent residue mordanted with Cr) and Co-EDTA as dual-phase markers. Concentration of total P in the ileal digesta (P less than .01) and feces (P less than .001) of pigs fed microbial phytase was lower than without this enzyme, irrespective of the diet. Ileal digestibility of total P was 18.5 and 29.8 percentage units higher (which was a 1.7- to 2.9-fold increase) due to added Aspergillus niger phytase (P less than .05). Also, total tract (overall) digestibility increased by 27.0 to 29.7 percentage units (P less than .01). Phytic acid concentration in the duodenal and ileal digesta of pigs receiving microbial phytase was lower (P less than .01 or .001), resulting in its higher ileal digestibility (dephosphorylation rate) by 50.1 percentage units for Diet A and by 75.4 percentage units for Diet B. Irrespective of the treatment, no phytase activity could be detected in the ileal digesta of pigs.     

The effects of phytase and phytic acid on the loss of endogenous amino acids and minerals from broiler chickens

1. The effects of myo-inositol hexaphosphate (IP6) and phytase (EC 3.1.3.26) on the excretion of endogenous compounds were investigated using growing broiler chickens. 2. A total of 32 female Ross broilers were used in a precision feeding assay involving a 2 x 2 factorial arrangement of treatments. The materials administered were glucose, glucose + 1000 units of phytase activity (FTU), glucose + 1 g of IP6 and glucose + 1 g of IP6 + 1000 FTU. Excreta were collected quantitatively over a 48-h period following intubation of the test materials. The excretion of nitrogen, amino acids, minerals, sialic acid and phytate phosphorus was determined. 3. The ingestion of 1 g of IP6 by broilers increased the excretion of endogenous nitrogen, amino acids, iron, sodium, sulphur and sialic acid compared with birds fed on glucose. Supplementation of IP6 with exogenous phytase reduced the excretion of endogenous amino acids, calcium, sodium, phytate phosphorus and sialic acid compared with birds fed IP6. 4. It can be concluded that IP6 increases the excretion of endogenous minerals and amino acids in broiler chickens. Part of the beneficial effects of the addition of exogenous phytases to the diets of poultry appears to be mediated through a reduction in endogenous losses of these nutrients.     

Degradation of phytate in the gut of pigs--pathway of gastro-intestinal inositol phosphate hydrolysis and enzymes involved.

The present study gives an overview on the whole mechanism of phytate degradation in the gut and the enzymes involved. Based on the similarity of the human and pigs gut, the study was carried out in pigs as model for humans. To differentiate between intrinsic feed phytases and endogenous phytases hydrolysing phytate in the gut, two diets, one high (control diet) and the other one very low in intrinsic feed phytases (phytase inactivated diet) were applied. In the chyme of stomach, small intestine and colon inositol phosphate isomers and activities of phytases and alkaline phosphatases were determined. In parallel total tract phytate degradation and apparent phosphorus digestibility were assessed. In the stomach chyme of pigs fed the control diet, comparable high phytase activity and strong phytate degradation were observed. The predominant phytate hydrolysis products were inositol phosphates, typically formed by plant phytases. For the phytase inactivated diet, comparable very low phytase activity and almost no phytate degradation in the stomach were determined. In the small intestine and colon, high activity of alkaline phosphatases and low activity of phytases were observed, irrespective of the diet fed. In the colon, stronger phytate degradation for the phytase inactivated diet than for the control diet was detected. Phytate degradation throughout the whole gut was nearly complete and very similar for both diets while the apparent availability of total phosphorus was significantly higher for the pigs fed the control diet than the phytase inactivated diet. The pathway of inositol phosphate hydrolysis in the gut has been elucidated.     

微生物植酸酶对猪钙、磷利用率及骨骼发育的影响

本试验旨在研究从断奶到肥育结束,在饲粮中添加微生物植酸酶对猪钙、磷代谢及骨骼发育的影响。试验采用完全随机设计,将72头35日龄断奶的仔猪分为4组,每组3个重复,每个重复6头。试验在相同钙水平基础上设4个组,分别为:1)玉米-豆粕基础日粮(对照组);2)基础日粮 植酸酶-50%磷酸氢钙(处理1);3)基础日粮 植酸酶-75%磷酸氢钙(处理2);4)基础日粮 植酸酶-100%磷酸氢钙(处理3)。试验期按体重分3个阶段:8~20 kg、20~50 kg、50~90 kg,3个阶段植酸酶的添加量分别为750、500和250 U/kg。试验结果表明:(1)在8~20 kg和20~50 kg阶段,添加植酸酶的3个处理,钙、磷消化率显著高于对照组(P<0.05);血钙和血磷浓度,处理1和处理2与对照组无显著差异(P>0.05),处理3显著低于对照组(P<0.05)。在50~90 kg阶段,添加250 U/kg植酸酶代替100%磷酸氢钙,血磷、血钙浓度显著低于对照组(P<0.05);(2)在试验的3个阶段添加植酸酶同时降低饲粮无机磷水平,对血清碱性磷酸酶活性均无显著影响(P>0.05);(3)添加植酸酶使胃中植酸磷消化率显著提高(P<0.0 1),粪中磷排出量显著降低(8~2 0 kg、2 0~5 0 kg阶段,P<0.0 1;5 0~9 0 kg阶段,P<0.0 5);(4)添加植酸酶代替50%或75%磷酸氢钙,猪的掌骨灰分与采食正常磷日粮的对照组无显著差异,但代替100%磷酸氢钙组,猪的掌骨灰分和蹠骨强度则显著低于对照组(P<0.05);添加植酸酶代替50%磷酸氢钙组,猪的蹠骨强度显著高于对照组(P<0.05),代替75%磷酸氢钙组与对照组差异不显著(P>0.05)。总之,在断奶和生长阶段,在猪玉米-豆粕日粮中添加植酸酶可代替部分磷酸氢钙,促进了钙、磷消化利用,促进了骨骼生长,也促进了植酸磷的利用,降低了粪磷排出。     

Distribution of supplemental Escherichia coli AppA2 phytase activity in digesta of various gastrointestinal segments of young pigs

The objective of this study was to determine the functional location and disappearance of activity of a supplemental Escherichia coli AppA2 phytase and its impact on digesta P and Ca concentrations in the gastrointestinal tract of pigs. In Exp. 1, 18 pigs (8.3 +/- 0.2 kg of BW) were allotted to 3 groups (n = 6 each) and fed a low-P (0.4%) corn-soybean meal, basal diet (BD), BD + phytase [500 units (U)/kg of feed], or BD + inorganic P (iP, 0.1%) for 4 wk. In Exp. 2, 30 pigs (14.5 +/- 0.2 kg of BW) were allotted to 3 groups (n = 10 each) and fed BD, BD + 500 U of phytase/kg of feed, or BD + 2,000 U of phytase/kg of feed for 2 wk. Five or six pigs from each treatment group were killed at the end of both experiments to assay for digesta phytase activity and soluble P concentration in 6 segments of the digestive tract and digesta total P and Ca concentrations in stomach and colon. Compared with pigs fed BD, pigs fed BD + 500 U of phytase/kg of feed in Exp. 1 and BD + 2,000 U of phytase/kg of feed in Exp. 2 had greater (P < 0.05) phytase activities in the digesta of the stomach and upper jejunum (2 m aborally from the duodenum). No phytase activity was detected in the digesta of the lower jejunum (2.12 m cranial to the ileocecal junction) or ileum from any of the treatment groups in either trial. Concentrations of digesta-soluble P peaked in the upper jejunum of pigs fed BD in Exp. 1 and 2, but showed gradual decreases between the stomach and the upper jejunum of pigs fed BD + phytase or BD + iP. In both experiments, pigs fed only BD had greater (P < 0.05) colonic digesta phytase activity and soluble P concentrations than those fed phytase. In Exp. 2, total colonic digesta P or Ca concentrations, or both, of pigs displayed a phytase-dose-dependent reduction (P < 0.05). In conclusion, supplemental dietary AppA2 mainly functioned in the stomach and was associated with a reduced phytase activity in colonic digesta of weanling pigs.     

Phytase improves iron bioavailability for hemoglobin synthesis in young pigs.

Dietary phytase supplementation improves bioavailabilities of phytate-bound minerals such as P, Ca, and Zn to pigs, but its effect on Fe utilization is not clear. The efficacy of phytase in releasing phytate-bound Fe and P from soybean meal in vitro and in improving dietary Fe bioavailability for hemoglobin repletion in young, anemic pigs was examined. In Exp. 1, soybean meal was incubated at 37 degrees C for 4 h with either 0, 400, 800, or 1,200 units (U) of phytase/kg, and the released Fe and P concentrations were determined. In Exp. 2, 12 anemic, 21-d-old pigs were fed either a strict vegetarian, high-phytate (1.34%) basal diet alone, or the diet supplemented with 50 mg Fe/kg diet (ferrous sulfate) or phytase at 1,200 U/kg diet (Natuphos, BASF, Mt. Olive, NJ) for 4 wk. In Exp. 3, 20 anemic, 28-d-old pigs were fed either a basal diet with a moderately high phytate concentration (1.18%) and some animal protein or the diet supplemented with 70 mg Fe/kg diet, or with one of two types of phytase (Natuphos or a new phytase developed in our laboratory, 1,200 U/kg diet) for 5 wk. In Exp. 2 and 3, diets supplemented with phytase contained no inorganic P. In Exp. 1, free P concentrations in the supernatant increased in a phytase dose-dependent fashion (P<.05), whereas free Fe concentrations only increased at the dose of 1,200 U/kg (P<.10). In Exp. 2 and 3, dietary phytase increased hemoglobin concentrations and packed cell volumes over the unsupplemented group; these two measures, including growth performance, were not significantly different than those obtained with dietary supplemental Fe. In conclusion, both sources of phytase effectively degraded phytate in corn-soy diets and subsequently released phytate-bound Fe from the diets for hemoglobin repletion in young, anemic pigs.     

Interactions of phytate and myo-inositol phosphate esters (IP1-5) including IP5 isomers with dietary protein and iron and inhibition of pepsin

Phytic acid (IP(6)) and myo-inositol phosphate esters (IP(1-5)), including IP(5) isomers prepared chemically and enzymatically with bacterial and fungal phytases, were examined for their effects on protein aggregation of soy protein and β-casein, interaction with Fe(3+), and pepsin activity. The results indicated that the aggregating capabilities of IP esters (IP(1-6)) on the 2 proteins decreased dramatically from IP(6) to IP(5) and became negligible with IP(1-4). Among the IP(5) isomers tested, InsP(5)(1,2,3,4,5) produced by 6-phytase was slightly less powerful in aggregating protein than InsP(5)(1,2,4,5,6) produced by 3-phytase (P = 0.001). For protein hydrolysis, IP esters of IP(3-4) still showed inhibition of pepsin though to a lesser extent than IP(5-6). The in vitro data with IP(1-5) generated with microbial 3- and 6-phytases indicate that, for complete alleviation of pepsin inhibition, IP(6) needs to be broken down to IP(1-2.) In contrast to the aggregation with protein, the reactivity of IP(1-6) toward Fe(3+) decreased proportionally from IP(6) to IP(3.) Based on the radical decrease in turbidity of IP(6) -protein complex observed, as a result of IP(6) dephosphorylation to IP(5), a novel qualitative and semi-quantitative phytase plate assay was established using IP(6)-protein complex incorporated into an agarose petri-dish as substrate. Phytase activity was shown as the development of clear halos on the agarose plate with time. This simple phytase plate assay method can be used at animal farms, control laboratories, and even for the screening of engineered phytase variants. The current study, thus, stresses the importance of the efficient hydrolysis of IP(6) at lower pH range to alleviate the negative effect of phytic acid and its degradation products on protein and Fe(3+) digestion.     

Comparison of phytase from genetically engineered Aspergillus and canola in weanling pig diets

Ninety-six crossbred pigs with an average weight of 9.0 kg were used in a 5-wk trial to compare the efficacy of genetically engineered Aspergillus ficuum phytase, expressed in Aspergillus niger (Natuphos) or in canola seed (Phytaseed), for enhancing the utilization of phytate P in corn-soybean meal-based diets fed to young pigs and to evaluate the safety of Phytaseed phytase. Three levels of the two sources of phytase (250, 500, or 2,500 U/kg of diet) were added to a corn-soybean meal basal diet containing .35% total P, .09% available P, and .50% Ca. There were six pens per treatment (one barrow and one gilt/pen), except that the diet without added phytase was fed to 12 pens of pigs. Pen feed consumption and BW were recorded weekly. During wk 5, pen fecal samples were collected for determination of apparent digestibilities of DM, Ca, and P. At the end of wk 5, all barrows were killed, and the 10th rib on both sides was removed for determination of shear force and energy. Thirty pigs (six from the diet without added phytase and the diets with 500 and 2,500 U/kg phytase from both sources) were randomly selected for gross necropsy and histologic evaluation of liver, kidney, and bone tissues. Both sources of phytase were equally effective in increasing (P < .05) daily gain, gain:feed, apparent digestibilities of DM, P, and Ca, and 10th rib measurements. Fecal P excretion was reduced with phytase addition. Feed intake was increased by phytase levels during wk 4 to 5. No significant abnormalities were seen in any of the 30 pigs necropsied. The fit of a nonlinear function revealed that most measurements were reaching a plateau at 2,500 U/kg phytase. In summary, based on performance, bone measurements, and digestibilities of P, Ca, and DM of young pigs, the efficiency of Phytaseed was similar to that of Natuphos for enhancing the utilization of phytate P in corn-soybean meal-based diets. General necropsy and histologic examination of tissues indicated no toxic effect of phytase.     

Increased dietary sodium chloride concentrations reduce endogenous amino acid flow and influence the physiological response to the ingestion of phytic acid by broiler chickens

1. A total of 240 Ross 308 broilers were used to investigate the effect of sodium (1·5 or 2·5?g/kg), phytate-P (0 or 3·2?g/kg), and phytase (0 or 1000 FTU/kg; 2x2x2 factorial) on endogenous amino acid flow using the enzyme-hydrolysed casein method.

2. The ingestion of phytate increased endogenous amino acid flow (～30%) compared with the phytate-free control diets. Phytase reduced endogenous amino acid flow only when fed in concert with phytate, resulting in a significant phytate x phytase interaction.

3. Increasing dietary sodium concentration from 1·5 to 2·5?g/kg reduced endogenous amino acid flow by around 10%. This reduction of endogenous flow was particularly evident in diets which contained phytate, resulting in a significant sodium x phytate interaction for several amino acids, including Thr and Ser. Further, high sodium concentrations muted the effect of phytase resulting in a significant sodium x phytase interaction for some amino acids.

4. The concentration of Asp, Thr, Ser and some other amino acids was increased in the endogenous protein in response to the ingestion of phytate. Both sodium and phytase essentially restored the composition of endogenous protein to that of the phytate-free control. Further, as both sodium and phytase had similar effects there were significant interactions between sodium and phytase for most amino acids, such that one was only effective in the absence of the other.

5. These data confirm previous reports that phytate is a nutritional aggressor, causing quantitative and qualitative changes in endogenous protein flow. However, this is the first report which has shown that dietary sodium concentrations play a role in the severity of this antinutritional effect and consequently may blunt the efficacy of exogenous phytase. The mechanism is obscure, though it has been previously demonstrated that sodium can disrupt phytate:protein complexes, thus mitigating one of the mechanisms by which phytate exerts its antinutritional effect.     

The effects of citric acid on phytate-phosphorus utilization in young chicks and pigs

Several bioassays were conducted with young chicks and pigs fed phosphorus (P)-deficient corn-soybean meal diets. With diets for chicks containing .62% Ca and .42% P (.10% available P), graded doses of a citric acid + sodium citrate (1:1, wt:wt) mixture (0, 1, 2, 4, or 6% of diet) resulted in linear (P < .01) increases in both weight gain and tibia ash. Relative to chicks fed no citric acid, tibia ash (%) and weight gain (g/d) were increased by 43 and 22%, respectively, in chicks fed 6% citric acid. Additional chick trials showed that 6% citric acid alone or sodium citrate alone was as efficacious as the citric acid + sodium citrate mixture and that 1,450 U/kg of phytase produced a positive response in bone ash and weight gain in chicks fed a diet containing 6% citrate. Varying the Ca:available P ratio with and without citrate supplementation indicated that citric acid primarily affected phytate-P utilization, not Ca, in chicks. Moreover, chicks did not respond to citrate supplementation when fed a P-deficient (.13% available P), phytate-free casein-dextrose diet. Young pigs averaging 10 to 11 kg also were used to evaluate citric acid efficacy in two experiments. A P-deficient corn-soybean meal basal diet was used to construct five treatment diets that contained 1) no additive, 2) 3% citric acid, 3) 6% citric acid, 4) 1,450 U/kg phytase, and 5) 6% citric acid + 1,450 U/kg phytase. Phytase supplementation increased (P < .01) weight gain, gain:feed, and metatarsal ash, whereas citric acid addition increased only gain:feed (P < .05) and metatarsal ash (P < .08). A subsequent 22-d pig experiment was conducted to evaluate the effect of lower levels of citric acid (0, 1, 2, or 3%) or 1,450 U/kg phytase addition to a P-deficient corn-soybean meal diet. Phytase supplementation improved (P < .01) all criteria measured. Weight gain and gain:feed data suggested a response to citric acid addition, but this was not supported by fibula ash results (P > .10). The positive responses to phytase were much greater than those to citric acid in both pig experiments. Thus, dietary citric acid effectively improved phytate P utilization in chicks but had a much smaller effect in pigs.     

Effects of individual or combined xylanase and phytase supplementation on energy, amino acid, and phosphorus digestibility and growth performance of grower pigs fed wheat-based diets containing wheat millrun

The objective of these studies was to determine if dietary enzymes increase the digestibility of nutrients bound by nonstarch polysaccharides, such as arabinoxylans, or phytate in wheat millrun. Effects of millrun inclusion rates (20 or 40%), xylanase (0 or 4,375 units/kg of feed), and phytase (0 or 500 phytase units/kg of feed) on nutrient digestibility and growth performance were investigated in a 2 x 2 x 2 factorial arrangement with a wheat control diet (0% millrun). Diets were formulated to contain 3.34 Mcal of DE/kg and 3.0 g of true ileal digestible Lys/Mcal of DE and contained 0.4% chromic oxide. Each of 18 cannulated pigs (36.2 +/- 1.9 kg of BW) was fed 3 diets at 3x maintenance in successive 10-d periods for 6 observations per diet. Feces and ileal digesta were collected for 2 d. Ileal energy digestibility was reduced (P < 0.01) linearly by millrun and increased by xylanase (P < 0.01) and phytase (P < 0.05). Total tract energy digestibility was reduced linearly by millrun (P < 0.01) and increased by xylanase (P < 0.01). For 20% millrun, xylanase plus phytase improved DE content from 3.53 to 3.69 Mcal/kg of DM, a similar content to that of the wheat control diet (3.72 Mcal/kg of DM). Millrun linearly reduced (P < 0.01) ileal digestibility of Lys, Thr, Met, Ile, and Val. Xylanase improved (P < 0.05) ileal digestibility of Ile. Phytase improved ileal digestibility of Lys, Thr, Ile, and Val (P < 0.05). Millrun linearly reduced (P < 0.05) total tract P and Ca digestibility and retention. Phytase (P < 0.01) and xylanase (P < 0.05) improved total tract P digestibility, and phytase and xylanase tended to improve (P < 0.10) P retention. Phytase improved Ca digestibility (P < 0.05) and retention (P < 0.01). The 9 diets were also fed for 35 d to 8 individually housed pigs (36.2 +/- 3.4 kg of BW) per diet. Millrun reduced (P < 0.05) ADFI, ADG, and final BW. Xylanase increased (P < 0.05) G:F; phytase reduced (P < 0.05) ADFI; and xylanase tended to reduce (P = 0.07) ADFI. In summary, millrun reduced energy, AA, P, and Ca digestibility and growth performance compared with the wheat control diet. Xylanase and phytase improved energy, AA, and P digestibility, indicating that nonstarch polysaccharides and phytate limit nutrient digestibility in wheat byproducts. The improvement by xylanase of energy digestibility coincided with improved G:F but did not translate into improved ADG.     

Effect of phytase addition and dietary calcium and phosphorus levels on plasma metabolites and ileal and total-tract nutrient digestibility in pigs

Two experiments were conducted to determine the effect of phytase on plasma metabolites and AA and energy digestibility in swine. In Exp. 1, eight barrows (surgery BW = 52 kg) were fitted with steered ileocecal cannulas. The experiment was a Latin rectangle and the treatments were 1) corn-soybean meal diet adequate in Ca and P (0.5% Ca, 0.19% available P [aP]), 2) corn-soybean meal diet with reduced Ca and P (0.4% Ca, 0.09% aP), 3) Diet 1 with 500 phytase units/kg, or 4) Diet 2 with 500 phytase units/kg. Pigs were fed twice daily to a total daily energy intake of 2.6 x maintenance (106 kcal of ME/kg of BW(0.75)). For each ileal digesta sample, digesta samples were collected for two 24-h periods and combined for each pig. The combination of supplementing with phytase and decreasing the concentration of dietary Ca and P increased average ileal AA (P < 0.02), starch (P < 0.02), GE (P < 0.04), and DM (P < 0.03) digestibilities. In Exp. 2, a feeding challenge was conducted with barrows (eight per treatment; average BW of 53 kg). The treatments consisted of a corn-soybean meal diet or corn-soybean meal diet + 500 phytase units per kilogram of diet. In the diet with no phytase, Ca and aP were at 0.50% and 0.19%, respectively, and, in the diet with phytase, Ca and aP were each decreased by 0.12%. A catheter was surgically inserted into the anterior vena cava of each pig 6 d before the start of the feeding challenge. The barrows were penned individually, and the diets were fed for 3 d before the challenge. The pigs were held without feed for 16 h, and blood samples were obtained at -60, -30, and 0 min before the pigs were fed (2% of BW). Blood samples were then collected at 10, 20, 30, 40, 50, 60, 75, 90, 105, 120, 150, 180, 210, 240, 270, and 300 min after feeding. Glucose area under the response curve and plasma glucose, insulin, urea N, and total alpha-amino N concentrations were increased (P < 0.05) in pigs fed the diet with reduced Ca and P and the phytase addition. Area under the response curve for insulin, urea N, and total alpha-amino N; insulin:glucose; and plasma NEFA concentration, clearance, and half-life were not affected by diet. In conclusion, the combination of Ca and P reduction and phytase addition increased nutrient and energy digestibility in diets for pigs and increased plasma concentrations of glucose, insulin, urea N, and alpha-amino N.     

Histomorphology and small intestinal sodium-dependent glucose transporter 1 gene expression in piglets fed phytic acid and phytase-supplemented diets

An experiment was conducted to determine the effect of dietary phytic acid (PA) and phytase supplementation on small intestinal histomorphology and Na-dependent glucose transporter 1 (SGLT1) gene expression in piglets. Twenty-four piglets with an average initial BW of 7.60 ± 0.73 kg were randomly assigned to 3 experimental diets, to give 8 piglets per diet. The diets were a casein-cornstarch-based diet that was supplemented with 0 or 2% PA, or 2% PA (as Na phytate) plus an Escherichia coli-derived phytase at 500 phytase units/kg. The basal diet was formulated to meet the 1998 NRC energy, digestible AA, mineral, and vitamin requirements for piglets. After 10 d of feeding, the piglets were killed to determine small intestinal histomorphology and small intestinal SGLT1 gene expression. Phytic acid supplementation did not affect (P > 0.1) villus height (VH) and the VH-to-crypt depth (CD) ratio, but did decrease (P < 0.05) CD in the jejunum. Phytase supplementation did not affect (P > 0.1) VH, CD, and the VH-to-CD ratio. Phytic acid supplementation reduced SGLT1 gene expression in the duodenum, jejunum, and ileum by 1.1-, 5.4-, and 2.4-fold, respectively. Phytase supplementation increased SGLT1 gene expression in the jejunum by 2.6-fold, but reduced SGLT1 gene expression in the duodenum and ileum by 2.0- and 4.0-fold, respectively. In conclusion, PA reduced CD in the jejunum and SGLT1 gene expression in the duodenum, jejunum, and ileum, whereas phytase supplementation increased the expression of SGLT1 in the jejunum. The reduced SGLT1 gene expression by PA implies that PA reduces nutrient utilization in pigs partly through reduced expression of SGLT1, which is involved in glucose and Na absorption. The increased expression of SGLT1 in the jejunum by phytase supplementation implies that phytase alleviated the negative effects of PA partly through increased expression of SGLT1.     

The presence of inositol phosphates in gastric pig digesta is affected by time after feeding a nonfermented or fermented liquid wheat- and barley-based diet

The objective was to quantify the retention of digesta and evaluate the degradation of phytate or inositol hexakisphosphate (InsP(6)) and lower inositol phosphates (InsP?, InsP?, InsP?, and InsP?) in the stomach at different times after feeding pigs a fermented liquid diet with microbial phytase or a nonfermented diet with or without microbial phytase. Six barrows fitted with gastric cannulas were used. The experiment was a 3 × 3 Latin square with 3 pigs fed 3 diets during 3 wk in 2 replicates. Each experimental period lasted for 7 d, comprising 3 d of adaptation and 4 d of total collection of gastric digesta. For each pig, the digesta was collected once daily at 1, 2, 3, or 5 h after feeding the morning meal. A basal wheat- and barley-based diet was steam-pelleted at 90°C. The dietary treatments were a nonfermented basal diet (NF-BD), the NF-BD with microbial phytase (750 phytase units of phytase/kg, as-fed basis; NF-BD + phytase), and the NF-BD + phytase fermented for 17.5 h (F-BD + phytase). Gastric InsP?-P was not detected at all in pigs fed F-BD + phytase because of complete InsP? degradation during fermentation of the feed before feeding. Gastric InsP?-P decreased over time (P < 0.05) in pigs fed NF-BD and NF-BD + phytase. The decreases were 45, 54, 56, and 61 percentage points greater at 1, 2, 3, and 5 h, respectively, in pigs fed NF-BD + phytase compared with NF-BD. However, substantial amounts of InsP? still passed into the small intestine in pigs fed NF-BD + phytase, especially within the first hour (estimated to 17% of InsP?-P intake). The accumulation of lower inositol phosphates in gastric digesta was very small for all treatments and at all times because of a rapid and almost complete degradation. In conclusion, phytase addition to the nonfermented diet increased the degradation of gastric InsP?. However, considerable amounts of intact InsP? still passed into the small intestine because of a shortage of time for InsP? degradation in the stomach. Therefore, to increase the apparent digestibility of plant P in dry wheat- and barley-based diets, the development of phytases that can degrade InsP? effectively immediately after ingestion of the feed at an initial gastric pH from 6.5 to 5.0 is needed. Feeding F-BD + phytase compensated for the shortage of time because the InsP? degradation was completed during fermentation before feeding. The degradation of InsP? to InsP? is the bottleneck for plant P utilization in pigs because the degradation of the lower inositol phosphates is rapid and almost complete.     

Dietary phytate (inositol hexaphosphate) regulates the activity of intestinal mucosa phytase

The role of dietary phytate (inositol hexaphosphate) in the regulation of intestinal mucosa phytase was investigated in chicks. Seven-day-old chicks were grouped by weight into six blocks of three cages with six birds per cage. Three purified diets [a chemically defined casein diet, a chemically defined casein diet plus sodium phytate (20 g/kg diet) and a chemically defined casein diet plus sodium phytate (20 g/kg diet) and microbial phytase (1000 units/kg diet)] were randomly assigned to cages within each block. Chicks were fed experimental diets from 8 to 22 days of age then killed, and duodenal mucosa and left tibia removed. Phytase activity in duodenal mucosa, growth performance and bone ash content were determined. Addition of phytate to the chemically defined casein diet reduced (p < 0.05) the V(max) of the duodenal brush border phytase, but the K(m) of the enzyme was not affected. Addition of phytate also reduced (p < 0.05) weight gain, feed intake, feed efficiency and percentage ash. Addition of microbial phytase fully restored the feed efficiency (p < 0.05), but V(max) and body weight gain were only partially restored (p < 0.05). In conclusion, it would seem that dietary phytates non-competitively inhibit intestinal mucosa phytase.     

Non‐phytate phosphorus requirements and efficacy of a genetically engineered yeast phytase in male lingnan yellow broilers from 1 to 21 days of age

This experiment was conducted to investigate the requirement of non‐phytate phosphorus (nPP) and efficacy of a genetically engineered yeast phytase in performance and tibia characteristics by male Lingnan Yellow broilers from 1 to 21 days of age. A total of 2640 1‐day‐old male chicks were randomly allotted to one of 11 dietary treatments, which consisted of six replicate floor pens with 40 birds per pen. All treatments had the same levels of all nutrients except for phosphorus and phytase. The control group (treatment 1) was fed the basal diet without dicalcium phosphate or phytase supplementation. Dietary concentrations of nPP were 0.11%, 0.19%, 0.27%, 0.35%, 0.43%, 0.51% and 0.59% respectively for treatments 1, 2, 3, 4, 5, 6 and 7, through addition of dicalcium phosphate (chemistry grade) to the basal diet. Diets 8–11 were supplemented with a genetically engineered yeast phytase 250, 500, 750 U/kg and a commercial phytase product 500 U/kg in basal diet respectively. The results showed that 0.46% and 0.51% nPP were required for maximum body‐weight gain and optimum tibia development indicators respectively. However, 0.59% nPP had a negative effect on bird growth. The equivalency value of the genetically engineered yeast phytase was estimated to be 874 U/kg to liberate 0.1% nPP.     

Response of broiler chickens to microbial phytase supplementation as influenced by dietary phytic acid and non-phytate phosphorus contents. I. Effects on bird performance and toe ash

1. Seven-day old male broilers (n=900) were fed on wheat-sorghum-soyabean meal-based diets containing 3 concentrations of phytic acid (10.4, 13.2 and 15.7 g/kg; equivalent to 2.9, 3.7 and 4.4 g/kg phytate phosphorus), 2 of non-phytate phosphorus (2.3 and 4.5 g/kg) and 3 of microbial phytase (Natuphos 5000 L; 0, 400 and 800 FTU/kg) in a 19-d trial. The dietary phytic acid contents were manipulated by the inclusion of rice pollard. 2. Each dietary treatment was fed to 5 pens (10 birds/pen) from 7 to 25 d of age. Records of body weight, food intake and mortality were maintained. On d 25, all surviving birds were killed and toe samples were obtained for toe ash measurements. 3. Increasing dietary phytic acid negatively influenced body weight gain, food intake and food/gain. These adverse effects were partially overcome by the addition of microbial phytase. 4. Supplemental phytase caused improvements in weight gain and food efficiency of broilers but the magnitude of the responses was greater in low non-phytate phosphorus diets, resulting in significant non-phytate phosphorus x phytase interactions. 5. Toe ash contents were improved by phytase addition but the response was greater at the highest concentration of phytic acid, resulting in a significant phytic acid x phytase interaction. Responses were also greater in low non-phytate phosphorus diets as indicated by significant non-phytate phosphorus x phytase interaction. 6. In general, there was very little difference in the responses to phytase additions at 400 and 800 FTU/kg. 7. The performance responses to added phytase in birds receiving adequate non-phytate phosphorus diets provide evidence for the influence of the enzyme on animal performance independent of its effect on phosphorus availability.     

Response of broiler chickens to microbial phytase supplementation as influenced by dietary phytic acid and non-phytate phosphorous levels. II. Effects on apparent metabolisable energy, nutrient digestibility and nutrient retention

1. Male broilers (n=900) were fed on wheat-sorghum-soyabean meal based diets containing 3 concentrations of phytic acid (10.4, 13.2 and 15.7 g/kg; equivalent to 2.9, 3.7 and 4.4 g/kg phytate P), 2 concentrations of non-phytate (or available) phosphorus (2.3 and 4.5 g/kg) and 3 concentrations of microbial phytase (0, 400 and 800 FTU/kg) from day 7 to 25 post-hatch. The dietary concentrations of phytic acid were manipulated by the inclusion of rice pollards. All diets contained celite (20 g/kg) as a source of acid-insoluble ash. 2. The apparent metabolisable energy (AME) concentrations of the diets were determined using a classical total collection procedure during the 3rd week of the trial. On d 25, digesta from the terminal ileum were collected and analysed for phosphorus, nitrogen and amino acids. Nutrient digestibilities were calculated using acid-insoluble ash as the indigestible marker. 2. Ileal digestibilities of nitrogen and essential amino acids were negatively influenced by increasing dietary levels of phytic acid but these negative effects were overcome by the addition of phytase. 3. Supplemental phytase increased AME, ileal digestibilities of phosphorus, nitrogen and amino acids and the retention of dry matter, phosphorus and nitrogen in broilers. There were no differences in the phytase responses between additions of 400 and 800 FTU/kg. 4. The responses in all variables, except AME, were greater in low non-phytate phosphorus diets. 5. In the case of AME, the response to added phytase was greater in adequate non-phytate phosphorus diets. Supplemental phytase increased AME values from 13.36 to 13.54 MJ/kg dry matter in low non-phytate phosphorus diets and from 12.66 to 13.38 MJ/kg dry matter in adequate non-phytate phosphorus diets.