Effects of combining three fungal phytases with a bacterial phytase on plasma phosphorus status of weanling pigs fed a corn-soy diet

The objective of this study was to determine possible synergistic effects of supplementing one of three fungal phytases: Aspergillus fumitagus PhyA (AFP),A. niger PhyA (ANP), or Peniophora lyci phytase (PLP) with an Escherichia coli AppA phytase (EP) in diets for pigs. Three experiments, each lasting for 4 wk, were conducted with a total of 106 weanling pigs (5 wk old). The corn-soybean meal basal diet (BD) contained no supplemental inorganic P. In Exp. 1, 35 pigs (8.6 +/- 1.0 kg BW) were fed (as-fed basis) BD + AFP at 750 U/ kg of feed, BD + inorganic P (0.2% P), or BD + PLP at 500, 750, or 1,000 U/kg feed. Pigs fed BD + AFP or BD + 0.2% P had higher (P < 0.05) plasma inorganic P concentrations than those fed BD + PLP at the end of the trial (wk 4). In Exp. 2, 35 pigs (8.1 +/- 0.9 kg BW) were fed BD + AFP, EP, PLP, a 1:1 mix of AFP:EP, or a 1:1 mix of PLP:EP at 500 U/kg. Pigs fed the AFP:EP mixture had growth performance and plasma measures similar to those fed either enzyme alone. Pigs fed the PLP:EP mixture had lower (P < 0.05) plasma alkaline phosphatase activity than those fed BD + PLP. Pigs fed BD + PLP had lower (P < 0.05) plasma inorganic P concentrations than pigs fed BD + EP, and higher (P < 0.05) plasma alkaline phosphatase activity than all other groups at wk 4. In Exp. 3, 36 pigs (9.1 +/- 1.2 kg BW) were fed BD + ANP, EP, or a 1:1 mix of ANP:EP at 500 U/kg feed. Pigs fed the two enzymes together had lower (P < 0.05) plasma inorganic P concentration than those fed BD + EP and lower (P < 0.05) plasma alkaline phosphatase activity than pigs fed BD + ANP at wk 4. In conclusion, although the four phytases showed different effects on plasma P status of weanling pigs, there was no synergistic effect between any of the three fungal phytases and the bacterial phytase on the plasma measures or growth performance under the conditions of the present study.     

Effectiveness of an experimental consensus phytase in improving dietary phytate-phosphorus utilization by weanling pigs

Consensus phytase is a new biosynthetic, heat-stable enzyme derived from the sequences of multiple homologous phytases. Two experiments were conducted to determine its effectiveness, relative to inorganic P and a mutant enzyme of Escherichia coli phytase (Mutant-EP), in improving dietary phytate-P availability to pigs. In Exp. 1, 36 pigs (3 wk old, 7.00 +/- 0.24 kg of BW) were fed a low-P corn-soybean meal basal diet plus consensus phytase at 0, 250, 500, 750, 1,000, or 1,250 U/kg of feed for 5 wk. Plasma inorganic P concentration, plasma alkaline phosphatase activity, bone strength, and overall ADG and gain:feed ratio of pigs were improved (P < 0.05) by consensus phytase in both linear (R2 = 0.20 to 0.70) and quadratic (R2 = 0.30 to 0.70) dose-dependent fashions. In Exp. 2, 36 pigs (4 wk old, 9.61 +/- 0.52 kg BW) were fed the basal diet + inorganic P at 0.1 or 0.2%, consensus phytase at 750 or 450 U/kg of feed, Mutant-EP at 450 U/kg of feed, or 225 U consensus + 225 U Mutant-EP/kg of feed. Pigs fed 750 U of consensus phytase or 450 U of Mutant-EP/kg feed had plasma inorganic concentrations and bone strength that fell between those of pigs fed 0.1 or 0.2% inorganic P. These two measures were 16 to 29% lower (P < 0.05) in pigs fed 450 U of consensus phytase/kg of feed than those of pigs fed 0.2% inorganic P. Plasma inorganic P concentrations were 14 to 29% higher (P < 0.05) in pigs fed Mutant-EP vs. consensus phytase at 450 U/kg at wk 2 and 3. In conclusion, the experimental consensus phytase effectively releases phytate P from the corn-soy diet for weanling pigs. The inorganic P equivalent of 750 U of consensus phytase/kg of feed may fall between 0.1 and 0.2%, but this requires further determination.     

Efficacy and equivalency of an Escherichia coli-derived phytase for replacing inorganic phosphorus in the diets of broiler chickens and young pigs

Two studies were conducted to determine the efficacy of an Escherichia coli-derived phytase (ECP) and its equivalency relative to inorganic phosphorus (iP) from monosodium phosphate (MSP). In Exp. 1, one thousand two hundred 1-d-old male broilers were used in a 42-d trial to assess the effect of ECP and iP supplementation on growth performance and nutrient digestibility. Dietary treatments were based on corn-soybean meal basal diets (BD) containing 239 and 221 g of CP, 8.2 and 6.6 g of Ca, and 2.4 and 1.5 g of nonphytate P (nPP) per kg for the starter and grower phases, respectively. Treatments consisted of the BD; the BD + 0.6, 1.2, or 1.8 g of iP from MSP per kg; and the BD + 250, 500, 750, or 1,000 phytase units (FTU) of ECP per kg. Increasing levels of MSP improved gain, gain:feed, and tibia ash (linear, P < 0.01). Increasing levels of ECP improved gain, gain:feed, tibia ash (linear, P < 0.01), apparent ileal digestibility of P, N, Arg, His, Phe, and Trp at d 21 (linear, P < 0.05), and apparent retention of P at d 21 (linear, P < 0.05). Increasing levels of ECP decreased apparent retention of energy (linear, P < 0.01). Five hundred FTU of ECP per kg was determined to be equivalent to the addition of 0.72, 0.78, and 1.19 g of iP from MSP per kg in broiler diets based on gain, feed intake, and bone ash, respectively. In Exp. 2, forty-eight 10-kg pigs were used in a 28-d trial to assess the effect of ECP and iP supplementation on growth performance and nutrient digestibility. Dietary treatments consisted of a positive control containing 6.1 and 3.5 g of Ca and nPP, respectively, per kg; a negative control (NC) containing 4.8 and 1.7 g of Ca and nPP, respectively, per kg; the NC diet plus 0.4, 0.8, or 1.2 g of iP from MSP per kg; and the NC diet plus 500, 750, or 1,000 FTU of ECP per kg. Daily gain improved (linear, P < 0.05) with ECP addition, as did apparent digestibility of Ca and P (linear, P < 0.01). Five hundred FTU of ECP per kg was determined to be equivalent to the addition of 0.49 and 1.00 g of iP from MSP per kg in starter pigs diets, based on ADG and bone ash, respectively.     

Effect of low-phytate barley or phytase supplementation to a barley-soybean meal diet on phosphorus retention and excretion by grower pigs

Two studies were conducted to determine the effect of diets containing low-phytate barley or supplemented with phytase on P balance and excretion in grower pigs. In Exp. 1, eight 32-kg barrows were assigned to a repeated, 4 x 4 Latin square design and fed 4 diets that contained 96% barley: normal-phytate hulled barley (HB), low-phytate hulled barley (LPHB), normal-phytate hull-less barley (HLB), and low-phytate hull-less barley (LPHLB). The barley cultivars contained 0.16, 0.05, 0.24, and 0.03% phytate, respectively. Inorganic P (iP) was added to the HB and HLB diets to meet the 1998 National Research Council recommendation of available P (aP, 0.23%), whereas LPHB and LPHLB contained sufficient aP. The diets were fed at 2.5 times the maintenance requirement for ME. The apparent total tract digestibilities (ATTD) of P did not differ between the hulled and hull-less barley diets, but P retention (%) and excretion were greater in pigs fed the hull-less barley diets (P < 0.05). The ATTD of P was greater and P excretion was 35% lower in pigs fed the low-phytate compared with the normal-phytate diets (P < 0.001). The amount of P retained (g/d) was greater (P < 0.001) in pigs fed low-phytate barley, reflecting an ATTD of P of 65 and 49% for low-phytate and normal-phytate barley, respectively (P < 0.001). In Exp. 2, eight 21-kg barrows were assigned to a repeated, 4 x 4 Latin square design and fed 4 diets based on barley and soybean meal (SBM): HB-SBM, HB-SBM + iP, HB-SBM + phytase, and LPHB-SBM. The HB-SBM and HB-SBM + phytase diets were deficient in aP, whereas the HB-SBM + iP and LPHB-SBM diets had adequate aP. The feeding regimen was similar to that of Exp. 1. Adding iP to the HB-SBM diet did not affect the ATTD but increased the amount of P retained (g/d) and excreted (P < 0.001). The ATTD and amount of P retained (g/d) did not differ among pigs fed the HB-SBM + iP, HB-SBM + phytase, and LPHB-SBM diets. However, pigs fed the HB-SBM + phytase and LPHB-SBM diets excreted 32 and 29% less P, respectively, than pigs fed the HB-SBM + iP diet (P < 0.05), confirming that low-phytate barley is as effective as supplemental phytase in improving P digestibility and utilization and decreasing P excretion in grower pigs.     

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.     

A new phytase expressed in yeast effectively improves the bioavailability of phytate phosphorus to weanling pigs

We have recently expressed a new phytase enzyme in a yeast system. Three experiments with a total of 140 weanling crossbred pigs were conducted to examine the efficacy of this enzyme in improving the bioavailability of phytate-P in corn-soybean meal diets to young pigs. Experiment 1 compared the efficacy of this new phytase with a commercially available phytase (Natuphos, BASF) for 4 wk at an inclusion level of 1,200 U/kg of diet. Experiment 2 compared the responses of pigs to four doses of the new phytase supplementation (300, 600, 900, and 1,200 U/kg of diet) for 4 wk. Experiment 3 compared the efficacy of this new phytase and Natuphos at a marginally optimal dose (700 U/kg of diet) for 5 wk. A group of pigs were fed the P-deficient basal diet as a negative control in Exp. 1, and a group of pigs were fed the basal diet plus .17 or .22% inorganic P as a positive control in all experiments. In Exp. 1, pigs fed the two sources of phytase had similar ADG (564 vs 567 g), gain/feed (.597 vs .589), plasma inorganic P concentrations (8.9 vs 8.4 mg/dL), and mobility scores (4.25 vs 4.46) that were higher (P < .05) than those of the negative control. In Exp. 2, plasma inorganic P concentration was a fairly linear response to the phytase dose (r > .83) at wk 1 and 2. Overall ADG of pigs also tended to increase with the phytase dose (P = .15). In Exp. 3, pigs fed the two sources of phytase had ADG (483 vs 506 g) similar to that of the positive control (508 g). These two groups also had similar plasma inorganic P concentrations (7.7 vs 7.4 mg/dL) that were lower (P < .05) than those of the positive control group (9.7 mg/dL). There was no significant effect of dietary treatments on ADFI in all three experiments. In conclusion, our new phytase was as effective as Natuphos, at the inclusion level of 700 or 1,200 U/kg of a P-deficient, corn-soybean meal diet, in improving phytate-P utilization by young 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.     

Effect of microbial phytase on energy availability,and lipid and protein deposition in growing swine

Two experiments were conducted to determine the effect of phytase on energy availability in pigs. In Exp. 1, barrows (initial and final BW of 26 and 52 kg) were allotted to four treatments in a 2 x 2 factorial arrangement. Corn-soybean meal (C-SBM) diets were fed at two energy levels (2.9 and 3.2 x maintenance [M]) with and without the addition of 500 phytase units/kg of diet. The diets contained 115% of the requirement for Ca, available P (aP), and total lysine, and Ca and aP were decreased by 0.10% in diets with added phytase. Pigs were penned individually and fed daily at 0600 and 1700, and water was available constantly. Eight pigs were killed and ground to determine initial body composition. At the end of Exp. 1, all 48 pigs were killed for determination of carcass traits and protein and fat content by total-body electrical conductivity (TOBEC) analysis. Six pigs per treatment were ground for chemical composition. In Exp. 2, 64 barrows and gilts (initial and final BW of 23 and 47 kg) were allotted to two treatments (C-SBM with 10% defatted rice bran or that diet with reduced Ca and aP and 500 phytase units/kg of diet), with five replicate pens of barrows and three replicate pens of gilts (four pigs per pen). In Exp. 1, ADG was increased (P < 0.01) in pigs fed at 3.2 x M. Based on chemical analyses, fat deposition, kilograms of fat, retained energy (RE) in the carcass and in the carcass + viscera, fat deposition in the organs, and kilograms of protein in the carcass were increased (P < 0.10) in pigs fed the diets at 3.2 vs. 2.9 x M. Based on TOBEC analysis, fat deposition, percentage of fat increase, and RE were increased (P < 0.09) in pigs fed at 3.2 x M. Plasma urea N concentrations were increased in pigs fed at 3.2 x M with no added phytase but were not affected when phytase was added to the diet (phytase x energy, P < 0.06). Fasting plasma glucose measured on d 28, ultrasound longissimus muscle area (LMA), and 10th-rib fat depth were increased (P < 0.08) in pigs fed phytase, but many other response variables were numerically affected by phytase addition. In Exp. 2, phytase had no effect (P > 0.10) on ADG, ADFI, gain:feed, LMA, or 10th-rib fat depth. These results suggest that phytase had small, mostly nonsignificant effects on energy availability in diets for growing pigs; however, given that phytase increased most of the response variables measured, further research on its possible effects on energy availability seems warranted.     

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.     

An Escherichia coli phytase expressed in yeast effectively replaces inorganic phosphorus for finishing pigs and laying hens

Two experiments determined the efficacy of an Escherichia coli phytase (ECP) added to P-deficient, corn-soybean meal diets fed to finishing pigs and second-cycle laying hens. Sixty finishing pigs (49 +/- 0.9 kg) were formed into blocks within sex based on weight and ancestry and allotted to a P-deficient diet unsupplemented or supplemented with 0.10% inorganic P (iP) from KH2PO4 or ECP at 250, 500, 1,000, or 10,000 phytase units (FTU)/kg. Individually fed pigs were allowed ad libitum access to the experimental diets until a BW of 120 +/- 3 kg was achieved, at which time the pigs were euthanized and the left fibula and fourth metatarsal were excised for determination of bone ash. Pigs were fed a 2-phase diet program for early- and late-finishing pigs; available P in the basal diets was set 0.10% below the requirement. Dietary supplementation of iP or ECP increased weight gain (P < 0.10) and G:F (P < 0.01); performance was not different (P > 0.13) among the phytase-supplemented groups. Fibula ash was greatest (P < 0.01) for pigs fed diets containing 10,000 FTU of ECP/kg. Two hundred forty second-cycle hens were allotted to a P-deficient diet or a P-deficient diet supplemented with 0.10% iP or ECP at 150, 300, or 10,000 FTU/kg for a 12-wk experiment. The basal diet was a corn-soybean meal diet with no added iP (17% CP, 3.8% Ca, 0.10% available P). Hens fed the P-deficient diet were removed from the experiment after 4 wk due to poor egg production. Supplementation of iP or ECP resulted in increased (P < 0.01) feed intake, egg weight, and egg production during the first 4 wk. During the entire 12-wk period, there were no differences (P > 0.28) between the iP- and ECP-supplemented groups in feed intake, egg weight, or egg production. These experiments reveal that ECP was as efficacious as supplemental iP and that supplementation of an excess dose of ECP was efficacious and without negative effects in finishing pigs and laying hens.     

Effects of supplemental dietary phytase and pharmacological concentrations of zinc on growth performance and tissue zinc concentrations of weanling pigs

Three experiments were conducted to determine the effects of phytase, excess Zn, or their combination in diets for nursery pigs. In all experiments, treatments were replicated with five to seven pens of six to seven pigs per pen, dietary Ca and available P (aP) levels were decreased by 0.1% when phytase was added to the diets, excess Zn was added as ZnO, a basal level of 127 mg/kg of Zn (Zn sulfate) was present in all diets, and the experimental periods were 19 to 21 d. In Exp. 1, pigs (5.7 kg and 18 d of age) were fed two levels of phytase (0 or 500 phytase units/kg) and three levels of excess Zn (0, 1,000, or 2,000 ppm) in a 2 x 3 factorial arrangement. Added Zn linearly increased ADG and ADFI during Phase 1 (P = 0.01 to 0.06), Phase 2 (P = 0.02 to 0.09), and overall (P = 0.01 to 0.02). Gain:feed was linearly increased by Zn during Phase 1 (P = 0.01) but not at other times. Dietary phytase decreased ADG in pigs fed 1,000 or 2,000 ppm Zn during Phase 2 (Zn linear x phytase interaction; P = 0.10), did not affect (P = 0.27 to 0.62) ADFI during any period, and decreased G:F during Phase 2 (P = 0.01) and for the overall (P = 0.07) period. Plasma Zn was increased by supplemental Zn (Zn quadratic, P = 0.01) but not affected (P = 0.70) by phytase addition. In Exp. 2, pigs (5.2 kg and 18 d of age) were fed two levels of phytase (0 or 500 phytase units/kg) and two levels of Zn (0 or 2,000 ppm) in a 2 x 2 factorial arrangement. Supplemental Zn increased ADG and G:F during Phase 2 (P = 0.02 to 0.09) and overall (P = 0.07 to 0.08), but it had no effect (P = 0.11 to 0.89) on ADG during Phase 1 or ADFI during any period. Phytase supplementation increased ADG (P = 0.06) and G:F (P = 0.01) during Phase 2. Gain:feed was greatest for pigs fed 2,000 ppm Zn and phytase (Zn x phytase interaction; P = 0.01). Bone (d 20) and plasma Zn (d 7 and 20) were increased (P = 0.01) by added Zn but not affected (P = 0.51 to 0.90) by phytase. In Exp. 3, pigs (5.7 kg and 19 d of age) were fed a basal diet or the basal diet with Ca and aP levels decreased by 0.10% and these two diets with or without 500 phytase units/kg. Supplemental phytase had no effect (P = 0.21 to 0.81) on growth performance. Reduction of dietary Ca and aP decreased (P = 0.02 to 0.08) ADG, ADFI, and G:F for the overall data. These results indicate that excess dietary supplemental Zn increases ADG and plasma and bone Zn concentrations. Dietary phytase did not affect plasma or bone Zn concentrations.     

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.     

Supplementation of carbohydrases or phytase individually or in combination to diets for weanling and growing-finishing pigs

The overall objective of the studies reported here was to evaluate the growth and nutrient utilization responses of pigs to dietary supplementation of phytate- or nonstarch polysaccharide-degrading enzymes. In Exp. 1, growth performance and nutrient digestibility responses of forty-eight 10-kg pigs to dietary supplementation of phytase or a cocktail of xylanase, amylase, and protease (XAP) alone or in combination were evaluated. The growth response of one hundred fifty 23-kg pigs to dietary supplementation of phytase or xylanase individually or in combination was studied in Exp. 2 in a 6-wk growth trial, whereas Exp. 3 investigated the nutrient digestibility and nutrient retention responses of thirty 24-kg pigs to dietary supplementation of the same enzymes used in Exp. 2. In Exp. 1, the pigs were used in a 28-d feeding trial. They were blocked by BW and sex and allocated to 6 dietary treatments. The treatments were a positive control (PC) diet; a negative control (NC) diet marginally deficient in P and DE; NC with phytase added at 500 or 1,000 phytase units (FTU)/kg; NC with xylanase at 2,500 units (U)/kg, amylase at 400 U/kg, and protease at 4,000 U/kg; and NC with a combination of phytase added at 500 FTU/kg and XAP as above. In Exp. 2 and 3, the 5 dietary treatments were positive control (PC), negative control (NC), NC plus 500 FTU of phytase/kg, NC plus 4,000 U of xylanase/kg, and NC plus phytase and xylanase. In Exp. 1, low levels of nonphytate P and DE in the NC diet depressed (P < 0.05) ADG of the pigs by 16%, but phytase linearly increased (P < 0.05) ADG by up to 24% compared with NC. The cocktail of XAP alone had no effect on ADG of pigs, but the combination of XAP and phytase increased (P < 0.05) ADG by 17% compared with the NC treatment. There was a linear increase (P < 0.01) in Ca and P digestibility in response to phytase. In Exp. 2, ADG was 7% greater in PC than NC (P < 0.05); there were no effects of enzyme addition on any response. In Exp. 3, addition of phytase alone or in combination with xylanase improved (P < 0.05) P digestibility. Phosphorus excretion was greatest (P < 0.01) in the PC and lowest (P < 0.05) in the diet with the combination of phytase and xylanase. The combination of phytase and xylanase improved P retention (P < 0.01) above the NC diet to a level similar to the PC diet. In conclusion, a combination of phytase and carbohydrases improved ADG in 10-kg but not 23-kg pigs, but was efficient in improving P digestibility in pigs of all ages.     

Corn expressing an Escherichia coli-derived phytase gene: a proof-of-concept nutritional study in pigs

Two experiments were conducted to investigate the concept that the addition of corn expressing an Escherichia coli-derived gene (corn-based phytase; CBP) to a P-deficient diet would improve growth performance and P utilization in pigs. An E. coli-derived microbial phytase (expressed in Pichia pastoris) sprayed onto a wheat carrier (Quantum) was included for comparison. In Exp. 1, forty-eight 10-kg pigs were blocked by BW into 6 blocks and allotted to 8 dietary treatments such that the BW among dietary treatments was similar and given free access to feed for 28 d. The dietary treatments were a negative control (NC) with no inorganic P supplementation; NC + 2, 4, or 6 g of monosodium phosphate/kg; NC + 16,500, 33,000, or 49,500 phytase units (FTU) of CBP/kg; and NC + 16,500 FTU of Quantum/kg. In Exp. 2, twenty-four 13-kg barrows were assigned to the NC, NC + 16,500 or 33,000 FTU of CBP/kg, or NC + 16,500 FTU of Quantum/kg, in a nutrient- and energy-balance study consisting of 5 d of adjustment and 5-d collection periods. The total collection method was used to determine nutrient and energy balance. Addition of CBP to the low-P NC diet linearly increased (P < 0.01) ADG, G:F, and plasma P concentration of pigs during the 28-d study. There was no difference in ADG, G:F, or plasma P concentration between pigs fed the CBP or Quantum phytase at 16,500 FTU/kg. Weight gain, G:F, and plasma P concentration of pigs increased (P < 0.01) with monosodium phosphate supplementation, confirming P deficiency of the NC diet. Linear improvements (P < 0.05) in DM digestibility and energy retention were observed with CBP supplementation of the NC diet. Although there were linear (P < 0.01) and quadratic (P < 0.05) increases in N digestibility, N retention was unaffected by CBP supplementation of the NC diet in growing pigs. Phosphorus and Ca digestibilities and retentions improved linearly and quadratically (P < 0.01) with the addition of CBP to the NC diet. There was no difference in digestive utilization of P or Ca between pigs fed CBP and Quantum phytase at 16,500 FTU/kg. The data showed that the addition of a corn expressing an E. coli-derived gene to a P-deficient diet improved growth performance and indices of P utilization in pigs, and corn expressing phytase was as efficacious as Quantum phytase when supplemented in P-deficient diets for weanling pigs.     

Effect of microbial phytase addition with or without the trace mineral premix in nursery, growing, and finishing pig diets

Two experiments were conducted to determine the interactive effects of phytase with and without a trace mineral premix (TMP) in diets for nursery, growing, and finishing pigs on growth performance, bone responses, and tissue mineral concentrations. Pigs (initial and final BW of 5.5 and 111.6 kg [Exp. 1] or 5.4 and 22.6 kg [Exp. 2]) were allotted to treatments on the basis of BW with eight (Exp. 1) or six (Exp. 2) replications of six or seven pigs per replicate pen. Pigs were started on the diets the day of weaning (average of 18 d). In both experiments, the treatments were with or without 500 phytase units/kg of diet and with or without the TMP in a 2 x 2 factorial arrangement. The Ca and available P concentrations were decreased by 0.10% in diets with phytase. The nursery phase consisted of Phase I (7 d), Phase II (14 d), and Phase III (13 d) periods. In Exp. 1, 26 of 52 pigs fed the diet without the TMP and without phytase had severe skin lesions and decreased growth performance; therefore, pigs fed this diet were switched to the positive control diet. In Exp. 2, the treatment without the TMP and without phytase had 12 replications instead of six. At the end of Phase III, half these replications were switched to the positive control diet and half were switched to the diet without the TMP but with phytase. In Exp. 1 during Phases II and III and in the overall data, pigs fed the diet without the TMP had decreased ADG and ADFI, but the addition of phytase prevented these responses (phytase x TMP; P < 0.02). Growth performance was not affected by diet during the growing-finishing period. Coccygeal bone Zn and Na concentrations were decreased (P < 0.09) in pigs fed the diet without the TMP, and adding phytase increased (P < 0.03) Zn and Fe concentrations. In Exp. 2 during Phases I and II, pigs fed the diet without the TMP had decreased ADG, but the addition of phytase prevented this response (phytase x TMP; P < 0.10). Pigs fed the diet without the TMP had decreased (P < 0.10) ADG (Phase II and overall), ADFI (Phases II and III and in the overall data), and G:F (Phase III). Coccygeal bone Zn and Cu concentrations were decreased (P < 0.09) in pigs fed the diet without the TMP, and adding phytase increased (P < 0.03) Zn concentration in the bones. These data indicate that removing the TMP in diets for nursery pigs decreases growth performance and bone mineral content, and that phytase addition to the diet without the TMP prevented the decreased growth performance.     

Nutrient digestibility and performance responses of growing pigs fed phytase- and xylanase-supplemented wheat-based diets

Three experiments were conducted to evaluate the effect of supplementing phytase and xylanase on nutrient digestibility and performance of growing pigs fed wheat-based diets. In Exp. 1, 10 diets were fed to 60 pigs from 20 to 60 kg of BW to determine the effect of combining phytase and xylanase on apparent total tract digestibility (ATTD) of nutrients and growth performance. The 10 diets included a positive control diet (PC; 0.23% available P; 0.60% Ca) and a negative control diet (NC; 0.16% available P; 0.50% Ca) supplemented with phytase at 0, 250, and 500 fytase units (FTU)/kg and xylanase at 0, 2,000, and 4,000 xylanase units (XU)/kg in a 3 x 3 factorial arrangement. In Exp. 2, 6 ileally cannulated barrows (initial BW = 35.1 kg) were fed 4 wheat-based diets in a 4 x 4 Latin square design, with 2 added columns to determine the effect of combining phytase and xylanase on apparent ileal digestibility (AID) of nutrients. The 4 diets were NC (same as that used in Exp. 1) or NC supplemented with phytase at 500 FTU/kg, xylanase at 4,000 XU/kg, or phytase at 500 FTU/kg plus xylanase at 4,000 XU/kg. In Exp. 3, 36 barrows (initial BW = 55.5 kg) were fed 4 diets based on prepelleted (at 80 degrees C) and crumpled wheat for 2 wk to determine the effect of phytase supplementation on ATTD of nutrients. The 4 diets fed were a PC (0.22% available P; 0.54% Ca) and a NC (0.13% available P; 0.43% Ca) alone or with phytase at 500 or 1,000 FTU/kg. All diets in the 3 experiments contained Cr(2)O(3) as an indigestible marker. No synergistic interactions were detected between phytase and xylanase on any of the response criteria measured in Exp. 1 or 2. There were no dietary effects on growth performance in Exp. 1. In Exp. 1, phytase at 250 FTU/kg increased the ATTD of P and Ca by 51 and 11% at 20 kg of BW or by 54 and 10% at 60 kg of BW, respectively, but increasing the level of phytase to 500 FTU/kg only increased (P < 0.05) ATTD of P at 20 kg of BW. In Exp. 2, phytase at 500 FTU/kg increased (P < 0.05) the AID of P and Ca by 21 and 12%, respectively. In Exp. 3, phytase at 500 FTU/kg improved (P < 0.05) ATTD of P by 36%, but had no further effect at 1,000 FTU/kg. Xylanase at 4,000 XU/kg improved (P < 0.05) AID of Lys, Leu, Phe, Thr, Gly, and Ser in Exp. 2. In conclusion, phytase and xylanase improved P and AA digestibilities, respectively, but no interaction between the 2 enzymes was noted.     

Phosphorus bioavailability, growth performance, and nutrient balance in pigs fed high available phosphorus corn and phytase

Three experiments were conducted to evaluate P bioavailability, growth performance, and nutrient balance in pigs fed high available P (HAP) corn with or without phytase. The bioavailability of P in normal and HAP corn relative to monosodiumphosphate (MSP) for pigs was assessed in Exp. 1. In a randomized complete block design, 96 pigs (average initial BW 9.75 kg) were fed eight diets for 28 d. The reference and test diets were formulated by adding P as MSP, HAP, or normal corn at 0, 0.75, or 1.5 g/kg to a corn-starch-soybean meal basal diet (2.5 g/kg P) at the expense of cornstarch. Plasma inorganic P concentration responded linearly (P < 0.05) to supplemental P intake. Estimates of P bioavailability from HAP andnormal corn when plasma P was regressed on supplemental P intake were 46 and 33%, respectively. In Exp. 2 and 3, pigs were fed corn-soybean meal-based diets containing HAP corn or normal corn and 0 or 600 units of phytase per kilogram in a 2 x 2 factorial arrangement (two corn sources and two levels of phytase). In Exp. 2, 48 crossbred pigs (barrow:gilt, 1:1) averaging 9.25 kg were used to evaluate growth performance. There were no detectable interactions between corn source and phytase for any of the performance criteria measured. Pigs receiving normal corn had the lowest (P < 0.05) BW and rate of gain. Feed efficiency was lower (P < 0.05) in pigs fed normal compared with those fed the HAP corn phytase-supplemented diet. In Exp. 3, 24 crossbred barrows averaging 14.0 kg were used to evaluate nutrient digestibility. There were no detectable interactions between corn and phytase for any of the N and Ca balance criteria. Nitrogen and Ca retention were improved in pigs receiving HAP corn with phytase (P < 0.05). Retention and digestibility of P was lowest (P < 0.01) for pigs on normal corn diet without phytase. The percentage of P digested and retained was improved and fecal P excretion lowered (P < 0.05) by feeding HAP corn.The results of this study indicate that the bioavailability and balance of P in HAP corn is superior to that of normal corn. The addition of 600 phytase units (Natuphos 600, BASF) to HAP corn-based diets further improved P digestibility and reduced P excretion in pigs.     

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.     

Effect of an organic acid blend and phytase added to a rapeseed cake‐containing diet on performance,intestinal morphology,caecal microflora activity and thyroid status of broiler chickens

The experiment was carried out on 96 female broilers, allocated to eight groups of 12 birds kept in individual cages. Two basal wheat‐ and soyabean meal‐based diets containing 150 g/kg of rapeseed expeller cake were formulated, differing in the level of P: 7.1 g/kg in diet H or 5.9 g/kg in diet L. Rapeseed cake supplied 3.15 μmol alkenyl glucosinolates per gram of diet. The eight treatments were: basal diets only, basal diets + phytase (1000 U/kg), basal diets + organic acid blend (OA, 6 g/kg), or basal diets + both additives. Diets were fed from day 8 to 28 of life. The results showed that the lower dietary P content and OA supplementation did not significantly affect feed intake or BWG, while both increased (p < 0.001) after phytase supplementation. Tibia ash content as well as tibia ultimate strength were lower (p < 0.001) in birds fed diets L compared with diets H, and increased (p < 0.01) with phytase supplementation of diet L, while OA had no influence on either parameter. Dietary P levels and OA supplementation had no influence on the pH of gut digesta, but the pH of jejunal digesta increased following phytase supplementation (p < 0.01). Morphological measurements of the small intestinal mucosa of chicks indicated that OA added to diet L depressed villi height (p < 0.001) and crypt depth (p < 0.001); both parameters increased after phytase supplementation (p < 0.01). The lower total SCFA as well as acetic, propionic and butyric acid concentrations in caecal digesta indicated lower activity of caecal microflora in birds fed diets L compared with H. OA supplementation had no influence, while phytase supplementation increased the concentration of acetic acid in caecal digesta. Supplementation of diets with either phytase or OA increased thyroid weight by 16% (p < 0.01) and 11% (p < 0.05) respectively. The increase in thyroid weight because of phytase supplementation was greater at the lower dietary P level, and the greatest when both phytase and OA were added to the diet.     

Effect of diet composition on postweaning colibacillosis in piglets

The weaning of piglets is often associated with digestive disorders, particularly diarrhea--postweaning colibacillosis (PWC)--which is caused by infection with enterotoxigenic strains of Escherichia coli. It has been shown previously that a diet for newly weaned pigs based on cooked white rice and animal protein decreases the occurrence of PWC, whereas the addition of carboxymethylcellulose (CMC) to this diet enhances PWC. The aims of the current work were to 1) determine whether substitution of animal protein with plant proteins in the cooked-white-rice diet influenced its protective effects on PWC and 2) confirm that an increase in viscosity of the digesta by adding CMC to the diet favors the development of PWC--with (Exp. 1) or without (Exp. 2) experimental infection of piglets with E. coli. The diets were 1) cooked white rice and animal protein sources (RAP), 2) RAP + CMC added at 40 g of CMC/kg (air-dry basis) of diet, 3) cooked white rice and plant protein sources (RPP), and 4) wheat and plant protein sources (WPP). Experiments 1 and 2 were conducted using 32 and 24 piglets (eight and six per treatment), respectively. Piglets were weaned at 21 d (d 1), and fed ad libitum until slaughter on d 9. In Exp. 1, piglets were orally infected with enterotoxigenic E. coli on d 4, 5, 6, and 7. On d 8 of Exp. 1, the E. coli scores in feces of pigs fed RAP + CMC were higher than with RAP (P < 0.01). On d 9 after weaning, feces from pigs fed diet RAP were normal or moist, whereas feces from pigs fed RAP + CMC were wet to diarrheic. On d 7 of Exp. 2, pigs fed diets RAP + CMC and WPP had wetter feces than pigs fed diets RAP or RPP (P < 0.05). On d 8, the E. coli scores in feces were higher (P < 0.01) with pigs fed RAP + CMC than with all other diets. The E. coli scores in the digesta were also higher with pigs fed RAP + CMC, and to a lesser extent with diet WPP, than with pigs fed RAP or RPP (P < 0.01). The large intestine was heavier in pigs fed diets RPP and WPP, and the digesta were more acidic (P < 0.05). This study confirmed that diet RAP was protective against PWC, and that substitution of animal proteins with plant protein in a rice-based diet did not diminish its protective effects. The addition of CMC to cooked white rice increased digesta viscosity and enhanced PWC. Consequently, this diet represents a useful model for studying this condition.