Efficacy of an E. coli phytase expressed in yeast for releasing phytate-bound phosphorus in young chicks and pigs

Four chick trials and one pig trial were conducted to investigate the phosphorus-releasing efficacy oftwo commercial phytase enzymes (Natuphos and Ronozyme) and an experimental E. coli phytase enzyme (ECP) when added to corn-soybean meal diets containing no supplemental inorganic P (iP). In the 13- or 14-d chick trials, three or four graded levels of iP (0, 0.05,0.10,0.15%) from KH2PO4 were added to the basal diet to construct standard curves from which bioavailable P release could be calculated for the phytase treatments. In all cases, phytase supplementation levels were based on an assessment of phytase premix activity (i.e., P release from Na phytate at pH 5.5). Linear (P < 0.01) responses in tibia ash and weight gain resulted from iP supplementation in all assays. In the first chick trial, supplementation of 500 phytase units (FTU)/kg of ECP resulted in superior (P < 0.01) weight gain and tibia ash values compared with 500 FTU/kg of Natuphos. Results of the second chick trial revealed P-release values of 0.032 and 0.028% for 500 FTU/kg Natuphos and Ronozyme, respectively, and these were lower (P < 0.01) than the 0.125% P-release value for 500 FTU/kg of ECP. Tibia ash responded quadratically (P < 0.05) in response to graded levels of ECP up to 1,500 FTU/kg in the third chick trial. Combining Natuphos with either Ronozyme or ECP in Chick Trial 4 revealed no synergism between phytases with different initiation sites of P removal. The pig trial involved 10 individually fed weanling pigs per diet, and and phytase enzymes were supplemented to provide 400 FTU/kg in diets containing 0.60% Ca. Based on the linear regression of fibula ash on supplemental iP intake (r2 = 0.87), P-release values were 0.081% for Natuphos, 0.043% for Ronozyme, and 0.108% for ECP. These trials revealed an advantage of the E. coli phytase over the commercial phytases in young chicks.     

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.     

Pharmacological zinc levels reduce the phosphorus-releasing efficacy of phytase in young pigs and chickens

A pig trial and a chick trial were done to determine the effect of high levels of Zn and Cu on the P-releasing efficacy of phytase. Ninety-nine individually fed pigs (7.2 kg) were given ad libitum access to one of 11 experimental diets for a period of 21 d. Fibula ash (mg) was regressed against supplemental inorganic P (iP) intake (g) to establish the standard curve, from which phytase treatments were compared to determine P-releasing efficacy. The basal diet was a corn-soybean meal diet with no supplemental P (21% CP, 0.075% estimated available P, 130 mg of Zn/kg, as-fed basis). Diets included three graded levels of supplemental iP (0, 0.075, 0.150%) from reagent-grade KH2PO4, two levels of phytase (500 and 1,000 FTU/kg) from EcoPhos, 1,500 mg of Zn/kg from either Waelz ZnO or basic Zn chloride (Zn5Cl2(OH)8), and all combinations of phytase and Zn. One phytase unit (FTU) was defined as the amount of enzyme required to release 1 micromol of iP per minute from sodium phytate at 37 degrees C and pH 5.5. Phytase supplementation improved (P < 0.01) weight gain, G:F, and fibula ash (% and mg). Bone ash (mg) was highest (P < 0.01) for pigs fed diets containing 1,000 FTU/kg of phytase. Supplemental Zn had no effect (P > 0.50) on growth performance, but decreased (P < 0.05) fibula ash (mg). Comparison of the phytase treatments to the standard curve (r2 = 0.87) revealed P-release values of 0.130 and 0.195% for 500 and 1,000 FTU of phytase/kg, respectively, in the absence of Zn, whereas in the presence of Zn (pooled), P-release values were decreased (P < 0.01) to 0.092 and 0.132%, respectively. The effects of high levels of supplemental Zn (basic Zn chloride) and Cu (CuSO4 x 5H2O) on phytase efficacy also were investigated in a 12-d chick trial. Dietary treatments were arranged according to a 2(3) factorial, with two levels each of supplemental phytase (0 and 500 FTU/kg from EcoPhos), Zn (0 and 800 mg/kg), and Cu (0 and 200 mg/kg). There was a phytase x Zn interaction (P < 0.01) for tibia ash. Thus, Zn supplementation decreased tibia ash in the presence, but not in the absence, of phytase. Supplemental Cu did not affect (P > 0.30) the response to phytase. These results suggest that pharmacological levels of Zn chelate the phytate complex, thereby decreasing its availability for hydrolysis by phytase.     

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.     

High dietary phytase levels maximize phytate-phosphorus utilization but do not affect protein utilization in chicks fed phosphorus- or amino acid-deficient diets

Four trials investigated the effect of high levels of three phytase enzymes on P and protein utilization in chicks. The three phytases were derived from Aspergillus (Fungal Phytase 1), Peniophora (Fungal Phytase 2), and E. coli. Within each assay, 8-d-old male chicks were given ad libitum access to their experimental diet for 10 to 14 d. For Trials 1, 2, and 3, the basal diet was a corn-soybean meal diet deficient in P that was analyzed to contain 23% CP and 0.38% total P (0.10% estimated available P, as-fed basis). Phytase supplementation levels were based on the assessment of phytase premix activity (i.e., P release from Na phytate at pH 5.5 and 37 degrees C). In Trial 1, supplementation of inorganic P from KH2PO4 (0 to 0.20%) resulted in a quadratic (P < 0.05) response in weight gain, gain:feed, and tibia ash concentration but a linear (P < 0.01) increase in tibia ash weight. Tibia ash was higher (P < 0.01) for chicks fed E. coli phytase than for those fed Fungal Phytase 1 at 500, 1,000, and 5,000 phytase units (FTU)/kg, but did not differ between these two phytases at 10,000 FTU/kg. In Trial 2, E. coli phytase supplementation at 1,000 FTU/kg maximized growth and bone responses, whereas addition of either of the two fungal phytases resulted in increasing responses up to 5,000 and 10,000 FTU/kg. Dietary addition of Fungal Phytase 2 resulted in the poorest (P < 0.01) responses among the three phytases. Escherichia coli phytase supplementation at 10,000 FTU/kg in Trial 3 resulted in tibia ash (millligrams) responses that were greater (P < 0.05) than those resulting from either 0.35% inorganic P supplementation or 10,000 FTU/kg of Fungal Phytase 1 or 2. Trial 4 showed that E. coli phytase supplementation at either 500 or 10,000 FTU/ kg did not improve protein efficiency ratio (gain per unit of protein intake) of chicks fed low-protein soybean meal or corn gluten meal diets that were first-limiting in either methionine or lysine, respectively. These results demonstrate that high dietary levels of efficacious phytase enzymes can release most of the P from phytate, but they do not improve protein utilization.     

The efficacy of an Escherichia coli-derived phytase preparation

Five experiments were conducted to evaluate the effect of an Escherichia coli-derived phytase on phytate-P use and growth performance by young pigs. The first experiment involved time course, pH dependence, and phytase activity studies to investigate the in vitro release of P from corn, soybean meal, and an inorganic P-unsupplemented corn-soybean meal negative control diet. In Exp. 2, which was designed to determine the efficacy of the E. coli-derived vs. fungal phytase-added diets at 0, 250, 500, 750, 1,000, or 1,250 FTU/kg (as-fed basis; one phytase unit or FTU is defined as the quantity of enzyme required to liberate 1 micromol of inorganic P/min, at pH 5.5, from an excess of 15 microM sodium phytate at 37 approximately C) and a positive control diet, eight individually penned 10-kg pigs per diet (12 diets, 96 pigs) were used in a 28-d growth study. The third experiment was a 10-d nutrient balance study involving six 13-kg pigs per diet (four diets, 24 pigs) in individual metabolism crates. In Exp. 4, eight pens (four pigs per pen) of 19-kg pigs per treatment were used in a 42-d growth performance study to examine the effect of adding the E. coli-derived phytase to corn-soybean diets at 0, 500, or 1,000 FTU/kg (as-fed basis) and a positive control (four diets, 128 pigs). In Exp. 5, six 19-kg pigs per treatment were used in a 10-d nutrient balance study to investigate the effects of the E. coli-derived phytase added to diets at 0, 250, 500, 750, or 1,000 FTU/kg (as-fed basis) and a positive control diet (six diets, 36 pigs). The in vitro study showed that the E. coli-derived phytase has an optimal activity and pH range of 2 to 4.5. Inorganic phosphate release was greatest for soybean meal, least for corn, and intermediate for the negative control diet. Dietary supplementation with graded amounts of E. coli-derived phytase resulted in linear increases (P < 0.05) in weight gain, feed efficiency, and plasma Ca and P concentrations in 10-kg pigs in Exp. 2. Phytase also increased P digestibility and retention in the 13-kg pigs in Exp. 3. In Exp. 4, dietary supplementation with E. coli-derived phytase resulted in linear increases (P < 0.05) in weight gain and feed efficiency of 19-kg pigs. Supplementation of the diets of 19-kg pigs with the E. coli-derived phytase also improved Ca and P digestibility and retention in Exp. 5. In the current study, the new E. coli-derived phytase was efficacious in hydrolyzing phytate-P, both in vitro and in vivo, in young pigs.     

Effect of low doses of Aspergillus niger phytase on growth performance, bone strength, and nutrient absorption and excretion by growing and finishing swine fed corn-soybean meal diets deficient in available phosphorus and calcium

Two experiments were conducted to evaluate the efficacy of low doses of Aspergillus niger (AN) phytase for growing and finishing pigs fed corn-soybean meal (SBM) diets with narrow Ca:P ratios that were about 0.9 g/kg deficient in available P and Ca. Experiment 1 utilized 120 pigs with an early finisher period from 51.5 +/- 0.2 to 89.7 +/- 0.9 kg of BW and a late finisher period that ended at 122.5 +/- 2.0 kg of BW. During each period, treatments were the low-P diets with 0, 150, 300, or 450 units (U) of AN phytase added/kg of diet, and a positive control (PC) diet. There were linear increases (P < or = 0.001) in bone strength and ash weight, the absorption of P (g/d and %) and Ca (%), and overall ADG (P = 0.01) with increasing concentration of AN phytase. Pigs fed the diets with 150, 300, or 450 U of AN phytase/kg did not differ from pigs fed the PC diet in growth performance overall, and pigs fed the diets with 300 or 450 U of AN phytase did not differ in P and Ca absorption (g/d) or bone ash weight from pigs fed the PC diet. However, only pigs fed the diet with 450 U of AN phytase/kg had bone strength similar to that of pigs fed the PC diet. Experiment 2 utilized 120 pigs in a grower phase from 25.3 +/- 0.1 to 57.8 +/- 0.8 kg of BW and a finisher phase that ended at 107.6 +/- 1.0 kg of BW. Treatments were the low-P diet with AN phytase added at 300, 500, or 700 U/kg of grower diet, and 150, 250, or 350 U/kg of finisher diet, respectively, resulting in treatments AN300/150, AN500/250, and AN700/350. Growth performance and the absorption (g/d) of P and Ca for the grower and finisher phases were not different for pigs fed the diets containing AN phytase and pigs fed the PC diets. However, pigs fed the PC diets excreted more fecal P (g/d, P < or = 0.01) during the grower and more P and Ca (g/d, P < 0.001) during the finisher phases than the pigs fed the diets with phytase. There were linear increases (P < or = 0.05) in bone strength and bone ash weight with increasing concentration of AN phytase. However, pigs fed the PC diets had a greater bone strength and bone ash weight than pigs fed diets AN300/150, AN500/250 (P < or = 0.02), or AN700/350 (P < or = 0.08). There were no treatment responses for N or DM digestibility in either experiment. Phytase supplementation reduced fecal P excretion from 16 to 38% and fecal Ca excretion from 21 to 42% in these experiments. In conclusion, 450 U of AN phytase/kg was effective in replacing 0.9 g of the inorganic P/kg of corn-SBM diet for finishing swine based on bone strength, whereas 300 or 150 U of AN phytase/kg of diet maintained growth performance of grower or finisher pigs, respectively.     

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.     

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 an experimental phytase on performance, egg quality, tibia ash content and phosphorus bioavailability in laying hens fed on maize- or barley-based diets

A 24-week performance trial was conducted to evaluate the efficacy of an experimental phytase on performance, egg quality, tibia ash content and phosphorus excretion in laying hens fed on either a maize- or a barley-based diet. At the end of the trial, an ileal absorption assay was conducted in order to determine the influence of phytase supplementation on the apparent absorption of calcium and total phosphorus (P). Each experimental diet was formulated either as a positive control containing 3.2 g/kg non-phytate phosphorus (NPP), with the addition of dicalcium phosphate (DCP), or as a low P one, without DCP addition. Both low P diets (containing 1.3 or 1.1 g/kg NPP) were supplemented with microbial phytase at 0, 150, 300 and 450 U/kg. The birds were housed in cages, allocating two hens per cage as the experimental unit. Each of 10 dietary treatments was assigned to 16 replicates. Low dietary NPP (below 1.3 g/kg) was not able to support optimum performance of hens during the laying cycle (from 22 to 46 weeks of age), either in maize or barley diets. Rate of lay, daily egg mass output, feed consumption, tibia ash percentage and weight gain were reduced in hens fed low NPP diets. The adverse effects of a low P diet were more severe in hens on a maize diet than in those on a barley diet. Low dietary NPP reduced egg production, weight gain, feed consumption and tibia ash content and microbial phytase supplementation improved these parameters. Hens given low NPP diets supplemented with phytase performed as well as the hens on positive control diets containing 3.2 g/kg of NPP. A 49% reduction of excreta P content was achieved by feeding hens on low NPP diets supplemented with phytase, without compromising performance. Phytase addition to low NPP diets increased total phosphorus absorption at the ileal level, from 0.25 to 0.51 in the maize diet and from 0.34 to 0.58 in the barley diet. Phosphorus absorption increased linearly with increasing levels of dietary phytase. Mean phosphorus absorption was higher in barley diets than in maize diets (0.49 vs 0.39).     

Effects of microbial phytase, low calcium and phosphorus, and removing the dietary trace mineral premix on carcass traits, pork quality, plasma metabolites, and tissue mineral content in growing-finishing pigs

An experiment was conducted to determine the effects of phytase addition, reduced Ca and available P (aP), and removing the trace mineral premix (TMP) on growth performance, plasma metabolites, carcass traits, pork quality, and tissue mineral content in growing-finishing swine. One hundred twenty cross-bred pigs (initial and final BW of 22 and 109 kg, respectively) were allotted to five dietary treatments on the basis of weight within gender in a randomized complete block design. There were three replications of barrows and three replications of gilts, with four pigs per replicate pen. The dietary treatments were as follows: 1) corn-soybean meal (C-SBM), 2) C-SBM with reduced Ca and aP, 3) C-SBM with reduced Ca and aP plus 500 phytase units/kg of diet, 4) Diet 1 without the TMP, and 5) Diet 3 without the TMP. The Ca and aP were reduced by 0.10% in the low Ca and aP diets and the diets with added phytase. Daily gain, hot carcass weight, dressing percent, kilograms of carcass lean, bone ash percent, and bone strength were decreased (P = 0.10), but liver and kidney weight were increased (P = 0.10) in pigs fed diets with reduced Ca and aP; adding phytase reversed these responses (P = 0.10). The Commission Internationale de I'Eclairage L* was decreased (P = 0.09) in pigs fed the low Ca and aP diet plus phytase relative to those fed the control diet. Removing the TMP had no effect on overall growth performance, but it increased (P = 0.03) 10th-rib backfat thickness and fasting glucose and decreased (P = 0.03) carcass length and ham weight. Liver weight and liver weight as a percentage of final BW were not affected when phytase was added to the control diet, but removing the TMP increased liver weight and liver weight as a percentage of final BW; adding phytase reversed these responses (phytase x TMP, P = 0.06). Removing the TMP decreased (P = 0.08) Zn concentrations in the bone, muscle, and liver, and Cu and Fe concentrations in the bile but increased (P = 0.08) Mn concentrations in the bile and liver of pigs. The addition of phytase reversed the negative effects of the reduced Ca and aP diets. These data indicate that removing the TMP in diets for growing-finishing pigs has no negative effects on growth performance or pork quality, but it had negative effects on carcass traits and had variable effects on tissue mineral content.     

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.     

Effect of reduced energy, protein and entire substitution of inorganic phosphorus by phytase on performance and bone mineralisation of laying hens

1. The objective of this study was to evaluate the effects of total removal of dietary inorganic phosphorus and reduced energy and protein, without and with phytase supplementation, on the performance, egg quality and bone composition of laying hens. 2. Lohmann pink-shell hens were randomly assigned at 56 weeks of age to 5 treatments for 20 weeks as follows: (1) a positive control (PC) with 155 g CP/kg, 11·09 MJ ME/kg, calcium (Ca) 3·40% and non-phytic phosphorus (NPP) 0·26%, (2) a negative control (NC1) diet based on PC diet with Ca decreased to 3·30% and NPP to 0·14%, (3) NC2 diet was formulated on the basis of NC1 diet with 152·7 g CP/kg, 10·90 MJ/kg, (4) NC1 and (5) NC2 supplemented with phytase (300 FTU/kg) each. 3. Feed intake, hen-day or hen-housed egg production, egg number per hen-housed, and final body weight were depressed with NC1 and NC2 diets, but restored by phytase inclusion. There were no significant differences between the dietary treatments for feed conversion efficiency, rates of cracked and broken eggs, egg-shell thickness or egg-shell strength. Mortality was significantly increased by NC2 diet without phytase. Tibia ash was significantly decreased by both NC1 and NC2 diets. Bone strength, and Ca and P contents in tibia ash were significantly increased by phytase inclusion in the NC1 diet. 4. In conclusion, the NC1 and NC2 diets significantly depressed performance and tibia quality, but the addition of phytase (300 FTU/kg) significantly improved performance and tibia integrity.     

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.     

Water-soluble phosphorus excretion in pigs fed diets supplemented with microbial phytase

Three experiments were conducted to compare the excretion of water-soluble phosphorus (P) of starter, grower, and finisher pigs fed corn-soybean meal-based P-deficient basal diet containing no added inorganic P (B), P-adequate diet (the basal diet with added dicalcium phosphate; B + P), the basal diet plus 500 units of microbial phytase/kg (B + 500), or the basal diet plus 1000 units of microbial phytase/kg (B + 1000). There were 6 barrows per diet, in a randomized complete block design, with an average initial body weight of 10.4, 20.0, or 51.1 kg for each of starter, grower, and finisher pigs in the three phosphorus balance experiments, respectively. In the experiments, the addition of dicalcium phosphate or phytase to the basal diet increased ( P  < 0.05) the digestibility and retention of P, and there were linear reductions ( P  < 0.05) in water-soluble P excretion as a result of supplementing the basal diet with phytase. In the starter pig experiment, phytase addition at 500 or 1000 units/kg reduced ( P  < 0.05) water-soluble P excretion by 28 or 42%, respectively when compared with the B + P diet. In the grower pig experiment, adding phytase at 500 or 1000 units/kg reduced ( P  < 0.05) water-soluble P excretion by 24 or 34%, respectively when compared with the B + P diet. The use of phytase at 500 or 1000 units/kg reduced ( P  < 0.05) water-soluble P excretion by 11 or 30%, respectively in the finisher pig experiment. The proportion of water-soluble phosphorus in total phosphorus was not affected by dietary treatment in any of the three experiments. In conclusion, adding phytase at 1000 units/kg to a corn-soybean meal-based P-deficient diet basal diet containing no added inorganic P compared with B + P diet reduced the daily excretion of water-soluble P in starter, grower, and finisher pigs by 42, 34, and 30%, respectively.     

Effects of Source of Inorganic Phosphorus and Phytase Supplementation on Performance, Physiological Parameters and Bone Mineralization in Broilers

The objective of this study was to eval- uate the effects of inorganic phosphorus source and phytase addition on performance, nutrient digestibility and bone mineralization in broiler chickens. In Exp. 1,150 two-day old, male broiler chicks were fed a corn-soybean meal basal diet supplemented with phos- phorus provided by dicalcium phosphate, tricalcium phosphate or defluorinated rock phosphate. Five cages containing 10 birds were allotted to each of the three treatments. In Exp. 2,120 three-day old, male broiler chicks were fed the basal diet from Exp. 1 supplemen- ted with 0,250,500 ,or 1,000 P-＇rU phytase per kg of diet. Six cages containing five chicks were allotted to each of the four treatments. In Exp. 1, there was no difference in weight gain, feed intake or feed conver- sion as a result of feeding the different sources of in- organic phosphorus. The digestibility of phosphorus was significantly lower （P =0.01 ） for chicks fed di- ets supplemented with tricalcium phosphate than for chicks fed the other two diets. However, despite the lower digestibility, serum phosphorus levels did not differ among the three treatments. For Exp. 2, feedconversion showed a linear improvement （P = 0.03 ） with increasing levels of phytase inclusion （ days 0 to 33 ）. Phytase supplementation resulted in linear increa- ses in the digestibility of dry matter （P = 0.02 ）, crude protein （ P --- 0.04 ） and energy （ P 〈 0.01 ）. Chicks fed 1,000 FTU/kg phytase had significantly higher bone calcium （ P = 0.05 ） and bone breaking strength （P = 0.04 ） than chicks fed the basal diet on day 33. In conclusion, the results of the current study indicated that the performance of birds fed diets sup- plemented with dicalcium phosphate, tricalcium phos- phate or defluorinated phosphate was similar and therefore production costs could be lowered by choo- sing the cheapest inorganic phosphorus source when formulating diets for poultry. When diets were formu- lated to meet dietary phosphorus requirements, the growth of broilers was not enhanced with phytase sup- plementation. However, increases in feed conversion and bone breaking strength and its potential to impact culling and mortality in broiler operations may be suf- ficient justification for the routine inclusion of phytase in diets fed to broilers.     

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.     

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.     

植酸酶对产蛋高峰期蛋鸡生产性能及主要营养素存留的影响

在分别含非植酸磷为０．３７％,０．３７％,０．２７％和０．１７％的四种（ＣＬ,Ｔ１,Ｔ２和Ｔ３）等钙（３．７５％）蛋鸡（２５￣３５周龄）日粮中,Ｔ１,Ｔ２和Ｔ３均添加５００Ｕ／ｋｇ植酸酶。结果,主要营养素（干物质、氮、灰分、钙、磷）的存留率显著提高。当非植酸磷水平降到０．１７％时,不影响产蛋量和蛋品质。     

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.