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%).     

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. 3. Hydrolysis of phytic acid (IP6) and occurrence of hydrolysis products (IP5, IP4, IP3 and IP2)

Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (Aspergillus niger) were investigated with six minipigs (40-50 kg initial body weight) 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 dry matter (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 07 30 h and 19 30 h, each animal was fed 350 g diet mixed with 1050 ml de-ionized water. Digesta were collected continuously and completely during a 12-h period after feeding. Mean hydrolysis rates of IP6 in the stomach as measured at the proximal duodenum of animals 1, 4, 5 and 6 were 9.0, 77.2 and 66.2% for diet 1, 2 and 3, respectively. Microbial phytase was much more effective in phytate hydrolysis than wheat phytase. Mean IP6 hydrolysis rates of the respective diets in the stomach and small intestine as measured at the distal ileum of animals 1, 2 and 3 were 19.0, 62.6 and 64.6% and were lower than treatment means of the stomach only. Differences existed between experimental animals with respect to their ability to hydrolyse IP6 in the stomach independent of the presence and source of dietary phytase. Considerable amounts of hydrolysis products occurred in both the duodenal and ileal digesta when diets 2 and 3 were fed; however, only traces were determined after ingestion of diet 1. Independent of dietary treatment, four IP5 isomers were detected, but in different amounts.     

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.     

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 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.     

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.     

Assessment of dietary zinc requirement of weaned piglets fed diets with or without microbial phytase

Fifty-four pigs, weaned at 26 days of age at an average body weight of 7.74 kg were used in a 26-day experiment to assess the zinc requirement of piglets, using diets based on maize and soybean meal, with or without microbial phytase. The nine experimental diets were the basal diet containing 33 mg of zinc/kg supplemented with 10, 25, 40, 60 or 80 mg of zinc as sulphate (ZnSO(4), 7H(2)O)/kg and the basal diet supplemented with 0, 10, 25 or 40 mg of zinc as sulphate/kg and 700 units (U) of microbial phytase (Natuphos)/kg. Pigs were fed the basal diet for a 7-day adjustment period prior to the 19-day experimental period. Microbial phytase enhanced plasma alkaline phosphatase (AP) activity, plasma zinc and bone zinc concentrations. These parameters increased linearly with zinc intake, with a similar slope with and without phytase. The response of bone zinc-to-zinc added did not plateau. Without microbial phytase, plasma AP activity and zinc concentration were maximized when dietary zinc reached 86 and 92 mg/kg respectively. With microbial phytase they were maximized when dietary zinc concentration reached 54 and 49 mg/kg respectively. Accounting for a safety margin, the recommended supply of zinc for weaned piglets up to 16 kg fed maize-soybean meal diets supplemented with zinc as sulphate is thus of 100-110 mg/kg diet. This supply may be reduced by around 35 mg if the diet is supplemented with 700 U of microbial phytase.     

Nitrogen metabolism in the large intestine of ruminants. 7. Metabolism of intracecally-infused 15N urea with supplement of starch in bulls

Three bulls with an average live weight of 228 kg were fitted with ileo-caecal reentrant cannulas for the experiment. The rations were composed of 3 kg maize silage and 3 kg wheat straw pellets per animal and day. In a previous period 50% of the digesta was collected over 12 hours and stored deep-frozen. In the main period the digesta flow was interrupted for 30 hours. The digesta flow was collected quantitatively. In the caecal part of the re-entrant cannula previously collected digesta and starch (over 30 hours) as well as 15N urea (over 24 hours) were supplemented. The amount of starch corresponded to about 10% of the DM of the digesta. Analyses of the urine, faeces, ileum digesta and blood plasma were carried out. The quota of starch clearly stimulates bacterial processing in the large intestine so that 20.5% of the supplemented 15N was excreted in faeces within 24 hours. 91.2% of the 15N in the faeces was localised in the bacteria fraction. Individual differences of the animals distinctly show the connection between the excretion of the 15N in faeces and urine. A decreased isotope excretion in faeces of 17.2% for animal 3 in contrast to the 23% for animals 1 and 2 showed an increased elimination of the 15N through the kidney with 32.7% instead of 25.2%. The largest proportion of the ileum digesta, i.e. 46%, can be localised in the 15N urea fraction; the NH3-fraction is also distinctly labelled. With time progressing, the 15N quota flowing from the rumen to the small intestine increases.     

Effect of protein source and fumaric acid supplementation on apparent ileal digestibility of nutrients by young pigs

Two experiments were conducted to determine the apparent ileal digestibility of DM and N by young pigs fed diets supplemented with different protein sources or organic acids. Pigs were surgically fitted with silicone cannulas at 2 wk of age. Following surgery, pigs were allowed to recuperate with their dams while suckling normally. After weaning at 24 d, pigs were assigned to treatment diets at 28 d of age. A 3-d adjustment and 4-d collection sequence was followed for the duration of the 4-wk experiment. Four treatment diets were fed in each experiment in a weekly rotation until each diet had been fed to each pig. Diet samples and digesta collected through the ileal cannulas were analyzed for chromic oxide (used as an indigestible marker), DM, and N. Pigs in Exp. 1 were fed isolysinic (1.0%) corn-based diets supplemented with casein, soybean meal, soy protein concentrate, or isolated soy protein. Casein addition resulted in improved DM (P less than .001) and N (P less than .05) digestibility but reduced gain (P less than .05) compared with the average of the soy protein sources. Nitrogen from diets formulated with soybean meal was digested more completely (P less than .05) than N from diets based on soy protein concentrate and isolated soy protein. Experiment 2 was an evaluation of the effect of dried skim milk (25%) and fumaric acid (2%) addition on apparent ileal digestibility of N and DM in corn-soybean meal diets. Addition of dried skim milk improved DM (P less than .01) and N (P less than .05) digestibility and daily gain (P less than .001). Fumaric acid supplementation did not affect nutrient digestibility or gain (P greater than .10).     

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.     

Pre-treatment of dietary plant feedstuffs with phytase and its effect on growth and mineral concentration in common carp (Cyprinus carpio L.)

Effect of phytase pre-treatment of dietary plant feed stuffs (PF) on growth and mineral concentration in common carp (C. carpio L.) was investigated. Diets used were: C0, diet with no supplement; CI, diet with incubated PF; CP0, diet supplemented with 3 g P/kg; CPI, diet with incubated PF supplemented with 3 g P/kg; Phyt0, diet fortified with 4000 U phytase/kg; PhytI, diet with PF incubated with 4000 U phytase/kg. Mean weight gain, SGR and FCR were the same (p < 0.05) in fish fed CP0, CPI or PhytI but less (p < 0.05) in other fish. Bone P was similar in fish fed CP0 (74.9), CPI (75.9) or PhytI (71.5 mg/g DM) but higher (p < 0.05) than in fish fed C0. Bone Ca and Mg were similar in fish fed CP0, CPI or PhytI but less (p < 0.05) in other fish. Bone Zn in fish fed C0, Phyt0 or PhytI was higher (p < 0.05) than that in fish fed CP0 or CPI. In conclusion, CP0, CPI and PhytI had the same effect, suggesting phytase pre-treatment as the most effective method. CPI did not have any advantage over CP0. Phytase increased mineral availability and utilization, which would minimize cost of mineral supplementation and discharges into the environment.     

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.     

A genetically engineered Escherichia coli phytase improves nutrient utilization, growth performance, and bone strength of young swine fed diets deficient in available phosphorus

A 28-d experiment was conducted using 126 crossbred barrows to evaluate the addition of a genetically engineered Escherichia coli phytase to diets that were 0.15% deficient in available P. Growth performance, bone strength, ash weight, and the apparent absorption of P, Ca, Mg, N, energy, DM, Zn, Fe, and Cu were the response criteria. The pigs (2 pigs/pen) averaged 7.61 kg of BW and 30 d of age initially. The low-P basal diet was supplemented with 0, 100, 500, 2,500, or 12,500 units (U) of E. coli phytase/kg of diet, or 500 U of Peniophora lycii phytase/kg of diet. The positive control (PC) diet was adequate in available P. Pigs were fed the diets in meal form. Fecal samples were collected from each pig from d 22 to 27 of the experiment. There were linear and quadratic increases (P < 0.001) in 28-d growth performance (ADFI, ADG, and G:F), bone breaking strength and ash weight, and the apparent absorption (g/d and %) of P, Ca, and Mg (P < or = 0.01 for quadratic) with increasing concentrations of E. coli phytase. Pigs fed the low-P diets containing 2,500 or 12,500 U/kg of E. coli phytase had greater (P < or = 0.01 or P < 0.001, respectively) values for growth performance, bone breaking strength and ash weight, and the apparent absorption (g/d and %) of P, Ca, and Mg than pigs fed the PC diet. The addition of E. coli phytase did not increase the apparent percentage absorption of N, GE, DM, Zn, Fe, or Cu. There were no differences in the efficacy of the E. coli or P. lycii phytase enzymes at 500 U/kg of low-P diet for any criterion measured. In conclusion, there were linear increases in growth performance, bone breaking strength and ash weight, and the apparent absorption of P, Ca, and Mg with increasing addition of E. coli phytase up to 12,500 U/kg of diet. Also, all of these criteria were greater for pigs fed the low-P diets containing 2,500 or 12,500 U of E. coli phytase/kg than for pigs fed the PC diet. The addition of 500, 2,500, or 12,500 U of E. coli phytase/kg of low-P diet reduced P excretion (g/d) in manure by 35, 42, and 61%, respectively, compared with pigs fed the PC diet.     

The nitrogen metabolism in the large intestine of ruminants. 8. Metabolism of intracecally infused 15N-urea with a supplement of pectin in heifers]

Three heifers with live weights of 255, 261 and 300 kg were supplied with ileo-caecal re-entrant cannulas, jugular vena catheters and bladder catheters. The ration consisted of 4 kg maize silage and 4 kg wheat straw pellets. In a previous period 50% of the digesta flow was collected over 12 h/d on 5 consecutive days and stored in a deep-freeze. During the main period the re-entrant cannulas were disrupted and the flowing digesta was quantitatively collected. Precollected digesta and pectin were infused into the distal part of cannula hourly for about 30 hours. During the first 24 hours the digesta was also supplemented with 15N-labelled urea. The amount of pectin corresponded to about 10% of digesta dry mater. An analysis of urine, faeces, digesta and blood plasma were carried out. The application of pectin increased the 15N-incorporation in the bacterial protein of faeces from 4.7% (without supplementation in an earlier experiment) to 10.5% of the introduced 15N. The ammonia-fraction of faeces was markedly higher than the bacterial fraction. The 15N-utilization of urea by the microbes of large intestine was lower in the actual trial evident than with supplementation of starch in the anterior experiment. During the pectin administration the amount of urine increased in comparison with earlier experiments and according to the literature to about the 4.5 fold. The amount of passage of 15N at the ileum cannula (recycled 15N) was 3.8% of the 15N intake. It is the same amount as in experiments in which starch was applied.     

Estimating equivalency values of microbial phytase for amino acids in growing and finishing pigs fitted with steered ileo-cecal valve cannulas

Ten crossbred barrows (48.3 +/- 2.3 kg of initial BW) fitted with steered ileo-cecal valve cannulas were used to investigate the effects of supplemental microbial phytase on the apparent ileal digestibilities (AID) of AA, Ca, P, N, and DM, and the apparent total tract digestibilities of Ca, P, N, and DM. All diets were corn-soybean meal-based, and contained 0.44% Ca and 0.40% total P. Diets 1, 2, and 3 contained 12.0, 11.1, and 10.2% CP, respectively. Diets 4 and 5 had the same ingredient composition as diet 3, plus 250 and 500 U/kg phytase (Natuphos), respectively. Pigs were randomly allotted to 1 of 5 dietary treatments in a paired 5 x 5 Latin square with an extra period to test for carryover effects. Each 14-d period consisted of a 7-d adjustment followed by a 3-d total collection, a 12-h ileal digesta collection, a 3-d readjustment, and a second 12-h ileal digesta collection. Pigs were housed individually in metabolism pens (1.2 x 1.2 m). Water was supplied ad libitum, and feed was supplied at a level of 9% of the metabolic BW (BW(0.75)) per day in 2 equal daily feedings. As the dietary CP concentration increased, the AID of CP and all AA measured increased linearly (P < 0.05) with the exception of proline. In addition, the apparent total tract digestibilities (grams per day) and retention of N (grams per day) increased linearly (P < 0.01) with increasing CP levels. Supplementing diets with phytase increased the AID of Ca (P < 0.01), P (P < 0.001), CP (P = 0.07), and the AA (P < 0.10) Gly, Ala, Val, Ile, Thr, TSAA, Asp, Glu, Phe, Lys, and Arg. Protein and phytase response equations were generated for those AA affected (P < 0.10) by both CP level and phytase supplementation. Based on these equations, 500 U/kg of phytase can replace 0.52 percentage units of the dietary CP, which includes a 0.03 percentage unit improvement in Lys AID. The results of this study show that supplementing pig diets with microbial phytase improves CP and AA digestibilities in addition to Ca and P digestibilities.     

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 starch application into the proximal duodenum of ruminants on starch digestibility in the small and total intestine.

Four Slovakian Black-and-white bulls (LW 410 +/- 12 kg; Exp. 1) and four Slovakian Black-and-white non lactating dairy cows (LW 475 +/- 14 kg; Exp. 2) with permanent ruminal cannulas, duodenal T-cannulas and ileal re-entrant cannulas were used in a 4 x 4 Latin square design to determine the postruminal capacity of starch digestion. In Exp. 1 bulls received 5.4 kg DM from corn silage and 3.6 kg DM from alfalfa hay, in Exp. 2 cows consumed only 2.1 kg DM corn silage and 1.9 kg DM alfalfa hay. Additionally, either 750 or 1500 g (Exp. 1) or resp. 1000 or 2000 g (Exp. 2) gelatinized corn or wheat starch per animal and day were applied as pulse doses or as infusion into the proximal duodenum. In both experiments the duodenal and ileal nutrient flow, as well as the faecal excretion without starch application, were measured in a pre-period. After starting starch application ileal digesta and faeces were sampled over 120 h after 9 or 23 days of adaptation respectively. Cr2O3 was used as a flow marker. It was shown, that the capacity of starch utilisation in the small intestine was limited. The effect of different doses of bypass-starch was more pronounced than the effect of different starch sources. Starch digestibility decreased with increasing amounts of starch in the intestine (Exp. 1: corn starch: from 74.3 to 68.0%, P < 0.001; wheat starch: from 76.7 to 67.4%, P < 0.001; Exp. 2: corn starch: from 71.4 to 50.3%. P < 0.001; wheat starch: from 73.8 to 53.1%, P < 0.001). Corn starch was 0.6 to 2.4% units (P < 0.05) and 2.4 to 2.8% units (P < 0.001) less digested than wheat starch in Exp. 1 and Exp. 2, respectively.     

Influence of phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digestive tract measurements and gut morphology in broilers fed wheat-based diets containing adequate level of phosphorus

1. The aim of the present study was to examine the influence of microbial phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digesta viscosity, digestive tract measurements and gut morphology in broilers fed on wheat-soy diets containing adequate phosphorus (P). The wheat-soy basal diet was formulated to contain 4.5 g/kg non-phytate P and the experimental diets were formulated by supplementing the basal diet with xylanase (1000 xylanase units/kg diet), phytase (500 phytase units/kg diet) or a combination of phytase and xylanase. 2. Supplemental phytase improved the weight gains and feed efficiency by 17.5 and 2.9%, respectively. Corresponding improvements due to the addition of xylanase were 16.5 and 4.9%, respectively. The combination of phytase and xylanase caused no further improvements in broiler performance. 3. Individual additions of xylanase or phytase resulted in numerical improvements in apparent metabolisable energy (AME), but the differences were not significant. The combination of the two enzymes significantly increased AME. Addition of xylanase and the combination of the two enzymes reduced the viscosity of digesta in all sections of the intestine. Phytase supplementation reduced digesta viscosity in the duodenum and ileum, but not in the jejunum. 4. Enzyme supplementation lowered the relative weight and length of the small intestine. Additions of xylanase and phytase reduced the relative weight of the small intestine by 15.5 and 11.4%, respectively, while the corresponding reductions in the relative length of the small intestine were 16.5 and 14.1%, respectively. The combination of phytase and xylanase had no further effects on the relative weight and length of the small intestine compared with the xylanase group. 5. The addition of phytase increased villus height in the duodenum and decreased the number of goblet cells in the jejunum compared with those on the unsupplemented basal diet. Xylanase supplementation tended to increase goblet cell numbers in the duodenum and decreased crypt depth in thejejunum. The combination of phytase and xylanase increased villus height in the ileum and crypt depth in thejejunum and ileum. 6. In summary, the present results showed that the addition of a microbial phytase, produced by solid state fermentation and containing significant activities of beta-glucanase and xylanase, was as effective as xylanase in improving the performance of broiler chickens fed on wheat-based diets containing adequate levels of P. Improved performance with enzyme supplementation was generally associated with reduced digesta viscosity, increased AME, and reduced relative weight and length of small intestine.     

Effect of a novel microbial phytase on production performance and tibia mineral concentration in broiler chickens given low-calcium diets

1. In a 42-d feeding trial, 264 one-d-old, as hatched, Cobb 400 broiler chickens (6 pens per group, n = 11 per pen in a 2?×?2 factorial arrangement) were fed on two concentrations of dietary calcium (Ca) (9.0 and 7.5 g/kg in starter, 7.5 and 6 g/kg in grower phases) and supplemental phytase (0 and 500 U/kg diet).

2. During d 0–21, the high Ca + phytase diet improved body weight. During d 0–42, feed intake was increased by the low Ca diet and decreased by phytase supplementation. Feed conversion ratio during d 0–21 was improved by the high Ca + phytase diet.

3. At d 42, Ca in duodenal digesta was reduced by low dietary Ca and supplemental phytase. High dietary Ca reduced P in duodenal and jejunal digesta. Phytase reduced digesta P and increased serum P concentration.

4. Relative tibia length decreased with low dietary Ca and increased with phytase. The robusticity index of tibia was improved by the low Ca diet and phytase supplementation. Phytase supplementation increased tibia ash and concentrations of Ca, magnesium (Mg), manganese (Mn), copper (Cu), zinc (Zn) and iron (Fe) in tibia. The low Ca diet increased Mg, Mn and Fe and reduced Cu and Zn in tibia.

5. It was concluded that 7.5 g Ca/kg during weeks 0–3 and 6 g Ca/kg during weeks 3–6 sustained broiler performance and bone ash, while phytase supplementation facilitated tibia mineralisation, particularly during the grower phase.     

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.