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.     

Sampling duration and freezing temperature influence the analysed gastric inositol phosphate composition of pigs fed diets with different levels of phytase

This experiment was conducted to determine the effects of time and freezing temperature during sampling on gastric phytate (myo-inositol [MYO] hexakisphosphate [InsP₆]), lower inositol phosphates (InsP_2–5) and MYO concentrations in pigs fed diets containing different levels of phytase. Forty pigs were fed 1 of 4 wheat-barley diets on an ad libitum basis for 28 d. The diets comprised a nutritionally adequate positive control (PC), a similar diet but with Ca and P reduced by 1.6 and 1.24 g/kg, respectively (NC), and the NC supplemented with 500 (NC + 500) or 2,000 (NC + 2000) FTU phytase/kg. At the end of the experiment, chyme were collected from the stomach, thoroughly mixed and 2 subsamples (30 mL) were frozen immediately: one snap-frozen at −79 °C and the other at −20 °C. The remaining chyme were left to sit at room temperature (20 °C) and further subsamples were collected and frozen as above at 5, 10 and 15 min from the point of mixing. There were linear reductions in gastric InsP₆ concentration over time during sampling (P < 0.001), irrespective of diet or freezing temperature. Moreover, InsP₆ concentration was influenced by a diet × freezing temperature interaction (P < 0.05), with less InsP₆ measured in chyme frozen at −20 °C than at −79 °C; however, this difference was greater in the control diets than the phytase supplemented diets. Freezing chyme at −79 °C recovered more ∑InsP_2–5 + MYO than freezing at −20 °C in pigs fed phytase supplemented diets; however, this difference was not apparent in the diets without phytase (diet × freezing temperature, P < 0.01). It can be concluded that significant phytate hydrolysis occurs in the gastric chyme of pigs during sampling and processing, irrespective of supplementary phytase activity. Therefore, to minimise post-slaughter phytate degradation and changes in the gastric inositol phosphate profile, chyme should be snap-frozen immediately after collection.     

Consequences of calcium interactions with phytate and phytase for poultry and pigs

Despite increasing practical experience and cascades of scientific reports on exogenous microbial phytases, several issues associated with their use remain unresolved because of the ambiguous and, at times, conflicting data that has been generated. One possible cause of these inconsistent outcomes is dietary calcium (Ca) levels, which are mainly derived from limestone. Thus the purpose of this review is to examine Ca interactions with dietary phytate and phytases, particularly exogenous, microbial phytases, and their consequences for poultry and pigs. The polyanionic phytate molecule has a tremendous capacity to chelate cations and form insoluble Ca–phytate complexes, which are refractory to phytase activity. Thus Ca–phytate complex formation along the gastrointestinal tract, where one phytate (IP₆) molecule binds up to five Ca atoms, assumes importance and approximately one third of dietary Ca may be bound to phytate in digesta. Consequently, phytate limits the availability of both P and Ca as a result of insoluble Ca–phytate complex formation, the extent of which is driven by gut pH and molar ratios of the two components. It is accepted that Ca–phytate complexes are mainly formed in the small intestine where they have a substantial negative influence on the efficacy of mucosal phytase. However, exogenous phytases are mainly active in more proximal segments of the gut and lower pH levels, so their efficacy should not be influenced by Ca–phytate complexes in the small intestine. There is, however, data to indicate that Ca and phytate interactions occur under acidic conditions with the formation of soluble and insoluble Ca–phytate species, which could negatively impact on exogenous phytase efficacy. Also, Ca will tend to elevate gut pH because of limestone's very high acid binding capacity, which will favour Ca–phytate interactions and may influence the activity of exogenous phytases depending on their pH activity spectrum. The de novo formation of binary protein–phytate complexes that are refractory to pepsin hydrolysis may be fundamental to the negative impact of phytate on the digestibility of protein/amino acids. However, high dietary Ca levels may disrupt protein–phytate complex formation by interacting with both phytate and protein even at acidic pH levels, thereby influencing the outcomes of phytase amino acid digestibility assays. Finally, it is increasingly necessary to define the Ca and nonphytate-P requirements of pigs and poultry offered phytase-supplemented diets.     

The effectiveness of an Escherichia coli phytase in improving phosphorus and calcium bioavailabilities in poultry and young pigs.

The effectiveness of an Escherichia coli phytase in comparison with a commercially available Aspergillus phytase in improving the bioavailability of phosphorus in broilers, layers and young pigs was studied in three separate experiments. Three basal diets, marginally deficient in dietary P mainly provided as phytate, were formulated. Both phytases were added to the diets at the rate of 500 U/kg diet. The phytases significantly (P < or = 0.05) improved the availability of phytate P to broilers, layers and young pigs. Aspergillus and E. coli phytases enhanced the pre-caecal digestibility of P by 11 and 29% for broilers and 18 and 25% for layers, respectively. Total tract digestibility of P (P balance) was also enhanced but with smaller magnitude. In pigs, total tract digestibility of P was improved by 33 and 34% by Aspergillus and E. coli phytases, respectively. Under the conditions of this study, it was observed that E. coli consistently, though with small magnitude in layers and pigs, enhanced the availability of phytate P at the same range or slightly better than Aspergillus phytase. It was only in pigs that the availability of Ca was significantly (P < or = 0.05) improved by addition of both phytases. It can be concluded that E. coli phytase is highly effective in improving the bioavailability of phytate P to broilers, layers and young pigs. This seems to be based on the high proteolytic stability of the enzyme in the digestive tract, as shown recently.     

Influence of phytate level on broiler performance and the efficacy of 2 microbial phytases from 0 to 21 days of age

Phytate is an antinutrient in animal feeds, reducing the availability and increasing the excretion of nutrients. Phytases are widely used to mitigate the negative influences of phytate. This trial was designed to compare the efficacy of 2 Escherichia coli-derived phytases on broiler performance and bone ash as influenced by dietary phytate level. A total of 1,024 Arbor Acres male broilers were used with 8 replicate pens of 16 birds/pen. Experimental diets were based on low available phosphorus (avP; 1.8 g/kg) with low (6.40 g/kg) or high (10.65 g/kg) phytate. The low-avP diets were then supplemented with mono-dicalcium phosphate to increase the avP level to 4.5 g/kg, 500 phytase units/kg of phytase A, or 500 phytase units/kg of phytase B to create 8 experimental diets. Feed intake, BW gain, FCR, and livability were influenced by a P source × phytase interaction. Feed intake, BW gain, and livability were reduced and FCR was higher in broilers fed low-avP diets, particularly in the presence of high phytate. Phytase A or phytase B improved feed intake, BW gain, and FCR, particularly in the high-phytate diet. However, broilers fed phytase A ate more and were heavier than broilers fed phytase B. Tibia ash was lowest in broilers fed the low-avP diet and highest in broilers fed the diet supplemented with mono-dicalcium phosphate. Phytase increased tibia ash, and broilers fed phytase A had an increase in tibia ash compared with broilers fed phytase B. In conclusion, high dietary phytate reduced broiler performance. Phytase A and phytase B improved bone ash and growth performance, especially in the high-phytate diets. However, phytase A was more efficacious than phytase B, regardless of the level of phytate.     

植酸酶的研究进展

植酸酶是能降解饲料中植酸及其盐的酶。它能提高磷利用率,解除植酸对一些矿物元素如钙、锌、铁、铜等的抗营养效应,不仅对动物具有良好的增重效果,同时可降低动物排泄磷量,有利于环境保护。植酸酶作为饲料添加剂其作用效果受到饲料中钙、磷水平和钙磷比例以及维生素Ｄ含量的影响。植酸酶的运用需要降低饲料中钙、磷水平,维生素Ｄ与植酸酶之间可能存在协同效应。对植酸酶运用的经济分析表明,使用植酸酶能代替饲料中需添加的无机磷。     

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.     

Phytate-degrading enzymes in pig nutrition

Phytate, the mixed salt of phytic acid (myo-inositol hexaphosphate), derived from plant-sourced feed ingredients is invariably present in practical diets for pigs. Typically, swine diets contain in the order of 3.0 g kg^− 1 phytate-bound phosphorus (phytate-P) but phytate concentrations are subject to variation. Importantly, phytate-P is only partially utilised by pigs because they do not generate sufficient endogenous phytase activity. Phytate-degrading enzymes, via step-wise dephosphorylation of phytate, have the capacity to liberate phytate-P, thus enhancing P absorption and reducing P excretion, which are both nutritionally and ecologically beneficial consequences. The commercial introduction of microbial phytases in 1991 has greatly magnified the interest in the roles of phytate and phytase in pig nutrition.

The capacity of microbial phytases to enhance growth performance of pigs offered diets with inadequate P levels is well documented. However, in some instances, phytase has been shown to improve performance of pigs offered P adequate diets thus phytase-induced improvements in growth performance should not be attributed entirely to increased P availability. This raises the possibility that phytase is increasing the utilisation of nutrients other than P. These so-called ‘extra-phosphoric’ effects of phytase remain controversial, particularly in relation to protein and amino acid availability. There are conflicting opinions that are reflected in the inconsistent outcomes of studies to determine the effect of phytase on ileal digestibility of amino acids and protein utilisation in pigs. In phytase amino acid digestibility assays, it seems likely the choice of chromic oxide as the dietary marker has contributed to these ambiguous results, which may be further complicated when ileal digesta samples are taken from cannulated pigs fed on a restricted, twice-daily basis. In order to resolve this critical issue, there is an urgent need to assess the impacts of selection of dietary markers, methods of ileal digesta collection and feeding regimen relative on the outcomes of phytase amino acid digestibility assays in pigs.

However, inconsistent results from phytase studies in pigs are not confined to amino acid digestibility assays. Arguably, insufficient attention has been paid to dietary substrate levels in relation to phytase inclusion from both scientific and practical standpoints. Phytate analyses are not straightforward and there is a real need to develop more accurate and rapid methods to facilitate phytate determinations. The properties of phytate vary between (and within) feed ingredients where solubility of phytate may be critical; which, in turn, is a function of gut pH in pigs. Contemporary phytases have the capacity to degrade approximately 50% of dietary phytate at the level of the ileum, which may mean higher inclusion rates are warranted. Consequently, there is scope for the development of more effective ‘second-generation’ phytate-degrading feed enzymes and their possible introduction, coupled with a better scientific understanding of relevant fundamental issues, will ensure that phytate-degrading enzymes will contribute to viable and sustainable pig production to an even greater extent in the future.     

Enhancement of phosphorus utilization in growing pigs fed phytate-rich diets by using rye bran.

Some cereal by-products, such as bran, exhibit a high phytase activity that may enhance phytate P digestibility. This was studied in growing pigs fed a phytase-rich (1,200 IU/kg) diet containing 20% rye bran. The trial involved 12 animals; six were fed a control diet and six were fed a diet containing rye bran for 2 mo. Both diets contained the same levels of energy, protein, Ca (.7%) and total P (.4%). No inorganic P was added; thus, the dietary P was mainly phytic. Pigs fed the control diet, in contrast to those fed the diet containing rye bran, developed a P deficiency, as indicated by hypophosphatemia, hypophosphaturia, hyperhydroxyprolinuria, hypercalcemia, and hypercalciuria. Phosphorus from the rye bran diet was more completely absorbed (55 vs 36%) and retained (50 vs 36%) than that from the control diet. Calcium absorption was equal for the two diets, but Ca retention was higher in pigs fed rye bran than in controls. Pigs fed the rye bran diet showed greater bone density, ash content, and bending moments than controls. In conclusion, high dietary phytase levels or phytase-rich by-products increased phytate P availability and consequently improved bone scores.     

Comparison of different levels and sources of microbial phytases

Phytases catalyse the hydrolysis of phytate rendering phosphorus (P) available for absorption. Endogenous plant phytases are to some extent present in cereals (depending on species and varieties) while microbial phytases are added to cereal based diets to increase the digestibility of phytate bound P. The present study compared two different microbial phytases. The basal diet was composed of wheat, barley, soybean and rapeseed meal without feed phosphate. The diet was initially expanded, pelleted at 90 °C and crumbled. Phytases were added at 250, 500 and 750 FTU kg^− 1 diet (Aspergillus niger; Phytase 1) and 375 and 750 FYT kg^− 1 diet (Peniophora lycii; Phytase 2). The experiment comprised 6 treatment groups of 6 pigs each kept in metabolism crates and fed one of the 5 test diets or a diet with no added microbial phytase. The diets were fed for 12 days, 5 days for adaptation and 7 days for total collection of faeces and urine. Phosphorus digestibility of the basal diet averaged 43% and increased to 55, 61 and 66% following addition of 250, 500 and 750 FTU/kg of Phytase 1 and 54 and 60% following addition of 375 and 750 FYT/kg of Phytase 2, respectively. In conclusion, equivalent effects were obtained when Phytase 2 was given at 1.5 times the doses of Phytase 1.     

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.     

Recovery of intact IgG in the gastrointestinal tract of the growing rat following ingestion of an ovine serum immunoglobulin

The aim of this study was to determine whether orally ingested ovine serum IgG partly resists digestion in the growing rat. Fifteen Sprague‐Dawley male rats were allocated to one of three diets for a 3‐week study: a control diet (CON) and two test diets containing either freeze‐dried ovine serum immunoglobulin (FDOI) or inactivated ovine serum immunoglobulin (IOI). Samples of stomach chyme and intestinal digesta from the ad libitum‐fed rats were subjected to ELISA and Western blot analysis. Amounts of intact ovine IgG for the FDOI diet were found to be 13.9, 20.0, 34.1, 13.0 and 36.9 μg in the total wet digesta from the stomach chyme, duodenal, jejunal, ileal and colonic digesta respectively. Qualitative detection by Western blot revealed the presence of intact ovine serum IgG with a ~150 kDa MW. This was detected in all of the gut segments (stomach chyme, duodenal, jejunal, ileal and colonic digesta) for growing rats fed the FDOI diet. No ovine IgG was detected in the chyme or digesta from rats fed the CON or the IOI diets. Ovine serum IgG partly resisted digestion in the growing rat fed the FDOI diet and was found throughout the digestive tract. These results provide a basis to explain the reported biological effects of orally administered immunoglobulin.     

Effects of dietary calcium:phosphorus ratios on apparent absorption of calcium and phosphorus in the small intestine, cecum, and colon of pigs

Thirty-two crossbred barrows were used to investigate the effects of dietary Ca:total P (tP) ratios in phytase-supplemented diets on the apparent absorption of P and Ca in the small intestine, cecum, and colon. Three Ca:tP ratio treatments (1.5:1, 1.3:1, or 1.0:1) were created by adjusting the amount of ground limestone added to the basal low-P grower (.39% tP including .07% added inorganic P) and finisher (.32% tP without added inorganic P) diets. All low-P ratio diets were supplemented with Natuphos phytase at 500 units/kg. A positive control diet without phytase supplementation contained adequate P and Ca to meet dietary requirements. At 123 kg, the pigs were slaughtered and the contents of ileum, cecum, and colon were collected. Lowering the dietary Ca:tP ratio in the diets containing phytase linearly increased (P < .01) the apparent absorption (% and g/d) of P in the small intestine, but Ca absorption was not affected. Pigs fed the low-P diet with a Ca:tP ratio of 1.0:1 had an apparent absorption (g/d) of P or Ca similar to that of pigs fed the control diet, which was adequate in Ca and P. Averaged across all diets, the apparent absorption of P was highest when measured at the cecum, and the apparent absorption of Ca was highest when measured at the colon. In conclusion, lowering the dietary Ca:tP ratio to 1.0:1 in a low-P diet containing phytase increased the apparent absorption of P in the small intestine. Furthermore, a significant amount of P was absorbed in the cecum.     

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

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

Effect of exposure to dietary nivalenol on activity of enzymes involved in glutamine catabolism in the epithelium along the gastrointestinal tract of growing pigs.

Changes in the activity of enzymes involved in oxidative metabolism of glutamine, and in protein content, in the epithelial tissue along the gastrointestinal (GI) tract of growing pigs exposed to nivalenol (NIV) in the diet were investigated. The epithelial tissue was taken from the stomach, small intestine and colon of three groups of animals fed diets without NIV (control), with inclusion of 2.5 mg NIV/kg diet (low dose) and with inclusion of 5.0 mg NIV/kg diet (high dose). The activities of glutaminase, glutamate dehydrogenase, oxoglutarate dehydrogenase, isocitrate dehydrogenase and alanine aminotransferase were determined. In the control pigs the activities of oxoglutarate dehydrogenase and alanine aminotransferase were higher (P < 0.05) in the epithelium of the small intestine as compared with the stomach and colon, while there were no differences in the activities of glutaminase, glutamate dehydrogenase and isocitrate dehydrogenase. With increasing inclusion of NIV in the diet the activity of oxoglutarate dehydrogenase decreased (P < 0.05) in the epithelium of the small intestine and colon, and the activity of alanine aminotransferase tended (P = 0.07) to increase in the epithelium of the small intestine. The activities of glutaminase, glutamate dehydrogenase and isocitrate dehydrogenase remained unaffected by the inclusion of NIV in the diet. In the control pigs the protein content in the epithelium of the small intestine was higher (P < 0.05) than in the stomach and colon, while there were no effects of NIV inclusion in the diet on the protein content. It can be concluded from the present study that the epithelial tissue of the small intestine and colon of pigs exposed to a diet containing NIV will have a reduced enzymatic capacity to utilise alpha-ketoglutarate in the tricarboxylic acid cycle (TCA-cycle), suggesting an impaired energy supply to these organs.     

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.     

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.     

In vitro degradation of phytate and lower inositol phosphates in soaked diets and feedstuffs

A series of in vitro experiments simulating liquid feeding were performed to evaluate the effect of microbial phytase addition, heat-treatment and soaking time on degradation of phytate and lower inositol phosphates when soaking compound wheat/soybean meal diets or the single feedstuffs wheat or soybean meal. The effect of phytase addition on phytate degradation was greatest in soybean meal, almost intermediate for wheat/soybean meal diets and not detectable in wheat, which might be due to a better accessibility to phytate in soybean meal compared with wheat. Heat-treatment seemed to enhance the accessibility between phytase and phytate, whereby phytate degradation was stimulated. Additionally, it was shown that wheat phytase is able to stimulate degradation of phytate in soybean meal. Independent of treatment, the amount of IP₅–IP₂ was extremely small in relation to phytate in both wheat and soybean meal, indicating that when one phosphate group is removed from the phytate complex, degradation of IP₅–IP₂ is completed. Consequently, it is anticipated that liquid feeding might result in a higher digestibility of plant P compared with dry feeding of 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.     

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%).