InsP6 Degradation Research Articles

Effects of feed particle size, calcium concentration and phytase supplementation on InsP6 degradation in broiler chickens fed pelleted diets

ABSTRACT 1. The objective of the trial was to study the single and interactive effects of feed particle size in pelleted feed, dietary calcium (Ca) concentration and microbial phytase supplementation in broiler chickens. The studied traits were myo-inositol (1,2,3,4,5,6) hexakis (dihydrogen phosphate) (InsP6) degradation, pre-caecal digestibility of phosphorus (P), Ca and amino acids (AA) and retention of P, Ca and nitrogen (N). 2. Male Ross 308 broiler chickens were housed in metabolism units in groups of 10 and allocated to one of eight diets with seven pen replicates per diet. The 2 × 2 × 2-factorial arrangement included coarse and fine feed particle size (309 or 222 µm), low and high Ca concentration (4.9 and 7.2 g/kg) and without or with phytase supplementation (1,000 FTU/kg). 3. Pre-caecal InsP6 disappearance was higher with coarse than fine feed particle size when no phytase was added (54 vs. 48%) but not when phytase was added (74%; p = 0.046). High dietary Ca feeds decreased pre-caecal InsP6 disappearance (67 to 59%) and P digestibility (65 to 55%; p < 0.001). Gizzard pH was lower with coarse than fine feed particle size and higher with high Ca than low Ca (p < 0.001). Pre-caecal digestibility of most AA was approximately 3.5%-points lower with high Ca without phytase compared to the other treatments (p ≤ 0.047). Coarse feed particle size caused higher pre-caecal AA digestibility than fine particle size (~2%-points; p ≤ 0.031). InsP6 disappearance in the crop increased at high Ca concentration when phytase was added (22 vs. 37%; p = 0.011). 4. Coarser feed particle size in pellets increased gastrointestinal InsP6 degradation and nutrient digestibility, likely owing to effects on the gizzard functions. Additional Ca supply exerted antinutritive effects that was not compensated for by using coarser feed particles.

Effect of dietary calcium source, exogenous phytase, and formic acid on inositol phosphate degradation, mineral and amino acid digestibility, and microbiota in growing pigs.

The choice of the calcium (Ca) source in pig diets and the addition of formic acid may affect the gastrointestinal inositol phosphate (InsP) degradation and thereby, phosphorus (P) digestibility in pigs. This study assessed the effects of different Ca sources (Ca carbonate, Ca formate), exogenous phytase, and chemical acidification on InsP degradation, nutrient digestion and retention, blood metabolites, and microbiota composition in growing pigs. In a randomized design, 8 ileal-cannulated barrows (24kg initial BW) were fed 5 diets containing Ca formate or Ca carbonate as the only mineral Ca addition, with or without 1,500 FTU/kg of an exogenous hybrid 6-phytase. A fifth diet was composed of Ca carbonate with phytase but with 8g formic acid/kg diet. No mineral P was added to the diets. Prececal InsP6 disappearance and P digestibility were lower (P ≤ 0.032) in pigs fed diets containing Ca formate. In the presence of exogenous phytase, InsP5 and InsP4 concentrations in the ileal digesta were lower (P ≤ 0.019) with Ca carbonate than Ca formate. The addition of formic acid to Ca carbonate with phytase diet resulted in greater (P = 0.027) prececal InsP6 disappearance (87% vs. 80%), lower (P = 0.001) InsP5 concentration, and greater (P ≤ 0.031) InsP2 and myo-inositol concentrations in the ileal digesta. Prececal P digestibility was greater (P = 0.004) with the addition of formic acid compared to Ca carbonate with phytase alone. Prececal amino acid (AA) digestibility of some AA was greater with Ca formate compared to Ca carbonate but only in diets with phytase (P ≤ 0.048). The addition of formic acid to the diet with Ca carbonate and phytase increased (P ≤ 0.006) the prececal AA digestibility of most indispensable AA. Exogenous phytase affected more microbial genera in the feces when Ca formate was used compared to Ca carbonate. In the ileal digesta, the Ca carbonate diet supplemented with formic acid and phytase led to a similar microbial community as the Ca formate diets. In conclusion, Ca formate reduced prececal InsP6 degradation and P digestibility, but might be of advantage in regard to prececal AA digestibility in pigs compared to Ca carbonate when exogenous phytase is added. The addition of formic acid to Ca carbonate with phytase, however, resulted in greater InsP6 disappearance, P and AA digestibility values, and changed ileal microbiota composition compared to Ca carbonate with phytase alone.

Mucosal phosphatase activity, phytate degradation, and mineral digestibility in 6-week-old turkeys and broilers at different dietary levels of phosphorus and phytase and comparison with 3-week-old animals

Female turkeys (B.U.T. 6) and broilers (Ross 308) were compared at 6 wk of age to evaluate the effects of species, dietary P, Ca, and phytase levels on myo-inositol hexakisphosphate (InsP6) degradation along the digestive tract, gut mucosal phosphatase activity, P and Ca digestibility, and myo-inositol concentrations in the digesta and blood. The environmental conditions and experimental corn-soybean meal-based diets were the same for both species. Four diets with either combination of 2 levels of P and Ca (CaP-: 4.0 g P/kg, 5.4 g Ca/kg and CaP+: 6.0 g P/kg, 8.0 g Ca/kg) and 2 levels of phytase supplementation (0 and 1,500 FTU/kg) were fed to the animals for 7d at their sixth wk of age. Each diet was randomly assigned to 6 pens per species, with 10 birds each. After slaughter, blood, digesta from the crop, gizzard, duodenum, lower ileum, and jejunal mucosa were collected. Endogenous mucosal phosphatase activity in the jejunum was higher in turkeys than in broilers. Prececal InsP6 disappearance was also higher in turkeys than in broilers when phytase was not supplemented. Phytase supplementation led to a higher prececal InsP6 disappearance in broilers than in turkeys, likely due to different crop conditions such as moisture content. However, prececal P digestibility was higher in turkeys than broilers. Different relationships between myo-inositol concentration in the ileum digesta and blood were found, depending on the species. A comparison of the results with those obtained in 3-wk-old birds of a companion study showed that in diets with low Ca and P levels, prececal InsP6 disappearance increased with age in turkeys, but not in broilers. This coincided with changes in the conditions of the digestive tract, such as the water content in the crop, gizzard pH, and mucosal phosphatase activity. In conclusion, occurrence of differences in phytate degradation between turkeys and broilers, fed the same feed, depended on age and can be explained by different physiological development of the digestive tract.

Research Note: Influence of monocalcium phosphate and phytase in the diet on phytate degradation in cecectomized laying hens

This study investigated the effects of phytase and monocalcium phosphate supplementation on the dephosphorylation of phytic acid [myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate); InsP6] in cecectomized laying hens using total excreta collection. Four corn-soybean meal-rapeseed meal-based diets were mixed with or without 6 g of monocalcium phosphate/kg, with or without supplementation of 1,500 FTU phytase/kg, and had the same calcium concentration at 39 g/kg of feed. Each diet was tested in 5 replicates using a row-column design with 10 cecectomized laying hens in 2 periods. The hens received 120 g/d of feed while being housed individually in metabolism units, and total excreta were collected for a period of 4 d. The monocalcium phosphate × phytase interaction was not significant for InsP6 degradation (P = 0.054). Phytase increased InsP6 disappearance from 13% to 83% (P < 0.001), whereas monocalcium phosphate had no effect. Concentrations of most of the lower inositol phosphate isomers in excreta were higher when monocalcium phosphate was added to the diets. The concentration of Ins(1,2,5,6)P4 in excreta was the highest among the studied partially dephosphorylated inositol phosphates with phytase supplementation and was higher than in diets without phytase supplementation (P < 0.001). Supplementation with phytase increased myo-inositol concentration in excreta (P = 0.002), whereas monocalcium phosphate had no effect. Phosphorus utilization ranged from 4% to 18% and was not significantly affected by the treatments. These results suggest that phytase supplementation markedly increased InsP6 degradation in laying hens. The cecectomized laying hen assay may be suitable for studying the effects of phytase supplementation on phytate dephosphorylation under dietary conditions when performance and phosphorus excretion are unlikely to be affected.

Effect of dietary calcium concentration and exogenous phytase on inositol phosphate degradation, mineral digestibility, and gut microbiota in growing pigs.

Variations in the dietary Ca concentration may affect inositol phosphate (InsP) degradation, and thereby, P digestibility in pigs. This study assessed the effects of dietary Ca concentration and exogenous phytase on InsP degradation, nutrient digestion and retention, blood metabolites, and microbiota composition in growing pigs with ileal cannulation. In a completely randomized row-column design with four periods, eight ileal-cannulated barrows (initial body weight 27 kg) were fed four corn-soybean- and rapeseed meal-based diets containing 5.5 or 8.5 g Ca/kg dry matter (DM), with or without 1,500 FTU of an exogenous hybrid-6-phytase/kg diet. No mineral P was added and the P concentration in the feed was 4.8 g P/kg DM. Prececal InsP6 disappearance in pigs fed diets containing exogenous phytase was lower (P = 0.022) with additional Ca than without. Concentrations of InsP2-4 isomers and myo-inositol in the distal ileal digesta and prececal P digestibility were greater (P < 0.001) with exogenous phytase than without exogenous phytase. In feces, InsP6 disappearance was lower (P < 0.002) and concentration of InsP5 and InsP4 isomers was higher (P ≤ 0.031) with additional Ca compared to without additional Ca. The prececal amino acid digestibility, energy digestibility, and hindgut disappearance of energy did not differ. The Shannon diversity index of the microbiota in the distal ileal digesta and feces was similar among the diets but was lower in the distal ileal digesta than in the feces (P < 0.001). Permutation analysis of variance revealed no dietary differences between the bacterial groups within the ileal digesta and fecal samples (P > 0.05). In conclusion, additional Ca reduced the effect of exogenous phytase on prececal InsP6 degradation. Endogenous InsP degradation was impaired by additional Ca only in the hindgut but the abundance of bacterial genera in feces was not affected.

Comparison of mucosal phosphatase activity, phytate degradation, and nutrient digestibility in 3-week-old turkeys and broilers at different dietary levels of phosphorus and phytase

A comparison between 3-wk-old female turkeys (B.U.T. 6) and broilers (Ross 308) was performed to study the effects of species, dietary P, Ca, and phytase levels on gut mucosal phosphatase activity, myo-inositol hexakisphosphate (InsP6) degradation along the digestive tract, digestibility of P, Ca, and amino acids, and concentrations of myo-inositol in the digesta and blood. The experimental diets were corn-soybean meal-based and identical for both species. Two dietary P and Ca concentrations (CaP-: 4.1 g P/kg, 5.5 g Ca/kg and CaP+: 9.0 g P/kg, 12.0 g Ca/kg) and 2 levels of phytase supplementation (0 and 1,500 FTU/kg) were used in a 2×2 factorial design and fed to the animals for 7 d in their third week of age. Each diet was randomly assigned to 6 broiler and 6 turkey pens, with 10 birds each. After slaughter, blood, digesta from the crop, gizzard, duodenum, lower ileum, and mucosa from the jejunum were collected. When fed CaP- without phytase supplementation, there were no differences between species in gut mucosal phosphatase activity, prececal InsP6 disappearance, and P and Ca digestibility, indicating a similar intrinsic capacity for phytate degradation in both species. When fed CaP+ without phytase supplementation, turkeys showed higher prececal InsP6 disappearance than broilers. Phytase supplementation increased prececal InsP6 disappearance and digestibility of P and Ca in both species. However, the phytase-induced increase in prececal InsP6 disappearance was more pronounced in broilers than in turkeys, possibly due to more adequate conditions for phytase activity in the broiler crop. In broilers, phytase supplementation increased amino acid digestibility overall, whereas, in turkeys, it increased with CaP+ and decreased with CaP-. In addition, the relationship between myo-inositol concentration in the ileum and blood differed between species, indicating differences in myo-inositol metabolism. It was concluded that 3-week-old turkeys and broilers differ in nutrient digestibility and InsP degradation in some segments of the digestive tract but have similar endogenous InsP6 degradation when fed low P and Ca diets.

Ruminal and post-ruminal phytate degradation of diets containing rapeseed meal or soybean meal

ABSTRACT This study aimed to investigate ruminal and post-ruminal degradation of phytic acid (InsP6) in diets containing either rapeseed meal (RSM) or soybean meal (SBM). In Experiment 1, the effective degradability of crude protein (CPED) and InsP6 (InsP6ED) was evaluated by incubating RSM and SBM in situ in three rumen-fistulated lactating Jersey cows for 2, 4, 6, 8, 16, 24, 48 and 72 h, and calculating effective degradability at rumen passage rates of 2% and 5%/h. In Experiment 2, eight wethers were assigned for 8 weeks to two dietary treatments (Diet RSM and Diet SBM) containing 150 g of either meal and 100 g of maize silage per feeding time and had free access to hay and water. Titanium dioxide (TiO2) was added to the diets for the last 5 days of the study. The wethers were then stunned, exsanguinated and digesta from the reticulo-rumen, omasum, abomasum, jejunum, colon, and rectum were sampled. In Experiment 1, the InsP6ED of RSM (InsP6ED2: 83%; InsP6ED5: 64%) decreased almost identically to that of CPED with increasing passage rate (CPED2: 78%; CPED5: 63%) and was significantly lower than that of SBM (InsP6ED2: 93%; InsP6ED5: 85%). In Experiment 2, ruminal InsP6 disappearance was significantly higher in wethers fed Diet SBM (89%) than in those fed Diet RSM (76%). Total post-ruminal InsP6 degradation was 6% for Diet RSM and 4% for Diet SBM (p = 0.186). The total tract InsP6 disappearance was higher in Diet SBM (93%) than in Diet RSM (82%). Considering higher InsP6 contents in RSM, Diet RSM resulted in significantly higher amounts of ruminally (Diet RSM: 4.5 g/d; Diet SBM: 3.4 g/d) and total tract (Diet RSM: 4.9 g/d; Diet SBM: 3.5 g/d) degraded InsP6. InsP5 was quantified in most of the digesta samples after feeding Diet RSM but was not detectable in the majority of digesta samples for Diet SBM. Concentrations of myo-inositol (MI) tended to be higher (p = 0.060) in the blood plasma of wethers fed Diet RSM. The consistency between ruminal InsP6 disappearance in wethers and in situ calculated InsP6ED2, along with the very low extent of post-ruminal InsP6 degradation, suggests that at a low rumen passage rate, InsP6-P from the feed becoming available to ruminants is almost entirely from InsP6 degradation in the rumen.

Phytase dose-dependent response of kidney inositol phosphate levels in poultry.

Phytases, enzymes that degrade phytate present in feedstuffs, are widely added to the diets of monogastric animals. Many studies have correlated phytase addition with improved animal productivity and a subset of these have sought to correlate animal performance with phytase-mediated generation of inositol phosphates in different parts of the gastro-intestinal tract or with release of inositol or of phosphate, the absorbable products of phytate degradation. Remarkably, the effect of dietary phytase on tissue inositol phosphates has not been studied. The objective of this study was to determine effect of phytase supplementation on liver and kidney myo-inositol and myo-inositol phosphates in broiler chickens. For this, methods were developed to measure inositol phosphates in chicken tissues. The study comprised wheat/soy-based diets containing one of three levels of phytase (0, 500 and 6,000 FTU/kg of modified E. coli 6-phytase). Diets were provided to broilers for 21 D and on day 21 digesta were collected from the gizzard and ileum. Liver and kidney tissue were harvested. Myo-inositol and inositol phosphates were measured in diet, digesta, liver and kidney. Gizzard and ileal content inositol was increased progressively, and total inositol phosphates reduced progressively, by phytase supplementation. The predominant higher inositol phosphates detected in tissues, D-and/or L-Ins(3,4,5,6)P4 and Ins(1,3,4,5,6)P5, differed from those (D-and/or L-Ins(1,2,3,4)P4, D-and/or L-Ins(1,2,5,6)P4, Ins(1,2,3,4,6)P5, D-and/or L-Ins(1,2,3,4,5)P5 and D-and/or L-Ins(1,2,4,5,6)P5) generated from phytate (InsP6) degradation by E. coli 6-phytase or endogenous feed phytase, suggesting tissue inositol phosphates are not the result of direct absorption. Kidney inositol phosphates were reduced progressively by phytase supplementation. These data suggest that tissue inositol phosphate concentrations can be influenced by dietary phytase inclusion rate and that such effects are tissue specific, though the consequences for physiology of such changes have yet to be elucidated.

Phytate and mineral profile evolutions to explain the textural hardening of common beans (Phaseolus vulgaris L.) during postharvest storage and soaking: Insights obtained through a texture-based classification approach

During adverse postharvest storage of Red haricot beans, the inositol phosphate content, particularly InsP6, decreased significantly, along with a significant increase in InsP5. Using a texture-based classification approach, the InsP6 content in cotyledons was shown an indicator for the extent of hard-to-cook (HTC) development during bean aging. This textural defect development was predominated by storage-induced InsP6 degradation, rather than phytate interconversions during soaking. Ca cations, released during storage, did not leach out significantly during subsequent soaking, suggesting that they were bound with the cell wall pectin in cotyledons, while Mg cations were mostly leached out into the soaking water due to their weak binding capacity to the pectin, and the cell membrane damages developed during HTC. Results obtained herein provide evidence for the pectin-cation-phytate mechanism in textural hardening (and its distribution after cooking) of common beans, and call for a more detailed Ca-relocation study during postharvest storage, soaking and cooking.

Effect of Fiber Fermentation and Protein Digestion Kinetics on Mineral Digestion in Pigs.

Simple SummaryThere is increasing interest in improving nutrient utilization in pigs and poultry and thereby reduce nutrient excretion into the environment. The present review aims to provide an overview on interactions between fermentable substrates (e.g., starch, fiber, and protein) and selected minerals on nutrient digestion and absorption to determine nutritional solutions to maximize animal performance, principally in the grower–finisher phase. Using in vitro models, the site and rate (kinetics) of nutrient digestion or fermentation of a feed ingredient or diet can be estimated. However, for minerals, no standardized methodology to assess in vitro mineral digestion exists. In vivo, the diet total tract digestibility of phosphorus might be underestimated in diets with fermentable ingredients because of increased diet-specific endogenous phosphorus losses and requires further clarification to better calculate the true total tract digestibility of phosphorus in pigs. The quantification of fiber type, composition of fiber fractions within individual raw materials, their influence on digestion kinetics, and effects on digesta pH and nutrient solubility related to fermentation should be considered. In conclusion, applications of nutrient kinetic data should be considered as part of an integrated approach to support nutrient digestion and absorption in the gastrointestinal tract of pigs, thereby helping to reduce nutrient excretion.Nutrient kinetic data and the timing of nutrient release along the gastrointestinal tract (GIT), are not yet widely used in current feed formulations for pigs and poultry. The present review focuses on interactions between fermentable substrates (e.g., starch, fiber, and protein) and selected minerals on nutrient digestion and absorption to determine nutritional solutions to maximize animal performance, principally in the grower–finisher phase, with the aim of minimizing environmental pollution. For phosphorus (P), myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6), copper (Cu), and zinc (Zn), no standardized methodologies to assess in vitro mineral digestion exist. The stepwise degradation of InsP6 to lower inositol phosphate (InsP) forms in the GIT is rare, and inositol phosphate4 (InsP4) might be the limiting isomer of InsP degradation in diets with exogenous phytase. Furthermore, dietary coefficients of standardized total tract digestibility (CSTTD) of P might be underestimated in diets with fermentable ingredients because of increased diet-specific endogenous P losses (EPL), and further clarification is required to better calculate the coefficients of true total tract digestibility (CTTTD) of P. The quantification of fiber type, composition of fiber fractions, their influence on digestion kinetics, effects on digesta pH, and nutrient solubility related to fermentation should be considered for formulating diets. In conclusion, applications of nutrient kinetic data should be considered to help enhance nutrient digestion and absorption in the GIT, thereby reducing nutrient excretion.

Phosphorus digestibility and phytate degradation in pigs fed wheat-based diets with different intrinsic phytase activity and added microbial phytase

ABSTRACT The objective of this study was to investigate the effect of variation in wheat-derived phytase activity on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation, inositol phosphate (InsP) isomer concentration and phosphorus (P) digestibility in pigs fed wheat-based diets. Additional effects of a microbial phytase supplementation were also studied. Three wheat genotypes (W1-W3) with an analysed phytase activity between 2760 and 3700 FTU/kg were used to formulate four experimental diets that included soybean meal and rapeseed meal but did not contain a mineral P supplement. DietW1-DietW3 only differed in the included wheat genotypes (W1-W3) at an inclusion level of 400 g/kg. DietW3+ contained W3 and a commercial 6-phytase supplementation at 500 FTU/kg diet. Eight barrows with an initial body weight of 27 kg were fitted with a simple T-cannula at the distal ileum and assigned to the four dietary treatments in a completely randomised row column design. The experiment included four periods of 12 d each. The first 5 d of each period were for diet adaptation, followed by collection of faeces (4 d), ileal digesta (2 d), and blood (last day). In DietW1-DietW3, the mean precaecal (pc) InsP6 disappearance was 48% and the mean pc P digestibility was 37% without a significant effect of the wheat genotype. The InsP6 disappearance measured in the faeces was close to complete in all treatments, and faecal P digestibility was not significantly affected by the wheat genotype (36% overall). The addition of microbial phytase caused a significant increase in pc InsP6 degradation (to 79%) and pc and total tract P digestibility (to 53% and 52%, respectively). The concentration of InsP6 degradation products in ileal digesta was not significantly affected by the wheat genotype, except for that of Ins(1,2,3,4,6)P5 and myo-inositol, which were higher in DietW3 than in DietW1 and DietW2. The added microbial phytase significantly reduced the concentration of InsP5 isomers in the ileal digesta and increased the concentrations of lower InsP isomers and myo-inositol. There were no significant effects of the added microbial phytase on pc amino acid digestibility; however, the wheat genotype exerted significant effects on the pc digestibility of Cys, Gly and Val. It was concluded that an increase in the intrinsic phytase activity of wheat achieved by crossbreeding was not reflected in InsP6 degradation and P digestibility in pigs fed wheat-based diets.

Phytate degradation and phosphorus utilisation by broiler chickens fed diets containing wheat with increased phytase activity

ABSTRACT 1. The objective of this study was to investigate wheat genotypes bred for increased intrinsic phytase activity for InsP6 disappearance and the formation of lower inositol phosphates in such wheat-fed broiler chickens. The influence of monocalcium phosphate (MCP) supplementation on these characteristics and the utilisation of P and Ca were also determined. A three-step in vitro assay and a broiler trial were performed. 2. In the 63 wheat genotypes tested in vitro, phytase activity varied from 1900 FTU/kg to 5200 FTU/kg, and InsP6 disappearance increased with higher phytase activity of wheat in a linear manner. The addition of MCP significantly reduced in vitro InsP6 disappearance by one-third, independent of the inclusion level of wheat in the feed. When exogenous phytase was added to wheat, in vitro InsP6 disappearance increased independently of the phytase activity of the wheat used. 3. In the broiler trial, four wheat genotypes with phytase activities between 2400 and 3700 FTU/kg were included at 400 g/kg in diets with and without MCP. The diets were not pelleted. Separately, wheat 1, without MCP, was tested with the addition of exogenous phytase. Unsexed Ross 308 broiler chickens were allocated to 72 metabolic units of 10 birds each and assigned one of the nine diets. Mineral utilisation was measured based on excreta collection from 20 to 23 d of age. Digesta from the crop and terminal ileum were collected on d 24. 4. In the crop and ileum, InsP6 disappearance was not affected by the wheat genotypes, but the addition of MCP significantly decreased InsP6 disappearance. Precaecal P disappearance was significantly reduced by the addition of MCP, with wheat genotypes also exerting an effect. Wheat genotypes and the addition of exogenous phytase significantly affected P utilisation. Exogenous phytase had no effect on InsP6 disappearance in the crop but did up to the terminal ileum, the precaecal InsP6 and P disappearance increased with the addition of exogenous phytase. 5. Although the intrinsic wheat phytase activity exerted distinct effects on in vitro InsP6 disappearance, no such effect was found in the broiler trial. The addition of MCP significantly inhibited InsP6 degradation in vitro and in vivo.

Effects of calcium level and source, formic acid, and phytase on phytate degradation and the microbiota in the digestive tract of broiler chickens

BackgroundDiet acidification, dietary calcium (Ca) level, and phytase supplementation are known influences on the microbial community in the digestive tract and on phosphorus (P) utilization of broiler chickens. Effects of dietary factors and microbiota on P utilization may be linked because microorganisms produce enzymes that release P from phytate (InsP6), the main source of P in plant feedstuffs. This study aimed to detect linkages between microbiota and InsP6 degradation by acidifying diets (i.e., replacing Ca carbonate (CaCO3) by Ca formate or adding formic acid to CaCO3-containing diets), varying Ca levels, and supplementing phytase in a three-factorial design. We investigated i) the microbial community and pH in the digestive tract, ii) prececal (pc) P and Ca digestibility, and iii) InsP6 degradation.ResultsAll factors under investigation influenced digesta pH and the microbiota composition. Predicted functionality and relative abundance of microorganisms indicated that diets influenced the potential contribution of the microbiota on InsP degradation. Values of InsP6 degradation and relative abundance of the strains Lactobacillus johnsonii and Lactobacillus reuteri were correlated. Phytase supplementation increased pc InsP6 disappearance, with differences between Ca levels, and influenced concentrations of lower inositol phosphate isomers in the digestive tract. Formic acid supplementation increased pc InsP6 degradation to myo-inositol. Replacing CaCO3 by Ca-formate and the high level of these Ca sources reduced pc InsP6 disappearance, except when the combination of CaCO3 + formic acid was used. Supplementing phytase to CaCO3 + formic acid led to the highest InsP6 disappearance (52%) in the crop and increased myo-inositol concentration in the ileum digesta. Supplementing phytase leveled the effect of high Ca content on pc InsP6 disappearance.ConclusionsThe results point towards a contribution of changing microbial community on InsP6 degradation in the crop and up to the terminal ileum. This is indicated by relationships between InsP6 degradation and relative abundance of phosphatase-producing strains. Functional predictions supported influences of microbiota on InsP6 degradation. The extent of such effects remains to be clarified. InsP6 degradation may also be influenced by variation of pH caused by dietary concentration and solubility of the Ca in the feed.

Phytate degradation, myo-inositol release, and utilization of phosphorus and calcium by two strains of laying hens in five production periods

The objective of this study was to compare 2 laying hen strains in 5 production periods regarding phytase activity, phytate (InsP6) degradation, and myo-inositol (MI) release in the digestive tract and phosphorus (P) and calcium (Ca) utilization. One offspring of 10 nonrelated roosters per strain (Lohmann Brown-classic (LB) or Lohmann LSL-classic (LSL)) was placed in one of 20 metabolic units in a completely randomized block design in week 8, 14, 22, 28, and 58 of life. All hens were fed the same corn and soybean meal–based diet at one time, but the diet composition was adjusted to the requirements in the respective period. For 4 consecutive days, excreta were collected quantitatively at 24-hour intervals. In week 10, 16, 24, 30, and 60, the blood plasma, digesta of crop, gizzard, jejunum, ileum, and ceca, and mucosa of the jejunum was collected. The concentration of inorganic P in the blood plasma was higher in LB than in LSL hens (P = 0.026). Plasma Ca concentrations increased with each period (P < 0.001) in both strains. In jejunum digesta, the MI concentration did not differ between strains, but InsP6 concentration was higher in LB than in LSL hens (P = 0.002) and the highest in week 30 and 60. Total phosphatase and phytase activities were higher in LB than in LSL hens (P ≤ 0.009). Period effects were also significant for these enzymes. Concentrations of some constituents of the cecal content were different between the strains. The MI concentration in the egg albumen and yolk was higher in LB than in LSL hens. Differences in InsP6- and MI-related metabolism of the 2 hen strains existed. These differences were partly dependent of the period. Especially, week 24 was a period of remarkable change of metabolism. Great differences also existed among individuals, making it worth to have a closer look at the metabolism of individuals in addition to evaluating treatment means. Further studies on metabolic, genetic, and microbiome level may help explain these differences.

Phytase supplementation effects on amino acid digestibility depend on the protein source in the diet but are not related to InsP6 degradation in broiler chickens

The objective was to determine phytase effects on prececal amino acid (AA) digestibility and phytate (InsP6) breakdown when different oilseed meals were used in broiler chicken diets. The study included 14 diets: a corn-soybean meal (SBM) basal diet and 6 diets that contained SBM, rapeseed meal (RSM), and sunflower meal (SFM) with 2 inclusion levels at the expense of corn starch (150 and 300 g/kg SBM or SFM, or 100 and 200 g/kg RSM). Each diet was mixed with or without a phytase supplement of 1,500 FTU/kg. Diets were provided to broilers for 5 D. Digesta from the posterior half of the ileum were collected on day 21. The average essential AA digestibility, calculated by a regression approach, without and with phytase was 84 and 85% (SBM), 74 and 77% (SFM), and 66 and 73% (RSM), respectively. In the diets, phytase effects on AA digestibility were lower owing to other protein sources also present in the diet, but significant. Prececal InsP6 disappearance was significantly affected by interactions between oilseed meal, inclusion level, and phytase supplementation. Overall, prececal InsP6 disappearance was higher in SBM diets (52%) than in SFM diets (38%) and intermediate in RSM diets (43%). Across diets, phytase supplementation effects on prececal InsP6 degradation linearly increased with the InsP6 concentration of the diet up to 12 g/kg DM. The only exception from linearity was the diet with the high inclusion of SFM, which contained 15.9 g InsP6/kg DM. In the ileal content, the concentration of myo-inositol was significantly increased by phytase supplementation, and this effect was highest in the diets that contained SBM as the only oilseed meal. Concentrations of lower inositol phosphates were increased by phytase supplementation, and this effect was most remarkable for Ins(1,2,3,4)P4 and inositol tetrakisphosphates. The study showed that phytase effects on AA digestibility varied among the 3 tested oilseed meals, but these differences were not detectable in the diets containing these meals. Although phytase effects on ileal content of InsP6 and its degradation products were substantial, they were not related to the effects on AA digestibility.

Phytate degradation cascade in pigs as affected by phytase supplementation and rapeseed cake inclusion in corn-soybean meal-based diets.

Two experiments (Exp.) with ileally cannulated growing barrows were conducted. The concentrations of positional inositol phosphate (InsP) isomers in ileal digesta and feces were determined, as well as the prececal and total tract phytate (InsP6) hydrolysis, and digestibility of dry matter, P, Ca, nitrogen, and gross energy. Prececal amino acid (AA) digestibility and digestive enzyme activities in ileal digesta were also studied. In both Exp., pigs had an initial body weight (BW) of 28 kg and were completely randomized to a Double Latin Square Design with eight pigs, four diets, and three periods of 12 d each. Feces and ileal digesta were collected for 5 d and 2 d, respectively. Pigs were housed individually in stainless steel metabolic units. Water was available ad libitum and feed was provided two times daily at an amount of 4% of mean BW. In Exp. 1, pigs received a corn–soybean meal (SBM)-based diet that was supplemented with 0, 750, 1,500, or 3,000 FTU of a microbial phytase/kg diet. In Exp. 2, pigs were allotted to a 2 × 2 arrangement of diets based on corn and SBM or an SBM-rapeseed cake (RSC) mix and phytase supplementation at 0 or 1,500 FTU/kg of diet. In ileal digesta of pigs fed without the phytase supplement, the dominating InsP isomers beside InsP6 were InsP5 isomers. The InsP pattern in ileal digesta changed with the inclusion of microbial phytase in both Exp., as there was a remarkable increase in Ins(1,2,5,6)P4 concentration (P < 0.001). In both Exp., the myo-inositol concentration in ileal digesta was greater upon phytase addition (P < 0.001). Without phytase supplementation, prececal and total tract P digestibility were low, whereas hardly any InsP6 was excreted in feces. There was no difference between prececal and total tract P digestibility values. For most AA studied in Exp. 2, prececal digestibility was lower (P < 0.01) when the diet contained RSC. However, phytase supplementation did not significantly affect prececal AA digestibility in both Exp. The present study showed that InsP6 disappearance by the end of the ileum can be increased up to around 90% in SBM- and SBM–RSC-based diets when microbial phytase is supplemented, but prececal P digestibility hardly exceeded 60%. The study confirms that pigs cannot benefit from a remarkable InsP6 degradation in the hindgut.

Determination of in situ ruminal degradation of phytate phosphorus from single and compound feeds in dairy cows using chemical analysis and near-infrared spectroscopy

The ruminal degradation of P bound in phytate (InsP6) can vary between feeds, but data on ruminal degradation of InsP6 from different feedstuffs for cattle are rare. One objective of this study was to increase the data base on ruminal effective degradation of InsP6 (InsP6ED) and to assess if InsP6ED of compound feeds (CF) can be calculated from comprising single feeds. As a second objective, use of near-infrared spectroscopy (NIRS) to predict InsP6 concentrations was tested. Nine single feeds (maize, wheat, barley, faba beans, soybeans, soybean meal (SBM), rapeseed meal (RSM), sunflower meal (SFM), dried distillers’ grains with solubles (DDGS)) and two CF (CF1/CF2), consisting of different amounts of the examined single feeds, were incubated for 2, 4, 8, 16, 24, 48 and 72 h in the rumen of three ruminally fistulated Jersey cows. Samples of CF were examined before (CF1/CF2 Mash) and after pelleting (CF1/CF2 Pellet), and InsP6ED was calculated for all feeds at two passage rates (InsP6ED5: k = 5%/h; InsP6ED8: k = 8%/h). For CF1 and CF2, InsP6ED was also calculated from values of the respective single feeds. Near-infrared spectra were recorded in duplicate and used to establish calibrations to predict InsP6 concentration. Besides a global calibration, also local calibrations were evaluated by separating samples into different data sets based on their origin. The InsP6ED8 was highest for faba beans (91%), followed by maize (90%), DDGS (89%), soybeans (85%), wheat (76%) and barley (74%). Lower values were determined for oilseed meals (48% RSM, 65% SFM, 66% SBM). Calculating InsP6ED of CF from values of single feeds underestimated observed values up to 11 percentage points. The NIRS calibrations in general showed a good performance, but statistical key data suggest that local calibrations should be established. The wide variation of InsP6ED between feeds indicates that the ruminal availability of P bound in InsP6 should be evaluated individually for feeds. This requires further in situ studies with high amounts of samples for InsP6 analysis. Near-infrared spectroscopy has the potential to simplify the analytical step of InsP6 in the future, but the calibrations need to be expanded.

Phytate degradation in gnotobiotic broiler chickens and effects of dietary supplements of phosphorus, calcium, and phytase

ABSTRACTGnotobiotic broiler chickens were used to study interactive effects of supplemented phosphorus, calcium (PCa), and phytase (Phy) on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and release of myo-inositol in the digestive tract. In 2 subsequent runs, the chickens were subjected to 1 of 4 dietary treatments with and without PCa and Phy supplementation. Sanitized eggs were hatched in 8 germfree isolators, and a minimum of 9 male Ross 308 chickens were placed in each pen (total 16 pens). Treatments implemented on day 10 included gamma-irradiated diets without (PCa−; 4.1 g P and 6.2 g Ca/kg DM) or with (PCa+; 6.9 g P and 10.4 g Ca/kg DM) monosodium phosphate and limestone supplementation and without (Phy−) or with (Phy+) 1,500 FTU Phy/kg feed in a factorial arrangement. On day 15, digesta was collected from different sections of the intestinal tract and analyzed for InsP isomers and myo-inositol. The isolators did not remain germfree, but analysis of contaminants and results of InsP degradation indicated no or minor effects of the identified contaminants. Prececal InsP6 disappearance was 42% with the PCa−Phy− treatment and 17% with PCa+Phy−. No InsP3–4 isomers were found in the digesta of the terminal ileum in PCa−Phy−. The concentration of myo-inositol in the ileal digesta from PCa−Phy− (6.1 μmol/g DM) was significantly higher than that from PCa+Phy− (1.7 μmol/g DM), suggesting rapid degradation of the lower InsP isomers by mucosal phosphatases and their inhibition by PCa. Phytase supplementation increased InsP6 disappearance and prevented inhibitory effects of PCa supplements (72% in PCa−Phy+ and 67% in PCa+Phy+). However, PCa supplementation reduced the degradation of lower InsP isomers mainly in the posterior intestinal sections in the presence of Phy, resulting in significantly lower myo-inositol concentrations. It is concluded that mucosa-derived phosphatases might significantly contribute to InsP6 degradation in broiler chickens. The potential of mucosa-derived phosphatases to degrade InsP6 and lower InsP is markedly reduced by dietary PCa supplementation.

Prececal amino acid digestibility and phytate degradation in broiler chickens when using different oilseed meals, phytase and protease supplements in the feed

ABSTRACTThe purpose of this study was to investigate the effects of phytase and protease supplementation on prececal (pc) amino acid (AA) digestibility, phytate (InsP6) degradation, and MEn concentration in diets using 3 oilseed meals as main protein sources in broiler chicken feed. The broiler chicken diets, which lacked mineral phosphorus, contained either soybean meal (SBM), SBM and rapeseed meal (SBM/RSM), or SBM and sunflower meal (SBM/SFM) as main protein sources. Diets were not supplemented with enzymes or supplemented with 1,500 or 3,000 FTU phytase/kg, or with 1,600 mg protease/kg. For diets containing SBM as the main protein source, the effects of phytase supplementation with and without monocalcium phosphate were also investigated. Data were obtained during 2 subsequent runs from days 14 to 22 and from days 23 to 31. Each diet was tested using 8 replicates with 4 replicates per run. For pc AA digestibility, no significant interactions were observed between main protein sources, enzyme supplementation, or addition of monocalcium phosphate except for Cys. Supplementation of 1,500 FTU phytase/kg increased pc digestibility of all AA. No differences in pc AA digestibility were observed between 1,500 and 3,000 FTU phytase/kg supplementation treatments. Prececal disappearance of InsP6 and pc P digestibility were greater in the high phytase supplementation treatment. Protease supplementation increased pc digestibility of all AA except for Cys when SBM/RSM was the main protein source. Supplementation of protease and 3,000 FTU phytase/kg increased MEn concentrations. The effect of phytase on pc AA digestibility was fully expressed at a lower supplementation level than needed for a maximized pc InsP6 disappearance and MEn concentration.

Effects of phosphate and phytase supplementation on phytate degradation in rainbow trout (Oncorhynchus mykiss W.) and Atlantic salmon (Salmo salar L.)

The objectives of the present study were to investigate the single and interactive effects of mineral phosphorus (P) (monoammonium phosphate; MAP; 1 g P/kg DM of diet) and Aspergillus oryzae 6-phytase (2800 FTU/kg DM diet) supplements on the disappearance of phytic acid (InsP6) and its lower inositol phosphate isomers (InsPs) and related P digestibility effects in Atlantic salmon (S. salar) and rainbow trout (O. mykiss). For each species, one experiment was conducted applying species-specific conditions but using the same diets in both experiments. Faeces were obtained by stripping the fish. The faecal InsP6 disappearance was similar in Atlantic salmon (8.6%) and rainbow trout (8.1%) when diets did not contain the MAP or phytase supplement. Phytase supplementation significantly increased InsP6 disappearance in both species, but to a larger extent in rainbow trout. The hydrolysis of lower inositol phosphate isomers progressed to a greater extent in rainbow trout. Supplementation of MAP had no significant effect on InsP6 disappearance in rainbow trout but slightly decreased InsP6 disappearance in Atlantic salmon. No significant interactive effect of the supplements on InsP6 disappearance was detected in both experiments. The analysis of InsPs in the faeces of the two species suggested that the degradation pathway of InsP6 differed between the two species. In both species, each supplement increased the amount of digested P. This effect was additive in rainbow trout but not in Atlantic salmon. In Atlantic salmon, the MAP supplementation decreased the efficacy of phytase by reducing the amount of digested InsP-P to negligible amounts. However, no significant interactive effect of MAP and phytase supplementation on P digestibility, digested P, and digested InsP-P was detected in either species. The present study provided insight into details of InsP6 degradation in the digestive tract and revealed similarities and differences between the two fish species. The effects of phytase in Atlantic salmon when using low-P diets may not be the same for commercial diets that contain more inorganic P.