Digestible calcium equivalency of phytase and nutrient utilization of broiler chickens fed graded levels of limestone or phytase during the starter phase.

The current study aimed to quantify the digestible calcium (Ca) equivalency of a new phytase (HiPhorius) in broiler chickens. A total of 1,152 male Cobb 500 broiler chickens were used in an experiment in which 8 diets consisting of graded levels of Ca supplied with limestone or graded levels of the phytase were fed. The 8 dietary treatments were based on a corn-soybean meal diet containing 5.1 g/kg of Ca and 5.1 g/kg of phosphorus (P) as negative control (NC); the NC + 1.3, 2.6, or 3.9 g/kg of Ca from limestone; and the NC + 500, 1,000, 2,000, or 4,000 FYT/kg of phytase. Birds were fed the experimental diets for 3 d (from d 7 to 10) or 14 d (from d 7 to 21) to determine apparent ileal digestibility (AID) and apparent total tract retention (ATTR) of dry matter (DM), CP, Ca, and P. In the 10-day-old birds, increasing the levels of Ca decreased the AID of P and Ca (linear, P < 0.05). Increasing the levels of phytase quadratically improved (P < 0.05) the AID of Ca and P. The AID of DM and CP in the younger birds quadratically increased (P < 0.05) as the levels of Ca increased. There were linear and quadratic effects (P < 0.05) of increasing phytase level for the AID of DM and a linear effect (P < 0.05) for the AID of CP. In the 21-day-old birds, increasing Ca levels resulted in a linear decrease (P < 0.05) in the AID of CP and P, and a quadratic decrease (P < 0.05) in the ATTR of CP. Increasing phytase levels linearly and quadratically (P < 0.05) improved the AID and ATTR of CP, P, and the ATTR of Ca. The analyzed phytase activity in the diets supplemented with phytase ranged from 1,520 to 7,661 FYT/kg. The digestible Ca equivalence for dietary phytase at 1,520 to 7,661 FYT/kg ranged from 1.55 to 2.02 g/kg in the 10-day-old birds fed for 3 d and from 0.52 to 0.64 g/kg in 21-day-old birds fed for 14 d. The results showed the reduction in Ca level that could be accommodated by phytase supplementation, which is markedly different between the younger and older birds. Feeding duration influenced the impact of phytase supplementation on Ca and P digestibility, with better efficacy of phytase observed in the 10-day-old birds fed for 3 d. Also, the results showed the extra-phosphoric effects of phytase on the utilization of other essential nutrients such as protein and Ca.

Dietary muramidase improved growth performance, feed efficiency, breast meat yield, and welfare of turkeys from hatch to market.

Muramidase is an enzyme that hydrolyzes peptidoglycans of bacterial cell walls and improves performance of broilers in a dose-dependent manner. An experiment was conducted to evaluate muramidase supplementation, at a high or step-down dose, in turkeys from hatch to market. Male, B.U.T. 6 turkey poults were placed in 24 floor pens at 32 birds per pen. Poults were fed 1 of 3 diets from d 1 to 126 of age. There were 8 replicate pens per treatment. The treatments were a control (CTL) diet, the CTL plus muramidase at 45,000 LSU(F)/kg from phase 1 to 6 (BAL45), and the CTL plus muramidase at 45,000 LSU(F)/kg from phase 1 to 3 and decreased to 25,000 LSU(F)/kg from phase 4 to 6 (BAL45-25). Data were analyzed using SAS. The model included treatment and block and means were separated by Fisher LSD test. Birds fed BAL45 were heavier (P < 0.05) and had a greater (P < 0.05) average daily gain compared with birds fed the CTL from hatch to d 126 of age. Birds fed BAL45-25 had a final BW and average daily gain intermediate to or equivalent to birds fed BAL45 at the same phases. Feed conversion ratio was improved (P < 0.05) in birds fed BAL45 compared with birds fed the CTL and intermediate in birds fed BAL45-25. Breast meat yield was greater (P < 0.05) in turkeys fed muramidase, regardless of dose, compared with birds fed the CTL. There was no effect of treatment on muramic acid content in the jejunum digesta or litter scores. The frequency of pododermatitis score 1 was greater (P < 0.05) and score 2 was lower (P < 0.05) in birds fed muramidase, regardless of dose, compared with birds fed the CTL diet. In conclusion, muramidase supplementation improved performance, breast meat yield, feed efficiency and some markers of welfare, proportional to the dose in the diets.

Determination of the standardized ileal digestible calcium requirement of male Arbor Acres Plus broilers from day 25 to 42 post-hatch.

An experiment was conducted to determine the standardized ileal digestible (SID) Ca requirement of Arbor Acres Plus male broilers from d 25 to 42 post-hatch. Broilers were obtained at hatch, placed in floor pens, and fed a nutrient adequate diet until d 24 post-hatch. On d 25, twelve hundred birds were weighed and allocated to one of 4 treatments. There were 25 birds per pen and 12 pens per diet. The diets were formulated to contain 0.46, 0.35, 0.24, or 0.13% SID Ca. Available P (avP) was 0.39% in all diets, including 0.16% avP expected from 2,500 FYT/kg of phytase. The SID Ca requirement was estimated using nonlinear models, including quadratic, straight broken-line, and quadratic broken-line. There was no effect of SID Ca on feed intake, body weight gain, feed conversion ratio, or livability. Tibia ash percent was greatest in birds fed 0.35% SID Ca and lowest in birds fed 0.13% SID Ca (quadratic, P = 0.063). Apparent ileal digestibility (AID) of Ca was highest in birds fed the diets containing 0.13% SID Ca and decreased (quadratic, P = 0.014) as dietary SID Ca increased to 0.46%. Apparent digested Ca was highest in birds fed 0.35% SID Ca and lowest in birds fed 0.13% SID Ca (quadratic, P = 0.005). Decreasing the concentration of SID Ca in the diet from 0.46 to 0.13% (P < 0.0001) increased the AID of P and apparent digested P. Litter N or P were lowest in birds fed 0.35% SID Ca and increased (quadratic, P ≤ 0.05) as dietary SID Ca decreased to 0.13%. Non-linear equations, developed using tibia ash percent, digested Ca, or litter P, estimate the SID Ca requirement of Arbor Acres Plus broilers from d 25 to 42 was 0.37, 0.35, or 0.35%, respectively. This corresponds to an SID Ca to available P ratio of 0.95 to 0.90.

Research Note: Formulating broiler diets using digestible calcium significantly improved growth performance but reduced apparent ileal digestibility of calcium and phosphorus.

An experiment was conducted to compare the use of a total and digestible Ca formulation system in broilers from hatch to d 37 post-hatch. Ross 308 (n = 288) male broilers were obtained and allocated to one of 2 dietary treatments in floor pens. There were 18 birds per pen and 8 replicate pens per diet. One diet was formulated using ingredient and nutrient recommendations for total Ca and the second diet was formulated using ingredient and nutrient recommendations for standardized ileal digestible (SID) Ca. All diets were formulated to be nutrient adequate using a 2-phase feeding program and including 0.19% available P (avP) and 0.209% total Ca or 0.073% digestible Ca from 1,000 FYT/kg of phytase. On d 17 and 37, tibias and ileal contents were obtained. From hatch to d 37, birds fed diets formulated using digestible Ca gained (P < 0.05) more and were more efficient (P < 0.05) compared with birds fed diets formulated using total Ca. There was no impact of formulation system on tibia ash or minerals. Litter pH (P < 0.05) was greater and litter dry matter (P < 0.05) was lower in birds fed the diets formulated using digestible Ca compared with those fed diets formulated using total Ca. Apparent ileal digestibility (AID) of Ca (P < 0.05), AID of P (P < 0.05) and digestible P intake (P < 0.05) were lower in birds fed diets formulated using digestible Ca compared with those formulated using total Ca at d 17 or 37. However, apparent ileal digested Ca and digestible Ca intake were not different between the experimental diets on d 17 or 37. In conclusion, formulating diets using digestible Ca improved weight gain and feed conversion ratio, but reduced the AID of Ca and P compared with birds fed diets formulated using total Ca. These findings might be reflective of the higher total Ca concentration in the diets formulated using digestible Ca compared with those formulated using total Ca.

Determination of the standardized ileal digestible calcium requirement of male Arbor Acres Plus broilers from day 11 to 24 post-hatch.

Male broilers (n = 576) were obtained and allocated to 96 cages with 6 birds per cage. From hatch to d 24, 16 randomly allocated cages were fed a nutrient adequate reference diet. The reference diet was formulated to contain 0.96 and 0.87% total Ca from hatch to d 10 and d 11 to 24, respectively. The remaining 80 cages were fed a nutrient adequate diet, formulated to contain 0.50% standardized ileal digestible (SID) Ca from hatch to d 10. On d 11, five diets containing 0.56, 0.46, 0.36, 0.26, or 0.16% SID Ca were randomly assigned to 16 cages per diet. Means were separated using polynomial contrasts and Dunnett's Multiple Comparison tests. From hatch to d 10, birds fed diets formulated to contain 0.50% SID Ca gained more (P < 0.05) compared with birds fed the reference diet. From d 11 to 24, there was no effect of diet on feed intake or BW gain. Birds fed 0.46 or 0.16% SID Ca were less (P < 0.05) efficient compared with birds fed the reference diet. Birds fed 0.26 or 0.36% SID Ca were more efficient compared with birds fed all other levels of SID Ca (quadratic, P < 0.05). Tibia ash percent was greatest in birds fed 0.56 or 0.46% SID Ca, decreased (quadratic, P < 0.05) as dietary SID Ca decreased and was lower (P < 0.001) in birds fed the 0.26 or 0.16% SID Ca diets compared with birds fed the reference diet. Apparent ileal digestibility (AID) of Ca or retention of P was greater (P < 0.05) in birds fed the diets formulated using SID Ca compared with birds fed the reference diet. Apparent P retention was greatest in birds fed 0.36% SID Ca (quadratic, P < 0.05). Regression equations developed using P retention, AID of Ca, and percent tibia ash estimated the SID Ca requirement of 11 to 24-d-old broilers was 0.380, 0.488, and 0.515%, respectively. This corresponds to a SID Ca to available P ratio of 0.86 to 1.17.

Determination of the standardized ileal digestible calcium requirement of male Arbor Acres Plus broilers from hatch to day 10 post-hatch.

Arbor Acres Plus male broilers (n = 1,152) were obtained at hatch and allocated to 1 of 6 diets from hatch to d 10 post-hatch. There were 16 replicate cages per diet and 12 birds per cage. Five of the diets were formulated to contain graded concentrations of standardized ileal digestible (SID) Ca at 0.60, 0.50, 0.40, 0.30, or 0.20%. A sixth reference diet was formulated using total Ca coefficients for each ingredient and contained 0.96% total Ca. Available P (avP) was maintained at 0.48% in all 6 diets. Data were subjected to an analysis of variance and the model included diet and block. Means were separated using contrasts to determine linear or quadratic effects of SID Ca and using the Dunnett's test to compare the reference diet to all SID Ca diets. There was no effect of graded levels of SID Ca on intake or gain. Birds fed diets containing 0.60, 0.50, 0.30, or 0.20% SID Ca ate (P < 0.05) or gained (P < 0.05) more compared with birds fed the reference diet. Mortality corrected FCR improved (linear, P < 0.05) as the SID Ca concentration in the diet increased from 0.20 to 0.60%. Tibia ash percent was greatest in birds fed 0.50% SID Ca and lowest in birds fed 0.20% SID Ca (quadratic, P < 0.05). Tibia ash percent was lower (P < 0.05) in birds fed diets formulated to contain 0.20% SID Ca compared with birds fed the reference diet. No other differences in tibia ash were reported. Apparent ileal digestibility (AID) or retention of P was greater (P < 0.05) in birds fed diets formulated using SID Ca compared with birds fed the reference diet. The AID of P increased (linear, P < 0.05) as the SID Ca content in the diet decreased from 0.60 to 0.20%. The AID or retention of Ca was similar in birds fed 0.60 or 0.50% SID Ca and increased as SID Ca decreased to 0.20% (quadratic, P < 0.05). Regression equations developed using bone ash and apparent P retention estimate the SID Ca requirement of Arbor Acres Plus broilers from hatch to d 10 post-hatch was 0.53 and 0.49%, respectively. This corresponds to a SID Ca to available P ratio of 1.1 to 1.02.

Contrasting the effects of phytase and pure myo-inositol on the performance, digestibility, blood and egg yolk inositol levels and digestion physiology of laying hens.

ABSRACT1. An experiment was designed to compare the effects of supplementing laying hen diets with phytase and myo-inositol (inositol).2. Five diets were formulated: high balanced protein (HBP - 840 mg of Dlys/hen/day), HBP with inositol (HBP+I - 0.16%), reduced balance protein (RBP - 672 mg of Dlys/hen/day), RBP with inositol (RBP+I - 0.16%) and RBP with phytase (RBP+P - 3000 FTU/kg).3. Laying hen production, inositol concentrations, digestive tract morphology, amino acid digestibility and intestinal inositol transporters transcript abundance were evaluated. Data were analysed with a one-way ANOVA in SAS 9.4. Contrasts were used to assess the effect of protein, inositol, phytase and phytase vs. inositol. Differences were accepted when P ≤ 0.05.4. No effect on hen-day egg production or feed efficiency was found. However, feed intake and the incidence of abnormally shaped eggs were 0.77 g/h/d and 0.17% higher, respectively, in inositol treatments. Inositol decreased egg specific gravity from 1.088 to 1.0865.5. Inositol concentration in egg yolk was similar among HBP+I, RBP+I and RBP+P, and higher than for the HBP and RBP diet groups. Both gizzard and ileal digesta were enriched in inositol in all supplemented treatments, and phytase supplementation decreased the level of IP5 and IP6 in the gizzard and ileum. Generally, neither phytase or inositol affected amino acid digestibility.6. Inositol increased transcript abundance of alkaline phosphatase in the ileum, while phytase upregulated duodenal alkaline phosphatase and SMIT1, jejunal SMIT2 and reduced ileal HMIT and SMIT1 abundance.7. In conclusion, no effect of phytase or inositol was found for laying hen production performance or amino acid digestibility, but egg quality was reduced by inositol supplementation. Inositol concentration in egg yolk was similar among supplemented treatments.

Inositol and gradient phytase supplementation in broiler diets during a 6-week production period: 1. effects on growth performance and meat yield.

An experiment was conducted to evaluate effects of inositol and gradient phytase supplementation on growth performance and meat yield of broilers from 1 to 41 d of age. A total of 1,920 Yield Plus × Ross 708 male chicks were placed in 64 floor pens (30 birds per pen). Each pen received one of the 8 dietary treatments (8 replicate pens) from 1 to 15, 16 to 29, and 30 to 40 d of age. Treatment 1 was formulated to contain 0.165 and 0.150% lower calcium and phosphorus, respectively, than treatment 7 (positive control). Phytase was added to treatment 1 at concentration of 500, 1,500, 4,500, 13,500, and 40,500 phytase units (FTU)/kg to establish treatments 2 to 6, respectively. Treatment 8 was formulated by adding inositol to treatment 7 based on the expected inositol liberation in treatment 6. Feed and birds were weighed at 1, 15, 29, and 40 d of age to determine BW gain, feed intake, and feed conversion. Twelve birds per pen were processed at 41 d of age to determine carcass characteristics. From 1 to 40 d of age, log-quadratic effects of phytase (treatments 1-6) were observed for BW gain (P = 0.002) and feed conversion in broilers (P = 0.018), whereas feed intake increased log-linearly (P = 0.045). The addition of 40,500 FTU/kg of phytase increased cumulative BW gain (P = 0.001) and decreased cumulative feed conversion (P = 0.005) by 4.7 and 2.6%, respectively, compared with birds subjected to treatment 8. Log-quadratic effects of phytase additions were observed for carcass (P < 0.001) and breast meat weights (P = 0.004). Growth performance and carcass characteristics of broilers subjected to treatment 7 were similar (P > 0.05) to those of birds subjected to treatment 8. These data demonstrate that the extraphosphoric effects of phytase may be associated with increased feed intake of broilers. Inositol supplementation did not provide additional benefits to broilers in this study.

Inositol and gradient phytase supplementation in broiler diets during a 6-week production period: 2. Effects on phytate degradation and inositol liberation in gizzard and ileal digesta contents.

An experiment was conducted to evaluate effects of dietary phytase and inositol supplementation on phytate degradation in gizzard and ileal digesta contents of broilers from 1 to 43 d of age. One thousand nine hundred and twenty Yield Plus × Ross 708 male chicks were placed in 64 floor pens (30 birds/pen). Each pen received 1 of 8 dietary treatments from 1 to 43 d of age. Treatment 1 was formulated to contain 0.165 and 0.150% lower calcium and phosphorus, respectively, than treatment 7. Treatments 2 to 6 were formulated by adding phytase at 500, 1,500, 4,500, 13,500, and 40,500 phytase units (FTU)/kg, respectively, to treatment 1. Treatment 8 was established by adding inositol to treatment 7 based on the maximum inositol liberation in treatment 6. At 15, 29, and 43 d of age, gizzard contents, ileal digesta, and blood were collected for analysis of inositol and inositol phosphate (IP) 2, 3, 4, 5, and 6 concentrations. Increasing phytase from 0 to 40,500 FTU/kg resulted in log-quadratic reductions (P < 0.01) of inositol pentaphosphate and inositol hexakisphosphate concentrations in the gizzard and ileal digesta contents of broilers at 15, 29, and 43 d of age. The increase in phytase doses in treatments 1 to 6 reduced IP3 and IP4 concentrations in a log-quadratic manner (P < 0.05) at each collection period in gizzard contents but only at 43 d of age in the ileal digesta of broilers (P < 0.01). Log-quadratic increases (P < 0.05) of inositol concentrations were observed in gizzard and ileal digesta contents when increasing phytase activity from 0 to 40,500 FTU/kg at each collection period, which translated to a log-quadratic increase (P < 0.01) in plasma inositol concentration of broilers at 15, 29, and 43 d of age. Phytase supplementation up to 40,500 FTU/kg may benefit broilers by reducing phytate concentrations in the gizzard and ileal digesta contents. Moreover, inositol release in the ileal digesta may translate to increased plasma inositol concentration.

Influence of exogenous phytase supplementation on phytate degradation, plasma inositol, alkaline phosphatase, and glucose concentrations of broilers at 28 days of age.

Inositol is the final product of phytate degradation, which has the potential to serve as an indicator of phytase efficacy. An experiment was conducted to evaluate effects of supplementing broiler diets with phytase on phytate degradation and plasma inositol concentrations at 28 d of age. Twenty-four Ross × Ross 708 male chicks were placed in battery cages (4 birds per cage) from 1 to 21 d of age and individually from 22 to 28 d of age. At 27 d of age, a catheter was placed in the brachial vein of broilers to avoid repeated puncture of the vein during blood collection. At 28 d of age, broilers received 1 of 3 experimental diets formulated to contain 0, 400, or 1,200 phytase units (FTU)/kg, respectively, in diet 1, 2, and 3. Blood was collected 1 h before feeding experimental diets and from 20 to 240 min after feeding experimental diets at 20-min intervals with a final blood collection at 480 min to determine plasma inositol concentrations. Inositol phosphate (IP) ester degradation was determined in gizzard contents and ileal digesta. Broilers provided the 1,200 FTU/kg phytase diet had 60% less (P < 0.01) IP6 concentration in gizzard content (1,264 vs. 4,176 nmol/g) and ileal digesta (13,472 vs. 33,244 nmol/g) than birds fed the 400 FTU/kg diet. Adding phytase at 1,200 FTU/kg increased (P < 0.01) inositol concentrations in gizzard content and ileal digesta of broilers by 2.5 (2,703 vs. 1,071 nmol/g) and 3.5 (16,485 vs. 4,667 nmol/g) fold, respectively, compared with adding 400 FTU/kg. Plasma inositol concentration of broilers was not different (P = 0.94) among the dietary treatments at each collection time. Inositol liberation in the digesta of broilers fed diets with 1,200 FTU/kg phytase did not translate to increased plasma inositol concentrations, which warrants further investigation.

Dietary phytate has a greater anti-nutrient effect on feed conversion ratio compared to body weight gain and greater doses of phytase are required to alleviate this effect as evidenced by prediction equations on growth performance, bone ash and phytate degradation in broilers.

Cobb 400, male broilers (n=4,752) were fed one of 12 diets, with 12 pens/diet and 33 birds/pen. Treatments consisted of 3 levels of phytate P (0.24, 0.345, or 0.45%) and 4 doses of phytase (0, 500, 1,000, or 2,000 phytase units (FTU)/kg). Diets were formulated with reduced Ca (0.22%), available P (0.20%), energy (80 to 120 kcal/kg), and amino acids (1 to 5%) when compared with breed requirements. Prediction equations suggested feeding dietary phytate P > 0.275, 0.295, or 0.319% reduced feed intake (FI) and body weight gain (BWG) and increased feed conversion ratio, respectively, from day 0 to 21. Supplementing phytase at 561, 1,285, or >2,000 FTU/kg resulted in the maximum FI, BWG, or feed efficiency, respectively. From day 0 to 42, maximum BWG or feed efficiency were achieved at phytate P concentrations <0.281 or 0.25%, respectively. Supplementing phytase at 449 or 2,000 FTU/kg maximized BWG or feed efficiency, respectively. Tibia ash weight, percent or Ca concentration were maximized at phytate P concentrations <0.24, 0.296, or 0.24%, respectively and phytase supplementation at 822 or >2,000 FTU/kg maximized tibia ash weight or percent respectively. In the absence of phytase, phytate (IP6) concentration in the gizzard was greatest in birds fed 0.45% phytate P and phytase supplementation between 1,132 to 1,285 FTU/kg resulted in the lowest IP6 concentration in the gizzard. There was no effect of dietary phytate P on the concentration of phytate esters (IP5 or IP4) in the gizzard, which were minimized at 1,208 FTU/kg of phytase. In the absence of phytase, the concentration of phytate ester (IP3) or inositol in the gizzard was greatest in birds fed 0.345% phytate P and phytase supplementation at ∼500 FTU/kg minimized IP3, whereas 2,000 FTU/kg maximized inositol, except in birds fed 0.45% phytate P, which was maximized at 202 FTU/kg of phytase. Prediction equations can be useful to determine the influence of phytase and phytate P on broiler performance, phytate degradation and bone ash.

Increasing dietary phytate has a significant anti-nutrient effect on apparent ileal amino acid digestibility and digestible amino acid intake requiring increasing doses of phytase as evidenced by prediction equations in broilers.

Cobb 400, male broilers (n = 4,752) were housed in 12 pens/diet and 33 birds/pen. There were 3 levels of phytate P (0.24, 0.345, or 0.45%) and 4 phytase doses (0, 500, 1,000 or 2,000 phytase units (FTU)/kg) to evaluate the influence of phytate and phytase dose on apparent ileal digestibility (AID) and digestible nutrient intake. Diets were formulated with reduced Ca (0.22%), available P (0.20%), energy (80 to 120 kcal/kg) and amino acids (1 to 5%). On day 21, digesta was collected from 8 birds/pen. Prediction equations determined the linear or non-linear influence of phytate P, log phytase dose, and the interaction. The AID of amino acids, Ca or P and digestible amino acid or Ca intake were influenced by linear or non-linear phytate P × log phytase dose (P < 0.0001). Increasing the dietary phytate P from 0.24 to 0.345 or 0.45% was predicted to reduce the AID of amino acids in a non-linear manner by an average of 6 to 7 percentage points, respectively. This corresponded to a non-linear decrease in digestible amino acid intake of an average of 80 to 90 mg/D. The negative effect of increasing dietary phytate P from 0.24 to 0.45% on AID was greatest for cysteine (-14 percentage points), aspartic acid or glycine (-9 percentage points) and lowest for methionine, tryptophan, serine, or glutamic acid (-5 percentage points). The predicted digestible intake of lysine (-120 mg/D), aspartic acid (-180 mg/D), or glutamic acid (-290 mg/D) were reduced in birds fed diets containing 0.345% vs. 0.24% phytate P. Phytase supplementation was predicted to increase the AID of amino acids, Ca, or P in a non-linear-log or log-linear manner at all levels of phytate P, with the greatest response at higher doses of phytase in diets containing 0.345 or 0.45% phytate P. The effect of phytase on digestible nutrient intake was less clear. Prediction equations can be useful to determine the influence of phytase and phytate P on AID and digestible nutrient intake in broilers.

Studies on the measurement of ileal calcium digestibility of calcium sources in broiler chickens.

Results from 3 experiments relating to the measurement of ileal calcium (Ca) digestibility in 4 different Ca sources for broiler chickens are presented herein. The first experiment was conducted to determine the effect of basal diet composition on the true Ca digestibility of limestone, meat and bone meal (MBM), monocalcium phosphate (MCP), and dicalcium phosphate (DCP). Eight experimental diets were developed based on 2 basal diets (corn-based or corn starch-based) with each of the 4 Ca sources. Two Ca-free diets representing both basal diets were used to determine the endogenous Ca losses. Each diet was randomly allotted to 6 replicate cages (6 birds per cage) and fed from 21 to 24 D post-hatch. Calcium digestibility of corn-based diet was higher (P < 0.05) than the corn starch-based purified diet. The average true Ca digestibility coefficients of limestone, MBM, MCP, and DCP were determined to be 0.51, 0.41, 0.43, and 0.32, respectively. The second experiment was conducted to examine the effect of indicator type on the apparent Ca digestibility of limestone. Two experimental diets with either titanium dioxide or acid insoluble ash (Celite) were developed. Each diet was randomly allotted to 6 replicate cages (8 birds per cage) and fed from 21 to 24 D post-hatch. Total tract Ca retention was also measured using the indicator ratios. Indicator type had no influence (P > 0.05) on the digestibility measurements. Ca retention determined using acid insoluble ash was higher (P < 0.05) compared to that determined using titanium dioxide. The third experiment was conducted to determine the effect of dietary adaptation length on apparent Ca digestibility of limestone. The experimental diet was offered from day 21 to 6 replicates (6 birds per cage) each for 24, 72, 120, or 168 h and the ileal digesta were collected. Calcium digestibility at 24 h was higher (P < 0.05), and increasing the adaptation length from 72 to 120 h had no effect (P > 0.05) on the digestibility.

Evaluation of novel protease enzymes on growth performance and nutrient digestibility of poultry: enzyme dose response.

Two experiments were conducted to evaluate 3 novel proteases in broilers. In experiment 1, 600 male, Cobb 500 broilers were allocated to 1 of 12 experimental diets (5 birds/pen and 10 replicates/diet). A control (C) diet was formulated to be adequate in all nutrients. Proteases 1, 2, or 3 were added to this diet at 3 doses (1x, 3x, or 9x) in a 3 × 3 factorial arrangement of treatments. The factorial was augmented with 2 treatments of phytase at 500 or 1,500 FTU/kg added to the C diet. In experiment 2, 2,050 male Ross 308 broilers were allocated to 1 of 10 experimental diets (25 birds/pen and 9 replicates/diet). A C diet was formulated to be adequate in all nutrients. Protease 1, 2, or 3 was then added to the C diet at 3 doses (1x, 2x, or 4x) in a 3 × 3 factorial arrangement of treatments plus the C. In experiment 1, birds fed phytase gained more (P < 0.05) than birds fed protease, but neither were different than birds fed the C. Supplementation of 9x dose of any protease resulted in a reduction (P < 0.05) in BWG when compared with birds fed 1x dose of protease or phytase at 500 or 1,500 FTU/kg. Feed conversion ratio was improved (P < 0.05) in birds fed phytase compared with birds fed the C diet. Nitrogen digestibility was greater (P < 0.05) in birds fed protease 1 when compared with birds fed protease 2. Birds fed the 1x dose of protease or 500 FTU/kg of phytase had a greater (P < 0.05) N digestibility than birds fed 3x dose of protease. In experiment 2, protease supplementation significantly reduced (P < 0.05) BWG when compared with birds fed the C from hatch to 35 D post-hatch. Protease supplementation did not improve broiler growth performance or N digestibility above that of a nutrient adequate control diet or a diet supplemented with 500 FTU/kg of phytase.

Influence of graded concentrations of phytase in high-phytate diets on growth performance, apparent ileal amino acid digestibility, and phytate concentration in broilers from hatch to 28 D post-hatch.

An experiment was conducted to evaluate graded doses of phytase in high-phytate diets. Ross 308, male broilers (n = 600) were assigned to one of 4 diets, with 10 replicate pens/diet and 15 birds/replicate pen. Diets were a nutrient adequate positive control (PC), a negative control (NC) diet with a reduction of Ca by 0.22%, available P by 0.20%, energy by 120 kcal/kg, and amino acids by 1 to 5% compared with the PC. The NC diet was supplemented with 0, 2,000, or 4,000 phytase units (FTU)/kg. Phytase increased (linear, P < 0.05) weight gain from hatch to day 18. Birds fed the NC + 4000 FTU/kg ate and gained more (P < 0.05) than birds fed the PC. The apparent ileal digestibility (AID) of all nutrients and amino acids were reduced (P < 0.05) in birds fed the NC compared with birds fed the PC. Phytase increased (linear, P < 0.10) AID of most nutrients. Digestibility was lower (P < 0.10) in birds fed the NC + 4000 FTU/kg compared with birds fed the PC. Using daily intake and AID to determine digestible nutrient intake resulted in no differences between birds fed the PC or NC + 4000 FTU/kg diets. Digestible intake of methionine or glutamate was better correlated with BW gain (P < 0.0001) than AID (P > 0.10). Phytase reduced (linear, P < 0.01) phytate concentration and increased inositol (linear, P < 0.01), phytate hydrolysis (linear, P < 0.05), and jejunal expression (linear, P < 0.05) of SNAT-1 and LAT-4 transporters. Supplementation of increasing doses of phytase in high-phytate, low-nutrient dense diets improved gain and digestibility through nearly complete phytate destruction. Digestible nutrient intake may be a better indication of broiler gain than AID alone.

High doses of phytase on growth performance and apparent ileal amino acid digestibility of broilers fed diets with graded concentrations of digestible sulfur amino acids.

Two experiments of the same design were conducted to determine the influence of phytase on performance and apparent ileal digestibility (AID) of amino acids in broilers fed graded concentrations of digestible sulfur amino acids (dgM+C). Cobb 400 male broilers were allocated to 1 of 10 diets consisting of 5 basal diets formulated at 75, 82, 89, 96, or 103% of the Cobb 400 dgM+C requirements for each feeding phase. Phytase was included in each basal diet at 0 or 2,000 FTU/kg. In experiment 1, 33 birds/pen from hatch to day 42 were fed a 2-phase feeding program with 10 replicate pens/diet. In experiment 2, there were 26 birds/pen from hatch to day 21 and 8 replicate pens/diet. Data were analyzed as a 5 × 2 factorial and means separated using orthogonal contrasts. In experiment 1, overall (hatch to day 42) feed intake (FI) decreased (linear, P < 0.05), body weight gain (BWG) increased (quadratic, P < 0.05), and feed conversion ratio (FCR) was improved (quadratic, P < 0.05) as dgM+C increased to ≥96% in the diet. Phytase increased (P < 0.05) BWG and improved (P < 0.05) FCR, regardless of the percent dgM+C in the diet. In experiment 2, overall (hatch to day 21) BWG increased (quadratic, P < 0.05), and FCR was improved (quadratic, P < 0.05) as dgM+C increased to ≥96% in the diet. Phytase increased FI (P < 0.05) and BWG (P < 0.05) and improved FCR (P < 0.05), regardless of the percent dgM+C in the diet. In the absence of phytase, the AID of all amino acids was greatest (quadratic, P < 0.05) in birds fed between 89 and 96% dgM+C. However, in the presence of phytase the AID of all amino acids was greatest (quadratic, P < 0.05) in birds fed 82% dgM+C and greater at all levels of dgM+C than birds fed diets without phytase (dgM+C × phytase, P < 0.05). In conclusion, phytase improved AID of all amino acids and improved growth performance regardless of the level of dgM+C in the diet.

Evaluation of novel protease enzymes on growth performance and apparent ileal digestibility of amino acids in poultry: enzyme screening.

Three experiments were conducted to evaluate eight neutral and six acid proteases on growth performance and apparent ileal amino acid digestibility (AID) of poults (Experiment 1) or chicks (Experiments 2 and 3). Two basal diets were formulated: a nutrient adequate positive control (PC), which met or exceeded the nutrient requirements for poults (Experiment 1) or chicks (Experiments 2 and 3) and a negative control (NC) formulated to achieve 85% (Experiments 1 and 2) or 80% (Experiments 3) of the requirement for protein and amino acids. Phytase was included in all diets to provide 500 phytase units (FTU)/kg and xylanase was included in all diets to provide 10,000 (Experiments 1 and 2) or 16,000 (Experiments 3) xylanase units (BXU)/kg. Proteases were supplemented in the NC diet at an equivalent amount of enzyme protein to create 16 experimental diets. There were five birds/pen and 10 replicate pens per treatment in each experiment. In experiment 1, birds fed the PC diet gained more (P < 0.05) than birds fed the NC. There were no differences in growth performance in birds fed the PC or NC in experiments 2 or 3. In all three experiments, birds fed the NC supplemented with neutral protease 1 had reduced (P < 0.05) feed intake (FI) or body weight gain (BWG) and increased (P < 0.05) feed conversion ratio (FCR) compared with birds fed the NC. Birds fed the NC diet supplemented with neutral protease 3, 7 (Experiment 1), or acid protease 4 (Experiment 3) had increased (P < 0.05) FCR and birds fed neutral protease 6 (Experiment 2) had reduced (P < 0.05) BWG compared with birds fed the NC. Apparent ileal amino acid digestibility was improved (P < 0.05) with protease supplementation to the NC diets (Experiment 1 or 3), but this was dependent on the protease and the amino acid. In conclusion, novel protease supplementation improved AID of amino acids but this was not reflected in improvements in growth performance of poults or chicks.

Effect of phytase on growth performance, phytate degradation and gene expression of myo-inositol transporters in the small intestine, liver and kidney of 21 day old broilers.

An experiment was conducted to evaluate phytase supplementation on growth, phytate degradation, and the gene expression of myo-inositol transporters in 21-day old broilers. Ross 308, male broilers (n = 240) were assigned to one of four diets, with 10 pens/diet and six birds/pen from day one to 21. The diets consisted of a negative control (NC) formulated to meet or exceed Ross 308 nutrient requirements, with the exception of calcium (Ca) and available P (avP), which were reduced by 0.16 and 0.15%, respectively. The NC diet was supplemented with 0, 500, 1,500, or 4,500 units/kg of phytase (FTU) to create four experimental diets. On day 21, all birds per pen were euthanized to obtain digesta and tissue samples for phytate degradation and gene expression. Data were analyzed as an analysis of variance using the fit model platform in JMP v 13.0. The model included phytase and significant means were separated using orthogonal linear and quadratic contrasts. Phytase supplementation increased gain (linear, P < 0.05). Phytate (iP6; quadratic, P < 0.05), phytate ester (iP5, iP4, iP3; quadratic, P < 0.05), and inositol (linear, P < 0.05) concentration in the gizzard was influenced by phytase supplementation. Phytate concentration decreased (linear, P < 0.05), iP5 or iP4 concentration increased and then decreased (quadratic, P < 0.05), and inositol concentration increased (quadratic, P < 0.05) in the ileal digesta as phytase supplementation increased in the diet. There was a tendency for the gene expression of the H+-dependent myo-inositol transporter, HMIT, to increase (linear, P < 0.05) in the ileum as phytase dose increased. Gene expression of the sodium-dependent myo-inositol transporter, SMIT2, increased in the jejunum (quadratic, P < 0.05) as phytase dose increased. Intestinal alkaline phosphatase expression increased (linear, P < 0.05) in the ileum as phytase supplementation increased in the diet. The influence of phytase on phytate, phytate esters, and inositol may influence intestinal alkaline phosphatase activity and the gene expression of myo-inositol transporters in the small intestine.

Meta-analysis: explicit value of mono-component proteases in monogastric diets.

A meta-analysis was conducted to investigate the effect of mono-component proteases on performance and apparent ileal amino acid digestibility (AIAAD, %) in monogastrics. A total of 67 experimental trials were included in the meta-analysis from published and internal reports, contributing 467 lines of data. Poultry and swine data accounted for 81 and 19% of the dataset, respectively. Forty-four different proteases were included in the meta-analysis, accounting for commercial and non-commercial products. Mixed Model analysis was used to assess protease effect and the influence of inherent characteristics of the control on protease response. The mean performance response to protease was a reduction in feed conversion ratio (FCR) for poultry (1%, P < 0.05) and swine (4%, P > 0.05). The mean relative effect of protease on AIAAD over the control was 1.6 ± 0.3%, ranging from 1.2% for Arg, Phe and Trp to 2.6% for Cys. For the majority of amino acids, inherent AIAAD of control diets influenced (P < 0.05) the magnitude of the protease response such that, as inherent digestibility increased, the effect of protease on amino acid digestibility decreased. The dataset was subsequently divided into 2 subgroups: diets with and without other enzymes, namely non-starch polysaccharide degrading enzymes (NSPase) and phytase. Addition of protease in diets containing no other enzymes significantly (P < 0.05) increased AIAAD for the majority of amino acids and tended (P < 0.10) to improve Met, Trp, Pro, Gly, and Tyr. However, when other enzymes were included in the experiment, the beneficial effect of protease on AIAAD was lost (P > 0.05). These findings suggest that when other enzymes are already included in the diet, addition of protease requires further justification for use in monogastric diets.

Effects of a high level of phytase on broiler performance, bone ash, phosphorus utilization, and phytate dephosphorylation to inositol.

An experiment was conducted to evaluate how the addition of microbial phytase influenced growth performance, bone mineralization, tissue P content, apparent digestibility and retention, and inositol phosphate (IP) concentrations in broilers fed diets with varying mineral matrices from 2 to 23 d of age. At 2 d of age, chicks were randomly allotted to receive 1 of 6 experimental diets arranged as a 3 × 2 factorial of mineral matrix [control diet with 1.0% Ca and 0.5% non-phytate phosphorus (NPP); mineral matrix 1 with 0.84% Ca and 0.35% NPP; and mineral matrix 2 with 0.77% Ca and 0.29% NPP] and phytase supplementation (0 or 1,500 FTU/kg). Feed intake was influenced (quadratic, P = 0.012) by the mineral matrix, but no interaction or main effect of phytase were observed. Phytase increased (P = 0.011) BW gain regardless of the mineral matrix applied. Feed efficiency was not influenced (P > 0.05) by mineral matrix, phytase, or their interaction. Phytase increased bone ash content differentially across matrices (interaction, P < 0.01), and tibia P content was lowest in birds fed matrix 2 and highest in the control (linear, P < 0.05). Concentrations of P in muscle, spleen, and liver were not affected by treatment. An interactive effect (P < 0.01) was observed for apparent ileal digestibility (AID) of P, where phytase increased AID in matrix 1. An interactive effect (P < 0.01) was observed for apparent retention of P and Ca, where phytase reduced P and Ca retention in the control diet. A main effect (P < 0.01) of mineral matrix was observed for AID of Ca, with birds fed matrix 1 having the lowest AID of Ca compared with control and matrix 2 treatments. Phytase influenced (P < 0.05) IP concentrations differently across matrices. Overall, phytase and the mineral matrix, either as main effects or in an interactive manner, influenced growth performance, apparent nutrient digestibility and retention, bone and IP concentration responses in broiler chicks.