Novel sfericase protease improves amino acid digestibility of soybean meal and rapeseed meal in broiler chickens.

1. Two experiments were conducted to explore the effects of an exogenous sfericase protease on the apparent ileal nutrient digestibility of soybean meal (SBM) and rapeseed meal (RSM) in broiler chickens.2. In each experiment, a total of 256 sixteen-day-old male Cobb 500 broilers were fed one of four semi-purified experimental diets, comprising two different batches (A and B) of samples for either SBM (Exp. 1) or RSM (Exp. 2) without or with an exogenous sfericase (0 or 30,000 NFP/kg). Each experimental diet was fed to eight replicate pens of broiler chickens from 16 to 21 d of age (eight birds per cage), and ileal digesta were collected for measuring the digestibility coefficients.3. In Exp. 1, the amino acid digestibility was greater (P < 0.05) in SBM B compared with SBM A for Arg and Val, and a similar trend (P < 0.1) was observed for Tyr, Leu and Thr. Exogenous sfericase increased (P < 0.10) digestibility of most of amino acids except Gly, His and Trp. There was an interaction between SBM source and sfericase, whereby digestibility of P, N and Asp was increased by sfericase for SBM B but not for SBM A. In Exp. 2, there was no interaction (P > 0.05) between RSM source and sfericase for ileal nutrient digestibility. Digestibility was greater in RSM A compared to RSM B for all non-essential AA and most essential AA (except for Trp), while the reverse was noted for Ca and P (P < 0.05). Exogenous sfericase increased (P < 0.1) digestibility for all amino acids except Cys and Met.4. In conclusion, the current studies showed that both SBM and RSM batches influenced amino acid digestibility. Sfericase protease supplementation increased amino acid digestibility for both SBM and RSM. The digestibility effects were greater in the SBM batch with low digestibility for N and Asp which was in line with an increase in P digestibility.

Effect of day-old chick weight on the response of broilers to high doses of exogenous phytase.

A total of 360 Ross 708 male broiler chicks were used in an 8 treatment and 9 replicate cage study to explore the influence of day-old chick weight on the efficacy of exogenous phytase. Treatments were arranged as a 2 × 4 factorial with the factors being diet (a positive (PC) and negative control (NC) varying in nutrient density fed without or with 2 concentrations of exogenous phytase) and chick weight (light; <38.5 g or heavy; >42 g). Chicks were sourced from the same breeder flock, with light and heavy chicks being selected from the naturally occurring heterogeneity in the population. The diets were corn-soybean meal based and the PC was formulated to meet the nutrient requirements of male broiler chicks. The NC was formulated to contain 120 kcal/kg, 0.5, 0.18, and 0.18% less apparent metabolizable energy (AME), crude protein (CP), calcium (Ca), and phosphorus (P), respectively, than the PC. Amino acid (AA) density in the NC was also reduced in line with the reduction in CP and the manufacturers' recommendations for the effect of phytase on amino acid digestibility. Phytase at either 1,000 FYT/kg or 3,000 FYT/kg was added only to the NC to create a total of 4 diets. Water and mash feed were available ad libitum and were offered to 8 replicate cages, each containing 5 chicks. The experiment was conducted over a period of 3 wk and diets were offered in 2 phases (starter from d 1 to 10 and grower from d 11 to 21). Growth performance was monitored at the end of each diet phase and on d 21 ileal digesta were collected for estimation of apparent digestibility of energy (DE), nitrogen (N), Ca, P, dry matter (DM), and AA. There were no statistically significant interactions between diet and day-old chick weight for any of the measured parameters. Light chicks had significantly lower weight gain (approx. 5%) at both d 10 and d 21 compared with heavy chicks. This effect was principally associated with reduced feed intake and there was no significant effect of chick weight on feed conversion ratio (FCR). Chick weight had no effect on ileal nutrient digestibility. The reduction in nutrient density from the PC to the NC generated a significant reduction in weight gain (around 12%) and a significant increase in FCR (1.68 vs. 1.83). This effect was associated with a significant reduction in ileal DE (approximately 150 kcal/kg) and in the digestibility of several AA. Exogenous phytase significantly increased weight gain, reduced FCR and generated a significant increase in the ileal digestibility of energy, N, P, and several AA. Although chick weight and diet did not interact statistically, heavy chicks benefited more than light chicks from high doses of exogenous phytase across almost all measured end points which was confirmed by regression analysis. In conclusion, light chicks have inferior performance outcomes than heavy chicks principally because of reduced feed intake, but putatively not in digestive capacity per se. Exogenous phytase is effective in improving performance and nutrient digestibility in nutrient deficient diets. The effect of chick weight per se, and also breeder flock age, on the utility of supra-nutritional inclusion concentrations of exogenous phytase warrants further study.

Dietary muramidase increases ileal amino acid digestibility of wheat and corn-based broiler diets without affecting endogenous amino acid losses.

The objective of these studies was to evaluate the impact of dietary muramidase (MUR) on endogenous amino acids (AA) losses and digestibility of nutrients in wheat and corn-based broiler diets. In experiment 1, the effect of dietary MUR on the flow of endogenous AA (EAA) at the jejunum and terminal ileum of broilers were assessed using either the nitrogen (N) free diet method (NFD) or the highly digestible protein diet method (HDP; 100 g casein/kg diet). Sialic acid and muramic acid concentrations were measured in the jejunal content. In experiment 2, a 2x2x2 factorial arrangement of treatments with 2 base grains (wheat or corn), with low or high metabolizable energy (ME) levels, and without or with MUR supplementation was implemented. All diets contained phytase, xylanase, and cellulase. Apparent ileal digestibility (AID) of dry matter (DM), protein (CP), amino acids (AA), crude fat, and energy, as well as the apparent total tract metabolizability (ATTM) of DM, CP, and gross energy (GE) were determined. The standardized ileal digestibility (SID) of AA was obtained by correcting AID values for basal ileal EAA obtained from chicks fed with NFD or HDP in experiment 1, jejunal EAA flow of all AA was higher (P < 0.001) compared to the ileum, but this effect was method dependent. Jejunal, but not ileal, EAA flow measured with HDP was higher compared to NFD, as well as sialic acid (P < 0.001) and muramic acid (P < 0.004) concentrations. Muramidase inclusion had no effect on basal EAA flow, independently of the segment and the method used. In experiment 2, dietary MUR supplementation increased the AID of CP (P < 0.05), all AA, and tended (P = 0.07) to increase the AID of GE, independently of the cereal type used. However, ATTM of DM and GE, but not CP, increased with MUR inclusion compared with the control treatments, especially in wheat and low ME diets (P < 0.05). In conclusion, MUR supplementation improved AID of CP and AA without affecting EAA losses and increases energy utilization.

Research Note: Amylase supplementation improves starch and amino acids digestibility of faba bean for broilers.

This study aimed to investigate the effect of amylase on the nutritional value and nitrogen-corrected apparent metabolizable energy (AMEN) of faba bean (FB) seeds in broiler chicken nutrition. The study was conducted on a total of 84, one-day-old male broiler chickens (Ross 308). The birds were fed a reference corn-soybean meal diet form d 1 to 16, and they were divided into three groups (1st day). The first group (1) was fed the same reference diet as in the first part, whereas 50% of the reference diet was replaced by FB seeds in experimental groups (2 and 3). Group 3 was supplemented with amylase, which contained 0.14 g/kg dry matter (DM). At the end of the study period, excreta and digesta samples were collected from all groups to estimate the nutritional value. Some fraction of starch found in the seeds was part of starch which is resistant to digestion (143.9 g/kg DM). The addition of exogenous amylase to the diet increased the apparent ileal digestibility of DM, CP, and starch in FB seeds. The AMEN value increased as a result of enzyme addition (P < 0.05). The group 3 showed higher apparent ileal digestibility of essential amino acids (Lys, Leu, Phe, His) as well as nonessential amino acids (Tyr, Ala, Pro) in FB seeds. It could be concluded that FB seeds supplemented with exogenous amylase have a greater nutrition value for broiler chickens.

Varying apparent metabolizable energy concentrations and protease supplementation affected broiler performance and jejunal and ileal nutrient digestibility from 1 to 35 d of age.

An experiment was conducted to evaluate the effects of varying AME concentrations and protease supplementation on broiler performance and jejunal and ileal nutrient digestibility from 1 to 35 d of age. Ross × Ross 308 male broilers (n = 1,008) were equally distributed into 48 floor pens and offered 1 of 6 dietary treatments (8 replicate pens/treatment). Dietary treatments consisted of a factorial arrangement with AME concentration (low-, moderate-, or high-AME) and supplemental protease (without or with) as the main factors. Birds and feed were weighed on 1, 15, 29, and 35 d of age to determine body BW, BW gain (BWG), feed intake (FI) and feed conversion ratio (FCR). At 15 and 29 d of age, jejunal and ileal digesta contents were collected to determine nutrient digestibility. From 1 to 15 d of age, broilers offered moderate-AME diets (P < 0.05) had 6.7, 7.1, 4.8% higher BW, BWG, FI, respectively, and a 2.1% lower FCR compared with those offered low-AME diets. Likewise, protease increased (P < 0.05) BW and BWG by 4.3 and 4.7%, respectively, and decreased (P < 0.05) FCR by 3.4%, compared with those offered the diets without protease. From 1 to 29 d of age, broilers offered high-AME diets had 2.9% lower (P < 0.05) FCR compared with those offered low-AME diets. Protease increased (P < 0.05) BW, BWG, and FI by 3.1, 3.2, and 4.2%, respectively, compared with the unsupplemented diets. From 1 to 35 d of age, broilers receiving high-AME diets had 2.9% lower (P < 0.05) FCR compared with those offered low-AME diets. Protease increased (P < 0.05) FCR by 1.0% compared with those offered unsupplemented diets. Jejunal (15 and 29 d of age) and ileal (29 d of age) starch digestibility and jejunal nitrogen digestibility (29 d of age) were lower (P < 0.05) in broilers offered high-AME diets compared with those offered low-AME diets. Both AME concentration and supplemental protease independently affected broiler performance, with responses being most apparent during early growth phases whereas digestibility measures were mainly influenced by AME concentration.

Influence of barley inclusion method and protease supplementation on growth performance, nutrient utilisation, and gastrointestinal tract development in broiler starters.

The influence of the method of barley inclusion (fine, coarse and whole barley) in a wheat-based diet and protease supplementation (0 and 0.20 g/kg) on growth performance, nutrient utilisation and gastrointestinal tract development of broilers (d 1 to 21) was evaluated in a 3 × 2 factorial arrangement. Whole barley (WB) grains were ground in a hammer mill to pass through the screen sizes of 2.5 and 8.0 mm to achieve fine (FB) and coarse (CB) barley particle sizes, respectively. A total of 288, one-day-old male broilers were allotted to 36 cages (6 cages/treatment; 8 birds/cage). There was no significant (P > 0.05) interaction between barley inclusion method and protease for any growth performance or nutrient utilisation parameters. Birds fed diets containing CB and WB showed higher (P < 0.05) weight gain, and digestibility of dry matter, nitrogen, calcium, gross energy, and ileal digestible energy compared to those fed FB diets. Compared to the birds fed FB diets, feed per gain was lower (P < 0.05) in birds fed diets made of WB. Fat digestibility of the birds fed CB was higher (P < 0.05) than those fed FB and WB birds. Compared to FB and CB diets, inclusion of WB resulted in heavier (P < 0.05) gizzards but reduced (P < 0.05) gizzard pH. Supplemental protease, however, had no effects (P > 0.05) on growth performance and nutrient utilisation, most likely due to the well balanced digestible amino acids and high inherent digestibility of protein in the basal diet, and/or the presence of exogenous carbohydrase and phytase. In conclusion, the present results showed that the inclusion of coarsely ground and whole barley in a wheat-based diet can enhance nutrient and energy utilisation and is beneficial to the growth performance of young broilers.

Assessment of crop usage in ad libitum fed birds and short-term phytase efficiency as affected by acid addition.

1. A field assessment was performed to map the extent of crop usage and thus retention time in broiler chickens. In addition, a broiler experiment was carried out to study the short-term effect of acid addition on phytase efficacy in the crop.2. In the field assessment, the crop content of 40 ad libitum fed broiler chickens from 4 different farms were sampled at 10, 20 and 30 d of age. The dry matter (DM) content varied from 0 to 32 g.3. From 11 d of age, 120 individually caged chickens were intermittently fed a high phytate-P diet with either no addition or 500 FYT C. braakii-derived phytase added or both phytase and 1.4% formic acid added. Excreta were collected for assessment of phosphorus (P) retention. At 20 and 21 d of age, starved birds were fed for 1 h, and thereafter crop and gizzard contents were collected every 20 min until 140 min after start of the feeding. At 60 and 140 min, the contents from the jejunum and ileum were collected.4. All diets reduced the concentration of phytate in the crop, however the combination of acid and phytase resulted in a higher degradation (P < 0.05) than the other diets from 20 min after the start of feeding. Simultaneously, the concentration of the smaller inositol phosphate isomers, such as inositol-5-phosphate, increased (P < 0.05). Phytase increased (P < 0.05) P retention, and the combination of acid and phytase increased jejunal P digestibility (P < 0.05) compared to the other diets.5. The results indicated that lower pH in the crop due to acid addition improved phytase efficacy and increased P digestibility in the anterior digestive tract, even with short retention times.

Effects of protease supplementation and diet type on jejunal and ileal digestibility and total tract metabolisability of nitrogen, starch, and energy in broilers.

1. A study was conducted to assess the effects of supplemental protease (0 or 15,000 units/kg) and diet type (maize- or wheat-based) on apparent jejunal and ileal digestibility and apparent total tract metabolisability of nutrients in Cobb 500 mixed-sex broilers from 6 to 31 d of age.2. Birds were randomly distributed into 56 metabolism cages (6 birds/cage; 14 replicates/treatment). At 22 d of age, jejunal and ileal digesta contents were collected and pooled from 4 birds/cage to determine apparent digestibility coefficients (DC) and digestible energy (DE). Feed intake was measured, and total excreta were collected from 8 to 11 and 18 to 21 d of age to determine apparent metabolisable energy (AME) and total tract nitrogen and starch metabolisability coefficients.3. Broilers offered the maize-based diet with protease had greater (P < 0.05) jejunal nitrogen DC, starch DC, and DE (8.2, 6.5, and 14.9%, respectively) and ileal nitrogen DC and DE (5.1 and 6.8%, respectively) than those offered the maize-based diet without protease. Ileal starch DC was increased (P < 0.05) by 1.1% with protease supplementation.4. Broilers offered maize-based diets had greater (P < 0.05) nitrogen (7.3%) and starch (0.6%) metabolisability coefficients and AME (4.7%) from 8 to 11 d of age, and nitrogen (4.3%) metabolisability coefficients and AME (2.0%) from 18 to 21 d of age compared with those offered wheat-based diets.5. Thus, protease supplementation and diet type can affect digestive dynamics and nutrient utilisation in broilers.

Contribution of purified soybean trypsin inhibitor and exogenous protease to endogenous amino acid losses and mineral digestibility.

The primary objective of the current study was to evaluate the impact of trypsin inhibitor (TI) and exogenous protease supplementation on endogenous loss of amino acids (AA) in broiler chickens. A total of 384 Cobb-500 broiler chicks were allocated to 4 nitrogen-free diets, each with 8 replicate cages and 12 birds per replicate. The diets were arranged as a 2 × 2 factorial with factors being dietary TI (0 or 8,000 TIU/g) and exogenous protease (0 or 15,000 PROT/kg). Desired dietary TI concentration was achieved by addition of commercially available, purified soybean TI. There was no effect of TI or exogenous protease or their interaction on growth performance of birds. However, the endogenous loss of nitrogen (N) and all AA increased (P < 0.05) due to dietary TI concentration except for Cys. The increase in endogenous AA due to TI ranged from 17% for Thr to 52.2% for Trp. Exogenous protease had no effect on endogenous loss of N and all AA. There was no effect of TI or exogenous protease or their interaction on the AID of P, however AID of Ca, Fe, Mg, Mn, and Cu was reduced (P < 0.05) due to dietary TI. The AID of Cu (P < 0.01) and K (P < 0.05) improved with exogenous protease supplementation. Significant interactions (P < 0.05) between exogenous protease and TI existed for Zn, Mg, Cu, and Na. The concentration (g/kg DM intake) of crude mucin and sialic acid increased (P < 0.05) with increased dietary TI. Decreased trypsin (P < 0.001) and increased chymotrypsin (P < 0.001) activity in the pancreas were observed as a result of exogenous protease supplementation. In conclusion, the current study showed that TI increases the endogenous loss of AA and reduced the digestibility of minerals in broiler chickens. Furthermore, exogenous protease did not affect endogenous AA flow, irrespective of added purified dietary TI.

Exogenous α-amylase improves the digestibility of corn and corn-soybean meal diets for broilers.

Starch is the main energy source in broiler diets. However, endogenous amylase secretion in young broilers is suboptimal to completely digest dietary starch, so exogenous α-amylase supplementation may help increase starch digestibility. The objective of this study was to assess the supplementation of increasing doses of an exogenous α-amylase (0, 40, 80, 120, and 160 kilo-novo α-amylase units (KNU)/kg) on corn and on a complete corn-soybean meal diet for 25-day-old broilers. Jejunal and ileal apparent digestibility coefficients of available starch, resistant starch, total starch, and DM, DM total tract retention, as well as dietary AME levels were evaluated. Interactions (P < 0.05) between diets and α-amylase showed that the enzyme had a more evident effect on increasing DM jejunal digestibility and AME on corn compared with the complete diet. Corn DM digestibility increased to a maximum of 67.84% with up to 47 KNU/kg, whereas 89 KNU/kg led to a maximum of 53.92% in the complete diet A maximum increase of 64 kcal AME/kg was obtained with 80 KNU/kg on the complete diet, whereas 109 KNU/kg generated 327 kcal AME/kg on corn (P < 0.05). Increasing the α-amylase dose linearly increased ileal digestibility of resistant starch (P < 0.05), and the effect on DM total tract retention was quadratic (P < 0.05). Corn showed a higher digestibility for DM, resistant and total starch, as well as DM total tract retention and AME, compared with the complete diet (P < 0.05). Treatments had no influence on available starch. The inclusion of exogenous α-amylase improves starch, DM, and energy utilization of corn-based and corn-soybean meal-based diets for broilers.

Research Note: The effect of sequential displacement of dietary dextrose with myo-inositol on broiler chicken growth performance, bone characteristics, ileal nutrient digestibility, and total tract nutrient retention.

A total of 480 male Cobb 500 broiler chicks were assigned to one of 6 dietary treatments to explore the energy equivalence of myo-inositol compared with dextrose. The 6 dietary treatments included a corn and soy-based control ration formulated with 5% anhydrous dextrose and 5 further diets that were generated by the sequential displacement of increments of 1% dextrose with myo-inositol. Each diet was fed to 8 replicate cages of 10 chicks per cage from day 8 to day 18 after hatch. The BW gain, feed intake, and feed conversion ratio (FCR) were measured, and on day 15 to day 17, excreta were collected to estimate the total tract nutrient retention. Ileal digestibility of nutrients and tibia mineral content was assessed on day 18. The displacement of dextrose with myo-inositol generated a significant linear reduction in the FCR that did not reach a plateau at 5% dietary inclusion of myo-inositol. There was no effect of the displacement of dextrose with myo-inositol on bone mineral concentration. However, supplemental myo-inositol linearly reduced ileal digestibility of DM, calcium, and ileal digestible energy. Myo-inositol addition resulted in a significant linear increase in the total tract retention of CP. It can be concluded that myo-inositol has an energy equivalence equal to approximately 78% of that of dextrose for young broiler chicks but exerts a range of extra caloric effects that improve feed efficiency and may influence nitrogen (N) retention and the uric acid cycle. Future work should focus on the role of phytase and myo-inositol on uric acid, creatine kinase, and other metabolites involved in renal function and biochemical flows of N in urine and feces in nonruminants.

Growth phase and dietary α-amylase supplementation effects on nutrient digestibility and feedback enzyme secretion in broiler chickens.

Growth performance, nutrient digestibility, intestinal health, and endogenous enzyme secretion responses to dietary α-amylase supplementation during 4 growth phases of broiler chickens fed corn-soybean meal-based diets were evaluated in the present study. A total of 1,136 male broiler chicks were assigned at day 0 after hatching to 8 treatments in a 2 × 4 factorial arrangement. There were 2 dietary levels of α-amylase supplementation of 0 or 80 kilo-Novo alpha amylase units per kg diet and 4 posthatching growth phases of day 0 to 11, day 11 to 21, day 21 to 42, or day 42 to 56 in a randomized complete block design. Each treatment comprised 8 replicate pens, with either 25 (day 0-11), 20 (day 11-21), 16 (day 21-42), or 10 (day 42-56) birds per pen. Body weight gain and feed efficiency of birds improved (P < 0.01) with α-amylase supplementation. There were main effects of α-amylase, growth phase, and interaction (P < 0.01) on apparent ileal digestibility (AID) of starch. This ranged from 0.8% during day 11 to 21 to 2.8% during day 0 to 11 after hatching. The total tract retention of starch increased (P < 0.05) with amylase supplementation but was not different across growth phases. Amylase supplementation increased (P < 0.05) AID of gross energy, AME (kcal/kg), and AMEn (kcal/kg). Villus height in the jejunal tissue was increased (P < 0.01) by α-amylase supplementation. During day 11 to 21 after hatching, the viscosity of jejunal digesta and pancreatic amylase activity increased (P < 0.01) with amylase supplementation. In conclusion, dietary amylase supplementation improved growth performance, apparent nutrient digestibility, and digestive enzyme activity of broiler chickens fed a corn-soybean diet. The study indicates that the growth phase of birds may affect response to exogenous amylase.

Possible role of available phosphorus in potentiating the use of low-protein diets for broiler chicken production.

A total of 945 male Ross 308 broiler chicks were used in a growth study to explore the interaction between dietary crude protein concentration and available phosphorus. Nine experimental treatments were constructed factorially by offering low, medium, or standard protein concentrations without or with low, standard, or high available phosphorus. Diets were based on corn, wheat, and soybean meal and all nutrients other than protein/amino acids and available phosphorus were maintained at or above breeder guidelines. Additional synthetic amino acids were used in the diets with low protein concentration in attempt to maintain digestible amino acid supply. Diets were offered to 7 replicate pens of 15 chicks per pen from day 8 to 35. Growth performance was measured during the grower (day 8-24) and finisher (day 25-35) periods. On day 35 carcass composition was determined, blood was drawn for various biochemical measurements and the tibia was excised for mechanical and compositional analyses. Birds that received the low-protein diet had lower terminal body weight and higher feed conversion ratio compared with those that received diets with adequate crude protein content. However, addition of available phosphorus to the low-protein diet resulted in significant reductions in weight-corrected feed conversion that were not evident in the diet with adequate protein content. Bone architecture was only moderately influenced by dietary treatment but birds that ingested the diets containing low and medium protein concentrations had relatively heavier abdominal fat pad weight. Blood biochemistry, especially ammonia, uric acid, and phosphorus, was influenced by both dietary protein and available phosphorus and trends suggested that both axes are involved in protein accretion and catabolism. It can be concluded that performance losses associated with feeding low protein diets to broiler chickens may be partially restored by additional available phosphorus. The implications for use of exogenous enzymes such as protease and phytase and protein nutrition per se warrants further examination.

Growth performance and amino acid digestibility responses of broiler chickens fed diets containing purified soybean trypsin inhibitor and supplemented with a monocomponent protease.

Trypsin inhibitors (TI) resident in soybeans affects protein utilization. While heat treatment influences residual TI, it simultaneously affects the structure and solubility of the soybean proteins and confounds any response to exogenous proteases. Using purified TI, the effect of exogenous protease to TI can be dissociated from changes in the soybean protein. Thus, the current study was designed to evaluate the growth performance and protein utilization responses of broiler chickens to purified TI and exogenous protease. Soybean meal (SBM) was preanalyzed for basal TI (2,996 TIU/g SBM), formulated into nutritionally adequate experimental diets to contain 1,033 TIU/g diet, and purified TI was added at 9,000 TIU/g diet. A total of 320 Cobb-500 broiler chicks were allocated to 4 diets, each with 8 replicate cages and 10 birds per replicate. The experimental diets were arranged as a 2 × 2 factorial with factors being dietary TI (1,033 or 10,033 TIU/g) and exogenous protease (0 or 15,000 PROT/kg). On day 7, 14, and 21 posthatching, protease supplementation improved the BW gain (P < 0.01) and gain to feed ratio (P < 0.05) of birds. On day 14 and 21 posthatching, the relative weight of pancreas increased (P < 0.05) with added TI but was reduced (P < 0.001) with protease supplementation. Apparent ileal digestibility of all amino acids, except methionine, decreased (P < 0.001) with added TI but increased (P < 0.05) with protease supplementation. Jejunal MUC-2 was downregulated (P < 0.01) and SCL7A-2 was upregulated (P < 0.05) by protease supplementation. Duodenal trypsin and chymotrypsin activities reduced (P < 0.05) with added TI but increased (P < 0.01) with protease supplementation. Exogenous protease produced longer villi (P < 0.05) and deeper crypts (P < 0.01) in the jejunal tissue. In conclusion, dietary addition of purified TI negatively affects nutrient utilization by broiler chickens. Furthermore, the study showed that the efficacy of the exogenous protease might be independent of dietary TI concentration.

Research Note: Delay in sampling influences the profile of phytate in gizzard digesta and ileal digestibility of phosphorus in broilers.

The objective of this study was to investigate the effect of different durations of time delay when sampling digesta from the gizzard and ileum of broilers on the degradation of myo-inositol hexakisphosphate (InsP6) and digestibility of phosphorus (P). There was 1 experimental diet with a supplemental phytase activity of 1,212 phytase units/kg feed, which was provided to birds from day 13 to 18 after hatching. The diet was formulated to provide 6.6 g/kg Ca and 1.9 g/kg nonphytate P and fed to 24 cages of 6 birds. The 24 cages of birds were further randomly divided into 6 subgroups of 4 cages from which the digesta samples in the gizzard and ileum were collected at 0, 5, 10, or 20 min postmortem. The results showed that the concentration of InsP6 decreased linearly (P = 0.002), InsP5 decreased quadratically (P = 0.038), and the summation of concentrations of P in InsP6-4 decreased linearly (P = 0.028) in the gizzard digesta with the increasing delay of sampling. In the ileum, the digestibility of phytate P tended to decrease linearly (P = 0.087), and the digestibility of total P decreased linearly (P = 0.026) with prolonged delay. In conclusion, delay in sampling could alter the measured profile of InsP esters in gizzard digesta probably because of a continued effect of supplemental phytase, while the ileal digestibility of total P could diminish. Therefore, standard sampling procedures should be implemented to minimize variance.

Toward standardized amino acid matrices for exogenous phytase and protease in corn-soybean meal-based diets for broilers.

A total of 468 male Ross 308 broilers were used in a digestibility study to determine the additivity of apparent or standardized amino acid (AA) digestibility values for corn, soybean meal (SBM), or a mixture of corn and SBM that were supplemented, or not, with either phytase, protease, or a combination of phytase and protease. These treatments generated a total of 12 experimental diets that were arranged in a 3 × 4 design. A nitrogen-free diet was also fed to estimate endogenous AA loss. Apparent and standardized AA digestibility values were assessed on day 28 posthatch. The apparent digestibility of AA in the complete diet was higher (P < 0.05) than expected based on the digestibility of the corn and SBM individually. However, this overestimation was corrected by the adjustment to standardized values. Importantly, addition of protease or the combination of protease and phytase increased (P < 0.05) the digestibility of AA in corn and SBM. Furthermore, these effects were arithmetically coherent with respect to the measured effects of the enzymes in the mixture of corn and SBM, even improving the additivity of AA digestibility values when assessed on an apparent basis. This study demonstrates that the effect of exogenous protease and phytase on AA digestibility in complete diets is predictable based on measurements made in individual ingredients. In addition to improving digestibility values per se, exogenous protease and phytase may enhance precision in least cost formulation systems.

The impact of age and feeding length on phytase efficacy during the starter phase of broiler chickens.

Phytase is of importance to the poultry industry because of its ability to hydrolyze phytate and release phosphorus (P) for use by poultry. However, the effect of age on phytase efficacy is not fully understood. A total of 864 day-old broiler chicks were used to investigate the effect of age and feeding length on phytase efficacy using growth performance, mineral utilization, and tibia ash as response criteria of evaluation. The experiment was arranged as a 3 × 2 × 2 factorial in a randomized complete block design with 3 diets including; a positive control (PC; 0.4% non-phytate P (nPP)), a negative control (NC; 0.2% nPP) and a NC diet supplemented with phytase at 2,000 FYT/kg; 2 ages (i.e., days 14 and 22); and 2 feeding lengths (i.e., 2 and 5 D) with 8 replicates each. Birds fed the NC had decreased (P < 0.01) body weight gain and feed efficiency compared with birds fed the PC regardless of age or feeding length. Similarly, birds fed the phytase-supplemented diet had improved (P < 0.01) performance as compared to birds fed the NC regardless of age. There were no significant differences in P utilization between birds fed for 2 to 14 D or 22 D and birds fed for 5 D to both ages. However, phytase was more efficacious at day 14 than day 22 when mineral utilization was considered because the super dose of phytase elicited greater response in birds fed the phytase supplemented diet for 2 D until day 14. In contrast, percentage tibia ash improved (P < 0.01) in birds fed phytase supplemented diet for 5 D at both ages as compared with birds fed for 2 D. In conclusion, testing phytase products, even at high doses, for 2 D during the second week in the life cycle of broiler chicks, can be recommended from the results of this study.

Utilization of corn-based diets supplemented with an exogenous α-amylase for broilers.

A study was conducted to evaluate the effects of a supplemental α-amylase on energy and nutrient utilization of broiler chickens fed diets with variable amounts of corn. A total of 480 slow feathering Cobb × Cobb 500 male broilers were randomly distributed into 10 treatments with 8 replicates of 6 birds each. Birds were fed a common starter to 14 D post-hatch. The experimental diets were provided subsequently until 25 D, which were a conventional corn-soy basal diet (CS) and a corn-soy diet displaced with 40% of corn (CN). These were fed as-is or supplemented with 40, 80, 120, or 160 kilo-Novo α-amylase units (KNU)/kg. Dietary treatments were distributed factorially as a 2 × 5 arrangement (diet type vs. amylase). Samples of feed, excreta, and ileal digesta were analyzed for determination of ileal digestible energy (IDE), ME, total tract retention, and digestibility of dry matter, protein, and fat. No interactions between diet type and amylase were observed. The CN diet had lower (P < 0.05) energy utilization and nutrient digestibility when compared to the CS diet. AMEn and IDE increased (P < 0.05) by 110 and 207 kcal/kg, respectively, when CS and CN diets were supplemented with 80 KNU/kg. The amylase added to the CS diet led to quadratic increases (P < 0.05) on growth performance, IDE, AME, and AMEn, as well as in dry matter, protein, and fat digestibility. Energy utilization and crude protein digestibility were linearly increased (P < 0.05) when amylase was added to the CN diet or the extrapolated 100% of corn. In conclusion, energy utilization, digestibility of crude protein, fat, and dry matter increased with amylase supplementation in corn-soy-based diets. When amylase was tested in a complete diet having 53.6% corn, 100 and 105 KNU/kg maximized AMEn and IDE, respectively; however, the maximum energy response in the CN diet or 100% of corn was not achieved until 160 KNU/kg, suggesting an association between amylase dose optimization and dietary starch concentration.

Influence of age and duration of feeding low-phosphorus diet on phytase efficacy in broiler chickens during the starter phase.

A total of 1,408 male broiler chickens were used to evaluate the impact of age and duration of feeding low-phosphorus (P) diet on the efficacy of phytase using growth performance, nutrient utilization, tibia ash, and plasma indices. Diets were formulated with 2 non-phytate P (nPP) concentrations (i.e., 0.20 and 0.40%) and 2 phytase concentrations (i.e., 1,000 and 2,000 FTU/kg) added to the 0.20% nPP diet. Four dietary treatments with 8 replicate cages each were fed to broiler chicks at different ages and for different duration. Specifically, these were days 6 to 8 (12 birds per replicate), 12 to 14, 9 to 14, 20 to 22, or 6 to 22 (8 birds per replicate). Parameters were measured on the last day of each period. Duration of feeding effect was examined by comparing responses of birds fed for 2 or 5 D at day 14 and for 2 or 16 D at day 22; age effect was determined by comparing responses of birds fed for 2 D at age 8, 14 and 22 D post-hatching. Body weight gain and gain-to-feed ratio were increased (P < 0.01) in birds fed diets supplemented with phytase; however, an increase in duration of feeding improved (P < 0.01) feed efficiency with birds fed for 16 D performing better than birds fed for 2 D at day 22. In addition, phytase supplementation improved (P < 0.01) apparent P and calcium digestibility and retention, but the age effect on phytase efficacy was more apparent at day 14 and the duration of feeding effect was evident in birds fed for 2 D due to the increased levels of mineral utilization at that age/duration of feeding as compared with the other groups. The results of this study show that phytase efficacy was at optimum in birds fed low-P diet for 2 D at day 14. This period can be recommended for further bioefficacy studies of phytase.

A mono-component microbial protease improves performance, net energy, and digestibility of amino acids and starch, and upregulates jejunal expression of genes responsible for peptide transport in broilers fed corn/wheat-based diets supplemented with xylanase and phytase.

A total of 90 male Ross 308 broiler chicks were used in a digestibility and performance bioassay to explore the effect of reduction in dietary protein and digestible amino acids and inclusion of an exogenous mono-component protease on amino acid digestibility, net energy, jejunal gene expression, and bird performance. Four dietary treatments were created by the supplementation, or not, of 2 control diets with a mono-component exogenous protease. The control diets were corn/wheat/soybean meal-based and were formulated to be either nutritionally adequate or reduced in protein and amino acids (around 3%). The 2 control diets were supplemented with xylanase and phytase (2000 FYT). Treatments were therefore arranged as a 2 × 2 factorial design. The reduction in diet nutrient density had no significant effect on various experimental outcomes (including bird performance, amino acid digestibility, and net energy [NE]) that were measured with the exception of a reduction in the expression of aminopeptidase N and glucose transporter 2. However, the addition of exogenous protease resulted in an increase in weight gain and a reduction in feed conversion ratio (around 4%; P < 0.05) and an increase in the digestibility of several amino acids (P < 0.05) and starch (P = 0.06). Protease addition also resulted in an increase in both apparent metabolizable energy (AME) (+73 kcal/kg; P < 0.05) and NE (+107 kcal/kg; P < 0.05). The addition of exogenous protease to the diet also increased the jejunal expression of genes responsible for peptide transport (PepT2; P < 0.01) and starch digestion (sucrase isomaltase; P = 0.06). These results confirm the efficacy of exogenous protease in broiler diets that contain both xylanase and phytase and suggest substantial beneficial effects that extend beyond protein and amino acid nutrition. The effect of exogenous protease on energy partitioning, starch digestibility and the efficiency of nitrogen cycling is an area for further study.