Novel 4th-generation phytase improves broiler growth performance and reduces woody breast severity through modulation of muscle glucose uptake and metabolism.

The objective of the present study was to determine the effect of a novel (4th generation) phytase supplementation as well as its mode of action on growth, meat quality, and incidence of muscle myopathies. One-day old male broilers (n = 720) were weighed and randomly allocated to 30 floor pens (24 birds/pen) with 10 replicate pens per treatment. Three diets were fed from hatch to 56- days-old: a 3-phase corn-soy based diet as a positive control (PC); a negative control (NC) formulated to be isocaloric and isonitrogenous to the PC and with a reduction in Ca and available P, respectively; and the NC supplemented with 2,000 phytase units per kg of diet (NC + P). At the conclusion of the experiment, birds fed with NC + P diet were significantly heavier and had 2.1- and 4.2-points better feed conversion ratio (FCR) compared to birds offered NC and PC diets, respectively. Processing data showed that phytase supplementation increased live weight, hot carcass without giblets, wings, tender, and skin-on drum and thigh compared to both NC and PC diets. Macroscopic scoring showed that birds fed the NC + P diet had lower woody breast (WB) severity compared to those fed the PC and NC diets, however there was no effect on white striping (WS) incidence and meat quality parameters (pH, drip loss, meat color). To delineate its mode of action, iSTAT showed that blood glucose concentrations were significantly lower in birds fed NC + P diet compared to those offered PC and NC diets, suggesting a better glucose uptake. In support, molecular analyses demonstrated that the breast muscle expression (mRNA and protein) of glucose transporter 1 (GLUT1) and glucokinase (GK) was significantly upregulated in birds fed NC + P diet compared to those fed the NC and PC diets. The expression of mitochondrial ATP synthase F0 subunit 8 (MT-ATP8) was significantly upregulated in NC + P compared to other groups, indicating intracellular ATP abundance for anabolic pathways. This was confirmed by the reduced level of phosphorylated-AMP-activated protein kinase (AMPKα1/2) at Thr172 site, upregulation of glycogen synthase (GYS1) gene and activation of mechanistic target of rapamycin and ribosomal protein S6 kinase (mTOR-P70S6K) pathway. In conclusion, this is the first report showing that in-feed supplementation of the novel phytase improves growth performance and reduces WB severity in broilers potentially through enhancement of glucose uptake, glycolysis, and intracellular ATP production, which used for muscle glycogenesis and protein synthesis.

Determination of the standardized ileal digestible calcium requirement of Ross broilers from hatch to day 14 post-hatch.

An experiment was conducted to determine the standardized ileal digestible (SID) calcium requirement of fast-growing broilers from hatch to d 14 post-hatch. Ross 308 (n = 360) male broilers were obtained on day of hatch and allocated to 1 of 5 treatments in battery cages. There were 6 birds per cage and 12 pens per treatment. Four treatments were formulated to contain 0.60%, 0.46%, 0.32% or 0.18% SID Ca. The final treatment was formulated using total Ca to meet or exceed all nutrient requirements, including 0.90% total Ca and 0.49% non-phytate P (nPP), using the same ingredients. This treatment was the reference diet for comparison and validation of the SID Ca diets. Birds and feed were weighed at placement and on d 14. Tibias and ileal contents were obtained on d 14 and excreta was collected per pen and pooled on d 14. Data were analyzed using JMP Pro and requirements were estimated using 3 different non-linear regression models. Increasing the SID Ca content in the diet from 0.18% to 0.60% improved (quadratic, P < 0.05) body weight gain and mortality corrected feed conversion ratio (mFCR). The estimated SID Ca requirement to optimize gain or mFCR was between 0.39% and 0.52%. Tibia ash percent and weight increased (quadratic, P < 0.05) as SID Ca content in the diet increased and the estimated SID Ca requirement was between 0.32% and 0.58%. Phosphorus utilization was improved in birds fed diets formulated using SID Ca compared with birds fed the reference diet. In conclusion, the SID Ca requirement of fast-growing broilers from hatch to d 14 was estimated between 0.534% and 0.398% when quadratic, straight-broken line, or quadratic-broken line regressions were used. These results agree with previously published data evaluating the SID Ca requirement of fast-growth broilers from hatch to d 10.

Phosphorus equivalency of exogenous phytase relative to phosphorus in monosodium phosphate in broiler chickens.

The objective of this study was to estimate phosphorus (P) equivalency of exogenous phytase relative to P from monosodium phosphate (MSP) in broiler chickens. To attain this objective, the impact of dietary MSP or exogenous phytase on growth performance, ileal digestibility of P, and bone characteristics in broiler chickens was assessed. Six experimental diets consisting of a P-deficient basal diet based on corn and soybean meal, basal diet plus 0.9 or 1.8 g/kg of inorganic P from MSP, and basal diet plus 500, 1,000, or 2,000 FYT/kg of exogenous phytase. Calcium to P ratio in all diets was maintained at 1.5:1. A total of 576 male broiler chickens (Cobb 500; initial BW = 190 ± 17 g) on d 8 post hatching were allotted to the 6 dietary treatments in a randomized complete block design using BW as a blocking factor. Each dietary treatment contained 8 replicate cages with 12 birds per cage. On d 11 post hatching, 7 birds from each cage were euthanized by CO2 asphyxiation and dissected for the collection of ileal digesta. On d 18, ileal digesta were also collected from the remaining 5 birds in each cage. The left femur and tibia were collected from the bird with the median BW on d 11 and 18 from each cage, and analyzed for bone breaking strength and bone ash. Weight gain and gain-to-feed ratio linearly or quadratically increased (P < 0.05) in every period as the inclusion rate of MSP or phytase increased. Ileal digestibility of P linearly increased (P < 0.05) on d 11 with increasing MSP, but there was no significant effect on d 18. Increasing phytase concentration linearly increased (P < 0.05) ileal digestibility of P on d 11 and 18. Increasing dietary MSP or phytase linearly increased ileal digestible P concentration in diets (P < 0.05). Bone breaking strength and bone ash linearly or quadratically increased (P < 0.05) with increasing inclusion rate of MSP or phytase on d 11 and 18. The equivalency of 1,000 FYT/kg of exogenous phytase based on dietary ileal digestible P concentration was 1.5 and 1.2 g/kg of inorganic P in diets on d 11 and 18, respectively. The current results showed that the supplementation of MSP or exogenous phytase can increase growth performance, ileal P digestibility, bone breaking strength, and bone ash in young broiler chickens.

Ileal mineral digestibility and expression of nutrient transporter genes of broiler chickens in response to variable dietary total Ca and phytase supplementation are influenced by time on experimental diet and age of the birds.

Two experiments were conducted to determine the impact of Ca, phytase, sampling time, and age on the digestibility (AID) of Ca and P and the expression of their transporters. Cobb 500 male chicks (N = 600) were used in each experiment and allocated to cages with 10 (Exp 1, 8-11 d) or 5 (Exp 2, 21-24 d) birds/cage and 10 (Exp 1) or 20 (Exp 2) reps/treatment. Treatments were a 2 × 3 factorial arrangement, with low (LOW) or standard (STD) Ca level and 3 phytase (PHY) levels (0, 300, or 3,000 FYT/kg). Ileal digesta were collected at 8, 12, 24, 48, and 72 h, and jejunum tissues at 12, 48, and 72 h after the start of feeding experimental diets. In Exp 1, there was no effect of Ca or phytase on the AID of Ca at 8, 12, or 24 h. Phytase increased the AID of P (P < 0.05) at all time points, and the magnitude was influenced by Ca. At 12 h, the mRNA level of P (NaPi-IIb) and Ca (CaSR) transporters was greatest in the LOW diets without phytase (Ca × PHY, P ≤ 0.06). In Exp 2, the STD diet decreased the AID of Ca and P (P < 0.05) at 8, 24, 48, or 72 h. Phytase increased the AID of Ca (P < 0.05) at 8, 12, and 24 h, and decreased the AID of Ca (quadratic, P < 0.05) in the STD diet (48 h). The AID of P (P < 0.05) increased with phytase at all sampling times. At 48 h, 3,000 FYT/kg decreased (P < 0.05) mRNA expression of NaPi-IIb and Ca transporter ATP2B1 in the STD diet (Ca × PHY, P < 0.05). In conclusion, to avoid adaptation of broilers to Ca and P deficiencies, the optimal time on experimental diets is ≤ 48 h for young broilers and ≤ 24 h in older birds due to up- or down-regulation of Ca and P transporters in response to dietary Ca, P, and phytase.

Description of 3 failed attempts to estimate the calcium equivalency of phytase for growth performance and tibia ash of broiler chickens when using graded dietary levels of limestone.

Three broiler experiments were conducted to estimate the Ca equivalency of a novel phytase using direct and indirect methods. All 3 experiments employed 4 concentrations of limestone to create 4 reference diets, deficient in nonphytate P, with increasing dietary Ca. Phytase was supplemented to the lowest Ca reference diet at 350, 700, 1,400, or 2,800 FYT/kg in experiment (Exp.) 1 and Exp. 2 and at 500, 1,000, 2,000, or 4,000 FYT/kg in Exp. 3. Broilers were fed from d 8 to 10 and 20 to 24, 19 to 21, or 7 to 10 and 7 to 21 posthatching in Exp. 1, 2, or 3, respectively. Diet did not affect growth performance or tibia ash in Exp. 1. Reducing the dietary Ca linearly (P < 0.05) increased body weight gain (BWG) and feed intake (FI) in Exp. 2 or Exp. 3. Feed conversion ratio (FCR) was decreased (linear or quadratic, P < 0.05) as dietary Ca was reduced in Exp. 2 or Exp. 3 (d 7-21). Tibia ash percent linearly (P < 0.05) decreased as dietary Ca decreased in Exp. 3 but only from d 7 to 21 and phytase increased (linear or quadratic, P < 0.05) FI and BWG, and decreased FCR. In Exp. 1 (d 8-10) and Exp. 2, apparent ileal digestibility (AID), total tract retention, and apparent digested and retained Ca or P increased (linear or quadratic, P < 0.05) as dietary Ca decreased. Phytase increased (linear or quadratic, P < 0.05) AID and apparent digested and retained Ca or P in Exp. 1 or Exp. 2. Due to the nature of the effect of dietary Ca on performance or tibia ash, it was not possible to use the indirect method to estimate the Ca equivalence of phytase in the current experiments. The total and digestible Ca equivalence of phytase could be estimated using the direct method. These experiments highlight challenges to consider when designing experiments to estimate the Ca equivalency for phytase in the future.

Influence of Dietary Phytase Inclusion Rates on Yolk Inositol Concentration, Hatchability, Chick Quality, and Early Growth Performance.

The aim of this study was to determine the influence of dietary phytase in breeder hens on yolk nutrients, hatchability, chick quality, and growth rate of their progeny, and their subsequent performance to 42 d post-hatch when fed diets with the same phytase concentrations. Breeder hens (n = 216) were divided into 3 groups receiving nutrient-adequate diets with reduced calcium (Ca) and phosphorus (P) (by 0.16% and 0.15%, respectively), supplemented with either 500, 1500, or 4500 FTU/kg phytase from 27 to 50 weeks of age. Eggs were collected at 38 weeks of age and incubated. On the day of hatch, the chick quality and hatchability were determined, and 18 chicks/group were euthanized for yolk sac collection and the determination of inositol and glycerol concentrations. The remaining chicks were divided into three groups, receiving different diets with reduced Ca and P (by 0.16% and 0.15%, respectively), supplemented with 0, 500, or 1500 FTU/kg phytase to 42 d post-hatch. Increasing the phytase concentration in the breeder hen diet linearly (p < 0.05) increased the number of early embryo deaths and decreased the number of late deaths and pips. The inositol concentration in the yolk sac at day of hatch increased (quadratic; p < 0.05) as the phytase dose increased in the breeder hen diet. The breeder hen diet (p < 0.05) influenced the body weight (BW), feed intake (FI), and feed conversion ratio (FCR) up to 21 days of age. The supplementation of breeder hen diets with 1500 FTU/kg phytase increased the concentration of sodium (Na), magnesium (Mg), potassium (K), manganese (Mn), and zinc (Zn) in the yolk sac. The inclusion of phytase doses up to 4500 FTU/kg appeared to influence embryo mortality, chick feed intake, and BW gain to 21 days and the FCR throughout the entire production phase.

The Growth Performance, Nutrient Digestibility, Gut Bacteria and Bone Strength of Broilers Offered Alternative, Sustainable Diets Varying in Nutrient Specification and Phytase Dose.

This study assessed the use of locally sourced sustainable feed ingredients, rapeseed meal (RSM) and maize dried distiller grains with solubles (DDGS) in diets over traditional ingredients on the growth performance, bone strength and nutrient digestibility of broilers. This work also investigated the effects of supplementing exogenous phytase in two doses (500 vs. 1500 FTU/kg). Using male Ross 308 chicks (n = 320) assigned to receive one of four experimental diets: (1) Positive control diet 1 (PC1), a wheat, soya-based diet + 500 FTU/kg phytase. (2) Positive control diet 2, RSM/DDGS diet + 500 FTU/kg phytase (PC2). (3) Negative control (NC) reduced nutrient RSM/DDGS diet, no phytase. (4) The NC diet plus 1500 FTU/kg phytase (NC+). PC1 birds displayed higher feed intake and body weight gain consistently throughout the trial (p < 0.001) as well as increased body weight by 28 d and 42 d (p < 0.001). Whole-body dual emission X-ray absorptiometry (DXA) analysis revealed PC1 birds also had higher bone mineral density (BMD), bone mineral content (BMC), total bone mass, total lean mass and total fat mass than birds offered other treatments (p < 0.01). Diet had no significant effect on bone strength. Phytase superdosing improved the digestibility of dry matter (DM), neutral detergent fibre (NDF), gross energy (GE), calcium (Ca), potassium (K) and magnesium (Mg) compared to birds in other treatment groups. The phytase superdose also improved performance in comparison to birds offered the NC diet. Phytase superdosing increased the IP6 and IP5 degradation and increased the ileal inositol concentration of the birds. N excretion was lower for birds offered the traditional wheat−soya diet and highest for those offered the high-specification RSM/DDGS diet with a commercial dose of phytase. The addition of a phytase superdose to the negative control diet (NC+) reduced P excretion of birds by 15% compared to birds offered NC.

Quantifying the effect of coccidiosis on broiler performance and infection outcomes in the presence and absence of control methods.

A systematic review and meta-analysis was conducted to investigate the role of Eimeria species, dose and inoculation time, on performance and infection outcomes of different broiler strains infected for different study durations. The meta-analysis addressed E. acervulina, E. maxima, E. tenella, and mixed species infections, and involved data from 72 peer-reviewed articles, corresponding to 521 treatments performed on 20,756 broilers. A secondary objective was to investigate the effects of synthetic anticoccidials, ionophores, and vaccination against Eimeria on the above outcomes. Performance during infection was scaled (%) to that of the uninfected birds. Infection reduced scaled ADFI and ADG (P < 0.001) and increased feed conversion ratio (FCR; P < 0.05); there was a significant interaction between dose and species on scaled ADFI and ADG, suggesting that different species affected these variables to different extents (P < 0.001). There was a tendency for an interaction between dose and broiler strain on scaled ADFI (P = 0.079), and a significant interaction between these variables on scaled ADG (P < 0.01). A tendency for an interaction between oocyst dose and Eimeria species (P = 0.067) on maximum number of oocysts excreted was observed. Lesion scores were significantly affected by dose, species, and their interaction (P < 0.05), the latter caused by an increase in the lesion scores during E. maxima and E. tenella infections. Control methods significantly affected scaled ADG and FCR (P < 0.05) and there was an interaction between dose and control methods on ADFI (P < 0.001). Synthetic anticoccidial use improved scaled ADG (P < 0.01), whereas ionophores improved FCR compared with untreated birds (P < 0.01). An interaction between dose and control method on scaled ADFI was caused by the higher ADFI of vaccinated compared to untreated birds, as dose increased. There was a significant effect of control methods on lesion scores (P < 0.01). All findings advance our understanding of the factors that influence the impact of coccidiosis and its controls in broilers.

Dietary Oxidative Distress: A Review of Nutritional Challenges as Models for Poultry, Swine and Fish.

The redox system is essential for maintaining cellular homeostasis. When redox homeostasis is disrupted through an increase of reactive oxygen species or a decrease of antioxidants, oxidative distress occurs resulting in multiple tissue and systemic responses and damage. Poultry, swine and fish, raised in commercial conditions, are exposed to different stressors that can affect their productivity. Some dietary stressors can generate oxidative distress and alter the health status and subsequent productive performance of commercial farm animals. For several years, researchers used different dietary stressors to describe the multiple and detrimental effects of oxidative distress in animals. Some of these dietary challenge models, including oxidized fats and oils, exposure to excess heavy metals, soybean meal, protein or amino acids, and feeding diets contaminated with mycotoxins are discussed in this review. A better understanding of the oxidative distress mechanisms associated with dietary stressors allows for improved understanding and evaluation of feed additives as mitigators of oxidative distress.

Research Note: Dietary phytase reduces broiler woody breast severity via potential modulation of breast muscle fatty acid profiles.

Woody breast (WB) myopathy is a major concern and economic burden to the poultry industry, and for which, there is no effective solution because of its unknown etiology. In a previous study, we have shown that phytase (Quantum Blue, QB) reduces the WB severity by 5% via modulation of oxygen homeostasis-related pathways. As WB has been suggested to be associated with lipid dysmetabolism, we aimed to determine the effect of QB on WB and breast muscle fatty acid profile. Male broilers were subjected to 6 treatments (96 birds/treatment): a nutrient adequate control group (PC), the PC supplemented with 0.3% myo-inositol (PC + MI), a negative control (NC) deficient in available P and Ca by 0.15 and 0.16%, respectively, the NC fed with QB at 500 (NC+500 FTU), and 1,000 (NC+ 1,000 FTU) or 2,000 FTU/kg of feed (NC+2,000 FTU). Woody breast and white striping scores were recorded, and fatty acid profiles were determined using gas liquid chromatography. Woody breast-affected muscles exhibited a significant higher incidence of white striping as liquid chromatography analysis reveals an imbalance of fatty acid profile in the breast of WB-affected birds with a significant higher percent of saturated fatty acids (SFA, myristic [14:0], pentadecanoic [15:0], and margaric [17:0]) and monounsaturated fatty acids (myristoleic [14:1], palmitoleic [16:1c], 10-trans-heptadecenoic [17:1t], oleic [18:1c9], and vaccenic [18:1c11]), and lower content of polyunsaturated fatty acids (PUFA) and omega-3 (P < 0.05). Quantum Blue at high doses (1,000 and 2,000 FTU) significantly reduces the percent of SFA and increases that of PUFA compared with the control group. In conclusion, WB myopathy seemed to be associated with an imbalance of fatty acid profile, and QB ameliorates the severity of WB potentially via modulation of SFA and PUFA contents.

How do pigs deal with dietary phosphorus deficiency?

Feeding strategies for growing monogastric livestock (particularly pigs) must focus on maximising animal performance, while attempting to reduce environmental P load. Achieving these goals requires a comprehensive understanding of how different P feeding strategies affect animal responses and an ability to predict P retention. Although along with Ca, P is the most researched macromineral in pig nutrition, knowledge gaps still exist in relation to: (1) the effects of P feed content on feed intake (FI); (2) the impact of P intake on body composition; (3) the distribution of absorbed P to pools within the body. Here, we address these knowledge gaps by gathering empirical evidence on the effects of P-deficient feeds and by developing a predictive, mechanistic model of P utilisation and retention incorporating this evidence. Based on our statistical analyses of published literature data, we found: (1) no change in FI response in pigs given lower P feed contents; (2) the body ash-protein relationship to be dependent upon feed composition, with the isometric relationship only holding for pigs given balanced feeds and (3) the priority to be given towards P retention in soft tissue over P retention in bones. Subsequent results of the mechanistic model of P retention indicated that a potential reduction in P feeding recommendations could be possible without compromising average daily gain; however, such a reduction would impact P deposition in bones. Our study enhances our current knowledge of P utilisation and by extension excretion and could contribute towards developing more accurate P feeding guidelines.

Dietary Inositol Reduces Fearfulness and Avoidance in Laying Hens.

Myo-inositol (inositol) affects memory, and the incidence of depression and anxiety in mammals. An experiment was designed to determine if pure inositol (0.16%), or high levels of phytase (3000 FTU/kg) affect the behaviour of fully beaked Lohmann LSL lite hens fed amino acid sufficient (19% crude protein (CP)) and deficient diets (16% CP), from 19 to 59 weeks of age. The data collected included live-scan behaviour observations and novel object (NO) tests (both at 1, 10 and 40 weeks of the trial); heterophil-to-lymphocyte (H/L) ratios (week 1 and week 40 of the trial); end of trial feather cover, and comb and skin lesions; and daily mortality. Reducing CP increased sitting by 2.5%. Inositol, but not phytase, reduced the latency to peck at the NO by 300 sec. Inositol reduced vent feather cover by 12% and tended to increase mortality by 13%. No effects on H/L ratio, and comb or skin lesions were found. In conclusion, regardless of the source, inositol reduced vent feather cover, while it tended to increase mortality. Only pure inositol reduced fearfulness in laying hens.

Quantum Blue Reduces the Severity of Woody Breast Myopathy via Modulation of Oxygen Homeostasis-Related Genes in Broiler Chickens.

The incidence of woody breast (WB) is increasing on a global scale representing a significant welfare problem and economic burden to the poultry industry and for which there is no effective treatment due to its unknown etiology. In this study, using diffuse reflectance spectroscopy (DRS) coupled with iSTAT portable clinical analyzer, we provide evidence that the circulatory- and breast muscle-oxygen homeostasis is dysregulated [low oxygen and hemoglobin (HB) levels] in chickens with WB myopathy compared to healthy counterparts. Molecular analysis showed that blood HB subunit Mu (HBM), Zeta (HBZ), and hephaestin (HEPH) expression were significantly down regulated; however, the expression of the subunit rho of HB beta (HBBR) was upregulated in chicken with WB compared to healthy counterparts. The breast muscle HBBR, HBE, HBZ, and hypoxia-inducible factor prolyl hydroxylase 2 (PHD2) mRNA abundances were significantly down regulated in WB-affected compared to normal birds. The expression of HIF-1α at mRNA and protein levels was significantly induced in breasts of WB-affected compared to unaffected birds confirming a local hypoxic status. The phosphorylated levels of the upstream mediators AKT at Ser473 site, mTOR at Ser2481 site, and PI3K P85 at Tyr458 site, as well as their mRNA levels were significantly increased in breasts of WB-affected birds. In attempt to identify a nutritional strategy to reduce WB incidence, male broiler chicks (Cobb 500, n = 576) were randomly distributed into 48 floor pens and subjected to six treatments (12 birds/pen; 8 pens/treatment): a nutrient adequate control group (PC), the PC supplemented with 0.3% myo-inositol (PC + MI), a negative control (NC) deficient in available P and Ca by 0.15 and 0.16%, respectively, the NC fed with quantum blue (QB) at 500 (NC + 500 FTU), 1,000 (NC + 1,000 FTU), or 2,000 FTU/kg of feed (NC + 2,000 FTU). Although QB-enriched diets did not affect growth performances (FCR and FE), it did reduce the severity of WB by 5% compared to the PC diet. This effect is mediated by reversing the expression profile of oxygen homeostasis-related genes; i.e., significant down regulation of HBBR and upregulation of HBM, HBZ, and HEPH in blood, as well as a significant upregulation of HBA1, HBBR, HBE, HBZ, and PHD2 in breast muscle compared to the positive control.

Influence of the concentration of dietary digestible calcium on growth performance, bone mineralization, plasma calcium, and abundance of genes involved in intestinal absorption of calcium in pigs from 11 to 22 kg fed diets with different concentrations of digestible phosphorus.

BACKGROUND: A 21-day experiment was conducted to test the hypothesis that Ca requirements to maximize growth performance expressed as the standardized total tract digestible (STTD) Ca to STTD P ratio is less than 1.40:1. The second hypothesis was that increasing dietary Ca increases plasma Ca concentration and downregulates abundance of genes related to Ca absorption (TRPV6, S100G, and ATP2B1) in the duodenum, and tight junction proteins (OCLN, CLDN1, and ZO1) in the duodenum and ileum. METHODS: Twenty corn-soybean meal diets were formulated using a 4 × 5 factorial design with diets containing 0.16%, 0.33%, 0.42%, or 0.50% STTD P, and 0.14%, 0.29%, 0.44%, 0.59%, or 0.74% STTD Ca. Six hundred and forty pigs (initial weight: 11.1 ± 1.4 kg) were allotted to 20 diets and 5 blocks in a randomized complete block design. On day 21, weights of pigs and feed left in feeders were recorded and blood, duodenal tissue, ileal mucosa, and the right femur were collected from 1 pig per pen. Abundance of mRNA was determined in duodenal and ileal tissue via quantitative RT-PCR. Data were analyzed using a response surface model. RESULTS: The predicted maximum ADG (614 g), G:F (0.65), and bone ash (11.68 g) was obtained at STTD Ca:STTD P ratios of 1.39:1, 1.25:1, and 1.66:1, respectively, when STTD P was provided at the requirement (0.33%). If dietary STTD P was below the requirement, increasing dietary Ca resulted in reduced (P < 0.05) ADG and G:F. However, if dietary STTD P was above the requirement, negative effects (P < 0.05) on ADG and G:F of increasing STTD Ca were observed only if dietary STTD Ca exceeded 0.6%. Plasma Ca concentration was positively affected by STTD Ca over the range studied (quadratic, P < 0.01) and negatively affected by increasing STTD P (linear, P < 0.01). There was a linear negative effect (P < 0.05) of STTD Ca on the abundance of S100G, TRPV6, OCLN, and ZO1 in duodenum, and CLDN and ZO1 in ileum. CONCLUSIONS: The STTD Ca:STTD P ratio needed to maximize growth performance of 11- to 25-kg pigs is less than 1.40:1, if P is at the estimated requirement. Increasing dietary Ca reduces transcellular absorption of Ca and increases paracellular absorption of Ca.

Standardized total tract digestibility of calcium varies among sources of calcium carbonate, but not among sources of dicalcium phosphate, but microbial phytase increases calcium digestibility in calcium carbonate1.

Two experiments were conducted to test the hypothesis that standardized total tract digestibility (STTD) of Ca and the response to microbial phytase is constant among different sources of Ca carbonate and that the STTD of Ca is constant among different sources of dicalcium phosphate (DCP) when fed to growing pigs. In Exp. 1, 80 pigs (initial BW: 19.0 ± 1.9 kg) were randomly allotted to 10 diets and 2 blocks with 4 pigs per diet in each block. Four sources of Ca carbonate were used, and each source was included in a diet without microbial phytase and a diet with microbial phytase (500 units/kg diet). Two Ca-free diets without or with microbial phytase were also formulated. Feed allowance was 2.7 times the maintenance energy requirement for ME and daily feed allotments were divided into 2 equal meals. The initial 4 d of each period were considered the adaptation period to the diets followed by 4 d of fecal collection using the marker-to-marker procedure. Pigs fed diets containing exogenous phytase had lower (P < 0.05) basal endogenous loss of Ca compared with pigs fed diets containing no phytase. There were no interactions between phytase and source of Ca carbonate. Values for STTD of Ca were greater (P < 0.05) for diets containing microbial phytase (77.3% to 85.4%) compared with diets without exogenous phytase (70.6% to 75.2%), and values for STTD of Ca differed (P < 0.05) among the 4 sources of Ca carbonate. In Exp. 2, 40 pigs (initial BW: 14.9 ± 1.3 kg) were allotted to a completely randomized design with 5 diets and 8 replicate pigs per diet. A basal diet in which all Ca was supplied by Ca carbonate was formulated. Three diets were formulated by adding 3 sources of DCP to the basal diet and a Ca-free diet was also used. Feeding and collection methods were as described for Exp. 1. Results indicated that values for STTD of Ca and ATTD of P were not different among diets, indicating that under the conditions of this experiment, the digestibility of Ca and P in DCP appears to be constant regardless of origin of DCP. In conclusion, use of microbial phytase reduces the basal endogenous loss of Ca and increases Ca digestibility in Ca carbonate. The STTD of Ca varies among sources of Ca carbonate, regardless of phytase inclusion, but that appears not to be the case for the STTD of Ca in different sources of DCP.

Basal endogenous loss, standardized total tract digestibility of calcium in calcium carbonate, and retention of calcium in gestating sows change during gestation, but microbial phytase reduces basal endogenous loss of calcium1.

The objective was to test the hypothesis that the standardized total tract digestibility (STTD) of Ca and the response to microbial phytase on STTD of Ca and apparent total tract digestibility (ATTD) of P in diets fed to gestating sows are constant throughout gestation. The second objective was to test the hypothesis that retention of Ca and P does not change during gestation. Thirty-six gestating sows (parity = 3.3 ± 1.5; d of gestation = 7 d) were allotted to 4 diets. Two diets containing 0 or 500 units of microbial phytase per kilogram were based on corn, potato protein concentrate, and calcium carbonate. Two Ca-free diets were also formulated without or with microbial phytase to estimate basal endogenous loss of Ca. Daily feed allowance was 1.5 times the maintenance energy requirement. Sows were housed individually in gestation stalls and fed a common gestation diet, but they were moved to metabolism crates from days 7 to 20 (early gestation), days 49 to 62 (midgestation), and again from days 91 to 104 (late gestation). When sows were in metabolism crates, they were fed experimental diets and feces and urine were quantitatively collected for 4 d after 4 d of adaptation. Results indicated that outcomes were not influenced by the interaction between period of gestation and dietary phytase. The basal endogenous loss of Ca was greater (P < 0.05) by sows in early gestation than by sows in mid- or late-gestation, but supplementation of microbial phytase to the Ca-free diet decreased (P < 0.01) the basal endogenous loss of Ca and tended (P = 0.099) to increase ATTD of P. Supplementation of microbial phytase did not affect ATTD of DM, STTD of Ca, Ca retention, ATTD of P, or P retention in sows fed the calcium carbonate-containing diet. The ATTD of DM was not affected by period of gestation, but the ATTD of Ca, the ATTD of P, and the retention of Ca were least (P < 0.05) in midgestation, followed by early and late gestation, respectively, and the STTD of Ca in midgestation was also reduced (P < 0.05) compared with sows in early or late gestation. Phosphorus retention was greater (P < 0.05) in late gestation than in the earlier periods. In conclusion, Ca retention was less negative and ATTD of P tended to increase with supplementation of microbial phytase to the Ca-free diet regardless of gestation period. The basal endogenous loss, STTD of Ca, ATTD of P, and retention of Ca and P in gestating sows change during gestation with the greatest digestibility values observed in late gestation.

Phytase as an alleviator of high-temperature stress in broilers fed adequate and low dietary calcium.

This study aimed to investigate the effect of phytase and dietary Ca level on performance, phytate (inositol hexa-phosphate, IP6) concentration, and anti-oxidant enzyme activity in broilers exposed to cycling high environmental temperatures. A total of 2,970 day-old Cobb 400 male broilers were randomly allocated among 6 treatments, with 15 replicate pens, and 33 birds per pen. Corn-soy diets low in available phosphorus (avP, 0.15% matrix applied) were fed in 2 phases; starter (0 to 21 d) and grower (22 to 42 d). Diets were factorially arranged by 2 × 3, with adequate (0.90% starter, 0.80% grower) or low (0.60% starter, 0.50% grower) dietary Ca and phytase (0, 500, or 3,000 phytase units (FTU)/kg). Temperature was not regulated, but followed environmental changes determined by the summer season in India; 22 to 39°C in the morning and 23 to 40°C at night. At 42 d of age, supplementing either dose of phytase increased (P < 0.001) weight gain (+128 g) and feed intake (+194 g) of birds, while only 3,000 FTU/kg phytase lowered (P < 0.01) feed conversion ratio (FCR) compared to the control. Birds fed low Ca diets had poorer (P < 0.01) weight gain (-33 g) and FCR (+0.04) than birds fed adequate Ca levels, suggesting that Ca was limiting in these diets. Phytase and low dietary Ca reduced the concentration of IP esters in the gastrointestinal tract of broilers. Lower IP6, IP5, IP4, and IP3 concentrations and increased myo-inositol provision was positively correlated to anti-oxidant enzyme activity in tissues. These results indicate that phytase can be used to alleviate the negative performance traits accredited to broilers under high temperature stress, potentially by reducing the anti-nutritive effect of IP6 and improving anti-oxidant status of birds. However, caution should be taken when reducing dietary Ca levels as improvements in IP6 degradation and anti-oxidant status cannot overcome the constraints on growth by diets that do not meet the Ca requirements.

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

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 lysine (dgLys). Cobb 400, male broilers were allocated to 1 of 16 diets consisting of 4 basal diets formulated at 80, 88, 96, or 104% of the Cobb 400 dgLys requirements for each feeding phase. Phytase was included in each basal diet at 0, 750, 1,500, or 3,000 phytase units (FTU)/kg. In Exp. 1, 33 birds/pen from hatch to day 42 were fed a 2-phase feeding program with 12 replicate pens/diet. In Exp. 2, there were 25 birds/pen from hatch to day 21 and 8 replicate pens/diet. Data were analyzed as a 4 × 4 factorial and means separated using orthogonal contrasts. In Exp. 1, feed intake (FI) increased (quadratic, P < 0.05) as dgLys increased in the diet. Body weight gain (BWG) increased (quadratic, P < 0.05) as dgLys concentration or phytase dose increased in the diet. As phytase dose increased in the diet, feed conversion ratio (FCR) was improved in a linear or quadratic (P < 0.05) manner depending on the dgLys concentration of the diet (dgLys × phytase, P<0.05). In Exp. 2, FI linearly (P < 0.05) increased as dgLys increased in the diet. Increasing the concentration of dgLys or phytase in the diet increased (quadratic, P < 0.05) BWG and improved (quadratic, P < 0.05) FCR. The AID of most amino acids was influenced by a dgLys × phytase interaction (P < 0.05), except threonine, valine, tryptophan, serine, cysteine, or leucine (linear or quadratic effect of phytase, P < 0.05), where phytase improved the AID in birds fed diets containing 80, 88, or 96% of the dgLys requirement, but not birds fed 104%. The predicted dgLys requirement to maximize performance, carcass, and digestible lysine intake was 97.6 to ≥ 104%. The predicted dose of phytase to maximize BWG or FCR was between 1,990 and 2,308 FTU/kg, regardless of the dgLys concentration in the diet. The predicted dose of phytase to maximize carcass weight was between 1,527 and 2,658 FTU/kg of diet and to maximize breast weight was 0 to ≥ 3,000 FTU/kg diet, depending on the dgLys concentration in the diet. In conclusion, optimal performance in the absence of phytase could be achieved at much lower levels of lysine in the presence of phytase.

Effects of microbial phytase on mucin synthesis, gastric protein hydrolysis, and degradation of phytate along the gastrointestinal tract of growing pigs.

An experiment was conducted to test the hypothesis that pigs fed diets supplemented with exogenous phytase reduce mucin synthesis in the small intestine, increase protein hydrolysis in the stomach, increase breakdown of phytate along the gastrointestinal tract, and increase mineral and AA digestibility. A diet based on corn, soybean meal, and canola meal was formulated to meet requirements for growing pigs except for Ca and P, which were lower than requirements. Three additional diets were formulated by adding 750, 1,500, or 3,000 units of phytase (FTU) per kilogram to the basal diet. Eight growing barrows (38.45 ± 3.06 kg) were prepared with a T-cannula in the duodenum and another T-cannula in the distal ileum. Pigs were housed individually and allotted to a replicated 4 × 4 Latin square design with four pigs and four periods in each square. Each period lasted 14 d with the initial 7 d being the adaptation period to the diets. Pigs were fed twice daily in combined amounts equal to 3.2 times the estimated requirement for maintenance energy. Results indicated that the apparent ileal digestibility (AID) and the apparent total tract digestibility (ATTD) of Ca and P increased (linear and quadratic, P ≤ 0.05) as phytase inclusion increased. However, values for AID of Ca and P were not different from values for ATTD of Ca and P, indicating that there is no net absorption of Ca and P in the hindgut. The apparent duodenal digestibility (ADD) of Ca and P was ~30% and 10% to 20%, respectively, indicating some digestion in the stomach of both Ca and P. A quadratic increase (P < 0.05) of the AID of GE was observed with the breakpoint around 1,500 FTU, but there was a negative linear (P ≤ 0.001) effect of dietary phytase on the ATTD of GE. Phytase did not affect mucin synthesis in the small intestine, protein hydrolysis in the stomach, or ileal digestibility of dispensable and indispensable AA. However, degradation of higher phytate esters (IP6 and IP5) into lower phytate esters (IP4 and IP3) and inositol increased as dietary phytase increased, indicating that it is possible to completely degrade dietary phytate if microbial phytase is included by at least 3,000 FTU in the diet. In conclusion, supplementing diets with phytase resulted in increased degradation of phytate and phytate esters and improved digestibility of Ca and P, but phytase did not change intestinal mucin synthesis, gastric protein hydrolysis, or the AID of AA.

Evaluating phosphorus release by phytase in diets fed to growing pigs that are not deficient in phosphorus.

Microbial phytase is widely used to enhance digestibility of phytate-P. By tradition, diets with P content well below requirement are used to quantify phytate-P release by phytase, but P-adequate diets may be more physiologically relevant. The objective of this study was to investigate the effects of phytase on P digestion and metabolism and develop a P release curve for phytase in P-adequate diets (above requirement according to NRC, 2012), and to compare these effects in a P-deficient diet. Three replicates of 24 barrows each (BW = 23.0 ± 1.8 kg) were randomly assigned to 1 of 8 dietary treatments, housed in individual pens for 21 d, then moved to metabolism crates for 5 d urine and fecal collections. A basal corn-soybean meal diet (P-adequate, A) was formulated at 0.36% standardized total tract digestible (STTD) P and total Ca:STTD P of 1.83. Phytase was added to A at 200 (A200), 400 (A400), 600 (A600), and 800 (A800) phytase units (FTU)/kg. A positive control diet (PC) was formulated using monocalcium phosphate (MCP) to increase STTD P by 0.16% to 0.52%, the expected STTD P release of 800 FTU/kg. A P-deficient diet (D) was formulated by reducing MCP to achieve 0.21% STTD P, and 200 FTU phytase/kg was added to D for D200. Pig was the experimental unit, and replicate and dietary treatment were fixed effects. Orthogonal polynomial contrasts were used to test linear and quadratic effects of phytase within A, A200, A400, A600, and A800. Phytase increased percent apparent total tract digestibility (ATTD) and STTD of P (quadratic P < 0.001), and quantity of absorbed P (linear P < 0.001; quadratic P = 0.069). Urinary P increased linearly with phytase (P < 0.001) and retained P also increased (linear P = 0.001, quadratic P = 0.094). Phytate-P release was estimated to be 0.049, 0.080, 0.093, and 0.09% STTD P for 200, 400, 600, and 800 FTU/kg, respectively. It appears that the effect of phytase may be lower in P-adequate diets as compared to P-deficient diets, given that there was a 12% improvement for A200 versus A, and a 28% improvement in STTD P for D200 versus D. In conclusion, phytase improved P digestibility and retention in P-adequate diets, and P digestibility was used to estimate the quantity of P released by phytase. Further research investigating P release by phytase in P-adequate diets, rather than P-deficient diets, may be preferable.