Exploratory transcriptomic analysis in muscle tissue of broilers fed a phytase-supplemented diet.

The effect of phytase on phosphorus retention, broiler (Gallus gallus) performance and bone mineralization in diets with reduced inorganic phosphate concentration is well documented. Furthermore, so-called 'extra-phosphoric' effects of phytase have been described in the literature that may be associated with changes in mineral and amino acid partitioning and requirements per se. In particular, the role of myo-inositol in phytase responses is implied but not well elucidated. It was the purpose of the experiment reported herein to explore the effect of phytase on broiler growth, nutrient digestibility, blood biochemistry and gene expression. A 5-week broiler floor pen trial was conducted to evaluate the effect of supplementation of a moderately phosphorus-deficient diet with 1000 U/kg of a 6-microbial phytase. Parameters measured were growth performance, phosphorus (P), calcium (Ca) and myo-inositol plasma concentrations, apparent ileal P digestibility, bone mineralization, breast meat weight and Pectoralis major muscle transcriptome. Supplementation of the diet with phytase improved weight gain during the starter period (18%) and the whole period (24%) compared with animals that received the control diet (p < 0.05). Improved feed conversion ratio, increased myo-inositol plasma concentration, tibia ash contents and breast meat weight were also observed in animals fed phytase. The transcriptomic analysis revealed that some differentially expressed genes (DEG) in broilers, receiving phytase in comparison with animals fed reduced phosphorus diet without phytase, were part of pathways involved in muscle development, via calmodulin/calcineurin and insulin-like growth factor. Microarray data confirmation was performed on six genes by quantitative PCR (qPCR): PI3K regulatory and catalytic subunit, Phospholipase C beta, Myocyte Enhancer Factors 2A and 2C, and calcineurin A. The results suggested that dietary supplementation with this phytase could generate low molecular weight phytate esters and indirectly myo-inositol, and could help us to understand how muscle metabolism may be affected at a gene level.

The interplay of dietary nutrient specification and varying calcium to total phosphorus ratio on efficacy of a bacterial phytase: 1. Growth performance and tibia mineralization.

A 14-d experiment was conducted to study the effects of 2 dietary variables on efficacy of a 6-phytase from Citrobacter braakii on broiler growth performance and tibia mineralization. Diets were formulated with or without nutrient matrix values for phytase as negative or positive control (NC or PC, respectively) and with 2 Ca:total P (tP; 2:1 or 2.5:1). The diets were supplemented with 0, 1,000, or 2,000 phytase units (FYT)/kg, thus producing a 2 × 2 × 3 factorial arrangement. Birds and feed were weighed on d 7 and 21, and tibia bones were collected from all the birds on d 21. The main effects of nutrient matrix, Ca:tP, and phytase supplementation were significant (P < 0.05) for all the growth performance responses (except for G:F for which there was no effect of matrix). The Ca:tP × phytase and matrix × phytase interactions were significant (P < 0.05) for weight gain. In the PC diets, phytase increased weight gain (P < 0.05) relative to the control only in diets with 2,000 FYT/kg, whereas in NC diets weight gain increased (P < 0.01) only from 0 to 1,000 FYT/kg levels. Broilers consuming diets with 2.5:1 Ca:tP had lower (P < 0.05) tibia ash, whereas phytase increased (P < 0.01) tibia ash, Ca, P, and Zn but decreased (P < 0.01) tibia K. Phytase supplementation of diets with 2:1 Ca:tP increased (P < 0.05) tibia P in birds receiving 1,000 FYT/kg relative to the control with no further increase at 2,000 FYT/kg, whereas each level of phytase supplementation increased (P < 0.05) tibia P in the diets with 2.5:1 Ca:tP. It was concluded that the best response to lower phytase supplementation (1,000 FYT/kg) was in NC diets with narrow Ca:tP, whereas the best response to higher level of phytase supplementation (2,000 FYT/kg) was achieved in diets in PC diets with wide Ca:tP.

The interplay of dietary nutrient level and varying calcium to phosphorus ratios on efficacy of a bacterial phytase: 2. Ileal and total tract nutrient utilization.

A 14-d broiler experiment was conducted to assess the effects of 2 dietary variables on efficacy of a bacterial 6-phytase from Citobacter braakii on nutrient and phytate P (PP) utilization. Diets were formulated with or without nutrient matrix values (matrix) for phytase as negative control (NC) or positive control (PC), respectively, and with 2 Ca:total P (tP) levels (2:1 or 2.5:1). The diets were supplemented with 0, 1,000, or 2,000 phytase units (FYT)/kg of diet, thus producing a 2 × 2 × 3 factorial arrangement. Excreta were collected on d 19 to 21 and ileal digesta on d 21. There was no 3-way interaction on digestibility of any nutrient. There was matrix × phytase (P < 0.01) interaction for Ca and DM digestibility and Ca:tP × phytase interaction (P < 0.05) for acid hydrolyzed fat and Ca and P digestibility. Prececal flow of Mn, Zn, and Na was greater (P < 0.05) in NC diets, whereas phytase increased (P < 0.05) prececal flow of Mg, Fe, Mn, and Zn but decreased (P < 0.05) prececal Na flow. Total tract PP disappearance and total tract Ca retention increased (P < 0.05) with phytase supplementation in diets with 2:1 Ca:tP, whereas there was no effect of phytase supplementation on PP disappearance or Ca retention in diets with 2.5:1 Ca:tP. Total P and Ca retention were reduced (P < 0.05) in PC and NC diets when Ca:tP increased to 2.5:1, but the depression was more pronounced in the NC diet. In addition, PP disappearance decreased (P < 0.05) with increasing Ca:tP in the PC diets, but there was no effect of widening Ca:tP on PP disappearance in NC diets. It was concluded from the current study that the effect of phytase supplementation on P utilization is reduced when diets contain adequate P as exemplified in the PC diets and that the negative impact of wide Ca:tP is more pronounced in diets with phytase matrix allowance as exemplified in the NC diets.

Assessment of a bacterial 6-phytase in the diets of broiler chickens.

Two 21-d broiler experiments were conducted to assess the efficacy of a bacterial 6-phytase expressed in Aspergillus oryzae on growth performance, nutrient utilization, and intestinal molecular markers. Two hundred forty birds in 5 treatments (experiment 1) or 256 birds in 4 treatments (experiment 2) were used. The treatments included a negative control diet that was marginally deficient in P (NC) or NC plus tricalcium phosphate, 500, 1,000, or 2,000 phytase units/kg (experiment 1), and NC or NC plus monocalcium phosphate, 500 or 1,000 phytase units/kg (experiment 2). In both experiments, excreta were collected on d 19 to 21, whereas birds and feed were weighed and ileal digesta collected on d 21. For experiment 1, mucosa scraping was collected from the duodenum, jejunum, and ileum from all birds for quantification of expression level of gut level inflammatory cytokines, Toll-like receptors, and phosphate transporter (NaPi-IIb). In both experiments, tricalcium phosphate, monocalcium phosphate, and phytase supplementation improved (P < 0.05) weight gain and percentage tibia ash. Phosphorus and Ca retention and phytic acid disappearance improved (P < 0.05) with phytase supplementation (experiment 1) and there was an increase (P < 0.01) in Ca and P retention in response to phytase supplementation (experiment 2). Diets did not affect the expression of gut level cytokines, Toll-like receptors, or the mucin gene. Phytase supplementation tended to decrease (P < 0.10) the expression of NaPi-IIb. It was concluded from these studies that the bacterial 6-phytase was effective in enhancing growth of broilers receiving low-P diets as well as in increasing efficiency of P utilization and phytic acid degradation.

True phosphorus digestibility of black-eyed pea and peanut flour without or with phytase supplementation in broiler chickens.

Two studies were conducted to determine the true P digestibility (TPD) of black-eyed pea (BEP) and peanut flour (PNF) and the TPD response to phytase supplementation using the regression method. Sequential diets containing 115, 230, or 345 g of BEP/kg (experiment 1) and 115, 230, or 345 g of PNF/kg (experiment 2) without or with 1,000 units of phytase/kg were formulated. Chromic oxide was added to the diets at the rate of 5 g/kg as an indigestible marker. At 20 d posthatch in each study, 384 male broiler chickens (Ross 708) were weighed and allotted to the diets with 8 replicates of 8 birds each in a randomized complete block design. The birds had free access to the experimental diets until d 26 posthatch. In both studies, dietary P increase and phytase supplementation improved (P < 0.001) growth performance of the broiler chickens. There were linear increases (P < 0. 001) in ileal and excreta P output but a linear decrease (P < 0.001) in apparent P digestibility with an increase in dietary P levels. Phytase supplementation reduced (P < 0.001) ileal P and excreta P output and increased (P < 0.001) apparent P digestibility and retention. Apparent Ca digestibility was affected (P < 0.01) by P level and phytase addition in BEP and by P level (P < 0.05) in PNF. Apparent Ca retention in BEP increased (P < 0.05) with phytase addition. The TPD in the BEP increased (P < 0.01) from 29 without phytase to 83% with the addition of 1,000 units of phytase/kg. There was an increase (P < 0.01) in TPD of PNF from 67 without phytase to 75% with phytase supplementation. There was a corresponding increase (P < 0.01) in true P retention from 10% without phytase to 61% with phytase in birds on BEP diets and an equivalent increase (P < 0.01) in true P retention from 74% without phytase to 84% with phytase in birds that received the PNF diets.

Effects of raw and heat-treated bambara groundnut (Vigna subterranea) on the performance and body composition of growing broiler chicks.

Bambara groundnut (Vigna subterannea)--a leguminous root nut, which grows in the tropics and subtropics--was used in an experiment to test its effect on the performance of growing broiler chicks. Performance was measured by monitoring growth, measuring nutrients and energy balances and also by determining nutrient accretion. Twelve-day old broiler chicks, kept in individual metabolic cages, were used in an experiment in which birds were equally allotted (n = 10) into a control (fed high performance diet made up principally of wheat and soybean meal) and 6 test groups fed diets containing 19, 76 and 95% raw or autoclaved bambara groundnuts. All diets were similar in energy, nitrogen and total lipid contents. Diets were also balanced for amino acids, vitamins and minerals in accordance to the specific requirements of the birds. Feed and water were provided ad libitum. Growth performance was monitored over 15 days. Nutrient and energy balances were measured by use of TiO2 as marker. The comparative slaughter technique was implored for the determination of energy and nutrient accretion. The results showed that the level of raw or autoclaved bambara had no significant influence on feed intake. Increasing the bambara level in the diets caused a general linear drop in the performance of the broilers. However, with up to 95% bambara in the diet, the general performance was still above 75% compared to the control. Increasing levels of bambara groundnut caused a decrease in the metabolizability of energy and efficiency of utilisation of ME for BWG. The animals retained more water with increasing levels of raw bambara in the diets while autoclaving increased their DM content to levels comparable to the control. The protein composition of gain was comparable at all levels of inclusion. In general autoclaving improved the performance of bambara.