Effect of phytase and limestone particle size on mineral digestibility, performance, eggshell quality, and bone mineralization in laying hens.

The effect of microbial phytase and limestone particle size (LmPS) was assessed in Lohmann Tradition laying hens from 31 to 35 wk of age. Seventy-two hens were used in a completely randomized trial according to a 2 × 2 factorial arrangement with 2 levels of phytase/basal available P (aP); 0 FTU/kg with 0.30% aP or 300 FTU/kg with 0.15% aP, and 2 limestone particle sizes; fine particles (FL, <0.5 mm) or a mix (MIX) of 75% coarse limestone (CL, 2-4 mm) and 25% FL. Diets contained equivalent levels of Ca (3.5%), phytic P (PP; 0.18%), and aP (0.30%) considering the P equivalency of phytase. Thus, dietary treatments were FL0 and MIX0 without phytase, and FL300 and MIX300 with 300 FTU/kg phytase. Performance were recorded daily and eggshell quality (eggshell weight proportion, weight, thickness, and breaking strength) was measured weekly. At the end of the trial, bone parameters (tibia breaking strength, elasticity, and ash) and the apparent precaecal digestibility (APCD) of P and Ca were determined. No differences were observed between treatments in feed intake, FCR and bone parameters. Addition of MIX increased the eggshell proportion, weight and thickness in groups receiving no phytase (+6.5, +6.9, and +4.5%, respectively) while no effect was observed in groups receiving phytase (Phytase × LmPS, P < 0.05). In hens receiving FL, the APCD of P was lower in diets supplemented with phytase (-14 percentage points; Phytase × LmPS, P < 0.001). A higher phytate disappearance was observed in hens fed diets with phytase in combination with MIX (Phytase × LmPS, P = 0.005). Phytase and MIX together increased the APCD of Ca by 7.3 percentage points (Phytase × LmPS, P < 0.001). In conclusion, addition of CL could limit the formation of Ca-phytate complex thus improving the response of the birds to phytase compared to FL.

Effect of the physicochemical properties of zinc oxide sources on their bioavailability in broilers.

1. The bioavailability of a trace mineral source is related to its intestinal solubility (bioaccessibility), which in turn is determined by its physicochemical properties. It is still not clear which characteristics are more relevant in affecting solubility and bioavailability of mineral sources. Zinc oxide (ZnO) is a common feed additive used to supplement zinc in the diet of monogastric animals. However, different sources have shown variable responses on animal bioavailability.2. This study hypothesised that different sources of feed grade ZnO have various physicochemical features that lead to distinct bioavailability values. Feed grade ZnO samples collected from the feed industry worldwide were characterised for their physicochemical features and tested in broilers to allow bioavailability determined.3. A total of 135 male Cobb broiler chickens were fed a standard starter diet from day 1 after hatching up to d 7. At d 8, animals were allocated in individual cages and fed one of the following dietary treatments during 15 days: a basal diet with 23.5 ppm of zinc and seven test diets with supplemented ZnO or zinc sulphate (ZnSO4) at 6 or 12 ppm.4. Different sources of ZnO showed an effect of solubility in the stomach and supplementation influenced total Zn levels in the ileum. The bioavailability of the different sources varied from 49% to 160% in relation to ZnSO4. Aggregate size of particles seems to explain most of the variability in the bioavailability of the different sources tested in broilers. In conclusion, physicochemical properties of ZnO can partly explain the variability observed in terms of biological value.

Increased expression of fibroblast growth factor 23 is the signature of a deteriorated Ca/P balance in ageing laying hens.

The present study concerned the effect of ageing in laying hens, from 23 to 90 weeks of age, on the regulation of Ca metabolism related to the requirement for eggshell mineralization. Samples were collected from parathyroid gland (PG), liver, jejunum, medullary bone (MB) and kidney for a quantitative study of candidate gene expression. Although parathyroid hormone (PTH) gene expression in the PG did not vary with age, a stronger challenge to Ca homeostasis was suggested in aged hens. Indeed gene expression of Ca transporters , Vitamin D Receptor (VDR) in the jejunum, and that of transient receptor potential channel subfamily V member 5 (TRPV5) in the kidney decreased. This could exacerbate bone resorption and impair bone accretion, as attested by a higher expression of the Carbonic Anhydrase 2 (CA2) gene and a lower expression of collagen type I alpha 1 chain (COL1A1) in the MB. The increased expression of Fibroblast Growth Factor 23 (FGF23) in the MB likely contributed to the decreased plasma levels of 1.25(OH)2D3 and the altered expression of target genes under its regulation. Our data highlights the molecular mechanisms underlying the osteoporotic syndrome previously documented in aged laying hens, thus providing new perspectives for future interventions.

Early-life conditioning strategies to reduce dietary phosphorus in broilers: underlying mechanisms.

Chickens adapt to P and Ca restriction during the very first days of life by improving P utilisation efficiency. The present study was built to identify the mechanisms underlying this adaptive capacity, and to identify the optimal window of application of the restriction (depletion). A total of 1600 Cobb 500TM male broilers were used. During each phase (from age 0 to 4 d, 5 to 8 d, 9 to 18 d and 19 to 33 d), the animals received either a control diet (H) or a restricted diet (L) with reduced levels of non-phytate P (nPP) and Ca (between -14 and -25 % for both) with four dietary sequences: HHHH, HLHL, LHHL and LLHL. None of the feeding strategies affected growth. Tibia ash content at day 4 and 8 was impaired when the L diet was fed from 0 to 4 and 5 to 8 d, respectively (P = 0⋅038 and P = 0⋅005). Whatever the early restriction period or length between 0 and 8 d of age, the mineralisation delay was compensated by day 18. This was accompanied by an increased mRNA expression of the Ca transporter, CALB1, and an increased apparent ileal digestibility of Ca at day 8 (P < 0⋅001). This adaptation was limited to the starter phase in restricted birds. No effect was seen on P transporters mRNA or protein expression. In conclusion, birds adapted to mineral restriction by increasing Ca and nPP utilisation efficiencies. Depletion-repletion strategies are promising in improving the sustainability of broiler production but need to be validated in phytase-supplemented diets.

Effects of dietary electrolyte balance and calcium supply on mineral and acid-base status of piglets fed a diversified diet.

Dietary electrolyte balance (dEB) is known to affect acid-base status and mineral metabolism, but is rarely considered in diet formulation for pigs. Yet, the use of a wide variety of local feedstuffs in Europe contributes to lowering the dEB and increasing the fibre content. Hence, mineral requirements may be modified and skeletal health affected. Therefore, the effects of a lower dEB and a higher dietary Ca level on acid-base balance and mineral status were assessed in young pigs fed a diversified diet. A total of twenty-four weaned pigs were fed a control moderate-dEB diet (C) or a diversified moderate-dEB (D), low-dEB (D-A) or low-dEB supplemented with Ca (D-CA) diet. Growth performance, venous blood gas and chemistry, urine pH, mineral balance and femur characteristics were determined. With an equivalent dEB compared with the C diet, the D diet caused an acidification of the urine and increased the excretion of P as a result of a higher dietary content of S. Low-grade metabolic acidosis occurred in piglets fed the D-A diet with changes at systemic and urine levels. A higher excretion of ammonia and P in urine was observed and some bone characteristics tended to be negatively affected. Ca supplementation partially counteracted the effects of low-grade acidosis. Urine excretion of P and ammonia was alleviated and bone characteristics improved. In conclusion, a higher Ca supply must be considered in more diversified diets to counteract the risk of evolving towards low-grade metabolic acidosis which can negatively affect bone.

Possible roles of parathyroid hormone, 1.25(OH)2D3, and fibroblast growth factor 23 on genes controlling calcium metabolism across different tissues of the laying hen.

This study provides an integrative description of candidate gene expression across tissues involved in calcium (Ca) metabolism during the egg laying cycle, using the well-defined model of Ca supply as fine or coarse particles of calcium carbonate (CaCO3). Plasma and tissue samples were collected from hens at the peak of laying at 0 to 1, 9 to 10, and 18 to 19 h postovulation (PO). After mRNA preparation from the parathyroid gland, medullary bone, liver, kidney, duodenum, and jejunum, gene expressions were quantified using RT-qPCR. The highest levels of parathyroid hormone (PTH) mRNA in the parathyroid gland (P < 0.05), and of the active form of vitamin D3 1.25(OH)2D3 in the plasma (P < 0.01) were observed at 18 to 19 h PO. During this active phase of eggshell formation, bone resorption was attested to high levels of plasma inorganic phosphorus (iP) and the receptor activation of nuclear factor-κB expression in the bone (P < 0.001 and P < 0.05, respectively). At this stage, 5 genes of the transcellular and the paracellular Ca absorption pathways in the intestine (P < 0.05) and the Ca channel transient receptor potential cation channel subfamily V member 5 (P < 0.05), involved in its reabsorption in the kidney, were overexpressed. At 0 to 1 h PO during the subsequent daylight period, 2 candidates of the transcellular and the paracellular Ca pathways (P < 0.05) remained at high levels in the intestine, while calbindin D 28K expression was the highest in the kidney (P < 0.05). As PTH mRNA and 1.25(OH)2D3 were low, bone accretion was likely active at this stage. The phosphaturic hormone fibroblast growth factor 23 (FGF23) was overexpressed at 18 to 19 h PO (P < 0.05) in the bone when plasma iP was high, which suggested a role in the subsequent reduction of P reabsorption in the kidney, as attested to the decreased expression of P cotransporters, leading to iP clearance from the plasma at 0 to 1 h PO (P < 0.05). The low levels of 1.25(OH)2D3 at this stage coincided with increased expression of the 24-hydroxylase gene in the kidney (P < 0.05). In hens fed fine particles of CaCO3, higher plasma levels of 1,25(OH)2D3 and higher expression of several genes involved in bone turnover reflected a stronger challenge to Ca homeostasis. Altogether, these data support the hypothesis that FGF23 could drive vitamin D metabolism in the laying hen, as previously documented in other species and explain the tight link between P and Ca metabolisms.

Phytase supplementation in diets rich in fiber from rapeseed enhances phosphorus and calcium digestibility but not retention in broiler chickens.

Two experiments were conducted on broilers to assess the effect of dietary fiber from 00-rapeseed meal (RSM) on phosphorus (P) and calcium (Ca) apparent ileal digestibility (AID) and retention (AR) during the growing (Exp1: 10 to 21 d) or finishing period (Exp2: 21 to 31 d) in diets supplemented or not with microbial phytase. Each experiment involved 144 male Cobb 500 fed one of 8 diets. Fiber content was modulated by incorporating whole RSM, RSM from dehulled rapeseeds, either raw or supplemented with 2 levels of defatted rapeseed hulls. Diets were supplemented or not with 750 phytase units of microbial phytase per kg. Excreta were collected from d 14 to d 17 (Exp1) and from d 27 to d 30 (Exp2) to measure AR. At the end of experiments, digestive tracts were sampled and weighed. The distal ileum and tibias were collected to measure AID and bone mineralization, respectively. Age did not significantly alter the response of birds to the addition of dietary fiber. Inclusion of hulls decreased growth performance (P < 0.05). The weight of the proventriculus-gizzard (PG) increased with the dietary fiber content in Exp1: The decreased weight observed using dehulled RSM was reversed following the inclusion of hulls. In both trials, while the presence of phytase increased the AID of P (P < 0.001) but not Ca, the inclusion of hulls with phytase improved the AID of P and Ca [linear (Lin), P < 0.05]. This effect could depend on the effect of fiber on PG development and physiology. Hulls decreased the moisture content of excreta (P < 0.01), suggesting higher water retention or lower water consumption with fiber. The AR of P was lower than AID of P with hulls, contrary to Ca, suggesting a metabolic imbalance. The decrease of AR together with the decrease of bone characteristics indicates a lack of Ca in diets with hulls and suggests that P and Ca provision should be adapted to the level and the origin of fiber inclusion.

Effects of rapeseed meal fiber content on phosphorus and calcium digestibility in growing pigs fed diets without or with microbial phytase.

The optimization of dietary phosphorus (P) and calcium (Ca) supply requires a better understanding of the effect of dietary fiber content of co-products on the digestive utilization of minerals. This study was designed to evaluate the effects of dietary fiber content from 00-rapeseed meal (RSM) on P and Ca digestibility throughout the gastrointestinal tract in growing pigs fed diets without or with microbial phytase. In total, 48 castrated male pigs (initial BW=36.1±0.4 kg) were housed in metabolic crates for 29 days. After an 8-day adaptation period, pigs were allocated to one of the eight treatments. The impact of dietary fiber was modulated by adding whole RSM (wRSM), dehulled RSM (dRSM) or dRSM supplemented with 4.5% or 9.0% rapeseed hulls (dRSMh1 and dRSMh2). Diets contained 0 or 500 phytase unit of microbial phytase per kg. From day 14 to day 23, feces and urine were collected separately to determine apparent total tract digestibility (ATTD) and apparent retention (AR) of P and Ca. At the end of the experiment, femurs and digestive contents were sampled. No effect of variables of interest was observed on growth performance. Microbial phytase increased ATTD and AR of P (P<0.001) but the P equivalency with the wRSM diet was lower than expected. Moreover, stomach inorganic P (iP) solubility was improved by microbial phytase (P<0.001). The ATTD of Ca was not affected by microbial phytase which increased AR of Ca and femur characteristics (P<0.05). Ileal recovery of P was not affected by microbial phytase but cecal recovery was considerably reduced by microbial phytase (P<0.001). The decrease in digesta pH between the distal ileum and cecum (7.6 v. 5.9) enhanced the solubility of iP and may have improved its absorption, as supported by the negative relationship between soluble iP and pH (R 2=0.40, P<0.001 without microbial phytase and R 2=0.24, P=0.026 with microbial phytase). The inclusion of hulls improved the solubility of iP (P<0.05). In conclusion, dehulling does not largely increase nutrient digestibility although dRSM seems to improve the efficacy of microbial phytase in releasing phosphate in the stomach. Moreover, dietary fiber may affect solubilization process in the cecum which potentiates the effect of microbial phytase on P digestibility.

Genetic variability of metabolic characteristics in chickens selected for their ability to digest wheat.

Improving feed efficiency remains crucial for poultry production. Birds have previously been selected on their ability to digest their diet, as assessed by AMEn (Apparent ME corrected for 0 nitrogen). Such selection, for either a high (D+) or a low AMEn (D-), affects energy, nitrogen, lipid, and starch digestibility. The aim of this study was to establish whether selection on the digestive ability of birds modified metabolic traits. A total of 630 broiler chickens of the eighth generation of a divergent selection experiment on AMEn were used for this purpose. A balance trial was performed to determine energy, nitrogen, and phosphorus retention. Growth performance was recorded and body protein and lipid deposition assessed by breast and abdominal fat yields. Tibia development and mineralization were also studied and heat production was indirectly assessed through the measurement of body temperature during fasting and feeding. Phenotypic correlations estimated within line showed that an increased efficiency was associated to fatter birds and more solid bones in D- but not in D+ line, whereas increased consumption was associated with more solid bones in D+ but not in D- line. The heritability estimates for metabolic traits were relatively high, except for temperature traits (from 0.08 to 0.12), ranging from 0.28 to 0.56 for body composition, and from 0.38 to 0.77 for bone characteristics. Breast meat yield did not differ between the 2 lines whereas a slight increase in abdominal fat yield was observed in the high-digestion line (D+). The relative dry tibia weights and ash weights were greater in D+ birds (+6.56 and +8.06%, respectively) but the lengths and the diameters of the tibia were lower (-7.89 and -3.77%, respectively). Finally, AMEn was poorly correlated with almost all metabolic traits (ranging from -0.10 to 0.20), indicating that the ability of the animal to digest its diet is genetically independent of post-digestion metabolic traits.

Reducing the environmental impact of poultry breeding by genetic selection.

Improving the sustainability of poultry production involves limiting its environmental impact and maintaining effectiveness. It has recently been shown that genetic selection on the ability of chickens to digest wheat at 23 d of age can decrease chicken excretion without decreasing BW at this age. The aim of this study was to check whether selection on digestibility modified excretion and growth performance over the whole production cycle. The 2 divergent lines selected for high (D+) and low (D-) apparent metabolizable energy corrected for 0 N balance (AMEn) values were compared with a reference line used at the beginning of the selection experiment (RL) to evaluate the potential excretion improvement that could be expected with such selection. These 3 lines were therefore compared for growth and excretion (raw and relative to feed intake, fresh and dry excreta weights, and moisture content of excreta) from 4 to 53 d. Between 4 and 7 d, 17 and 21 d, and 49 and 53 d, AMEn and N and P excretion rates were also compared between the 3 lines. Furthermore, body composition (breast meat and abdominal fat yields), bone breaking strength, and meat quality traits (lightness, redness, yellowness, and ultimate pH) were compared between lines at 53 d. Over the whole rearing period, D+ birds excreted significantly less fresh and dry excreta (-56 and -61%) than D- and RL birds (-6 and -26%). Similarly, N and P excretion rates of D+ birds were 13% to 30% less than those of D- birds and 12% to 19% less than RL birds, depending on age. These excretion differences may be related to the differential development of the gastrointestinal tract. Differences between lines were already present at 7 d for relative gizzard weight and the weight of the upper to the lower part of the gastrointestinal tract ratio. Anatomic differences were maximum at 23 d for all traits except for relative weight of the duodenum. At slaughter age, BW, breast and fat yields, and meat color did not differ between D+ and RL birds, but D- birds were fatter than D+ and RL birds. Finally, ultimate meat pH was 1% to 2% greater in RL birds than in the D+ and D- lines. In conclusion, this study showed that selection of chickens for AMEn is a possible way to reduce the environmental impact of production over the whole rearing period without a negative impact on growth, body composition, or meat quality.

Phosphorus utilization in finishing broiler chickens: effects of dietary calcium and microbial phytase.

A decrease in dietary P, especially in finishing broilers (21 to 38 d old), is a crucial issue in poultry production from an environmental and economic point of view. Nevertheless, P must be considered together with other dietary components such as Ca and microbial phytase. Different corn and soybean meal-based diets varying in Ca [low (LCa) 0.37, medium (MCa) 0.57, and high (HCa) 0.77%], and nonphytate P [nPP; low (LnPP) 0.18 and high (HnPP) 0.32%] content were tested with and without microbial phytase [0 or 500 phytase units (FTU)/kg]. Feed intake, BW gain, bone mineralization, and mineral retention were examined in 144 Ross PM3 broilers (22 to 38 d old) reared in individual cages. Growth performance was not significantly affected by the treatments. Nevertheless, a numerical decrease of ADG and ADFI was observed in HCa-LnPP and LCa-HnPP associated with an increase of feed conversion ratio. Decreased dietary Ca reduced tibia ash content (Ca, linear: P < 0.001; quadratic: P = 0.034) and tibia ash weight for the highest level of nPP (Ca × nPP; P = 0.035). In parallel, increasing dietary Ca reduced the flow of retained P (P = 0.022) but also tibia ash weight in LnPP diets (Ca × nPP; P = 0.035). The responses of the animals in terms of tibia ash content and P retention were improved by the addition of microbial phytase especially for the lowest P diets (nPP × phytase, P = 0.021 and P = 0.009; respectively). Phytase increased dry tibia weight, bone breaking strength, and tibia diameter in broilers fed the highest Ca diets (Ca × phytase; P < 0.05). We conclude that is possible to decrease P levels in finishing broilers, if the Ca content is appropriate. Nevertheless, decreasing the dietary P and Ca cannot allow a maximization of bone mineralization, but the optimal threshold remains to be determined.

Adaptation of wheat and protein-mineral concentrate intakes by individual hens fed ad libitum in sequential or in loose-mix systems.

1. Feed intake and performance of birds given sequential or loose-mix feeding was investigated from 19 to 42 weeks of age. A complete diet was fed as control (C). A balancer diet (50) was fed either sequentially (S50) or in a loose-mix (L50) with wheat. This diet was formulated to provide a similar nutritive value as C assuming a 50 : 50 diet and wheat intake. Another balancer diet (25) was fed sequentially (S25) or in a loose-mix (L25) with wheat. The diet was to provide a similar nutritive value as C assuming 75 : 25 diet and wheat intakes. In sequential feeding, only wheat was fed in the morning (4 h after lights-on) and the balancer diet in the late afternoon (4 h before lights-off). In the loose-mix treatment, a mixture of the two diets was fed throughout the 16-h daily light. Each treatment was given ad libitum to 25 birds in individual cages. 2. Birds fed on L25 had lower total feed intakes than those receiving C, S50 or S25. Protein intake was reduced with L25 compared to C, S50, S25 and L50. Metabolisable energy (ME) intake was, however, similar among all treatments. Egg production and weight were reduced with L25 compared to S50 and S25. Body weight (BW) was lowered with L25. However, there was high individual variation in all variables. 3. Feeding system (sequential vs loose-mix) had no effect on ME intake. However, the loose-mix treatment reduced feed and protein intake due to lower balancer diet intake. It also resulted in low egg production, egg and BWs compared to sequential feeding. The weights of pancreas and gizzard were heavier with sequential and loose-mix compared to the control. 4. The loose-mix treatment reduced egg-laying performance. Sequential feeding resulted in similar egg-laying performance to conventional feeding and thus could be used to advantage in situations where it is applicable.

Sequential feeding using whole wheat and a separate protein-mineral concentrate improved feed efficiency in laying hens.

The effect of feeding nutritionally different diets in sequential or loose-mix systems on the performance of laying hen was investigated from 16 to 46 wk of age. Equal proportions of whole wheat grain and protein-mineral concentrate (balancer diet) were fed either alternatively (sequential) or together (loose-mix) to ISA Brown hens. The control was fed a complete layer diet conventionally. Each treatment was allocated 16 cages and each cage contained 5 birds. Light was provided 16 h daily (0400 to 2000 h). Feed offered was controlled (121 g/bird per d) and distributed twice (4 and 11 h after lights-on). In the sequential treatment, only wheat was fed at first distribution, followed by balancer diet at the second distribution. In loose-mix, the 2 rations were mixed and fed together during the 2 distributions. Leftover feed was always removed before the next distribution. Sequential feeding reduced total feed intake when compared with loose-mix and control. It had lower wheat (-9 g/bird per d) but higher balancer (+1.7 g/bird per d) intakes than loose-mix. Egg production, egg mass, and egg weight were similar among treatments. This led to an improvement in efficiency of feed utilization in sequential compared with loose-mix and control (10 and 5%, respectively). Birds fed sequentially had lower calculated ME (kcal/bird per d) intake than those fed in loose-mix and control. Calculated CP (g/bird per d) intake was reduced in sequential compared with loose-mix and control. Sequentially fed hens were lighter in BW. However, they had heavier gizzard, pancreas, and liver. Similar liver lipid was observed among treatments. Liver glycogen was higher in loose-mix than the 2 other treatments. It was concluded that feeding whole wheat and balancer diet, sequentially or loosely mixed, had no negative effect on performance in laying hens. Thus, the 2 systems are alternative to conventional feeding. The increased efficiency of feed utilization in sequential feeding is an added advantage compared with loose-mix and thus could be employed in situations where it is practicable.