Effects of dietary sulfur amino acid levels on growth performance and intestinal immunity in broilers vaccinated and subsequently infected with coccidiosis.

Coccidia vaccination is a common practice in the poultry industry. However, research is lacking regarding the optimal nutritional support for coccidia vaccinated broilers. In this study, broilers were vaccinated with coccidia oocyst at hatch and were fed with a common starter diet from 1 to 10 d. On d 11, the broilers were randomly assigned to groups in a 4 × 2 factorial arrangement. Briefly, the broilers were fed one of four diets containing 0.6, 0.8, 0.9, and 1.0% of standardized ileal digestible methionine plus cysteine (SID M+C), respectively, from 11 to 21 d. On d 14, the broilers from each diet group were orally gavaged with either PBS (Mock challenge) or Eimeria oocysts. Compared to PBS-gavaged broilers and regardless of dietary SID M+C levels, the Eimeria-gavaged broilers had 1) decreased gain-to-feed ratio (15-21 d, P = 0.002; 11-21 d, P = 0.011); 2) increased fecal oocysts (P < 0.001); 3) increased plasma anti-Eimeria IgY (P = 0.033); and 4) increased intestinal luminal interleukin-10 (IL-10; duodenum, P = 0.039; jejunum, P = 0.018) and gamma interferon (IFN-γ; duodenum, P < 0.001; jejunum, P = 0.017). Regardless of Eimeria gavage, broilers fed 0.6% SID M+C had decreased (P<0.001) body weight gain (15-21 and 11-21 d) and gain-to-feed ratio (11-14, 15-21, and 11-21 d) when compared to those fed ≥ 0.8% SID M+C. Eimeria challenge increased (P < 0.001) duodenum lesions when the broilers were fed with 0.6, 0.8, and 1.0% SID M+C, and increased (P = 0.014) mid-intestine lesions when the broilers were fed with 0.6 and 1.0% SID M+C. An interaction between the two experimental factors was detected on plasma anti-Eimeria IgY titers (P = 0.022), as coccidiosis challenge increased plasma anti-Eimeria IgY titers only when the broilers were fed with 0.9% SID M+C. In summary, the dietary SID M+C requirement for grower (11-21 d) broilers vaccinated with coccidiosis was ranged from 0.8 to 1.0% for optimal growth performance and intestinal immunity, regardless of coccidiosis challenge.

A simple daily dynamic feeding regimen for reducing phosphorus consumption and excretion in laying hens.

Phosphorus metabolism in laying hens is a highly dynamic process over the course of the 24 h egg-laying cycle. Adjusting the phosphorus feeding regimen according to the daily egg-laying cycle may help to improve phosphorus utilization efficiency. Hy-Line Brown layers (n = 120; 70 wk old) were offered 4 different phosphorus daily regimens: (1) RR, fed regular phosphorus at both 09:00 and 17:00; (2) RL, fed regular phosphorus at 09:00 and low phosphorus at 17:00; (3) LR, fed low phosphorus at 09:00 and regular phosphorus at 17:00; (4) LL, fed low phosphorus at both 09:00 and 17:00. The regular and low phosphorus diets contained 0.32% and 0.14% non-phytate phosphorus, respectively. The feeding trial lasted for 12 wk. As a result, layers on the RL regimen had decreased laying rate (P < 0.05; 5 to 8, 9 to 12, and 1 to 12 wk) when compared to all other regimens. Layers on the LL regimen had decreased eggshell thickness and specific gravity (P < 0.05; wk 8) when compared to all other regimens, and had decreased egg shell strength (P < 0.05; wk 8) when compared to RL and LR regimens. When compared to the RR regimen (a common practice in the industry), layers on the LR regimen had: (1) identical laying performance and egg quality (P > 0.05); (2) decreased phosphorus excretion (P < 0.05) during the period of 09:00 to 17:00; (3) increased jejunal calbindin D28k protein expression (P < 0.05) 2 h after feeding in the morning; (4) decreased serum fibroblast growth factor 23 and calcitriol levels (P < 0.05), decreased jejunal type III sodium-phosphate cotransporter 2 gene and protein expression (P < 0.05), and decreased renal type III sodium-phosphate cotransporter 1 protein expression (P < 0.05), 2 h after feeding in the afternoon. In summary, when dietary phosphorus was supplemented in accordance with daily serum phosphorus rhythms (i.e., the LR regimen), laying performance and egg quality were well supported whilst significantly decreasing phosphorus consumption and excretion. Thus, serum phosphorus rhythms will need to be carefully maintained when developing dietary phosphorus-reduction strategies in laying hens.

Dietary supplementation of sulfur amino acids improves intestinal immunity to Eimeria in broilers treated with anti-interleukin-10 antibody.

Oral antibody to interleukin-10 (anti-IL-10) enhances the intestinal immune defense against Eimeria. The sulfur amino acids methionine and cysteine (M+C) play essential roles in inducing and maintaining protective immune responses during intestinal infections. Hence, increased dietary M+C may support the anti-IL-10-induced intestinal immunity to Eimeria. Broilers (n = 640) were arranged in a 2 × 2 × 2 factorial design with 2 levels of each of the 3 main factors: dietary standardized ileal digestible (SID) M+C levels (0.6% or 0.8%), dietary anti-IL-10 supplementation (with or without), and coccidiosis challenge (control or challenge). Briefly, the broilers were supplied with either 0.6% or 0.8% SID M+C, each with or without anti-IL-10 (300 µg/kg), from d 10 to 21. On d 14, broilers from each diet were gavaged with either PBS or Eimeria. The resulting Eimeria infection induced fecal oocyst shedding and intestinal lesions. Broilers fed 0.8% SID M+C (main effects, P ≤ 0.05) had decreased feed-to-gain ratio, increased duodenum and cecum luminal anti-Eimeria IgA titers, and decreased fecal oocyst counts, when compared to 0.6% SID M+C. The supplementation of anti-IL-10 (main effects, P ≤ 0.05) increased cecum luminal total IgA concentration and decreased cecum lesions. Interactions (P ≤ 0.05) were detected for growth performance and cecum luminal IFN-γ. Briefly, the highest body weight gain and feed intake were reached in PBS-gavaged broilers fed 0.8% SID M+C with no anti-IL-10 and in Eimeria-challenged broilers fed 0.8% SID M+C with anti-IL-10. In Eimeria-infected broilers, anti-IL-10 increased intestinal luminal IFN-γ and body weight gain only at 0.8% SID M+C. Collectively, anti-IL-10 increased intestinal luminal IFN-γ levels, decreased cecum lesions and restored growth only when fed with adequate amounts of sulfur amino acids. Our findings underscore the importance of providing sufficient essential nutrients to support the anti-IL-10 induced immunity against coccidiosis.

Estimation of phosphorus requirements of sows based on 24-h urinary phosphorus excretion during gestation and lactation.

Phosphorus requirements of reproducing sows were estimated using 24-h urinary P excretion. Thirty-six multiparous sows were fed one of six maize-soybean meal-based diets with total P ranging from 0·40 to 0·80 % in 0·08 % increments with a constant Ca:total P ratio (1·25:1). Diets were fed from day 7·5 ± 1 after breeding until the end of lactation (day 26 ± 1). Urine samples were collected in mid and late gestation (days 77·1 ± 2 and 112·4 ± 1) and early and late lactation (days 4·5 ± 1 and 18·2 ± 1). Phosphorus requirements were estimated using linear and nonlinear regression models. Based on a single 24-h urinary P excretion, estimated daily dietary total P requirements in mid and late gestation were 10·3 g (6·0 g standardised total tract digestible P, STTD P), and estimates for early and late lactation were 31·1 g (16·6 g STTD P) and 40·3 g (22·1 g STTD P), respectively. Plasma P and Ca concentrations were maintained within normal ranges at the estimated levels of P requirements. No differences among treatments were observed for plasma parathyroid hormone (P ≥ 0·06) and bone formation marker (P ≥ 0·16). In lactation, bone resorption marker decreased (P ≤ 0·001) as sows consumed more P. Among the analysed variables, urinary P was the most sensitive response to changes in dietary P intake. Urinary P excretion offers a practical method to estimate P requirements in sows. Our recommended daily total P requirements are 10·3 g for gestation and 35·7 g for lactation.

Evaluation of calcium to phosphorus ratio in spot urine samples as a practical method to monitor phosphorus intake adequacy in sows.

The objective of this study was to evaluate the reliability of using Ca to P ratio measured in spot urine samples to assess P intake adequacy in gestating and lactating sows. A total of 36 sows were fed one of six concentrations of dietary total P (0.40%, 0.48%, 0.56%, 0.64%, 0.72%, and 0.80%) from day 7.5 ± 1 after breeding until the end of lactation (day 26.6 ± 1). Dietary Ca to P ratio was maintained constant across treatments at 1.25:1. Total 24-h urine samples were collected in mid- and late gestation (days 77.1 ± 2 and 112.4 ± 1), and early and late lactation (days 4.5 ± 1 and 18.2 ± 1). In parallel to 24-h collections, spot urine samples were collected at three different times (early morning, late morning, and late afternoon) in late gestation and late lactation. Urine Ca and P concentrations were measured and Ca to P ratio was calculated. Sows were classified as P-adequate or P-deficient according to dietary P intake. Urine Ca to P ratio was greater in sows fed P-deficient diets than sows fed P-adequate diets (P < 0.001). Receiver operating characteristic (ROC) curves were used to determine the cutoff values for urine Ca to P ratio to predict P intake adequacy. Three different categories of P intake were defined according to urine Ca to P ratio: deficient, adequate, and excessive. The area under the ROC for Ca to P ratio was 0.88 (95% CI 0.81 to 0.95). Best cutoff value of urine Ca to P ratio was 1.5 (sensitivity 94% and specificity 68%) to identify sows fed P-deficient diets and 0.5 for P-excessive diets (sensitivity 82% and specificity 82%). A strong relationship between Ca to P ratio in 24-h and spot urine samples was determined (r = 0.93, P < 0.01), independent of physiological state and collection time of spot samples (adjusted-R2 = 0.86, P < 0.01). The degree of agreement between spot and 24-h urine for P intake adequacy, assessed by Cohen's weighted kappa analysis, was substantial (0.78, 95% CI 0.69 to 0.88). We conclude that urinary Ca to P ratio provides a reliable prediction of the adequacy of P intake in reproducing sows. Urinary Ca to P ratio measurements in random spot urinary offers a practical method to determine dietary P adequacy.

Alleged predisposing dietary factors fail to increase the incidence of osteochondrosis-like lesions in growing pigs at 14 and 24 wk of age.

Early lesions of osteochondrosis (OC) are exhibited by regions of cartilage retention along the growth plate and articular cartilage. Progression of OC lesions may impair locomotion and necessitate euthanasia in adherence to animal welfare guides. Little is known about the role of nutrition in the initiation and early stages of OC. However, dietary components are commonly implicated as predisposing factors. In this study, diets were altered as an attempt to induce early stage OC lesions under controlled conditions. At 8 wk of age, 96 crossbred gilts (body weight [BW] = 17.4 ± 0.18 kg) were randomly assigned to one of four corn-soybean meal-based diets (four pens per diet, six pigs per pen) to assess diet effects on the number and volume of OC lesions in the distal femur. Diets included a non-pelleted control diet (Ctl); Ctl plus 20% glucose (Glc); the Ctl with increased concentrations of lysine, Ca, and P (+CaP); and the +CaP diet in a pelleted form (PEL). Femurs were collected from pigs euthanized at either 14-wk (Wk 14) or 24-wk (Wk 14) of age for assessments of OC lesions. Based on a mixed model analysis with pen as the experimental unit, dietary treatments did not affect final BW (129.3 ± 3.8 kg) or average daily gain (ADG) (1.00 ± 0.03 kg/d) over the trial. As expected, pigs fed PEL and Glc diets were more efficient (P < 0.05) in feed conversion compared with Ctl and +CaP. Using femurs as the experimental unit at Wk 14 (collected from two of the six pigs per pen), bone mineral content, determined by dual-energy x-ray absorptiometry scans, was greater (P < 0.05) in pigs fed +CaP and PEL than Ctl or Glc diets; however, only +CaP group differed (P < 0.05) at Wk 24 (collected from four pigs per pen). Computed tomography (CT) scans of femurs were reconstructed as three-dimensional images to allow detection of the number, volume, and surface area of lesions in distal growth plates. At Wk 14, pigs fed Ctl had fewer number of lesions (P < 0.05); however, no differences were detected among dietary treatments in lesion volume or lesion surface area. Pigs had fewer lesions at Wk 24 than Wk 14; however, differences were not detected among dietary treatments. At Wk 24, pigs fed Ctl diets had the greatest lesion volume among dietary treatments (P < 0.05). In conclusion, none of the pigs exhibited symptoms of lameness regardless of dietary treatment or OC lesion traits. Diet modifications due to pelleting or inclusion of rapidly digestible ingredients, such as glucose, did not increase prevalence or size of OC lesions. Image analysis of CT scans was a reliable method to quantify the number, size, and location of OC lesions.

Impact of dietary vitamin D3 supplements in nursery diets on subsequent growth and bone responses of pigs during an immune challenge.

Limited evidence is available to validate beneficial responses from extra nutrient supplements for mediation of growth suppression that results from immune challenges. Extrarenal roles of vitamin D metabolites in immune function implicate vitamin D3 supplements as a nutrient for potential beneficial effects. The current objective was to assess growth and bone ash responses to dietary vitamin D3 (D) supplements for growing pigs undergoing an immune challenge. At weaning, 216 crossbred pigs (4 pigs/pen, 6 pens/treatment) were randomly allotted within sex and weight blocks to 1 of the 9 treatments. Treatments included D supplements (0, 100, or 800 IU/kg) in a factorial arrangement with 3 vaccine (V) protocols; no injection (0 × V), a single 2 mL injection of a Lawsonia intracellularis vaccine at day 14 (1 × V), or 2 mL injections of the same vaccine at days 0 and 7 (2 × V). An adjustment diet with no supplemental D was fed for 1 wk, then assigned D diets for 2 wk (P2). After P2, all pigs were phase-fed standard diets (D = 280 IU/kg) to assess subsequent growth to 115 kg. No differences due to D supplements or vaccination protocol were detected in ADG (0.233 ± 0.021 kg/d) or GF (0.642 ± 0.028 kg/d) over the 21-d nursery trial; however, ADFI was lower (P < 0.10) in pigs fed D levels of 0 vs. 100 and 800 (0.340 vs. 0.375, 0.372 ± 0.027 kg/d). Bone mineral content (g) from whole-body dual energy X-ray absorptiometry scans at 9 wk (n = 4 pigs/treatment) was lower in pigs fed 0 vs. 100 and 800 IU of D (287 vs. 325, 323 ± 34.1 g/pig). Growth from nursery to 115 kg was lower (P < 0.01) in pigs fed D levels of 0 vs.100 and 800 (0.828 vs. 0.876, 0.889 ± 0.021 kg/d). At market, approximately two-thirds of pigs showed positive L. intracellularis serology titers regardless of treatment. Limited evidence for D-mediation of an immune challenge using the vaccination protocols may be a consequence of limited vaccine effects on growth in the nursery and seroconversion of most pigs to L. intracellularis by market.

Effect of anti-fibroblast growth factor receptor 1 antibodies on phosphorus metabolism in laying hens and their progeny chicks.

Targeting fibroblast growth factor 23 (FGF-23) signaling pathway is of interest in controlling body phosphate metabolism. This study investigated the effect of anti-fibroblast growth factor receptor 1 (FGFR1, major FGF-23 receptor in the kidney) antibodies on phosphate metabolism. White Leghorn laying hens (65-wk-old) were vaccinated with either a FGFR1 peptide vaccine (five 8-amino-acid peptides were selected, CrZ-1:LPEDPRWE, CrZ-2:LDKDKPNR, CrZ-3:RRPPGMEY, CrZ-4:GSPYPGVP, and CrZ-5:RMDKPSNC) or adjuvant control. At peak antibody titer, hens were artificially inseminated. Chicks from control-vaccinated hens were fed either a non-phytate phosphorus (nPP) sufficient (nPP = 0.45%, positive control) or deficient (nPP = 0.20%, negative control) diet, while chicks from each of the FGFR1 peptide vaccinated hens were fed with the above nPP-deficient diet, for 14 D. When compared to control hens, plasma phosphate in CrZ-1, CrZ-2, CrZ-3, CrZ-4, and CrZ-5 vaccinated hens were decreased by 33, 30, 24, 20, and 26%, respectively (P < 0.05); egg weight in CrZ-2 and CrZ-5 vaccinated hens were increased by 6 and 7%, respectively (P < 0.05); egg production in CrZ-3, CrZ-4, and CrZ-5 vaccinated hens tended to decrease (P = 0.085; decreased by 14, 15, and 13%, respectively). When compared to positive control, chicks from all other groups had decreased body weight gain (BWG) and feed intake (FI) during 1 to 14 D, and had decreased plasma phosphate, tibiotarsus ash, and 24-h phosphorus excretion on day 14. When compared to negative control, BWG of CrZ-1, CrZ-2, CrZ-3, and CrZ-4 antibody chicks were decreased by 23, 28, 26, and 20%, respectively (P < 0.05); FI of CrZ-1, CrZ-2, and CrZ-3 antibody chicks were decreased by 15, 15, and 18%, respectively (P < 0.05); plasma phosphate of CrZ-5 antibody chicks were decreased by 26% (P < 0.05); plasma FGF-23 levels of CrZ-4 antibody chicks were increased by 18% (P < 0.05); tibiotarsus ash content of CrZ-2, CrZ-3, and CrZ-4 antibody chicks were decreased by 20, 20, and 21%, respectively (P < 0.05). In conclusion, anti-FGFR1 peptide antibodies decreased egg production of hens and growth performance of their progeny chicks probably by activating FGF-23 signaling and stimulating FGF-23 production.

Maternal Diets Deficient in Vitamin D Increase the Risk of Kyphosis in Offspring: A Novel Kyphotic Porcine Model.

BACKGROUND: The purpose of this study was to explore the role of perinatal vitamin-D intake on the development and characterization of hyperkyphosis in a porcine model. METHODS: The spines of 16 pigs were assessed at 9, 13, and 17 weeks of age with radiography and at 17 weeks with computed tomography (CT), magnetic resonance imaging (MRI), histology, and bone-density testing. An additional 169 pigs exposed to 1 of 3 maternal dietary vitamin-D levels from conception through the entire lactation period were fed 1 of 4 nursery diets supplying different levels of vitamin D, calcium, and phosphorus. When the animals were 13 weeks of age, upright lateral spinal radiography was performed with use of a custom porcine lift and sagittal Cobb angles were measured in triplicate to determine the degree of kyphosis in each pig. RESULTS: The experimental animals had significantly greater kyphotic sagittal Cobb angles at all time points when compared with the control animals. These hyperkyphotic deformities demonstrated no significant differences in Hounsfield units, contained a slightly lower ash content (46.7% ± 1.1% compared with 50.9% ± 1.6%; p < 0.001), and demonstrated more physeal irregularities. Linear mixed model analysis of the measured kyphosis demonstrated that maternal diet had a greater effect on sagittal Cobb angle than did nursery diet and that postnatal supplementation did not completely eliminate the risk of hyperkyphosis. CONCLUSIONS: Maternal diets deficient in vitamin D increased the development of hyperkyphosis in offspring in this model. CLINICAL RELEVANCE: This study demonstrates that decreased maternal dietary vitamin-D intake during pregnancy increases the risk of spinal deformity in offspring. In addition, these data show the feasibility of generating a large-animal spinal-deformity model through dietary manipulation alone.

Serum and tissue 25-OH vitamin D3 concentrations do not predict bone abnormalities and molecular markers of vitamin D metabolism in the hypovitaminosis D kyphotic pig model.

The hypovitaminosis D kyphotic pig provides a model to study maternal vitamin D (D) carryover on gross and molecular characteristics of bone abnormalities in offspring. Excess maternal D is proposed to protect offspring under nutritional challenges from developing bone abnormalities. Relationships between D sufficiency parameters and bone abnormalities were characterised. Sows (n 37) were fed diets with 0 (-D), 8·125 (+D) or 43·750 (++D) µg D3/kg throughout gestation and lactation. At weaning (3 weeks) pigs were fed diets with 0 (-D) or 7·0 (+D) µg D3/kg, each with 75 and 95 % (LCaP) or 150 and 120 % (HCaP) of the Ca and P requirements. Pigs were euthanised before colostrum consumption at birth (n 27), 3 weeks (n 27) or after the nursery period (7 weeks, n 71) for tissue analysis. At 7 weeks, differences due to maternal D were detected (P≤0·05) in pig growth, serum parameters and mRNA expression regardless of nursery diet. Prevalence of kyphosis in pigs at 13 weeks was affected by maternal D, but not prevented by only HCaP or +D nursery diets. Increased (P≤0·05) serum 25-OH-D3 concentrations in sows fed +D or ++D diets were not reflected by similar magnitudes of 25-OH-D3 in colostrum, 18-d milk, or serum and tissue concentrations in pigs. The mode of action by which maternal dietary D influences development of skeletal abnormalities warrants further investigation.

Maternal dietary vitamin D carry-over alters offspring growth, skeletal mineralisation and tissue mRNA expressions of genes related to vitamin D, calcium and phosphorus homoeostasis in swine.

Maternal dietary vitamin D carry-over effects were assessed in young pigs to characterise skeletal abnormalities in a diet-induced model of kyphosis. Bone abnormalities were previously induced and bone mineral density (BMD) reduced in offspring from sows fed diets with inadequate vitamin D3. In a nested design, pigs from sows (n 23) fed diets with 0 (-D), 8·125 (+D) or 43·750 (++D) µg D3/kg from breeding through lactation were weaned and, within litter, fed nursery diets arranged as a 2×2 factorial design with 0 (-D) or 7·0 (+D) µg D3/kg, each with 95 % (95P) or 120 % (120P) of P requirements. Selected pigs were euthanised before colostrum consumption at birth (0 weeks, n 23), weaning (3 weeks, n 22) and after a growth period (8 weeks, n 185) for BMD, bone mechanical tests and tissue mRNA analysis. Pigs produced by +D or ++D sows had increased gain at 3 weeks (P<0·05), and at 8 weeks had increased BMD and improved femur mechanical properties. However, responses to nursery diets depended on maternal diets (P<0·05). Relative mRNA expressions of genes revealed a maternal dietary influence at birth in bone osteocalcin and at weaning in kidney 24-hydroxylase (P<0·05). Nursery treatments affected mRNA expressions at 8 weeks. Detection of a maternal and nursery diet interaction (P<0·05) provided insights into the long-term effects of maternal nutritional inputs. Characterising early stages of bone abnormalities provided inferences for humans and animals about maternal dietary influence on offspring skeletal health.

Supplemental Escherichia coli phytase and strontium enhance bone strength of young pigs fed a phosphorus-adequate diet.

Young pigs represent an excellent model of youth to assess potentials of dietary factors for improving bone structure and function. We conducted 2 experiments to determine whether adding microbial phytase (2,000 U/kg, OptiPhos, JBS United) and Sr (50 mg/kg, SrCO3 Alfa Aesar) into a P-adequate diet further improved bone strength of young pigs. In Expt. 1, 24 gilts (8.6 +/- 0.1 kg body wt) were divided into 2 groups (n = 12), and fed a corn-soybean-meal basal diet (BD, 0.33% available P) or BD + phytase for 6 wk. In Expt. 2, 32 pigs (11.4 +/- 0.2 kg) were divided into 4 groups (n = 8), and fed BD, BD + phytase, BD + Sr, or BD + phytase and Sr for 5 wk. Both supplemental phytase and Sr enhanced (P < 0.05) breaking strengths (11-20%), mineral content (6-15%), and mineral density (6-11%) of metatarsals and femurs. Supplemental phytase also resulted in larger total bone areas (P < 0.05) and a larger cross-sectional area of femur (P = 0.06). Concentrations of Sr were elevated 4-fold (P < 0.001) in both bones by Sr, and moderately increased (P = 0.05-0.07) in metatarsal by phytase. In conclusion, supplemental phytase at 2000 U/kg of P-adequate diets enhanced bone mechanical function of weanling pigs by modulating both geometrical and chemical properties of bone. The similar benefit of supplemental Sr was mainly due to an effect on bone chemical properties.