Effects of sources and routes of administration of vitamins A, D and copper on postnatal status of these micronutrients in piglets.

Twenty-six nulliparous sows were fed conventional gestation and lactation diets supplemented (N = 13) or not (N = 13) with extra daily supplements of 25-hydroxy-cholecalciferol (25-OH-D3; 4 ĸIU), ß-carotene (24 ĸIU), and copper (Cu)-proteinate (45 mg) from day 90 of gestation to 21 d of lactation (L21). In each litter, 10 piglets were divided into 5 pairs received, at 2 (L2) and 8 d (L8) of age, one of the five combinations of micronutrient sources and routes of administration (N = 260 piglets total). These neonatal treatments (N = 26 pairs or 52 piglets each) consisted of oral vitamin D3, retinol acetate and CuSO4 (T1); oral 25-OH-D3, ß-carotene, and Cu proteinate (T2); exposure to ultraviolet light (UVB), oral retinol palmitate and Cu gluconate (T3); intramuscular vitamin D3 and retinyl propionate and oral Cu acetate (T4); oral saline (CTRL). Oral or intramuscular provisions corresponded to 12 mg of Cu and 70 and 12 ĸIU of vitamins A and D, respectively. Blood samples were collected from all piglets at L2, L8, and L21 for determination of serum Cu, retinol, and 25-OH-D3. Body weight was measured at birth, L2, L8, and L21. Piglets were weaned at L21, and liver and blood samples were collected 2 d later to evaluate oxidative enzymes in blood and liver and hepatic ATP concentrations and expression of genes associated with antioxidant status. Sow treatments had marginal or no impacts on Cu, retinol, 25-OH-D3, or antioxidant status in piglet blood serum and liver. However, when supplements were given to piglets, hepatic Cu was 38% greater in for all treated piglets compared to CTRL (P < 0.01), hepatic retinol was 3 times higher in T1 than in CTRL (P < 0.01) and intermediate for other treatments whereas serum 25-OH-D3 was markedly increased with T2 and T3 at L8 and L21, respectively, compared to CTRL (Piglet treatment × Age interaction, P < 0.01). Concerning antioxidant activities, glutathione peroxidase, and superoxide dismutase were increased (P < 0.03) in plasma of T2 piglets whereas the highest values (P < 0.03) for indicators of oxidative damage to proteins were observed in T4 piglets. The study revealed that oral Cu proteinate from T2, oral retinol acetate from T1, oral 25-hydroxy-cholecalciferol from T2, and UVB light exposure from T3 were the most efficient ways of increasing the postnatal status of these micronutrients in suckling piglets and this may have some impacts on their peri-weaning antioxidant status.

Over 100 years of vitamin E: An overview from synthesis and formulation to application in animal nutrition.

The groundbreaking discovery of vitamin E by Evans and Bishop in 1922 was an important milestone in vitamin research, inspiring further investigation into its crucial role in both human and animal nutrition. Supplementing vitamin E has been proved to enhance multiple key physiological systems such as the reproductive, circulatory, nervous and muscular systems. As the main antioxidant in the blood and on a cellular level, vitamin E maintains the integrity of both cellular and vascular membranes and thus modulates the immune system. This overview showcases important and innovative routes for synthesizing vitamin E on a commercial scale, provides cutting-edge insights into formulation concepts for successful product form development and emphasizes the importance and future of vitamin E in healthy and sustainable animal nutrition.

Application of multi-omic features clustering and pathway enrichment to clarify the impact of vitamin B2 supplementation on broiler caeca microbiome.

Background: The results of omic methodologies are often reported as separate datasets. In this study we applied for the first time multi-omic features clustering and pathway enrichment to clarify the biological impact of vitamin B2 supplementation on broiler caeca microbiome. Methods: The caeca contents of broilers fed +50 and +100 mg/kg vitamin B2 were analyzed by shotgun metagenomic and metabolomic. Latent variables extracted from NMR spectra, as well as taxonomic and functional features profiled from metagenomes, were integrated to characterize the effect of vitamin B2 in modulating caeca microbiome. A pathway-based network was obtained by mapping the observed input genes and compounds, highlighting connected strands of metabolic ways through pathway-enrichment analysis. Results: At day 14, the taxonomic, functional and metabolomic features in the caeca of tested broilers showed some degree of separation between control and treated groups, becoming fully clear at 28 days and persisting up to 42 days. In the caeca of birds belonging to the control group Alistipes spp. was the signature species, while the signature species in the caeca of broilers fed +50 and +100 mg/kg vitamin B2 were Bacteroides fragilis and Lactobacillus crispatus, Lactobacillus reuteri, Ruminococcus torques, Subdoligranum spp., respectively. The pathway enrichment analysis highlighted that the specific biochemical pathways enhanced by the supplementations of vitamin B2 were N-Formyl-L-aspartate amidohydrolase, producing Aspartate and Formate; L-Alanine:2-oxoglutarate amino transferase, supporting the conversion of L-Alanine and 2-Oxoglutarate in Pyruvate and L-Glutamate; 1D-myo-inositol 1/4 phosphate phosphohydrolase, converting Inositol 1/4-phosphate and water in myo-Inositol and Orthophosphate. The results of this study demonstrated that the caeca of birds fed +50 and + 100 mg/kg were those characterized by taxonomic groups more beneficial to the host and with a higher concentration of myo-inositol, formic acid, amino acids and pyruvate involved in glycolysis and amino acid biosynthesis. Conclusion: In this study we demonstrated how to perform multi-omic features integration to describe the biochemical mechanisms enhanced by the supplementation of different concentrations of vitamin B2 in the poultry diet. The relationship between vitamin B2 supplementation and myo-inositol production was highlighted in our study for the first time.

Influence of vitamin D metabolites on vitamin D status, immunity and gut health of piglets.

Immediately post-weaning, piglets are prone to gastrointestinal infectious diseases. The active metabolite of vitamin D 1,25-dihydroxyvitamin D has direct impact on immune cell function and responses. Thus, a low vitamin D status may compromise the immune responses during infectious diseases. The aim of this study was to examine the effect of supplementation of different forms of vitamin D (25-OH-D3 and vitamin D3) to suckling piglets' vitamin D status at weaning. In addition, to determine whether the vitamin D status could affect the immune development in piglets and their robustness against E. coli challenge. Genetically E. coli F4 susceptible litters of piglets were divided into two treatment groups: group 1 (n = 16) provided milk formula supplemented with vitamin D3 (CON), and group 2 (n = 16) provided milk formula supplemented with 25-OH-D3 (TREAT). Piglets were offered the experimental milk formulas from day 3 after farrowing until weaning (at day 28 of age). A commercial weaner diet with high protein content were provided to induce weaning stress. Milk formulas, sow and weaner diets as well as plasma and milk samples obtained from sows (n = 8) were analysed for vitamin D metabolites. Vitamin D status in piglets was investigated by collection of plasma samples on day 3, 15, 28 and 35 of age. Eight piglets randomly selected from each dietary group (in total 16 pigs) were inoculated with E. coli F4 O149 on day 2 and 3 post-weaning. Blood samples collected on day 2 and 9 post-weaning (pre- and post E. coli inoculation, respectively) were analysed for haematological and immunological parameters including immunoglobulins, antibodies specific to E. coli O149 K88, cytokines and C-reactive protein. In addition, intestinal samples were obtained one week after E. coli inoculation to study the influence of infection and vitamin D status on immune responses at different sites of the intestine. This was accomplished by gene expression of various cytokines and tight junction proteins. In general, vitamin D status of the piglets were low. However, piglets provided TREAT during the suckling period had increased vitamin D status at weaning compared to piglets provided CON. Vitamin D was used during activation of the immune system as pigs inoculated with E. coli had lower plasma concentrations of 25-OH-D3 than non-inoculated pigs possibly due to mobilising of vitamin D in the liver. Hence, increased vitamin D status at weaning might improve piglets' resistance to E. coli infection.

Effect of combined maternal and post-hatch dietary 25-hydroxycholecalciferol supplementation on broiler chicken Pectoralis major muscle growth characteristics and satellite cell mitotic activity.

Skeletal muscle growth is largely dependent on the proliferation and differentiation of muscle-specific stem cells known as satellite cells (SC). Previous work has shown that dietary inclusion of the vitamin D3 metabolite, 25-hydroxycholecalciferol (25OHD3), also called calcidiol, can promote skeletal muscle growth in post-hatch broiler chickens. Improving vitamin D status of broiler breeder hens by feeding 25OHD3 in addition to vitamin D3 has also been shown to positively impact progeny. Yet, whether combined pre- and post-hatch supplementation with 25OHD3 produces an additive or synergistic SC-mediated, skeletal muscle growth response remains unanswered. To evaluate the effect of combined maternal and post-hatch dietary 25OHD3 supplementation on the growth and SC mitotic activity of the Pectoralis major (PM) muscles in broiler chickens, a randomized complete block design experiment with the main effects of maternal diet (MDIET) and post-hatch diet (PDIET) arranged in a 2 × 2 factorial treatment structure was conducted. From 25 to 36 wk of age, broiler breeder hens were fed 1 of 2 MDIET formulated to provide 5,000 IU D3 (MCTL) or 2,240 IU of D3 + 2,760 IU of 25OHD3 per kg of feed (M25OHD3). Their male broiler chick offspring (n = 400) hatched from eggs collected from 35 to 36 wk of age were reared in raised floor pens. Broilers were fed 1 of 2 PDIET formulated to provide 5,000 IU of D3 per kg of feed (PCTL) or 2,240 IU of D3 + 2,760 IU of 25OHD3 per kg of feed (P25OHD3). Muscle was collected at days 4, 8, 15, 22, and 29 and stored until immunofluorescence analysis. Data were analyzed as a 2-way ANOVA with SAS GLIMMIX. Dietary 25OHD3 was effectively transferred from hen plasma to egg yolks (P = 0.002) and to broiler progeny plasma (days 4 to 22; P ≤ 0.044). Including 25OHD3 in either MDIET or PDIET altered PM hypertrophic growth prior to day 29 (P ≥ 0.001) and tended to reduce Wooden Breast severity (P ≤ 0.089). Mitotic SC populations were increased in PM of MCTL:P25OHD3 and M25OHD:PCTL-fed broilers at d 4 (P = 0.037). At d 8, the PM mitotic SC populations were increased 33% by P25OHD3 (P = 0.054). The results of this study reveal that combined maternal and post-hatch 25OHD3 supplementation does not produce additive or synergistic effects on SC-mediated broiler muscle growth. However, vitamin D status improvement through dietary 25OHD3 inclusion in either the maternal or post-hatch diet stimulated broiler breast muscle growth by increasing proliferating SC populations.

Skeletal muscle growth is largely dependent on the proliferation and differentiation of muscle-specific stem cells known as satellite cells (SC). Previous work has shown that dietary inclusion of the vitamin D3 metabolite, 25-hydroxycholecalciferol (25OHD3), also called calcidiol, can promote skeletal muscle growth in post-hatch broiler chickens. Improving vitamin D status of broiler breeder hens by feeding 25OHD3 in addition to vitamin D3 has also been shown to positively impact progeny. Yet, whether combined pre- and post-hatch supplementation with 25OHD3 produces an additive or synergistic SC-mediated, skeletal muscle growth response remains unanswered. The results of this study reveal that combined maternal and post-hatch 25OHD3 supplementation does not produce additive or synergistic effects on SC-mediated broiler muscle growth. However, vitamin D status improvement through dietary 25OHD3 inclusion in either the maternal or post-hatch diet stimulated broiler breast muscle growth by increasing proliferating SC populations. Overall, this work answers not only practical questions for the broiler industry regarding the possible benefits of combining maternal and post-hatch dietary 25OHD3 supplementation but also improves our understanding of vitamin D's role in pre- and post-hatch broiler skeletal muscle growth.

A multi-omics approach to elucidate the mechanisms of action of a dietary muramidase administered to broiler chickens.

A novel dietary muramidase has been shown to have positive effects on broiler chickens. However, very little is known about its mechanisms of action. The present multi-omics investigation sought to address this knowledge gap. A total of 2,340 day-old male broilers were assigned to 3 groups (12 replicates each) fed, from 0 to 42 d, a basal diet (control group-CON) or the basal diet supplemented with muramidase at 25,000 (low-dose group-MUL) or 45,000 LSU(F)/kg feed (high-dose group-MUH). MUH significantly outperformed CON in terms of cumulative feed intake (4,798 vs 4,705 g), body weight (2,906 vs 2,775 g), and feed conversion ratio (1.686 vs 1.729), while MUL exhibited intermediate performance. At caecal level, MUH showed the lowest alpha diversity, a significantly different beta diversity, a reduction in Firmicutes, and a rise in Bacteroidetes, especially compared with MUL. MUH also exhibited a considerable decrease in Clostridiaceae and an overrepresentation of Bacteroidaceae and Lactobacillaceae. At blood level, MUH had lower hypoxanthine-probably due to its drop at caecal level-histidine, and uracil, while greater pyruvate, 2-oxoglutarate, and glucose. This study sheds light on the mode of action of this muramidase and lays the groundwork for future investigations on its effects on the intestinal ecosystem and systemic metabolism of broiler chickens.

Research Note: Changes in eggshell quality and microstructure related to hen age during a production cycle.

We have studied in detail the changes that occur in eggshell structure and composition during a production cycle in order to better understand the deterioration of eggshell quality with hen age (at 33, 45, and 67 wk). To study changes in eggshell ultrastructure and microstructure characteristics (mammillary density, palisade layer thickness, size, and orientation of calcite crystals) and the cuticle composition, we used complementary analytical techniques such optical and electron microscopy, X-ray diffraction, and infrared spectrometry. The marked decrease in eggshell breaking strength from 5.8 Kg at 33 wk to 4.4 Kg at 67 wk (25% reduction) could not be solely explained by the modest reduction in eggshell thickness (6-10% reduction) and seems to be associate to abrupt changes in eggshell ultra- and microstructure characteristics (i.e., decreased mammillary density; increased size of crystal units), occurring in older hens. Particularly, the decrease in mammillary density reduces the attachment points of the eggshell mineral to the membranes and therefore should negatively impact eggshell mechanical properties. Also, the observed increase in the calcite crystal size making the shell could also reduce the cohesion of crystals and eggshell resistance against impacts. Additionally, there was a decrease in the amount of cuticle and internal egg quality parameters (egg albumen height) with hen age that could have a negative impact in egg safety and quality.

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.

Role of long-term supplementation of 25-hydroxyvitamin D3 on laying hen bone 3-dimensional structural development.

Egg-laying hens have a unique bone development pattern due to the medullary bone formation and high bone turnover rate. The role of long-term supplementation of an intermediate form of vitamin D3, 25-hydroxyvitamin D3 (25OHD), on skeletal development of pullets and laying hens is not well established. Exploring its effects on layer bone development will help develop a strategy for preventing laying hen osteoporosis. The purpose of this study was to investigate the role of long-term supplementation of 25OHD in layer diets on bone 3-dimensional structural development. A total of 390 1-day-old Hy-Line W36 pullets were randomly allocated to 3 treatments with 10 replicate cages and 13 birds/cage. Dietary treatments were 1) vitamin D3 at 2,760 IU/kg, 2) vitamin D3 at 5,520 IU/kg, and 3) vitamin D3 at 2,760 IU/kg plus 25OHD at 2,760 IU (69 µg)/kg. The level of 25OHD in the serum was tested throughout the whole experimental period (0-95 wk). Bone growth rate (BGR) was measured at 10 wk using a calcein injection technique. Femurs were scanned using Micro-CT for 3D structural analysis, and the whole-body composition analysis was performed using a dual-energy x-ray absorptiometry method at 17, 60, and 95 wk. Dietary supplementation of 25OHD significantly increased 25OHD level in the serum from 0 to 95 wk. During the rearing period (0-17 wk), 25OHD increased BGR, cortical tissue volume, and bone marrow area at 17 wk, simultaneously. 25OHD created more pores in cortical bone, which resulted in a lower cortical bone mineral density (BMD) but without alerting bone mineral content (BMC). This effect allowed more space for mineral deposition in bones during the later egg-laying period. At 60 wk, the 25OHD group had significantly greater BMD, which led to the highest total BMC, cortical volume, and trabecular bone connectivity. At 95 wk, the birds fed 25OHD had significantly higher cortical bone volume and lower porosity. The 25OHD group also had higher total BMD and medullary bone volume but a lower BMC and volume of trabecular bone than vitamin D3 or double dosage vitamin D3 treatment. This indicated that the bone resorption rate was lower in cortical bone than that in trabecular bone in the late laying period. In conclusion, supplementation with dietary 25OHD could stimulate bone growth and increase bone volume in pullets to provide more space for mineral deposition during the laying period with positive effects on laying hen bone quality.

Role of long-term supplementation of 25-hydroxyvitamin D3 on egg production and egg quality of laying hen.

A study was conducted to evaluate the effect of dietary 25-hydroxyvitamin D3 (25OHD) on pullet and egg-laying hen growth performance, egg production, and egg quality. Three hundred and ninety 1-day-old Hy-Line W36 pullets were randomly allocated to 3 treatments with 10 replicated cages and 13 birds per cage. Dietary treatments were vitamin D3 at 2,760 IU/kg (D); vitamin D3 at 5,520 IU/kg (DD), and vitamin D3 at 2,760 IU/kg plus 25OHD at 2,760 IU (69 µg)/kg (25D). Body weight and feed intake were recorded at the end of each stage: starter 1 (0-3 wk), starter 2 (4-6 wk), grower (7-12 wk), developer (13-15 wk), prelay (15-17 wk), peaking (18-38 wk), layer 2 (39-48 wk), layer 3 (49-60 wk), layer 4 (61-75 wk), and layer 5 (76-95 wk). Egg production was recorded daily. Egg quality was evaluated every 8 wk starting from 25 wk. There was no difference in growth performance during the rearing period (0-17 wk). In the laying period (18-95 wk), DD showed lower feed intake at layer 2, but higher intake at layer 3 along with lower hen day production (HDP) from 22 to 48 wk compared to the other treatments. During the same period, the DD group laid smaller eggs with higher specific gravity and shell thickness compared with the other treatments or D alone at 40 wk, which may be partly due to the lower body weight. In contrast, 25D had better feed conversion ratio (feed intake per dozen of eggs) at layer 2, and higher overall (22-60 wk) HDP compared with DD. For the egg quality analysis, at 25 and 33 wk, both DD and 25D had higher Haugh unit compared with D. However, 25OHD has no effects on eggshell quality during the entire production period and no beneficial effects on egg production during the later laying period (after 60 wk). In summary, long-term and early supplementation of 25OHD has positive effects on egg production and egg quality, and the beneficial effects were mainly observed during the early laying stage.

Effects of Vitamin B2 Supplementation in Broilers Microbiota and Metabolome.

The study of the microbiome in broiler chickens holds great promise for the development of strategies for health maintenance and performance improvement. Nutritional strategies aimed at modulating the microbiota-host relationship can improve chickens' immunological status and metabolic fitness. Here, we present the results of a pilot trial aimed at analyzing the effects of a nutritional strategy involving vitamin B2 supplementation on the ileum, caeca and litter microbiota of Ross 308 broilers, as well as on the metabolic profile of the caecal content. Three groups of chickens were administered control diets and diets supplemented with two different dosages of vitamin B2. Ileum, caeca, and litter samples were obtained from subgroups of birds at three time points along the productive cycle. Sequencing of the 16S rRNA V3-V4 region and NMR metabolomics were used to explore microbiota composition and the concentration of metabolites of interest, including short-chain fatty acids. Vitamin B2 supplementation significantly modulated caeca microbiota, with the highest dosage being more effective in increasing the abundance of health-promoting bacterial groups, including Bifidobacterium, resulting in boosted production of butyrate, a well-known health-promoting metabolite, in the caeca environment.