Dietary 25-Hydroxycholecalciferol Supplementation as a Vitamin D3 Substitute Improves Performance, Egg Quality, Blood Indexes, Jejunal Morphology, and Tibia Quality in Late-Phase Laying Hens.

This study aimed to investigate whether a dietary 25-OHD3 addition improved the performance, egg quality, blood indexes, antioxidant status, jejunal morphology, and tibia quality of aged laying hens compared to a dietary VD3 addition. A total of 270 Hy-Line Brown laying hens at 55 wk of age were randomly assigned to three dietary treatments with six replicates (15 birds per replicate with 3 birds per cage). Chickens were fed a corn-soybean meal diet supplementation of 4000 IU/kg VD3 (control group), 50 µg/kg 25-OHD3 and 2000 IU/kg VD3 (experimental group 1), or 50 µg/kg 25-OHD3 and 4000 IU/kg VD3 (experimental group 2) for 12 weeks. The results demonstrated that 25-OHD3 caused a significant increase in the laying rate, especially in the 50 µg/kg 25-OHD3 + 2000 IU/kg VD3 group; the laying rate reached the maximum compared with other groups after 12 weeks (p < 0.05). However, there were no significant effects on the average egg weight, average daily feed intake, or feed-to-egg ratio (p > 0.05). A dietary supplementation of 50 µg/kg 25-OHD3 and 2000 IU/kg VD3 provided an improved eggshell strength, thick albumen height, and Haugh unit after 12 weeks (p < 0.05). Further analysis of the blood indexes showed that alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, calcium, and phosphorus were enhanced significantly in the 50 µg/kg 25-OHD3 + 2000 IU/kg VD3 group, while the content of total bilirubin decreased significantly (p < 0.05). In addition, the 25-OHD3 addition in diets improved the calcium and phosphorus contents in the serum (p < 0.05). The concentrations of 25-OHD3, parathyroid hormones, follicle-stimulating hormone, and progesterone were increased in the 50 µg/kg 25-OHD3 + 2000 IU/kg VD3 group, and the levels of cortisol, calcitonin, bone gla protein, and endotoxin in the serum reached a minimum in the 50 µg/kg 25-OHD3 + 4000 IU/kg VD3 group (p < 0.05), which constitutes an advantage for the aged laying hens. The antioxidant enzyme activities and free radical scavenging abilities in the 50 µg/kg 25-OHD3 + 2000 IU/kg VD3 group increased markedly, and the MDA level decreased significantly in the 50 µg/kg 25-OHD3 + 4000 IU/kg VD3 group (p < 0.05). Improvements in jejunal morphology and intestinal integrity resulted in an increased villi-length-to-crypt-depth ratio in the 50 µg/kg 25-OHD3 + 2000 IU/kg VD3 group (p < 0.05). Dietary 50 µg/kg 25-OHD3 and 2000 IU/kg VD3 additions improved the tibia quality, including fresh tibia weight, strength, mineral content (Ca), and trabeculae area (p < 0.05). Taken together, compared with the dietary VD3 addition, dietary supplementation of 25-OHD3 supported a stable physiological status for sustained egg production, egg quality, and bone quality in late-phase laying hens, and the addition levels of 50 µg/kg 25-OHD3 and 2000 IU/kg VD3 had the best effect. Therefore, this could provide a theoretical basis for the use of 25-OHD3 as a substitute forVD3.

Inclusion of guanidinoacetic acid in a low metabolizable energy diet improves broilers growth performance by elevating energy utilization efficiency through modulation serum metabolite profile.

This study was aimed to explore the elevating energy utilization efficiency mechanism for the potentially ameliorative effect of guanidinoacetic acid (GAA) addition on growth performance of broilers fed a low metabolizable energy (LME) diet. A total of 576 d old broilers were randomly allocated to one of the six treatments: a basal diet (normal ME, positive control, PC), or an LME diet (50 kcal/kg reduction in ME, negative control, NC) supplemented with 0.02%, 0.04%, 0.06%, and 0.08% GAA from 1 to 42 d of age, respectively. The GAA fortification in LME diet linearly or quadratically dropped (P < 0.05) the feed conversion ratio (FCR) from 22 to 42 and 1 to 42 d of age, abdominal fat rate on day 42, serum alanine aminotransferase (ALT) on day 21, and serum creatinine (CREAN) on days 21 and 42, elevated (P < 0.05) breast muscle rate and leg muscle rate on day 42, serum creatine kinase (CK) on days 21 and 42, as well as alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) on day 21. The dietary optimal GAA levels were 0.03%-0.08% based on the best-fitted quadratic models (P < 0.03) of the above parameters. Thus, the PC, LME, and 0.04% GAA-LME groups were selected for further analysis. Serum essential amino acids (EAA) tryptophan, histidine and arginine, non-essential amino acids (NEEA) serine, glutamine and aspartic acid were significantly decreased (P < 0.05), compared to PC diet by LME or 0.04% GAA-LME diet. 0.04% GAA-LME group reversed (P < 0.05) the reduction of arginine, 3-methyhistidine, and 1-methylhistidine by LME diet. Besides, six birds at 28 d of age from LME and 0.04% GAA-LME groups were selected for energy utilization observation in calorimetry chambers. The results demonstrated that 0.04% GAA-LME group significantly improved (P < 0.05) the ME intake (MEI) and net energy (NE) compared to the LME diet. Overall, these findings suggest that 0.04% GAA is the ideal dose of broilers fed the LME diet, which can significantly improve the growth performance and carcass characteristics by modulation of creatine metabolism through elevating serum CK activity and arginine concentration.

Guanidinoacetic acid (GAA) has been found to elevate energy utilization efficiency in broilers; however, the underlying mechanisms remain unclear. We investigated the effects of GAA addition in low metabolizable energy (LME) diet on growth performance, carcass characteristics and serum biochemical indices of broilers, and found that GAA addition linearly or quadratically dropped the feed conversion ratio from 22 to 42 and 1 to 42 d of age, abdominal fat rate on day 42, serum alanine aminotransferase on day 21, and serum creatinine on days 21 and 42, elevated breast muscle and leg muscle rate on day 42, serum creatine kinase, alkaline phosphatase, as well as lactate dehydrogenase on days 21 or 22. The dietary optimal GAA levels were 0.03%-0.08% based on the best-fitted quadratic models of the above parameters. Thus, further analysis was conducted and found that 0.04% GAA reversed the reduction of arginine, 3-methyhistidine, and 1-methylhistidine and improved the ME intake and net energy compared to the LME diet. These findings suggested that 0.04% GAA is the ideal dose for enhancing the energy utilization of broilers fed the LME diet, GAA addition can significantly improve the growth performance by elevating energy utilization efficiency through modulation serum metabolite profile.

Improvement in growth performance and digestive function from amniotic injections of N-acetylglutamate in broiler chickens.

BACKGROUND: N-acetylglutamate (NAG) is the initial and essectial substrate in the process of de novo arginine synthesis, plays an important role in intestinal development. The aim of this study was to determine the effects of in ovo feeding of NAG, 1.5 mg/egg at 17.5 days of incubation (DOI) via amnion, on hatching performance, early intestinal histomorphometry, jejunal barrier, digestive function, and growth performance of broiler chickens between 1 and 14 days of age. RESULTS: Amniotic injection of NAG had no significant effect on hatching characteristics compared with the non-injected control group (NC group). Birds in the NAG solution-injected group (NAG group) exhibited lower average daily feed intake and better feed efficiency during a period of 1-14 days. In comparison with the NC group, the NAG group had decreased crypt depth (CD) in the ileum and increased villus height (VH) / CD in the jejunum at 7 days, and decreased CD in duodenum and significantly increased VH in the jejunum at 14 days. However, the effects of in ovo supplementation with NAG on the density of goblet cells, and gene expression of mucin 2 and alkaline phosphatase were not significant. Chicks in the NAG group had a significantly higher mRNA expression level of trypsin and maltase in jejunum at 7 days than the NC group but not at 14 days. CONCLUSION: Amniotic injections of NAG (1.5 mg/egg) at 17.5 DOI could improve early growth performance of broilers during 1-14 days after hatching by accelerating the development of the intestine and enhancing jejunal digestive function. © 2023 Society of Chemical Industry.

Uterine inflammation status modulates eggshell mineralization via calcium transport and matrix protein synthesis in laying hens.

This study explored the effects of uterine inflammation on eggshell mineralization, ultrastructure and mechanical properties in laying hens modified by a lipopolysaccharide (LPS) challenge or dietary essential oil (EO) addition. In trial 1, a total of 72 Hy-line Brown layers at 36 wk of age were randomly assigned to 3 treatment groups (n = 8), where they were intravenously injected with phosphate buffered saline, LPS at 1 mg/kg body weight, or LPS 3 times at 24-h intervals. In trial 2, a total of 288 Hy-line Brown layers at 60 wk of age were randomly divided into 4 groups (n = 8), where they were fed basal diets supplemented with EO at 0, 50, 100 and 200 mg/kg for 12 wk. A uterine inflammation model was constructed with LPS treatment, indicated by the elevated expression of IL-1ß and TNF-α (P < 0.05) and lymphocyte infiltration. Uterine inflammation caused remarkable decreases in eggshell thickness and mechanical properties with structure deteriorations (P < 0.05). Uterine inflammation stimulated the expression of matrix proteins ovotransferrin (TF) and ovalbumin (OVAL), while depressing the mRNA levels of calbindin-1 (CALB1) and osteopontin in uterine mucosa (P < 0.05). In contrast, EO addition alleviated uterine inflammation, evidenced by depressed levels of IL-1ß and IL-6 (P < 0.05). There was a significant elevation in shell thickness and breaking strength following EO intervention (P < 0.05), and these effects were maximized at addition of 100 mg/kg. Further, EO improved shell ultrastructure including more early fusion, less type B mammillae, and increased effective thickness (P < 0.05). The alleviated inflammation decreased the expression of OVAL and TF, whereas ion transport genes like CALB1 and solute carrier family 26 member 9 were upregulated (P < 0.05). Our findings suggest that inflammatory status can impact uterine functions in calcium transport and the synthesis of matrix proteins especially such as OVAL and TF, which in turn modulates calcium precipitation and ultrastructure formation, thereby determining eggshell mechanical properties. These findings provide a novel insight into the uterine inflammation-mediated modifications of eggshell quality.

Effects of Saccharomyces cerevisiae hydrolysate on growth performance, immunity function, and intestinal health in broilers.

The current study was performed to explore the effects of dietary supplementation of Saccharomyces cerevisiae hydrolysate (SCH) on growth performance, immune function, and intestinal health in broiler chicken. A total of 300 Ross 308 male broilers (1-day-old) were randomly assigned to 2 dietary treatments including a basal diet (control group), and a basal diet supplemented with SCH feed additive (500 mg/kg in starter and grower phase, and 250 mg/kg in finisher phase). Each treatment had 6 replicates with 25 birds each. The results showed that the addition of SCH promoted growth during d 15 to 28 (P < 0.05). Although the addition of SCH had no significant effect on the intestinal relative indexes, it significantly increased the jejunum villus height (VH) and the ratio of villus height to crypt depth (VCR) of jejunum, and decreased the crypt depth (CD) of ileum (P < 0.05). Furthermore, SCH addition significantly downregulated the mRNA expression of immunomodulatory genes (TNF-α, IL-1ß, and IL-6), and upregulated the tight junction genes (ZO-1 and Claudin-1) (P < 0.05). High throughput sequencing analysis of bacterial 16S rRNA revealed that dietary SCH supplementation altered cecum microbiota. Alpha diversity analysis showed that a higher bacterial richness in cecum of broilers fed with SCH. The composition of cecum microbiota regulated by SCH addition was characterized by an increased abundance of Firmicutes and a reduced abundance of Bacteroidetes. At the genus level, dietary SCH resulted in a decrease of Bacteroides and an increase of short-chain fatty acids (SCFA) -producing bacteria including Lactobacillus and Faecalibacterium. Taken together, dietary SCH supplementation can stimulate the growth of broilers by regulating the intestinal immunity and barrier function, and improving the intestinal morphology, which may be related to the enhancement of bacterial diversity and the changes of intestinal microbial composition.

Yeast cell-wall polysaccharides improve immunity and attenuate inflammatory response via modulating gut microbiota in LPS-challenged laying hens.

Effects of dietary supplementation of yeast cell-wall polysaccharides (YCWP) on production performance, ileal microbial composition, immunomodulatory and anti-inflammatory effects in LPS-challenged laying hens, were evaluated. A total of 288 35-week-old Hy-Line Brown layers were randomly assigned into 4 dietary treatments: 0, 250, 500 and 1000 mg/kg YCWP, respectively. After a 12-week feeding period, a total of 32 birds were selected from the control (n = 16) and 1000 mg/kg YCWP group (n = 16). For each group, half (n = 8) received Escherichia coli LPS and half (n = 8) received PBS at 1 mg/kg body weight, intravenously. Results showed that YCWP enhanced feed efficiency and egg production linearly, with optimal laying performance notable in the 1000 mg/kg YCWP group. Dietary YCWP enhanced serum IgM and expression of ileal avian ß-defensin, alleviated the LPS-induced elevated levels of serum IL-6 and IL-1ß and the up-regulated expression of IL-1ß, TNF-α, IFN-γ, and IL-6 in spleen and/or ileal mucosa. Furthermore, anti-inflammatory and immunomodulatory effects of YCWP were linked with its enhancement effect on microbial diversity, proliferation of Bifidobacteriaceae, Lactocillus, Candidatus_Arthromitus, Streptomyces, Bacillaceae, and Desulfovibrio, and reduced abundance of Shigella. Therefore, YCWP has the potentials to be utilized as safe prebiotics and gut enhancer in laying hens.

Effects of Dietary Selenium Sources on Physiological Status of Laying Hens and Production of Selenium-Enriched Eggs.

Developing new sources of organic selenium (Se) has potential benefits for animal production and human nutrition via animal-based foods enriched with Se. The objective of this study was to evaluate the effects of Se-enriched insect protein (SEIP) in comparison with other sources, such as sodium selenite (SS) and selenium-enriched yeast (SEY), on performance, egg quality, selenium concentration in eggs, serum biochemical indices, immune capacity, and intestinal morphology of laying hens. Four hundred and fifty 24-week-old Hy-Line Brown laying hens with 94.0 ± 1.5% laying rate were randomly allocated to five groups with six replicates of 15 hens each. The control diet was prepared without adding exogenous selenium (calculated basal Se content of 0.08 mg/kg). The normal group was fed basal diets supplemented with 0.3 mg/kg of Se provided by sodium selenite. Three treatment groups (SS, SEY, and SEIP, respectively) were fed basal diets supplemented with 2 mg/kg of Se provided by sodium selenite, Se-enriched yeast, and SEIP, respectively. The feeding trial lasted for 12 weeks. Results revealed that dietary supplementation of 2 mg/kg of Se increased egg weight, decreased feed conversion ratio, and enhanced the antioxidant capacity of eggs in laying hens relative to the control group, whereas no significant differences were observed among SS, SEY, and SEIP treatment groups for the same. The organic source of Se provided by SEY or SEIP showed higher bio efficiency, as indicated by higher selenium content in eggs of SEY and SEIP compared with SS, although higher content was observed in SEY compared with SEIP. Also, the organic Se source significantly improved antioxidant capacity and immune functions of laying hens than the inorganic Se source. Diets supplemented with SEIP and SS significantly improved jejunal morphology of the laying hens compared with SEY, whereas SEIP was more effective than SEY to improve the oviduct health of laying hens. The results of this work evidently points the additive effect and nontoxicity of SEIP. Thus, SEIP could be used as another organic source of Se in the diet of laying hens and production of selenium-enriched eggs for humans.

Vitamin E Supplementation Enhances Lipid Oxidative Stability via Increasing Vitamin E Retention, Rather Than Gene Expression of MAPK-Nrf2 Signaling Pathway in Muscles of Broilers.

Dietary vitamin E (VE) supplementation is a method to produce VE-enriched meat and improve meat lipid oxidative stability. We aimed to study the effect of the VE supplementation duration on meat lipid oxidative stability, VE retention, and antioxidant enzymes' activity, and explore its relationship with the mitogen-activated protein kinases (MAPK)-nuclear factor-erythroid 2-related factor 2 (Nrf2) signaling pathway in broilers slaughtered after electrical stunning. A total of 240 male 18-day-old Arbor Acres Plus broilers were distributed to four treatments, with six replicates in each treatment, and ten broilers per replicate. Broilers were fed with a basal diet (no supplementation of VE) or VE diet (200 IU/kg VE, DL-α- tocopherol) for one (W1), two (W2), or three (W3) weeks before electrical stunning (130 mA, 60 Hz, for 1s) and slaughter. The VE retention was positively and linearly affected (p < 0.01) by the VE feeding duration at one to three weeks before slaughter, and negatively (all p < 0.01) related to the thiobarbituric acid reactive substance (TBARS) content in both breast and thigh muscles at d 0, d 2, and d 6 postmortem. The VE retention was negatively (p < 0.05) related to the gene expression of c-Jun N-terminal kinases 1 (JNK1) and 2 (JNK2), Nrf2 in breast muscles, and JNK1 and p38 MAPK in thigh muscles. In conclusion, dietary vitamin E supplementation at 200 IU/kg for three weeks before electrical stunning and slaughter improved lipid oxidative stability via increasing VE retention, rather than the regulation by gene expression of the MAPK-Nrf2 signaling pathway in skeletal muscles of broilers.

Effects of Selenium Conjugated to Insect Protein on Pharmacokinetics of Florfenicol and Enrofloxacin in Laying Hens.

In the context of increasing awareness on the dietary supplementation of organic selenium in commercial poultry production and ensuring safe egg production, the present study investigated the effects of selenium on the pharmacokinetics of the therapeutic use of florfenicol and enrofloxacin from perspectives of laying performance, selenium deposition in eggs, and drug residue in plasma, organs, and eggs. A 2 × 3 factorial arrangement with two kinds of drugs (florfenicol vs. enrofloxacin, 200 mg/kg) and three levels of dietary organic selenium SCIP (selenium conjugated to insect protein) (0, 2, and 5 mg/kg) was designed together with a blank control group. Healthy Hy-Line Brown laying hens (n = 252, 40-week-old and 90.0 ± 1.7% of egg production rate) were randomly allocated into one of seven treatments with six replicates and six hens per replicate. The experiment lasted for 42 days and consisted of three periods (adjusted stage, depositional stage, and eliminating stage) of 14 days each. These stages entail feeding of the laying hens with basal diets, addition of drugs and selenium synchronously into the diets, drug withdrawal from diet, and supply of selenium uninterruptedly in the diet. Egg production and feed intake were recorded on daily and weekly bases, respectively. The selenium content in egg yolk, egg white, and whole eggs and the drug residues in eggs, plasma, liver, kidney, and breast muscle were determined on days 2, 3, 5, 6, 7, 9, 11, and 14 of the depositional and eliminating stages. There was no significant difference (p > 0.05) in egg production among the dietary treatments, but feed intake decreased significantly (p < 0.05) in the drug treatment group compared to other groups. Dietary organic selenium decreased the residue of drugs in tissues and eggs, while the metabolism and deposition of selenium in laying hens were suppressed due to drug effects. The results of the present study are of significance to enrich the knowledge of the pharmacokinetics of florfenicol and enrofloxacin in laying hens and ensure the quality of poultry products.

Supplemental L-Arginine Improves the Embryonic Intestine Development and Microbial Succession in a Chick Embryo Model.

Early colonization of intestinal microbiota plays an important role in intestinal development. However, the microbial succession at an embryonic stage and its assembly patterns induced by prenatal nutrition are unknown. In the present study, we used a chick embryo model to investigate the effects of in ovo feeding (IOF) of L-arginine (Arg) on the intestinal development and microbial succession of embryos. A total of 216 fertile eggs were randomly distributed into 2 groups including the non-injected control group and IOF of Arg group with 7 mg/egg. The results showed that IOF Arg increased the intestinal index, absolute weight of jejunum, and improved jejunal morphology in terms of villus width and surface area (p < 0.05). The relative mRNA expressions of mTOR and 4E-BP1 were up-regulated and accompanied by higher contents of Mucin-2 in the Arg group (p < 0.05). There was a significant elevation in contents of serum glucose and high-density lipoprotein cholesterol, whereas there was a decreased low-density lipoprotein cholesterol in the Arg group (p < 0.05). Additionally, Proteobacteria and Firmicutes were major intestinal bacteria species at the embryonic stage. However, Arg supplementation targeted to shape assembly patterns of microbial succession and then changed microbial composition (p = 0.05). Meanwhile, several short-chain fatty acids (SCFAs)-producing bacteria, such as Roseburia, Blautia, and Ruminococcus were identified as biomarkers in the Arg group (LDA > 3, p < 0.05). Accordingly, significant elevated concentrations of SCFAs, including lactic acid and formic acid, were observed in the Arg group (p < 0.05), accompanied by the higher concentration of butyric acid (0.05 < p < 0.10). In conclusion, prenatal Arg supplementation improved embryonic intestine development by regulating glucose and lipid homeostasis to supply more energy for chick embryos. The possible mechanism could be the roles of Arg in shaping the microbial assembly pattern and succession of the embryonic intestine, particularly the enrichment of potential probiotics. These findings may contribute to exploring nutritional strategies to establish health-promoting microbiota by manipulating prenatal host-microbe interactions for the healthy development of neonates.

Effect of dietary serine supplementation on performance, egg quality, serum indices, and ileal mucosal immunity in laying hens fed a low crude protein diet.

This study was designed to evaluate the effect of dietary Ser on performance, egg quality, serum indices, and ileal mucosal immunity in laying hens fed low crude protein (LCP), essential amino acids (EAA) balanced diets. A total of 480 Hy-Line Brown layers at 24 wk of age were randomly assigned into 5 dietary treatments with 8 replicates of 12 birds each. Treatments included a control diet (16.49% CP), and 4 LCP, EAA balanced diets (14.05% CP) supplemented with 0, 0.114%, 0.306%, 0.498% L-Ser, respectively. Dietary Ser supplementation linearly increased hen-day egg production (HDEP; P < 0.05) and decreased feed-to-egg ratio (P < 0.05) among LCP groups from wk 6 to 10, and the optimal HDEP of layers occurred at Ser level of 0.498%. At the end of wk 10, birds in the control had higher albumen height and thick white proportion than those fed the LCP diet without Ser addition (P < 0.05), and presented a lower yolk color score than all LCP groups (P < 0.05). Among LCP groups, serum total protein and globulin contents were significantly increased by dietary Ser addition at the levels of 0.306% and 0.498% (P < 0.05), and had a linear response to the supplemental Ser levels (P < 0.05). Furthermore, dietary 0.498% Ser supplementation significantly increased serum immunoglobulin G and immunoglobulin M contents (P < 0.05) and up-regulated the expression of mucin 2, secretory immunoglobulin A, and relevant glycosyltransferases of O-glycosylation in ileal mucosa (P < 0.05). The increased expression of proinflammatory cytokines IFN-γ and IL-1ß induced by LCP diets (P < 0.05) was reversed following 0.498% Ser addition (P < 0.05). Collectively, dietary CP reduction by 2.44% could maintain the productive performance of layers when it was fortified with certain EAA, though poor albumen quality, and ileal inflammation were occurred. The addition of Ser to LCP diets improved performance probably through enhancing humoral and ileal mucosal immunity and attenuating the ileal inflammation of layers.

Application of Selenium Conjugated to Animal Protein in Laying Hens' Diet for the Production of Selenium-Enriched Eggs.

The current experiment was conducted to investigate the application effects of selenium conjugated to insect protein (SCIP) in the production of selenium-enriched eggs. A total of 450 laying hens were randomly assigned to five dietary groups, each group consisting of six replicates. Hens in the control group received a diet without selenium supplementation, whereas hens in the other four groups received diets supplemented with either 1, 2, 5, or 10 mg/kg of selenium from SCIP. The productive performance, egg quality, antioxidant and immune capacity, biochemical indices, intestinal morphology, and oviduct health of laying hens were evaluated. The results showed that the supplementation of organic selenium provided by SCIP in the diets of laying hens enhanced performance and egg quality without any toxicity effect, even at the 10 mg/kg inclusion level. A level of 2 mg/kg of selenium provided by SCIP in diets tentatively improved the serum antioxidant and immune capacity, intestinal development, and oviduct health of laying hens in a conspicuous manner. Hence, the biosafety and positive effects of SCIP as a feed additive supplement in laying hens' diet have been demonstrated with the enhanced production of safe and selenium-enriched eggs.

The partial replacement of sodium chloride with sodium bicarbonate or sodium sulfate in laying hen diets improved laying performance, and eggshell quality and ultrastructure.

This study investigated the effects of dietary chloride (Cl) reduction on laying performance and eggshell quality by substitution of sodium bicarbonate (NaHCO3) or sodium sulfate (Na2SO4) for part of dietary sodium chloride (NaCl), and further explored its mechanism for improving eggshell quality. A total of 360 29-wk-old Hy-line Brown laying hens were randomly allocated to 5 dietary treatments, including a basal diet contained 0.33% NaCl (control group, 0.27% dietary Cl), and 4 experimental diets that contained 0.21% and 0.15% dietary Cl by substituting Na2SO4 or NaHCO3 for part of NaCl in the basal diet. No significant differences were observed in blood Na+, Cl-, K+ and Ca2+ levels and pH value as well as serum creatinine and uric acid contents among 5 treatments (P > 0.05). Dietary Cl reduction increased egg production and ADFI during wk 33 to 36, 37 to 40 and 29 to 40 of age and decreased feed conversion ratio during wk 37 to 40 of age (P < 0.05). The hens fed with diets containing 0.15% Cl increased eggshell breaking strength, thickness and weight ratio in wk 40 of age (P < 0.05). Birds fed with dietary 0.21% and 0.15% Cl exhibited higher effective layer thickness and lower mammillary layer thickness of eggshell than those fed with dietary 0.27% Cl (P < 0.05). Apparent Ca metabolizability of hens was increased with dietary Cl reduction (P < 0.05). Total Ca of eggshell of dietary 0.15% Cl group was higher than that of dietary 0.27% Cl group (P < 0.05). No significant differences in laying performance, eggshell quality and Ca metabolism of layers were observed between Na2SO4 and NaHCO3 replacement groups (P > 0.05). Overall, dietary Cl reductions could improve laying performance and eggshell quality by substitution of NaHCO3 or Na2SO4 for part of NaCl, and there were no differences in the improvements between these two substitutes. The improved eggshell quality may be attributed to improved eggshell ultrastructure and increased supply of eggshell Ca2CO3.

Effects of dietary Bacillus subtilis supplementation and calcium levels on performance and eggshell quality of laying hens in the late phase of production.

The objective of this study was to evaluate the effects of dietary Bacillus subtilis supplementation and calcium (Ca) levels on performance, eggshell quality, intestinal morphology, and relative calbindin-D28k (CALB1) mRNA level of laying hens in the late phase of production. An experiment employing a 2 × 3 factorial arrangement of 3 levels of Ca (3.5, 4.0, and 4.5%) and the absence or presence of B. subtilis was carried out with a total of 576 Hy-Line Brown laying hens aged 72 to 79 wk. Every group had 8 replicates of 12 birds each. The results showed that 4.0 and 4.5% Ca levels improved (P < 0.05) apparent retention and serum Ca content of aged laying hens. Compared with the 3.5% Ca level, the 4.0% Ca level in diets increased (P < 0.05) thickness, eggshell weight, shell ratio, and eggshell Ca content of aged laying hens. Moreover, breaking strength, thickness, eggshell weight, shell ratio, eggshell Ca content, apparent retention of Ca in g/day, apparent retention of Ca in percent, villus height, villus height/crypt depth, serum Ca level, and relative CALB1 mRNA level of aged laying hens were all increased (P < 0.05) by B. subtilis supplementation in diets. The supplemental B. subtilis decreased feed conversion ratio (P = 0.001) significantly. In addition, there was an interaction effect between increased Ca levels from 3.5 to 4.5% and B. subtilis supplementation on crypt depth in the duodenum (P < 0.05). In conclusion, we found that both the increase in dietary Ca level from 3.5 to 4.5% and B. subtilis supplementation could enhance intestinal Ca absorption and improve eggshell quality of laying hens in the late phase of production (72-79 wk of age). Dietary supplementation of B. subtilis accompanying the 4.0% Ca level was appropriate in enhancement of eggshell quality.

Cyst(e)ine fortification in low crude protein diet improves growth performance of broilers by modulating serum metabolite profile.

This study was aimed to explore the metabolomical mechanisms for the potentially ameliorative effect of cyst(e)ine (Cys) fortification on growth performance of broilers fed low crude protein (CP) diet. A total of 432 1-d-old broilers were randomly divided into 6 groups, each of which received one of the following diets: normal-CP diet (positive control, PC), low-CP diet (negative control, NC), NC diet fortified with 0.05%, 0.1%, 0.15% or 0.2% of Cys. Samples were collected on d 42. Results showed that increasing Cys fortification quadratically elevated (P < 0.05) the accumulative growth performance and leg muscle yield of broilers fed NC diet, with 0.1% being the optimal dose. Thus, samples from PC, NC and NC plus 0.1% Cys (NCC) groups were selected for further analysis. Both dietary CP reduction and fortification of 0.1% Cys in NC diet caused complex changes (P < 0.05) in serum amino acids and some other metabolites primarily involved in lipid metabolism. Multiple lipogenesis-related pathways were regulated (P < 0.05) following Cys fortification in NC diet, which could at least partially interpret the benefit of Cys fortification in NC diet on broiler performance. In conclusion, fortifying low-CP diet with 0.1% Cys promoted the growth performance of broilers probably through modulating serum metabolite profile.

The Response of Broiler Chickens to Dietary Soybean Meal Reduction with Glycine and Cysteine Inclusion at Marginal Sulfur Amino Acids (SAA) Deficiency.

The responses of broiler chickens to dietary protein reduction were investigated in the presence of glycine and cysteine inclusion at the marginal deficiency of sulfur-containing amino acids. A total of 432 broiler chickens were allotted to six dietary treatments; SP1 is standard protein diet with 100% total sulfur amino acids (TSAA), SP2 is standard protein diet with 85% TSAA, RP is reduced protein diet without glycine and cysteine supplementation, RPC is reduced protein diet with cysteine supplementation at 0.1%, and RPG is reduced protein diet with 1% glycine supplementation, while RPGC is reduced protein diet with 0.1% cysteine and 1% glycine supplementation. In this study, 4.5% protein is reduced in diets-thus, 17.5% CP (crude protein) for starter phase and 15.5% CP for the grower phase. Reduced protein diets contained 85% TSAA. Broiler chickens fed standard protein diet SP2 had superior bodyweight (BW) (p ≤ 0.05) in the starter and grower phase, average daily gain (ADG) (p ≤ 0.05) in the starter and entire feeding period, average daily feed intake (ADFI) (p ≤ 0.05) in the starter phase, and better feed conversion ratio (FCR) (p ≤ 0.05) in the starter, grower and entire feeding period; however, RPGC showed higher ADG (p ≤ 0.05) in the grower phase, and ADFI (p ≤ 0.05) in the grower and entire feeding period. RPC and RPG diet improved BW (p ≤ 0.05), ADG (p ≤ 0.05), ADFI (p ≤ 0.05), and better FCR (p ≤ 0.05) in starter, grower, entire feeding period compared to RP. The RPGC group had higher BW (p ≤ 0.05), ADG (p ≤ 0.05), ADFI (p ≤ 0.05) and better FCR (p ≤ 0.05) compared to the RPC group. Blood biochemical parameters showed that Broiler chickens fed on the SP2 diet had higher levels of total protein (TP) (p ≤ 0.05), albumin (ALB) (p ≤ 0.05), creatinine (CRE) (p ≤ 0.05), and aspartate aminotransferase (AST) (p ≤ 0.05) and, lower level of uric acid (UA) (p ≤ 0.05), blood urea nitrogen (BUN) (p ≤ 0.05), glucose (GLU) (p ≤ 0.05), and alanine aminotransferase (ALT) (p ≤ 0.05) in the starter phase; however, higher level of TP (p ≤ 0.05), GLU (p ≤ 0.05), CRE (p ≤ 0.05), and AST (p ≤ 0.05), and lower level of ALB (p ≤ 0.05), UA (p ≤ 0.05), and ALT (p ≤ 0.05) in the grower phase; RPGC had higher level of TP (p ≤ 0.05), UA (p ≤ 0.05), GLU (p ≤ 0.05), ALT (p ≤ 0.05) and AST (p ≤ 0.05), and lower level of ALB (p ≤ 0.05), BUN (p ≤ 0.05), and CRE (p ≤ 0.05) in the starter phase; however, in grower phase, RPGC had higher level of TP (p ≤ 0.05), and ALB (p ≤ 0.05), and lower level of UA (p ≤ 0.05), CRE (p ≤ 0.05), ALT (p ≤ 0.05), and AST (p ≤ 0.05). Free amino acids profile showed that broiler fed on standard protein diet SP2 had reduced the methionine (p ≤ 0.05) concentration; RPC increased the concentrations of taurine (p ≤ 0.05), phosphoethanolamine (p ≤ 0.05), threonine (p ≤ 0.05), valine (p ≤ 0.05), isoleucine (p ≤ 0.05), phenylalanine (p ≤ 0.05), ornithine (p ≤ 0.05), and lysine (p ≤ 0.05) and reduced the citrulline (p ≤ 0.05) concentration; RPG increased the concentration of glutamate (p ≤ 0.05), glycine (p ≤ 0.05), cysteine (p ≤ 0.05), and arginine (p ≤ 0.05), and decreased the concentration of tyrosine (p ≤ 0.05); and RPGC increased the concentration of serine (p ≤ 0.05) and reduced the concentration of hydroxyproline (p ≤ 0.05). Serum metabolites analysis showed that reduced protein downregulated the 54 metabolites; however, glycine fortification up-regulated the Benzamide, Pro-Ser, N-Carbamylglutamate, D-gluconate, and Gamma-Glutamylcysteine. Carcass quality showed that SP2 decreased the abdominal fat percentage (p ≤ 0.05). Nitrogen digestibility was higher by the diet RP (p ≤ 0.05). This study demonstrated that protein content could be reduced up to 4.5% with 1% glycine and 0.1% cysteine fortification in diet, which has the potential to inhibit the adverse effect of reduced protein and attain the standard growth performance.

Effects of in ovo feeding of N-acetyl-L-glutamate on early intestinal development and growth performance in broiler chickens.

The present study determined the effects of in ovo feeding (IOF) of N-acetyl-L-glutamate (NAG) on early intestinal development and growth performance of broilers. A total of 702 fertile broiler eggs were randomly divided into 3 treatments: 1) non-punctured control group, 2) saline-injected control group, and 3) NAG solution-injected group (1.5 mg/egg). At 17.5 D of incubation, 300 µL of each solution was injected into each egg of injected groups. Results indicated that the hatchability and healthy chicken rate were not affected by NAG injection (P > 0.05). Chicks from NAG solution-injected group had significantly decreased average daily feed intake and feed conversion ratio during 1-14 D than those in the non-punctured control group (P < 0.05). Compared with the non-punctured control group, IOF of NAG significantly increased the density of goblet cells in jejunum at hatch, duodenum at 7 D, and ileum at 14 D; decreased crypt depth in jejunum at hatch; and increased villus height in duodenum and jejunum and villus height:crypt depth ratio in duodenum at 7 D (P < 0.05). The intestinal mRNA expression of Na+-dependent neutral amino acid transporter, peptide transporter, and excitatory amino acid transporter 3 did not differ between groups at 7 or 14 D. However, the mRNA expression level of rBAT in jejunum significantly increased in the NAG solution-injected group than in the non-punctured control group at 7 D (P < 0.05). In conclusion, IOF of NAG (1.5 mg/egg) accelerated the early intestinal development by enhancing intestinal immune and absorption function, thereby positively affecting the feed efficiency for the first 2 wk post-hatch.

Dynamic alterations in early intestinal development, microbiota and metabolome induced by in ovo feeding of L-arginine in a layer chick model.

BACKGROUND: Prenatal nutrition is crucial for embryonic development and neonatal growth, and has the potential to be a main determinant of life-long health. In the present study, we used a layer chick model to investigate the effects of in ovo feeding (IOF) of L-arginine (Arg) on growth, intestinal development, intestinal microbiota and metabolism. The treatments included the non-injected control, saline-injected control, and saline containing 2, 6, or 10 mg Arg groups. RESULTS: IOF Arg increased early intestinal index and villus height, and enhanced uptake of residual yolk lipid, contributing to subsequent improvement in the early growth performance of chicks. Prenatal Arg supplementation also increased the early microbial α-diversity, the relative abundance of Lactobacillales and Clostridiales, and decreased the relative abundance of Proteobacteria of cecum in chicks. Furthermore, the shift of cecal microbiota composition and the colonization of potential probiotics were accelerated by IOF of Arg. Simultaneously, metabolomics showed that metabolisms of galactose, taurine-conjugated bile acids and lipids were modulated to direct more energy and nutrients towards rapid growth of intestine at the beginning of post-hatch when embryos received IOF of Arg. CONCLUSIONS: Prenatal Arg supplementation showed beneficial effects on the early intestinal development, cecal microbiota and host metabolism of layer chicks, contributing to subsequent improvement in the early growth performance. These findings provide new insight into the role of IOF of Arg in the establishment of the gut microbiota of newly-hatched layer chicks, and can expand our fundamental knowledge about prenatal nutrition, early bacterial colonization and intestinal development in neonate.

Comparative effects of dietary microalgae oil and fish oil on fatty acid composition and sensory quality of table eggs.

This study was conducted to investigate the comparative effects of dietary supplementation with microalgae oil or fish oil on fatty acid composition, sensory quality, and overall acceptability of table eggs. A total of six hundred thirty, 30-week-old, Hy-Line Brown laying hens were allocated to 7 dietary treatments. Layers were fed with a control diet or the control diet supplemented with graded levels of docosahexaenoic acid (DHA; 1.25, 2.50, and 5.00 mg/g feed) derived from microalgae oil or fish oil. The feeding trial lasted 10 wk. Enrichment of eggs with DHA by dietary supplementation with microalgae oil or fish oil enhanced yolk DHA and total n-3 polyunsaturated fatty acid (PUFA) enrichment and reduced n-6-to-n-3 ratio in a dose-response manner, whereas the efficiency decreased (P < 0.05). The overall efficiency of DHA incorporation into eggs was similar for the 2 sources (P > 0.05). In comparison with fish oil, dietary microalgae oil supplementation resulted in higher scores for egg flavor and overall acceptability, both of which declined linearly in response to DHA supplementation (P < 0.05). Among the aroma and flavor characteristics, fishy aroma and flavor scores increased linearly and quadratically (P < 0.05) in response to dietary DHA supplementation, and egg aroma and flavor and milky flavor scores decreased linearly (P < 0.05). The results from partial least squares analysis showed that fishy flavor and aroma were closely associated with DHA, α-linolenic acid, and total n-3 PUFA, and oleic acid, arachidonic acid, and dihomo-gamma-linolenic acid were more related to egg aroma and flavor. The results suggested that microalgae oil would be more promising for egg DHA enrichment owing to better sensory quality of the resultant eggs.

Standardized ileal digestible amino acid and metabolizable energy content of wheat from different origins and the effect of exogenous xylanase on their determination in broilers.

This study was conducted to determine the standardized ileal digestible amino acids (SID AA) and nitrogen-corrected apparent metabolizable energy (AMEn) contents of 6 wheats from different origins in China and incidentally to investigate the effects of exogenous xylanase addition on SID AA and AMEn determination in broiler chicks. A total of 480 chicks were divided into 48 cages of 10 birds each balanced for body weight and fed 8 types of diets in a completely randomized design (6 replicated cages per diet) from 21 to 26 d of age. The individual wheat constituted the only source of crude protein in a semi-purified experimental diet. A nitrogen-free diet was designed to estimate basal endogenous AA loss and determine the SID AA. Titanium oxide (0.3%) was used as an indigestibility marker, and nutrient digestibility and retention were determined by the substitution method. From day 24 to 26, excreta samples were collected for AMEn determination. On day 26, the birds were euthanized, and ileum contents were obtained for AA digestibility determination. Wheat from Gansu had greater (P < 0.05) SID AA contents except Lys, Thr, Phe, and Cys, with a higher (P < 0.001) AMEn (11.83 MJ/kg) than the other wheats. The SID content of mean indispensable amino acids and dispensable amino acids were 87.35% and 88.17%, respectively, and the average AMEn value of 6 wheats was 11.14 MJ/kg. Compared with the diet without xylanase, the added xylanase resulted in higher (P < 0.05) SID contents of Met, Lys, Trp, Arg, Ile, Leu, Val, Gly, Asp, Glu, Pro, and Ala; the SID AA values were raised by 1.96% (mean of all AA); and the AMEn content was significantly increased (+0.87 MJ/kg) (P < 0.05). In conclusion, origins of wheats have significant effects on SID AA and AMEn values which were positively correlated with crude protein content of wheat; exogenous xylanase addition to a wheat-based poultry diet could significantly improve SID AA and AMEn contents for broilers.