Effects of early weaning on intestinal morphology, digestive enzyme activity, antioxidant status, and cytokine status in domestic pigeon squabs (Columba livia).

The aim of this study was to explore the effects of early weaning on growth performance, intestinal morphology, digestive enzyme activity, antioxidant status, and cytokine status in domestic pigeon squabs (Columba livia). The conclusion is based on body weight (BW) and average daily gain (ADG), length index and weight index of small intestine, small intestinal morphology, activity of digestive enzymes in duodenum content, the concentrations of jejunal antioxidant status and cytokines. A completely randomized design with 2 treatments, the control group (CON) and early weaning (EW) group, was utilized. Eight squabs per treatment were sampled at the age of 25 d. The results showed that early weaning reduced BW (P < 0.05), ADG (P < 0.05), ileac length index (P < 0.05), and weight index (P < 0.01). Compared with the CON group, small intestinal morphology was altered in the EW group. Ileac crypt depth (CD) increased significantly (P < 0.01). The villus area was decreased in the duodenum (P < 0.05), jejunum (P < 0.01), and ileum (P < 0.05). The ileac ratio of villus height to crypt depth (VCR) in the EW group was lower than the ileac ratio of villus height to VCR in the CON group (P < 0.01). The activity of trypsin (P < 0.05), sucrase (P < 0.01) and aminopeptidase-N (APN) (P < 0.01) in the duodenum was reduced. Jejunal malondialdehyde (MDA) (P < 0.01) was increased and total superoxide dismutase (T-SOD) (P < 0.01) was reduced significantly. Early weaning decreased the concentrations of interferon-γ (IFN-γ) (P < 0.01), interleukin-4 (IL-4) (P < 0.05) and interleukin-10 (IL-10) (P < 0.01) but induced significant upregulation of interleukin-2 (IL-2) (P < 0.05). In conclusion, our results suggested that early weaning did harm the BW and ADG, intestinal length index and weight index, intestinal morphology, activity of digestive enzymes, and antioxidant and cytokine status.

Effects of parental dietary linoleic acid on growth performance, antioxidant capacity, and lipid metabolism in domestic pigeons (Columba livia).

The objective of this study was to evaluate the effects of dietary linoleic acid (LA) on growth performance, antioxidant capacity, and lipid metabolism in pigeon squabs by supplementing LA in their parental diets. A completely randomized design that consisted of a control group, 1% dietary LA addition group (LA1%), 2% dietary LA addition group (LA2%), and 4% dietary LA addition group (LA4%) was used. Six squabs from each treatment were randomly sampled at the day of hatch and days 7, 14, and 21 after hatch. The results showed that parental dietary LA had no significant influence (P > 0.05) on body weight (BW) gain or relative organ weights (% of BW) in squabs. The activities of superoxide dismutase, catalase, and glutathione peroxidase in the LA1% were significantly increased (P < 0.05) compared with those in the control group. The malondialdehyde content in the LA1% was significantly lower (P < 0.05) than that in the control group. The levels of serum triglyceride in the LA1% and LA2% were significantly decreased (P < 0.05) compared with those in the control group, whereas the serum high-density lipoprotein cholesterol level in the LA1% and LA2% and the free fatty acid level in the LA4% were significantly higher (P < 0.05) than those of the control group. The activities of lipoprotein lipase, hepatic lipase, and hormone-sensitive lipase in the LA1% were significantly higher (P < 0.05) than those in the control group. The 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in the LA1% and the hormone-sensitive lipase activity in the LA4% were significantly decreased (P < 0.05) compared with those in the control group. The mRNA expression of carnitine palmitoyltransferase 1, acyl-CoA 1, and peroxisome proliferator-activated receptor α was significantly upregulated (P < 0.05) in the LA1% compared with that in the control group. The Oil Red O staining area in the LA1% and LA2% was significantly reduced compared with that in the control group. The results indicated that although supplemental LA had negligible effects on growth and development in pigeon squabs, parental dietary LA at a concentration of 1% could have beneficial effects on maintaining squabs healthy as reflected by improved antioxidant capacity and lipid metabolism.

Excess dietary fluoride affects laying performance, egg quality, tissue retention, serum biochemical indices, and reproductive hormones of laying hens.

The present study aimed to evaluate the effects of excess dietary fluoride (F) on laying performance, egg quality, tissue retention, serum biochemical indices, and serum reproductive hormones of laying hens. A total of 384 Hy-Line Gray hens, 37 wk old, were treated with sodium fluoride added to a corn-soybean meal basal diet at 0, 400, 800, and 1200 mg fluorine/kg feed. The results showed that dietary F levels at 800 and 1200 mg/kg markedly decreased ADFI, laying rate, average egg weight, and increased feed conversion ratio (FCR) (P < 0.05). Dietary F levels at 800 and 1200 mg/kg dramatically decreased the egg quality of albumen height, yolk color, eggshell strength, and eggshell thickness, and on the 49th D, 400 mg/kg F group significantly decreased the eggshell strength, compared to those of control group. Fluoride residues in tissues of hens were increased significantly with the increase of dietary F supplemental levels (P < 0.05). Fluoride concentrations were generally high in feces, eggshell, tibia, kidney, and ovary, and the highest in feces, following with eggshell and tibia, lower in kidney and ovary, and the lowest in serum. Serum uric acid levels and alanine aminotransferase activity increased significantly (P < 0.05), and glucose, triglycerides, and phosphorus decreased significantly (P < 0.05) in response to dietary F concentration, compared to those of the control group, respectively. Dietary F supplementation at 1200 mg/kg significantly decreased (P < 0.05) the estrogen concentrations in serum, compared to those of the control group. Concentrations of progesterone in the fluoride-treated groups were significantly (P < 0.05) decreased relative to those of the control group. In conclusion, these results indicated that the excessive ingestion of F has had a detrimental effect on egg laying rate and quality of eggs by damaging the function of the liver, kidney, and ovary of laying hens.

Effects of in ovo feeding of L-lysine on hatchability, hatching time, and early post-hatch development in domestic pigeons (Columba livia)1.

This study was conducted to evaluate the effects of in ovo feeding (IOF) of L-lysine (Lys) on hatchability and development in pigeon neonates. At day 13 of incubation, fertile eggs were injected with 200 µL of sterilized saline (0.75%, SC group), 1% Conc. Lys solution (Lys group, 2.11 mg Lys dissolved in 200 µL of sterilized saline), with controls not injected (NC group). The results showed that IOF of Lys decreased the hatchability (P < 0.05) compared with other groups, whereas the hatching time was not affected among groups. On day 14 of post-hatch (D14), the body weight (BW) of squabs received Lys IOF was increased relative to the NC group (P < 0.05). Squabs in Lys group exhibited higher (P < 0.05) body weight gain (BWG) than other groups from D14 to day of hatch (DOH). Meanwhile, IOF of Lys increased the brain relative weight on DOH (P < 0.05), and organ index of heart, legs, and gizzard on day 7 of post-hatch (D7) compared with other groups. In addition, the length index of duodenum, jejunum, or ileum had no difference between groups, as well as the weight index except the weight index of duodenum in Lys group was higher than that of SC group on DOH. However, squabs received Lys IOF showed lower villus height, crypt depth, and villus surface area of jejunum than NC group (P < 0.05) on DOH, and lower crypt depth than NC and SC groups on D7. The situation was improved by D14, although there was no significant difference in morphometric trait of jejunum between Lys group and NC group, squabs received Lys IOF showed higher villus height, crypt depth, and villus surface area of jejunum than NC group. In conclusion, the present study demonstrates that IOF of Lys has negative effects on hatchability and development of early post-hatch squabs, but the situation will be improved with the growth of age.

Effect of iron glycine chelate supplementation on egg quality and egg iron enrichment in laying hens.

This study was conducted to evaluate the effects of iron glycine chelate (Fe-Gly) on egg quality of laying hens. A total of 810 laying hens (HyLine Variety White, 26 wk old) were randomly assigned to 6 groups, and each group consisting of 135 hens (5 replicates of 27 hens each). Hens in the control group received a diet supplemented with 60 mg Fe/kg as FeSO4, whereas hens in the other 5 groups received diets supplemented with 0, 20, 40, 60, and 80 mg Fe/kg from Fe-Gly, respectively. The study showed that dietary Fe-Gly treatments influenced (P < 0.05) the internal egg quality (egg weight, Haugh unit, albumen height), compared with the control group. However, dietary Fe-Gly supplementation showed few effects on the ultrastructure of eggshell in this study. The group of 60 mg Fe/kg as Fe-Gly was promoted (P < 0.05) in succinate dehydrogenase levels of liver and spleen compared with the 0 mg Fe-Gly/kg group, whereas the control (Fe/kg as FeSO4) group has no differences compared with the 0 mg Fe-Gly/kg group. The concentrations of Fe in the eggshell, yolk, and albumen were increased with increasing concentrations of Fe-Gly, where Fe-Gly (60, 80 mg Fe/kg) had higher (P < 0.01) Fe concentration than the control in yolk and albumen. The Fe-Gly groups (60, 80 mg Fe/kg) were influenced (P < 0.05) in transferrin, divalent mental transport 1, and ferroportin 1, compared with the control (FeSO4). In conclusion, Fe-Gly (60 mg Fe/kg) improved egg quality and egg iron enrichment. In general, there were no significant differences between Fe-Gly (40) and the control group in albumen height, Haugh unit, Fe concentration in eggshell and yolk. It revealed that FeSO4 could be substituted by a lower concentration of Fe-Gly and Fe-Gly may be superior to FeSO4 for egg quality in laying hens.

Effects of dietary Zn-methionine supplementation on the laying performance, egg quality, antioxidant capacity, and serum parameters of laying hens.

This study was to determine the effects of dietary Zn-methionine (Zn-Met) supplementation on the laying performance, egg quality, antioxidant capacity, and serum parameters of laying hens. Jingh ong-1 strain laying hens (n = 720, 49 wk of age) were randomly assigned to 6 treatments with 6 replications of 20 birds. The control was fed a basal diet supplemented with 80 mg of Zn/kg as Zn sulphate of diet and the 5 groups were fed a basal diet supplemented with 20, 40, 60, 80, and 100 mg of Zn/kg as Zn-Met of diet for 10 wk, respectively. At the terminal experiment, there were significant differences between control and 80 mg/kg Zn-Met group in feed intake (P < 0.05) and feed conversion ratio (FCR) (P < 0.01). Egg weight (P < 0.05) and albumen height (P < 0.01) reduced and were not significantly influenced by supplemental 80 mg/kg Zn-Met group until being stored 15 d as compared to the control. Zn-Met group in 100 mg/kg significantly increased haugh unit (P < 0.05) as compared to the control. The activity of MDA in serum had a linear decrease in 20 to 100 mg/kg Zn-Met. The activity of CAT in liver and GSH-Px in serum had quadratic effects in response to the Zn-Met treatments. Compared to the control, 60 mg/kg Zn-Met group increased the T-AOC, GSH-Px activity in serum (P < 0.01), and the T-AOC (P < 0.05), CuZnSOD (P < 0.01), GSH-Px (P < 0.01) activity in liver. Compared with the control, the concentration of serum ionic Ca in 80, 100 mg/kg Zn-Met treatments reduced (P < 0.01) significantly while the activity of serum alkaline phosphatase (AKP) increased in the Zn-Met groups of 40, 60, and 80 mg/kg (P < 0.01), and 100 mg/kg (P < 0.05). In conclusion, dietary Zn-Met supplementation at 60 to 80 mg/kg had more positive effects on performance, egg quality, and antioxidant capacity in laying hens as compared to 80 mg/kg ZnSO4.

Effects of dietary supplementation with Clostridium butyricum on laying performance, egg quality, serum parameters, and cecal microflora of laying hens in the late phase of production.

This experiment was conducted to evaluate the effects of dietary supplementation with Clostridium butyricum on laying performance, egg quality, serum parameters, and cecal microflora of laying hens in the late phase of production. Jinghong-1 strain laying hens (n = 960; 48 wk of age) were randomly allocated to 5 treatment groups with 6 replicates of 32 hens. Hens were fed with basal diet (control) and basal diet supplemented with 2.5 × 104 (CB1), 5 × 104 (CB2), 1 × 105 (CB3), and 2 × 105 (CB4) cfu/g C. butyricum for 10 wk. The results showed that egg production, egg mass, and eggshell strength increased quadratically as supplemental C. butyricum increased, and these responses were maximized in the CB2 group (P < 0.05). Compared with the control group, the addition of C. butyricum resulted in quadratic effects on serum total protein, uric acid, calcium, complement component C3 and catalase concentrations, and these responses were maximized or minimized in the CB2 group (P < 0.05). Linear and quadratic increases were observed in serum IgM, total superoxide dismutase, and glutathione peroxidase concentrations, and these responses were maximized in CB2 or CB3 group (P < 0.05). The addition of C. butyricum in the CB2 group resulted in linearly increasing levels of serum IgG concentration as compared with the control group (P < 0.05). Spleen index increased (P < 0.05) in the CB2 group. Hens fed with C. butyricum reduced (P > 0.05) the population of E. coli, while Bifidobacterium counts increased quadratically and maximized in the CB2 group (P < 0.05). In conclusion, the results indicated that dietary supplementation with C. butyricum (5 × 104 or 1 × 105 cfu/g) could improve laying performance and egg quality by promoting immune function, enhancing antioxidative capacity, and benefiting the cecal microflora of laying hens in the late phase of production.

Proteomic analysis of hypothalamus and liver proteins affected by dietary l-arginine supplementation in laying hens.

The goal of this study was to investigate the influence of l-arginine (l-Arg) supplementation on diet-induced changes in hypothalamus and liver proteome of laying hens. Layers were fed either an isonitrogenous control diet or a l-Arg diet. The test included a 2-week acclimation period and a 12-week experimental period. Eight layers per group were sacrificed at terminal of the experiment underwent 12 fasting. Blood and tissue samples of hypothalamus and liver were collected for further analysis. The levels of serum nitric oxide and hypothalamus neuropeptide Y of layers in l-Arg group were increased in comparison with those in control group. Quantitative proteomic analyses showed that a total of 3,715 hypothalamus proteins (235 differentially expressed) and 3797 liver proteins (373 differentially expressed) were detected between control and l-Arg-fed groups. A further enriched Gene Ontology term analysis of proteins found that 17 hypothalamus proteins (11 upregulated and six downregulated) and 29 liver proteins (14 upregulated and 15 downregulated) were altered differentially between the two groups. Our findings revealed the changes in metabolic and hormonal signals in central nervous system and peripheral tissues by responding to l-Arg feeding, which provides a possible way to gain a better understanding of l-Arg function in laying hens.

Effects of dietary Mn-methionine supplementation on the egg quality of laying hens.

This study was conducted to investigate the effects of dietary manganese-methionine (Mn-Met) supplementation on the egg quality of laying hens. A total of 480 Jinghong-1 strain layers aged 53 wk were divided into 5 groups with 6 replicates of 16 layers. Birds in the control group were fed a diet supplemented with 60 mg Mn/kg in the form of MnSO4; the birds in other 4 experimental groups were fed a diet supplemented with 20, 40, 60, and 80 mg Mn/kg as Mn-Met, respectively. Dietary Mn-Met treatments significantly affected (P < 0.05) the albumen height, yolk color, and Haugh unit compared to those of the control diet. The Mn contents in the eggshell increased (P < 0.01) significantly by increasing the Mn-Met supplementation, whereas Mn content in eggshell was triple that in the yolk or albumen. Compared with the 60 mg/kg Mn-Met group, the transverse surface in the control group had (P < 0.01) a greater width of mammillary cones, and there were obvious cracks on the outer surface in the control. There was no difference (P > 0.05) in the eggshell gland (ESG) in the expression of calbindin-D28k (CaBP-D28k) mRNA in response to any diet treatment. In conclusion, dietary Mn-Met supplementation increased internal egg quality and the ultrastructure of the eggshell. Compared to the control, 60 mg/kg Mn-Met treatment resulted in improving egg quality, and 20 mg/kg Mn-Met treatment had similar effects the control treatment had on the egg quality. This indicates that the inorganic Mn can be replaced by the lower concentration of Mn-Met.

Effects of in ovo feeding of l-arginine on hatchability, hatching time, early posthatch development, and carcass traits in domestic pigeons (Columba livia).

The objective of this research was to test the hypothesis that in ovo feeding of arginine (Arg) may improve hatchability and posthatch performance in domestic pigeons (). A completely randomized design ( = 3) with an Arg feeding treatment (Arg group, 1.14 mg Arg dissolved in 200 µL of 0.75% NaCl buffered saline as 1% concentration compared to total Arg in the egg), a buffered saline feeding treatment (SC group, 7.5 g NaCl dissolved in 1 L sterile distilled water as the concentration of poultry physiological saline), and a nonfeeding treatment (NC group) was used. Six squabs from each treatment were randomly sampled on day of hatch (DOH), posthatch d 7 (D7), and posthatch d 14 (D14), respectively. Hatchability, hatch time, BW, organ development, and carcass traits were examined. Results showed that in ovo feeding of the Arg solution increased ( < 0.05) the hatchability and advanced ( < 0.05) the hatching time in comparison with those of the other groups. Body weight of pigeon squabs that received Arg in ovo feeding was heavier ( < 0.05) on DOH and D14 than that of the NC group, and a greater ( < 0.05) BW gain from DOH to D14 and D7 to D14 was observed. Three clusters of 12 organs were classified according to the changes of organ indices. Squabs provided the Arg in ovo feeding treatment gained a priority in organ development. The heart index and gizzard index on D7 and the proventriculus index on D14 of squabs receiving Arg in ovo feeding were increased ( < 0.05) compared to those of the other groups. The brain index on DOH, the small intestine index and pancreas index on D7, and the liver index, pancreas index, and spleen index on D14 of squabs fed Arg were higher ( < 0.05) than those of the NC group. The spleen index on D7 and the small intestine index on D14 of squabs provided the Arg feeding treatment were enhanced ( < 0.05) compared with those of the SC group. The semieviscerated carcass weight of squabs receiving Arg was higher ( < 0.05) on D14 than that of other groups. The absolute weight of breast meat yield on D7 and breast meat yield percentage on D7 and D14 were improved ( < 0.05) in the Arg group compared with the NC group. The leg meat percentage on D7 and the carcass weight, eviscerated carcass weight, and absolute weight of breast meat yield on D14 were increased ( < 0.05) in the Arg group compared with those of the SC group. The results of this study indicate that in ovo feeding of pigeon embryos with Arg may have beneficial effects on squab hatch performance and early posthatch performance.

Effects of dietary threonine supplementation on intestinal barrier function and gut microbiota of laying hens.

The effects of supplemental dietary threonine (Thr) on laying performance, expression of intestinal mucin 2 (MUC2) and secretory IgA (sIgA), and intestinal microbiota of laying hens fed a low CP diet were investigated. A total of 240 Lohmann Brown laying hens, 28 wk of age, was allocated to 3 dietary treatments, each of which included 5 replicates of 16 hens. Hens were fed a control diet (16% CP), a low CP diet (14% CP), or a low CP diet supplemented with 0.3% L-Thr for 12 weeks. Chemical analyses of the diets for Thr are 0.49, 0.45, and 0.69%, respectively. Lowering dietary CP impaired egg production and egg mass of laying hens. Dietary Thr supplementation to the low CP diet increased (P < 0.05) egg production and egg mass. In addition, ileal sIgA contents and MUC2 and sIgA mRNA expression were increased (P < 0.05) by dietary Thr addition. Dietary CP reduction reduced (P < 0.05) intestinal bacterial diversity, whereas dietary Thr supplementation to the low CP diet recovered the bacteria diversity and significantly increased the abundance of potential beneficial bacteria. In conclusion, dietary Thr supplementation to a low CP diet could affect intestinal health and hence productivity via regulating intestinal mucin and sIgA expression, and microbial population of laying hens.

Effects of arsenic supplementation in feed on laying performance, arsenic retention of eggs and organs, biochemical indices and endocrine hormones.

1. The primary objective of this experiment was to estimate the toxic effects of arsenic (As) supplementation in feed on laying performance, As retention by eggs and organs, serum biochemical indices and endocrine hormones in laying hens. 2. A total of 320 "Jinghong Number 1" hens, 56-week-old, were randomly allocated into four treatments of four replicates with 20 layers in each. Graded arsenical was added to the basal diet in the experimental diets at As levels of 0, 17, 34 and 51 mg/kg, respectively. The trial lasted for 9 weeks including 1 week for acclimatisation. 3. Supplementation of dietary As for eight weeks had no effect on laying performance. As retention in albumen, yolk, egg, liver and kidney increased as As levels increased The level of serum phosphorus (P) was minimised at the 17 mg As/kg group. The activity of serum glutamic oxaloacetic transaminase (GOT) increased linearly. No differences were observed for levels of serum calcium (Ca), alkaline phosphatase (AKP) and serum glutamic pyruvic transaminase (GPT). Concentrations of estradiol (E2) and progesterone (PG) declined at 34 and 51 mg/kg As levels compared with the control group. As supplementation exerted no influence on levels of serum follicle stimulating hormone (FSH), luteinising hormone (LH), triiodothyronine (T3), thyroxine (T4) and the ratio between T3 and T4. 4. In conclusion, dietary As supplementation accelerated retention in tissues and eggs, and affected the laying rate by diminishing hormone levels of E2 and PG at 51 mg/kg.

Molecular cloning, characterization, and expression analysis of ghrelin and cholecystokinin in the pigeon (Columba livia).

Ghrelin and cholecystokinin (CCK) are multifunctional peptides. In the current study, complete sequences of ghrelin (800 bp) and CCK (739 bp) were firstly cloned in Columba livia by using rapid amplification of cDNA ends (RACE) method. The open reading frames of ghrelin (351bp) and CCK (393bp) encoded 116 amino acids and 130 amino acids, respectively. Sequence comparison indicated that pigeon ghrelin and CCK shared high identity with those reported in other avian species. Quantitative real-time PCR analysis found that ghrelin and CCK mRNAs expressed in three intestinal segments of pigeon during development. Both ghrelin and CCK showed generally higher expressions at days posthatch than embryonic periods regardless of intestinal segments. In duodenum and ileum, the expressions of ghrelin and CCK mRNA reached the peak values at 8 d posthatch. Jejunum CCK mRNA level increased linearly after hatching, and reached the highest point at posthatch 28 d. Based on documented effects of long chain fatty acids (LCFAs) on pigeon ghrelin and CCK expression were also investigated in vitro. Higher concentrations (50 µM or 250 µM) of linoleic acid, α-linolenic acid or arachidonic acid can significantly increase ghrelin mRNA level in pigeon jejunum. However, for oleic acid, the induction of ghrelin gene expressions needed a lower concentration (5 µM). 5 µM of linoleic acid, α-linolenic acid or arachidonic acid and 250 µM palmitic acid repressed CCK expression significantly. A higher concentration (250 µM) of oleic acid or α-linolenic acid can up-regulate CCK mRNA level significantly. Our results indicated that ghrelin and CCK may act key functions in pigeon intestine development and their expressions could be regulated by LCFAs.

Effects of dietary L-isoleucine on laying performance and immunomodulation of laying hens.

Isoleucine may be a limiting amino acid for laying hens fed diets with a lowered protein level. An experiment was conducted to examine laying performance and the immune function of laying hens provided diets varying in digestible isoleucine levels during the peak production period. A total number of 400 Lohmann Brown laying hens, 28 wk of age, were allocated to 5 dietary treatment groups, each of which included 5 replicates of 16 hens per replicate (4 cages / replicate; 80 hens / treatment). L-isoleucine was added to the experimental diet (14% CP) containing synthetic amino (methionine, lysine, threonine, tryptophan, and valine) by zero, 1.0, 2.0, 3.0, and 4.0 g/kg, corresponding to 0.54%, 0.64%, 0.74%, 0.84, and 0.94% digestible isoleucine, respectively. At the end of the experiment (wk 40), dietary isoleucine did not affect laying performance or egg quality. Serum albumin concentration increased quadratically (P < 0.05) in response to digestible dietary isoleucine at 0.74%. Serum free isoleucine and lysine increased (P < 0.05) in response to digestible dietary isoleucine at 0.74%. Digestible dietary isoleucine levels did not affect the serum concentrations of total antioxidative capability (T-AOC), total superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), and CuZn-superoxide dismutase (CuZn-SOD). There was no significant (P > 0.05) response of excess digestible isoleucine level on the serum level of IgG, IgA, or IgM. In addition, dietary isoleucine levels did not affect the concentrations of secretory immunoglobulin A (sIgA), tumor necrosis factor alpha (TNFα), or interleukin (IL-2 and IL-6) in the ileum. Also, expressions of ileal MUC2 mRNA, sIgA mRNA, and IL-1ß mRNA were not changed (P > 0.05) by excess digestible isoleucine level. Furthermore, excess digestible isoleucine level did not change mRNA expression of ileal tight junction protein (claudin-1 and occludin). No effect occurred when isoleucine was supplemented, suggesting that it is not a limiting amino acid in the low crude protein diet on laying performance and intestinal mucosal immune.

Dietary L-arginine levels affect the liver protein turnover and alter the expression of genes related to protein synthesis and proteolysis of laying hens.

Amino acids are considered to be anabolic factors that affect protein turnover. The aim of this study was to test the effects of dietary L-arginine (Arg) levels on protein metabolism in the liver of laying hens and the expression of genes related to protein synthesis and proteolysis. Xinyang black commercial laying hens (n = 864, 31 wk of age) were randomly allotted to 6 treatments with 4 replicates of 36 birds. The dietary treatments were corn-corn gluten meal based diets containing 0.64, 0.86, 1.03, 1.27, 1.42, and 1.66% L-Arg, respectively. Serum concentrations of total protein and albumin were maximized in the 1.27% L-Arg group, and serum concentration of urea acid was the lowest in the 1.27% L-Arg group. The 1.27% L-Arg group had the highest fractional protein synthesis rate and fractional protein gain rate in the liver. Consistent with the data on protein turnover, mRNA abundances of target of rapamycin (TOR) and ribosomal protein S6 kinase 1 increased in the liver of layers fed 1.27% L-Arg, while mRNA abundances of cathepsin B and 20S proteasome decreased at the same dietary L-Arg level. In conclusion, the dietary level of L-Arg increased the liver fractional protein synthesis rate and fractional protein gain rate of laying hens, and the action of an appropriate level of dietary L-Arg involves upregulating the gene expression of the TOR signaling pathway accompanied by suppressing the mRNA expression of cathepsin B and 20S proteasome in the liver.

L-arginine upregulates the gene expression of target of rapamycin signaling pathway and stimulates protein synthesis in chicken intestinal epithelial cells.

L-arginine (Arg) is an indispensable amino acid in avians and is required for growth. The aim of this study was to investigate the effects of L-Arg on protein synthesis and genes expression involved in target of rapamycin (TOR) signaling pathway in chicken enterocytes. Cells were cultured for 4 days in L-Arg-free Dulbecco's modified Eagle's medium containing 10, 100, 200, 400, or 600 µM L-Arg. Cell growth, cell cycle, protein synthesis, and protein degradation as well as mRNA expression levels of TOR, ribosomal protein S6 kinase 1 (S6K1), and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) were determined. The results showed that cell viability was enhanced by L-Arg with a maximal response at 10 to 400 µM. Increasing extracellular concentrations of L-Arg from 10 to 400 µM increased the cells in S and G2/M phase to a significant extent and decreased cell numbers in G0/G1 phase. Further more, addition of 100, 200, or 400 µM L-Arg to culture medium increased protein synthesis and reduced protein degradation in chicken intestinal epithelial cells. Consistent with the data on cell growth and protein turnover, supplementation of 100, 200, or 400 µM L-Arg increased the mRNA abundances of TOR, 4E-BP1, and S6K1. It was concluded the action of L-Arg involves in upregulating the genes expression of TOR cell signaling pathway which increases protein synthesis and reduces protein degradation.

Effect of excess dietary L-valine on laying hen performance, egg quality, serum free amino acids, immune function and antioxidant enzyme activity.

1. The aim of this study was to evaluate the tolerance of laying hens for an excessive L-valine (L-val) supply on laying performance, egg quality, serum free amino acids, immune function and antioxidant enzyme activities of laying hens. 2. A total of 720 HyLine Brown hens were allocated to 5 dietary treatment groups, each of which included 6 replicates of 24 hens, from 40 to 47 weeks of age. Graded amounts of L-val were added to the basal diet to achieve concentrations of 0 (control), 1, 2, 3 and 4 g/kg, respectively, in the experimental diets. 3. Supplementing the diet with L-val did not affect egg production, egg mass, egg weight, feed conversion ratio (FCR) or egg quality. The average daily feed intake response to supplemental L-val was quadratic and was maximised at 2.0 g L-val/kg diet. No differences were observed for total protein, total amino acids, blood urea nitrogen (BUN), uric acid, lactate dehydrogenase (LDH), alkaline phosphatase (AKP), Ca and P concentrations among the treatments. 4. Serum albumin concentration increased significantly in response to supplemental L-val and was also maximised at 2.0 g/kg. In addition, serum glucose increased quadratically to peak at 2.0 g L-val/kg diet. Serum free valine increased as L-val concentration increased to 2.0 g/kg diet and then decreased linearly. 5. Supplementation of L-val did not affect the serum concentrations of total antioxidative capability (T-AOC), superoxide dismutase (SOD) and malondialdehyde (MDA). L-val supplementation did not affect the concentrations of immunoglobulins IgG, IgA, IgM and complements (C3 and C4). Serum concentration of triiodothyronine (T3) increased significantly at 2.0 g L-val/kg diet. 6. It is concluded that high concentrations of L-val are tolerated and can be successfully supplemented into diets without detrimental effects on laying performance or immune function of laying hens.

Evaluating the impact of excess dietary tryptophan on laying performance and immune function of laying hens reared under hot and humid summer conditions.

1. Tryptophan (Trp), besides its role as an essential amino acid in protein synthesis, may also have other important effects on laying hens under summer conditions. 2. Babcock Brown layers (n = 768), 40 weeks of age, were allocated to 4 treatment groups, each of which included 6 replicates of 32 hens. Each group received the same basal diet, formulated with maize and soybean meal, for 8 weeks. Hens were fed on the basal diet with 0·0, 0·2, 0·4, and 0·8 g/kg L-Trp to achieve dietary concentrations of 1·7, 1·9 g/kg, 2·1 g/kg or 2·5 g/kg of Trp, respectively. 3. Supplementing L-Trp had no affect on laying performance. Adding 0·2 or 0·4 g/kg L-Trp improved egg shell strength compared with those fed on the control diet. Serum albumin concentration increased at 0·4 g/kg compared with those receiving 0·0 or 0·8 g/kg Trp. The addition of Trp at 0·4 g/kg increased serum IgM concentration quadratically. Serum superoxide dismutase (T-SOD) increased linearly and quadratically at 0·4 g/kg. 4. In conclusion, we suggest that 0·2 to 0·4 g/kg Trp may have beneficial effects on laying hens under conditions of high temperature and humidity.

Influence of L-threonine supplementation on goblet cell numbers, histological structure and antioxidant enzyme activities of laying hens reared in a hot and humid climate.

1. A previous experiment in our laboratory found that L-threonine supplementation at 0.2 and 0.3% increased egg production and the concentration of serum IgG, respectively. The objective of this current trial was to determine if both supplementation levels can positively influence histological structure, goblet cell numbers, or antioxidant enzyme activities. 2. Babcock Brown layers (n=576), 40 weeks of age, were allocated to three treatment groups, each of which included 6 replicates of 32 hens. Each group received the same basal diet formulated with maize, peanut meal and crystalline amino acids. L-threonine was added to the basal diet at 0 (control), 0.2, and 0.3%, respectively for 8 weeks. Chemical analysis of the diets for threonine values were 0.47, 0.66 and 0.74 %, respectively. 3. The numbers of goblet cells did not change due to L-threonine supplementation. Also, L-threonine had no affect on the villus height and mucosal thickness. No differences were found due to treatments among groups in the activity of alkaline phosphatase (ALP) in jejunum or ileum. L-threonine supplementation at 0.2% maximised the concentration of superoxide dismutases (SOD) in both serum and liver. 4. In conclusion, L-threonine supplementation had no affect on gut morphology but may have an antioxidant function at 0.2%.

Effect of supplemental L-threonine on mucin 2 gene expression and intestine mucosal immune and digestive enzymes activities of laying hens in environments with high temperature and humidity.

In addition to being an essential amino acid in protein synthesis, threonine is an integral component of gut function. To verify the effects of l-threonine on gut function, Babcock Brown layers (n = 960; 40 wk of age) were allocated to 5 dietary treatment groups, each of which included 6 replicates of 32 hens. Each group received the same basal diet formulated with corn, peanut meal, and crystalline amino acids. l-Threonine was added to the basal diet at 0 (control), 0.1, 0.2, 0.3, and 0.4% for 8 wk to achieve 0.47 (NRC), 0.57, 0.67, 0.77, and 0.87% threonine, respectively. Expressions of jejunal and ileal mucin 2 mRNA were increased linearly by increasing l-threonine (P < 0.01). At 0.4% l-threonine, the concentrations of IgA antibody in the mucosa of the ileum increased linearly (P < 0.01). No differences attributable to treatment were found among groups in the activity of digestive enzymes in the jejunum or ileum. It was concluded that dietary threonine requirements as reported in current NRC recommendations are insufficient for modern commercial laying hens raised in summer climates. The results suggest that threonine might function as a nutrient immunomodulator in maintaining intestinal barrier function.