Effects of different selenium sources and levels on the physiological state, selenoprotein expression, and production and preservation of selenium-enriched eggs in laying hens.

Selenium (i.e., Se) is a trace element that is vital in poultry nutrition, and optimal forms and levels of Se are critical for poultry productivity and health. This study aimed to compare the effects of sodium selenite (SS), yeast selenium (SY), and methionine selenium (SM) at selenium levels of 0.15 mg/kg and 0.30 mg/kg on production performance, egg quality, egg selenium content, antioxidant capacity, immunity and selenoprotein expression in laying hens. The trial was conducted in a 3 × 2 factorial arrangement, and a total of 576 forty-three-wk-old Hyland Brown laying hens were randomly assigned into 6 treatment groups, with diets supplemented with 0.15 mg Se/kg and 0.3 mg Se/kg of SS, SY and SM for 8 wk, respectively. Results revealed that SM increased the laying rate compared to SS and SY (P < 0.05), whereas different selenium levels had no effect. Organic selenium improved egg quality, preservation performance, and selenium deposition compared to SS (P < 0.05), while SY and SM had different preferences for Se deposition in the yolk and albumen. Also, organic selenium enhanced the antioxidant capacity and immune functions of laying hens at 0.15 mg Se/kg, whereas no obvious improvement was observed at 0.30 mg Se/kg. Moreover, SY and SM increased the mRNA expression of most selenoproteins compared to SS (P < 0.05), with SM exhibiting a more pronounced effect. Correlation analysis revealed a strong positive association between glutathione peroxidase 2 (GPx2), thioredoxin reductases (TrxRs), selenoprotein K (SelK), selenoprotein S (SelS), and antioxidant and immune properties. In conclusion, the use of low-dose organic selenium is recommended as a more effective alternative to inorganic selenium, and a dosage of 0.15 mg Se/kg from SM is recommended based on the trail conditions.

Linoleic acid ameliorates intestinal mucosal barrier injury in early weaned pigeon squabs (Columba livia).

The study aimed to investigate whether linoleic acid could improve the intestinal barrier function of squabs under weaning stress conditions. Totally 320 7-d-old weaned squabs were randomly divided into four treatment groups, including control group (CON), 0.7% linoleic acid addition group (LA007), 1.4% linoleic acid addition group (LA014) and 2.1% linoleic acid addition group (LA021). At 21 d, eight squabs were randomly selected from each treatment group for sampling and determination. The results showed that adding linoleic acid could improve (P < 0.05) the body weight of weaned squabs, and LA014 had the best effect. With the increase of linoleic acid dosage, villi height and villi area increased linearly or quadratically (P < 0.05), and reached the maximum in LA021 or LA014, respectively. The linoleic acid supplementation could improve the intestinal tight junction of weaned squabs, and the LA014 was the most significant (P < 0.05). With the linoleic acid increasing, the levels of intestinal IL-6 and TNF-α decreased linearly (P < 0.05), while intestinal IL-10 increased quadratically (P < 0.05) and reached the maximum in LA014. Serum endotoxin and diamine oxidase levels decreased linearly (P < 0.05) and reached the lowest level in LA014. The ultrastructure of villi revealed that the length of ileal microvilli in LA014 was significantly increased (P < 0.05) and the microvilli became dense, and the mitochondria in epithelial cells returned to normal state. Further exploring the mechanism of linoleic acid alleviating intestinal injury caused by weaning stress in squabs, it was found that linoleic acid down-regulated (P < 0.05) the relative protein expression of TLR4, MyD88, phosphorylated JNK, and phosphorylated p38, reducing secretion of pro-inflammatory factors IL-6 and TNF-α. This study indicated that linoleic acid could alleviate intestinal barrier injury of early weaned squabs by down-regulating TLR4-MyD88-JNK/p38-IL6/TNF-α pathway.

Artificial feeding of early weaned squabs can reduce the burden of breeding pigeons and shorten the breeding cycle. However, similar to early weaned mammals, early weaned squabs would also inevitably undergo severe physiological and psychological stress responses in the early stage. The growth performance and immunity of early weaned squabs were inferior to those of the parent feeding squabs. Previous studies suggest that linoleic acid played an important role in the growth and development of squabs. Therefore, the study aimed to investigate whether linoleic acid could improve the intestinal barrier function of squabs under weaning stress conditions. Totally 320 7-d-old weaned squabs were randomly divided into four treatment groups, including control group and linoleic acid addition groups with three different doses. At 21 d, eight squabs were randomly selected from each treatment group for sampling and determination. The results indicated that under weaning stress conditions, linoleic acid could weaken the inflammatory response, and alleviate the intestinal epithelial barrier damage of weaned squabs, specifically by promoting the development of intestinal villi, strengthening the tight junction, reducing intestinal permeability, and promoting the secretion of anti-inflammatory factors.

Parental dietary arachidonic acid altered serum fatty acid profile, hepatic antioxidant capacity, and lipid metabolism in domestic pigeons (Columba livia).

The aim of this study was to explore the effects of dietary arachidonic acid on serum fatty acid profile, hepatic antioxidant capacity, and lipid metabolism in pigeon squabs by supplementing arachidonic acid in their parental diets. A completely randomized design was conducted consisting of control group, 0.05% dietary arachidonic acid supplementation group, 0.1% dietary arachidonic acid supplementation group, and 0.2% dietary arachidonic acid supplementation group. Six randomly selected squabs from each group were sampled on Day 21 post-hatch. Results indicated that moderate level (0.05%) of arachidonic acid in parental diets for pigeon squabs improved lipid metabolism via regulation on serum lipid profile and fatty acid composition and tended to reduce hepatic lipid accumulation in the premise of negligible damage to antioxidant status. Unfortunately, excessive parental supplementation of dietary arachidonic acid might be harmful to squab health. The regulatory effects of arachidonic acid were sensitive to the arachidonic acid doses. In conclusion, parental dietary arachidonic acid at 0.05% could be beneficial for squabs to maintain health as reflective aspects in ameliorative serum lipid profile, fatty acid composition, and reduced hepatic lipid accumulation.

MAPK, AKT/FoxO3a and mTOR pathways are involved in cadmium regulating the cell cycle, proliferation and apoptosis of chicken follicular granulosa cells.

The occurrence of cadmium (Cd) in feed is a major problem in animal health and production. Studies have confirmed that Cd depresses egg production of laying hens, which is closely related to follicular atresia. This study aimed to assess the toxic impacts of Cd on the ovarian tissue, and to examine the mechanism of Cd-induced granulosa cell proliferation and apoptosis. Results from the nitric oxide (NO) and malondialdehyde (MDA) content, total superoxide dismutase (T-SOD), glutathione peroxide (GSH-Px), total nitric oxide synthase (T-NOS) and adenosine triphosphatase (ATPase) activities, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, and hematoxylin-eosin (H & E) staining indicated that excess Cd induced oxidative stress, granulosa cell apoptosis and follicular atresia in the layer ovary. Low-dose Cd exposure (1 µM) induced the granulosa cell proliferation, upregulated the mRNA levels of RSK1 and RHEB, activated FoxO3a, AKT, ERK1/2, mTOR and p70S6K1 phosphorylation, and promoted cell cycle progression from phase G1 to S. However, high-dose Cd exposure (15 µM) induced reactive oxygen species (ROS) generation and cell apoptosis, upregulated the mRNA levels of the inflammatory factors, ASK1, JNK, p38 and TAK1, downregulated the expressions of RSK1 and RHEB genes, and inhibited the phosphorylation of ERK1/2, mTOR and p70S6K1 proteins, and the cell cycle progression. Rapamycin pre-treatment completely blocked the phosphorylation of mTOR and p70S6K1 proteins, and the cell cycle progression induced by 1 µM Cd, and accelerated 15 µM Cd-induced cell apoptosis and cell cycle arrest. The microRNA sequencing result showed that 15 µM Cd induced differential expression of microRNA genes, which may regulate AKT, ERK1/2 and mTOR signaling and cell cycle progression by regulating the activity of G proteins and cell cycle-related proteins. Conclusively, these results indicated that Cd can cause the ovarian damage and follicular atresia, and regulate cell cycle, cell proliferation or apoptosis of granulosa cells through MAPK, AKT/FoxO3a and mTOR pathways in laying hens.

Effects of Zinc on Cell Proliferation, Zinc Transport, and Calcium Deposition in Primary Endometrial Epithelial Cells of Laying Hens In Vitro.

For birds, the uterus is an important part for eggshell mineralization, and the establishment of the endometrial epithelial cell (EEC) model was beneficial to the study of uterine function. This study was conducted to establish a culture model of primary EECs of laying hens and explore the effects of zinc on primary EEC proliferation, zinc transport, and calcium deposition in vitro. The EECs were isolated and cultured via type I collagenase digestion, and in the logarithmic phase during 2-5 days, and then reached the plateau phase on the 7th day of inoculation. Results showed that the proliferation of EECs treated by 50 µM ZnSO4 or zinc-methionine (Zn-Met) were markedly promoted at 24-h or 48-h treating time (P < 0.05). In later experiments, the EECs were divided into three groups, involving a control group (no zinc treated), ZnSO4 group (50 µM zinc treated) and a Zn-Met group (50 µM zinc treated). Results showed the relative fluorescence intensity of Ca2+ in the Zn-Met group was significantly higher than that in the control group (P < 0.05). As for zinc transporters, it was only observed that mRNA levels of metallothionein (MT) in EECs showed a significant difference (P < 0.05) between the Zn-Met group and the control. In conclusion, the EECs of laying hens isolated by scraping and digested collagenase I were with better adherent growth. Moreover, Zn-Met can increase intracellular Ca2+ concentration and upregulate expressions of MT mRNA in the EECs of laying hens.

Effect of Glycine Nano-Selenium Supplementation on Production Performance, Egg Quality, Serum Biochemistry, Oxidative Status, and the Intestinal Morphology and Absorption of Laying Hens.

The objective of this study was to investigate the feasibility of using glycine nano-selenium (NS-Gly) as a feed supplement and to evaluate its influence on production performance, egg quality, serum biochemistry, oxidative status, and the intestinal morphology and absorption of laying hens. A total of 864 hens at 40 weeks were randomly assigned into six groups including the basal diet (control, 0.13 mg Se/kg); basal diet + 0.30 mg Se/kg (Na2SeO3) diet; and basal diet + 0.15, 0.30, 0.45, and 0.60 mg Se/kg (NS-Gly) diet. After 8 weeks of Se supplementation, no difference was observed among the treatments on production performance and egg quality (P > 0.05). The levels of albumin (ALB) and alanine aminotransferase (GPT) were significantly influenced by dietary Se supplementation (P < 0.05). In the serum, the level of glutathione peroxide (GSH-Px) was significantly increased in the groups with the dietary NS-Gly supplementation (P < 0.05). The superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) levels in all groups of NS-Gly supplementation had a remarkable increase (P < 0.05). In the liver, GSH-Px was significantly increased in 0.45 and 0.60 mg/kg NS-Gly groups (P < 0.05). The activities of SOD and catalase (CAT) were significantly increased in the groups of 0.30 mg/kg NS-Gly diet (P < 0.05). The results of intestinal morphology showed that the crypt depth was affected by higher dose groups of NS-Gly diets in the duodenum, and the differences (P < 0.05) were obtained in villus height, the crypt depth, and the V/C in the jejunum. In the ileum, a significant increase (P < 0.05) of villus height was observed in 0.15 and 0.3 mg/kg Se-added groups. The V/C was the highest in the SS groups (P < 0.05). The mRNA levels of solute carrier family 3 member 1 (rBAT), solute carrier family 6 member 19 (B0AT1), and solute carrier family 15 member 1 (PepT1) increased at different degrees in the duodenum, especially in 0.15 and 0.60 mg/kg NS-Gly groups (P < 0.05). In the jejunum, the expression of B0AT1 was similar to that in the duodenum, and the expression of rBAT increased significantly in the 0.30 and 0.45 mg/kg NS-Gly groups (P < 0.05). The mRNA level of PepT1 increased significantly in the 0.30 mg/kg SS group. Conclusively, dietary NS-Gly supplementation could improve the antioxidant capacity, as well as the structure of small intestine in laying hens, although have no significant effects on the production performance and egg quality.

Maternal dietary linoleic acid altered intestinal barrier function in domestic pigeons (Columba livia).

Linoleic acid (LA) is predominantly essential for poultry. Poultry lacking LA show retarded growth and reduced disease resistance. Intestinal barrier function plays an important role in pigeon squab growth, whereas research on the effects of LA on intestinal health in altrices is scant. Considering that squabs are fed by their parents, the study aimed to explore the effects of maternal dietary LA on intestinal morphology, tight junction proteins, immune cytokines and microbial flora in squabs. A completely randomised design with a control group, 1 % LA supplementation group, 2 % LA supplementation group and 4 % LA supplementation group was used. Six squabs from each treatment were randomly sampled at 21 d post-hatching. The results indicated that LA supplementation improved intestinal morphology, as reflected by increased villus height, villus area and the ratio of villi to crypts. Also, 1 % LA supplementation elevated the density of goblet cells in the intestine and strengthened tight junctions by up-regulating claudin-3 and occludin gene expression but down-regulating claudin-2 gene expression. Moreover, 1 % LA supplementation reduced the secretion of proinflammatory cytokines and partly increased anti-inflammatory cytokines. The intestinal microbial diversity in the 1 % LA supplementation group was higher than that in the other groups. As beneficial bacteria, Butyrivibrio was the biomarker of 1 % LA supplementation. However, excessive (4 %) LA supplementation led to adverse impacts on intestinal immunity and microbiota. In conclusion, maternal dietary LA might alter intestinal barrier function in pigeon squabs in a dose-dependent manner. Supplementation with 1 % LA was suggested in parental pigeons.

Dietary cadmium chloride impairs shell biomineralization by disrupting the metabolism of the eggshell gland in laying hens.

In this study, we identified cadmium (Cd) as a potential endocrine disruptor that impairs laying performance, egg quality, and eggshell deposition and induces oxidative stress and inflammation in the eggshell glands of laying hens. A total of 480 38-wk-old laying hens were randomly assigned into 5 groups that were fed a basal diet (control) or a basal diet supplemented with Cd (provided as CdCl2·2.5 H2O) at 7.5, 15, 30, and 60 mg Cd per kg feed for 9 wk. The results showed that, when compared with the control group, a low dose of dietary Cd (7.5 mg/kg) had positive effects on egg quality by improving albumen height, Haugh unit, yolk color, and shell thickness at the third or ninth week. However, with the increase in the dose and duration of Cd exposure, the laying performance, egg quality, and activities of eggshell gland antioxidant enzymes (catalase [CAT], glutathione peroxide [GSH-Px]), and ATPase (Na+/K+-ATPase, Ca2+-ATPase, and Mg2+-ATPase) deteriorated, and the activity of total nitric oxide synthase (T-NOS) and the level of malondialdehyde (MDA) increased significantly (P < 0.05). The histopathology and real-time quantitative PCR results showed that Cd induced endometrial epithelial cell proliferation accompanied by upregulation of the mRNA levels of progesterone receptor (PgR) and epidermal growth factor receptor (EGFR), downregulation of the mRNA levels of estrogen receptor α (ERα) and interleukin 6 (IL6), and inflammation of the eggshell gland accompanied by significantly increased expression of complement C3 and pro-inflammatory cytokine tumor necrosis factor α (TNFα) (P < 0.05). In addition, the ultrastructure of the eggshell showed that dietary supplementation with 7.5 mg/kg Cd increased the palisade layer and total thickness of the shell, but with the increase in dietary Cd supplementation (30 and 60 mg/kg) the thickness of the palisade layer and mammillary layer decreased significantly (P < 0.05), and the outer surface of the eggshell became rougher. Correspondingly, the expression of calbindin 1 (CALB1), ovocalyxin-32 (OCX-32), ovocalyxin-36 (OCX-36), osteopontin (SPP1), and ovocledidin-17 (OC-17) decreased significantly (P < 0.05) with increasing dietary Cd supplementation. Conclusively, the present study demonstrates that dietary supplementation with Cd negatively affects laying performance, egg quality, and eggshell deposition by disturbing the metabolism of eggshell glands in laying hens but has a positive effect on egg quality at low doses.

Effects of iron glycine chelate on laying performance, antioxidant activities, serum biochemical indices, iron concentrations and transferrin mRNA expression in laying hens.

This study was conducted to evaluate the effects of iron glycine chelate (Fe-Gly) on laying performance, antioxidant enzyme activities, serum biochemical indices and iron concentrations in laying hens. A total of 810 laying hens (Hy-Line Variety White, 26 weeks old) were randomly assigned to six groups with five replicates of 27 layers. Hens in the control group received diet supplemented with 60 mg Fe/kg as FeSO4 , while hens in other five groups received the diet supplemented with 0, 20, 40, 60 and 80 mg Fe/kg from Fe-Gly respectively. The results showed that dietary Fe-Gly treatments significantly influenced (p < 0.01) the laying rate and egg weight of layers, compared with the control group. Concerning to CuZn-superoxide dismutase (CuZn-SOD) and total superoxide dismutase (T-SOD) activity, Fe-Gly groups (60, 80 mg Fe/kg) were promoted significantly (p < 0.01) compared with 0 mg Fe/kg group. The concentrations of Fe in serum, liver, kidney, spleen and ovary were increased significantly with the level dietary Fe-Gly raised where Fe-Gly groups (60, 80 mg Fe/kg) had observably higher Fe concentration than the control (p < 0.01) in serum, kidney and spleen. There was a trend that transferrin mRNA expression was decreased with the increase of Fe as Fe-Gly in diets, and compared with the control, the expression was lower in the group fed diet with 60 mg/kg Fe as Fe-Gly. In conclusion, Fe-Gly (60 mg Fe/kg) had improved laying rate, egg weight, SOD enzyme activity, Fe absorption and protein synthesis in body and promoted iron metabolism in laying hens. Moreover, Fe-Gly (40 mg/kg Fe) had the similar effect with control group. It revealed that FeSO4 could be substituted by lower concentration of Fe-Gly and Fe-Gly may be superior to FeSO4 for iron fortification to laying hens.

Effect of dietary γ-aminobutyric acid on laying performance, egg quality, immune activity and endocrine hormone in heat-stressed Roman hens.

This study was conducted to evaluate the effect of γ-aminobutyric acid (GABA) on laying performance, egg quality, digestive enzyme activity, hormone level and immune activities in Roman hens under heat stress. Roman hens (320 days old) were fed with 0, 25, 50, 75 and 100 mg/kg GABA, respectively during a 60-day experiment. Compared with control, supplementation of 50 mg/kg GABA improved the laying performance and egg quality by significantly increasing egg production, average egg weight and shell strength (P < 0.05), while decreasing the feed-egg ratio and cholesterol level. Anti-oxidation activity was improved by significantly increasing the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), but decreasing malondialdehyde level in serum (P < 0.05), while significantly increasing the glucose and total protein (TP) level, follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2) ), insulin, triiodothyronine (T(3) ) and free triiodothyronine (FT(3) ) levels, and IgG, IgA and complement (C3)activity in serum (P < 0.05). The results indicated that oral GABA improved laying performance and physical condition mainly by modulating hormone secretion, enhancing anti-oxidation and immune activity, and maintaining electrolyte balance. Fifty mg/kg was the optimum level for laying hens under heat stress in the present study.