Pu-erh tea theabrownin improves the ovarian function and gut microbiota in laying hens.

Studies have reported that theabrownin can moderate the lipid metabolism and intestinal microbiota, thereby affecting the health of humans and model animals, however the research on laying hens is scarce. The present study aimed to investigate the effects of dietary theabrownin supplementation on lipid metabolism, microbial composition and ovarian function in laying hens. A total of 80 laying hens (25 wk of age) were fed with normal diet (CON) and normal diet +100 mg/kg theabrownin (PT group) for 12 wk. The results showed that the addition of theabrownin enhanced villus height of duodenum and decreased crypt depth of jejunum (P < 0.05). At the same time, compared with CON, the concentration of IL-6 and the mRNA expression of IL-1ß and IL-6 were decreased significantly in PT group (P < 0.05). Dietary theabrownin reduced the concentration of total cholesterol and glycerol, while decreased lipid droplet optical density in liver (P < 0.05). Compared with CON group, the mRNA expression of PPARγ, HMG-CoAS, ACC were down-regulated and the mRNA expression of CYP8B1 was up-regulated in PT group (P < 0.05). The ACE, Chao1 and Observed_species indexes in cecum microbiota were increased by PT group intervention (P < 0.05). Dietary PT supplementation enhanced the relative abundance of Firmicutes (phylum), Lactobacillus (genus) and the Firmicutes to Bacteroidetes ratio, and reduced the relative abundance of Bacteroidetes (phylum) in cecum (P < 0.05). The organic acids and its derivatives were up-regulated by theabrownin intervention in serum metabolites (P < 0.05). Dietary theabrownin supplementation resulted in higher mRNA expression of Bcl-2 and SIRT1 in ovary and increased the concentration of estradiol in serum (P < 0.05). These discovering indicated that dietary theabrownin supplementation enhanced the intestinal function and influenced serum metabolism by improving intestinal morphology, microbiota community structure and reducing the concentration and expression of inflammatory cytokines in intestine. Dietary theabrownin reduced hepatic lipid deposition and it also decreased the cell apoptosis rate to improve ovarian function and egg weight which were associated with the SIRT1 pathway.

Extract of Scutellaria baicalensis and Lonicerae flos improves growth performance, antioxidant capacity, and intestinal barrier of yellow-feather broiler chickens against Clostridium perfringens.

In this study, we aimed to investigate the effect of Scutellaria baicalensis and Lonicerae Flos (SL) extract on the growth performance and intestinal health of yellow-feather broilers following a Clostridium perfringens challenge. In total, 600 one-day-old yellow-feather broilers were divided into five treatments (6 replicate pens of 20 birds per treatment), including a control (Con) group fed a basal diet and the infected group (iCon) fed a basal diet and infected with Clostridium perfringens, the other 3 groups receiving different doses of SL (150, 300, and 450 mg/kg) and infected with Clostridium perfringens. The total experimental period was 80 d. When the birds were 24-days-old, a subclinical necrotizing enteritis model was induced by orally inoculating the birds with 11,000 oocysts of mixed Eimeria species on d 24, followed by C. perfringens (108 CFU/mL) from d 28 to 30. The birds were evaluated for parameters such as average weight gain (AWG), average daily feed intake (ADFI), mortality, feed conversion ration (FCR), intestinal lesion score, intestinal C. perfringens counts, and villus histomorphometry. Results indicated that C. perfringens infection led to reduced AWG and the levels of tight junction proteins, increased the FCR, ileum E. coli load, and intestinal permeability, causing damage to the intestinal mucosal barrier (P < 0.05). Compared with the infected group, supplementing 300 mg/kg of SL significantly increased AWG at 43 to 80 d, the ratio of villus height to crypt depth in the jejunum and ileum at 35 d, and the activity of superoxide dismutase (SOD) in serum. It also significantly reduced the FCR at 22 to 42 d, intestinal lesion score, and the amount of C. perfringens in the ileum (P < 0.05). Additionally, compared with the infected group, the addition of 300 mg/kg SL significantly increased mRNA levels of claudin-2, claudin-3, mucin-2, and toll-like receptor 2 (TLR-2) in the ileum of infected birds at 35 d of age. In conclusion, supplementation with SL extract could effectively mitigate the negative effects of C. perfringens challenge by improving intestinal barrier function and histomorphology, positively influencing the growth performance of challenged birds.

Effects of Long-Term Coated Sodium Butyrate Supplementation on the Intestinal Health and Colonization of Cecal Salmonella of Laying Hens Infected with Salmonella enteritidis.

Salmonella enterica ser. Enteritidis (S. Enteritidis) is widely found in chickens and eggs, and it can potentially induce human illness. The investigation in this study centers on the impacts of long-term dietary supplementation with coated sodium butyrate (CSB) on intestinal well-being and the colonization of cecum Salmonella in laying hens infected with S. Enteritidis. We segregated a total of 120 Lohmann laying hens aged 51 weeks into four treatment categories: 0 (CON), 300 (CSB1), 500 (CSB2), and 800 (CSB3) mg/kg of CSB, supplemented with CSB from the first day of the experiment. A 24-week observation process was carried out for each laying hen. The S. Enteritidis was orally administered to all chickens on the morning of the first and third days of week 22 of the trial. After the S. Enteritidis challenge, egg production decreased the most in the CON group. Compared to the CON group, the three doses of CSB significantly improved egg production after the S. Enteritidis challenge (PANOVA < 0.05). S. Enteritidis challenge increased plasma DAO activity, but CSB supplementation reduced plasma DAO activity (Plinear < 0.05). The S. Enteritidis challenge disrupted intestinal villi morphology; compared to the CON group, the three dosages of CSB resulted in an increase in villus height (VH) and the ratio of villus height to crypt depth (V/C) in the duodenum, jejunum, and ileum of infected laying hens (Plinear < 0.05), with a significant increase in jejunal villus height (PANOVA < 0.05). A decrease in ileal crypt depth was also observed (Plinear < 0.05). CSB2 and CSB3 markedly increased the content of butyric acid in the cecum (PANOVA < 0.05). Additionally, in contrast to those in the CON group, the propionic acid content in the CSB supplementation group increased (Plinear < 0.05). Compared with those in the CON group, mRNA relative expression of the IL-6 and IL-1ß in jejunum (Plinear < 0.05) and mRNA relative expression of the IL-1ß in ileum (PANOVA < 0.05) were significantly lower, and mRNA relative expression of the IL-10 in ileum (Plinear < 0.05) were significantly higher in the CSB group. In addition, in contrast to the CON group, the CSB supplementation group significantly upregulated mRNA relative expression of the ZO-1 and CLDN1 (PANOVA < 0.05). Additionally, CSB supplementation reduced the number of Salmonella and increased the number of Lactobacilli in the cecum (Plinear < 0.05) and tended to increase the total bacteria count (Plinear = 0.069) and reduce the E. coli count (Plinear = 0.081). In conclusion, long-term dietary supplementation with coated sodium butyrate can alleviate intestinal injury and the colonization of cecum Salmonella in laying hens infected with S. Enteritidis.

Effects of stevia extract on production performance, serum biochemistry, antioxidant capacity, and gut health of laying hens.

In the present study, we aimed to elucidate the effects of stevia extract on production performance, serum immune indexes, intestinal structure, and cecum microbial structure. We randomly divided eight hundred 46-wk-old Roman hens into 5 groups, with 8 replicates in each group and 20 chickens in each replicate. The control group was fed a basal diet, whereas the 4 experimental groups were fed 50, 100, 200, and 400 mg/kg stevia extracts. The study period was 24 wk. The addition of different concentrations of the stevia extract to the diet resulted in significant secondary changes in the egg production rate at 1 to 12 wk (P < 0.05). Furthermore, the addition of 50 and 100 mg/kg stevia extract to the diet significantly increased serum IgM and IgG levels in laying hens (P < 0.05) but linearly decreased serum IL-1ß levels (P < 0.05). Serum T-SOD activity linearly increased (P = 0.057); however, serum biochemical indexes showed no significant differences. Stevia extract tended to increase the ratio of the duodenal villi height to the depth of the crypt (P = 0.067), with no obvious lesions in the duodenum, jejunum, and ileum. In addition, stevia extract increased the relative abundance of species at the phylum level, with the abundance of Bacteroides and Firmicutes exhibiting significant secondary changes (P < 0.05). The ACE and Chao1 indexes suggested that stevia extract addition significantly increased the alpha diversity of cecum microorganisms in laying hens. Furthermore, NMDS analysis based on operational taxonomic units revealed that stevia extract addition increased the beta diversity of cecum microorganisms in laying hens. Adding a certain amount of stevia extract to feed can improve the production performance, immune ability, and intestinal health of laying hens to some extent, and we recommend an effective level of 200mg/kg of stevia extract for laying hen diets.

Metabolomics analysis to interpret changes in physiological and metabolic responses to chronic heat stress in Pekin ducks.

Heat stress (HS) is a major environmental threat that affects duck production in subtropical and tropical regions, especially in summer. This study aimed to evaluate the physiological and metabolic responses of Pekin ducks to chronic HS conditions via liquid chromatography-mass spectrometry (LC-MS) using a paired-fed (PF) experimental design. On the basis of equivalent feed intake (HS vs. PF), HS significantly reduced growth performance and the percentage of leg and breast muscles, however, markedly increased the percentage of abdominal fat and breast skin fat. Serum metabolomics results revealed that heat-stressed ducks showed enhanced glycolysis and pentose phosphate pathways, as demonstrated by higher glucose 6-phosphate and 6-phogluconic acid levels in the PF vs. HS comparison. HS decreased hepatic mRNA levels of mitochondrial fatty acid ß-oxidation-related genes (MCAD and SCAD) compared to the PF group, resulting in acetylcarnitine accumulation in serum. Moreover, HS elevated the concentrations of serum amino acids and mRNA levels of ubiquitination-related genes (MuRF1 and MAFbx) in the skeletal muscle and amino acid transporter-related genes (SLC1A1 and SLC7A1) and gluconeogenesis-related genes (PCK1 and PCase) in the liver compared to the PF group. When compared to the normal control group (NC), HS further decreased growth performance, but it elevated the abdominal fat rate. However, increased mRNA levels of ubiquitination-related genes and serum amino acid accumulation were not observed in the HS group compared to the NC group, implying that reduced feed intake masked the effect of HS on skeletal muscle breakdown and is a form of protection for the organism. These results suggest that chronic HS induces protein degradation in the skeletal muscle to provide amino acids for hepatic gluconeogenesis to provide sufficient energy, as Pekin ducks under HS conditions failed to efficiently oxidise fatty acids and ketones in the mitochondria, leading to poor growth performance and slaughter characteristics.

Effects of Dietary Level of Corn Bran on Laying Performance and Cecum Microbial Communities in Laying Ducks.

The application of corn bran (CB) to laying ducks via iso-energy and iso-nitrogen diets is rarely reported. Six hundred laying ducks (49 weeks) were equally assigned to five treatments: the control group with 0% CB and the other four groups with 3%, 6%, 9%, and 12% CB. The experiment lasted for 11 weeks. With the increase in CB, the relative weight of the proventriculus, gizzard, and ileum and the content and proportion of butyric acid in the cecal digesta were quadratically changed (p < 0.05), and the highest value was observed in the 12% CB group. Compared with the control, the 12% CB group showed decreased Deferribacteres, Spirochaetota, and Fusobacteriota at the phyla level and showed increased Pediococcus and decreased Bifidobacterium and Rikenellaceae_RC9_gut_group at the genus level (p < 0.10); the 12% CB group also showed 46 different metabolites, which are related to Lactobacillus and Pediococcus (p < 0.05). The 12% CB group showed increased (p < 0.05) albumen height at week 8 and yolk color at weeks 4 and 8 compared with the control. Overall, dietary inclusion of 3% to 12% CB is a possible feeding strategy for laying ducks under iso-energy and iso-nitrogen conditions, and the 12% CB group was more effective.

Research Note: Effects of feeding corn naturally contaminated with aflatoxin B1, deoxynivalenol, and zearalenone on reproductive performance of broiler breeders and growth performance of their progeny chicks.

To evaluate the toxic effects of mycotoxin-contaminated corn (MC) on the breeders and their progeny chicks, a total of 480 fifty-wk-old Cobb broiler breeder hens were fed the following dies: 1) a corn-soybean meal diet (Control; containing 70.35% corn), 2) MC substituting for 50% of corn in Control (LM), 3) LM diet plus 2 g/kg 1 mycotoxin sequestrant, Toxy-Nil Plus (TNP) (LMT2.0), 4) MC substituting for 100% of corn in Control (HM), 5) HM diet plus 2 g/kg TN (HMT2.0), and 6) HM diet plus 2.5 g/kg TNP (HMT2.5). The MC contained 69.25 µg aflatoxin B1 (AFB1)/kg, 4,875 µg deoxynivalenol (DON)/kg, and 2,262 µg zearalenone (ZEN)/kg. At wk 4 after MC inclusion, all eggs laid were used for hatch, and all progeny chicks were fed the same mycotoxin-untreated diet for 14 d. Dietary MC inclusion decreased the hatchability of set eggs and increased embryo mortality during d 18 to 21.5. The TNP addition increased these aforementioned indices in MC-included diets. Maternal HM treatment decreased the BW of progeny chicks at age of 14 d and BWG of progeny chicks during d 1 to 14, whereas maternal LM treatment did not affect these indices. In parallel, maternal HM treatment decreased the concentrations of serum IgA, IgG, and lysozyme in the progeny chicks on d 14, but maternal LM treatment did not affect these indices. Overall, maternal dietary TNP treatments increased the growth of progeny chicks and had a trend to increase the concentrations of serum IgA and IgG on d 14 compared to maternal MC treatments. It was concluded that the feeding of relative high ratio of corn contaminated with low level of AFB1, DON, and ZEN negatively affected the reproductive performance of breeders and the growth performance of their progeny chicks, and TNP addition alleviated these toxic effects.

The improving effect of soybean isoflavones on ovarian function in older laying hens.

Emerging evidence suggests an association between estrogen levels and reduced egg-laying performance as the layer became old. Since soy isoflavones (SF) have estrogen-mimic effects, whether it can enhance production performance and ovarian function of older layers is still not known. A total of 160 Lohmann pink layers (66-wk-old) were used in a 2 × 2 factorial design, which included 2 egg-laying levels [low (76.89 ± 1.65%; LOW) and normal (84.96 ± 1.01%; NOR)] and 2 different dietary groups [0 mg/kg SF, 20 mg/kg SF] were used. The results showed the NOR group had higher egg-laying rate, egg mass, and feed efficiency during the all phases (P(laying) < 0.05). The unqualified egg rate was lower in NOR group (9-12 wk, 1-12 wk) (P(laying) < 0.05). Dietary supplementation with SF increased the egg-laying rate and feed efficiency (5-8 wk, 9-12 wk, 1-12 wk), increased egg mass (9-12 wk, 1-12 wk) (P(SF) < 0.05). The NOR layers presented higher eggshell quality (redness, yellowness, brightness, eggshell ratio) at 12 wk (P(laying) < 0.05). Eggshell quality was found to be improved by SF (eggshell strength and eggshell thickness), egg albumen quality (higher albumen height and Haugh unit) at 12 wk (P(SF) < 0.05). Supplementing with SF led to an increase in eggshell strength in LOW group (P(laying*SF) < 0.05). The higher serum lever of glucose (GLU) and lower serum lever of follicle stimulating hormone (FSH) were in NOR group (P(laying) < 0.05). Supplementing SF in diets increased serum of estradiol (E2) and insulin-like growth factors-1 (IGF-1), decreased serum of FSH (P(SF) < 0.05). The NOR layers presented lower estrogen receptor α (ERα), estrogen receptor ß (ERß), B lymphoma 2 associated X protein (Bax), cytochrome c (Cytc), interleukin 6 (IL-6), caspase3, caspase9, IKKα, P50, and P65 expression in the ovary (P(laying) < 0.05). Dietary SF supplementation decreased the anti-Müllerian hormone receptor (AMHR), Bax, caspase3, caspase9, Cytc, IL-6, IKKα, P50, P65 expression in the ovary (P(SF) < 0.05). These findings indicated that layers with NOR group had higher production performance, egg quality, and ovarian function, while dietary supplementation with SF improved production performance and ovarian function by reducing inflammation and apoptosis-related genes expression in ovary.

Effects of Thymol and Carvacrol Eutectic on Growth Performance, Serum Biochemical Parameters, and Intestinal Health in Broiler Chickens.

This study aimed to evaluate the effect of diets supplementing with various levels of thymol and carvacrol eutectic (TCE) on growth performance, serum biochemical parameters, intestinal morphology, and the expression of intestinal nutrient absorption, barrier function- and inflammation-related genes in broiler chickens. A total of 640 one-day-old Arbor Acres male broilers with similar body weights were randomly divided into four groups (8 replicates/group, 20 broilers/replicate). Birds in the four experimental groups were fed a basal diet with TCE at 0, 30, 60, or 120 mg/kg. The results showed that the growth performance of birds during 22-42 d or 1-42 d, serum IgE and IgG content at 21 d of age, jejunal and ileal morphology, ileal MUC2, OCLN, and IL-10 mRNA expression were significantly increased compared with the control group (p < 0.05), and the ileal IL-6 mRNA expression quadratically decreased (p < 0.05) with increasing dietary TCE supplemented dosage, and its expression showed a linear downward trend (0.05 < p < 0.1). Meanwhile, compared with the other three groups, birds fed diets with 30 mg/kg TCE presented better (p < 0.05) growth performance, intestinal morphology, and function. These results indicated that the optimal supplementation amount of TCE in the broiler diets was 30 mg/kg.

Effects of cadmium, lead, mercury, chromium, and selenium co-treatment on egg quality and fatty acids.

This study aimed to reveal the effect of selenium (Se) and heavy metals (chromium (Cr), cadmium (Cd), lead (Pb), and mercury (Hg)) on the quality, fatty acids, and 13 kinds of ions in the egg yolk and albumen. Four experimental groups were established, including a control group (control; basal diet), Se group (basal diet + Se), heavy metals group (basal diet + CdCl2 + Pb(NO3)2 + HgCl2 + CrCl3), and Se + heavy metal (HM) group (basal diet + Se + CdCl2 + Pb(NO3)2 + HgCl2 + CrCl3). Se supplementation significantly increased the experimental egg yolk percentage since Se accumulation mainly occurred in the yolks of the eggs. The Cr content in the yolks of the Se + heavy metal groups decreased at 28 days, while a significant reduction was evident in the Cd and Hg levels of the Se + heavy metal yolks compared to the heavy metal group at 84 days. The complex interactions between the elements were analyzed to determine the positive and negative correlations. Se displayed a high positive correlation with Cd and Pb in the yolk and albumen, while the heavy metals minimally affected the fatty acids in the egg yolk.

Effects of dietary theabrownins on production performance, egg quality, and ovarian function of laying hens with different ages.

This experiment was conducted to investigate the effect of theabrownins (TB) on production performance, egg quality, and ovarian function of laying hens at different ages. A total of 240 Lohmann laying hens were assigned in a 2 × 2 factorial design, which encompassed 2 layers ages (47-wk-old and 67-wk-old) and 2 dietary levels of TB (0 and 100 mg/kg) for 12 wk. Results showed that older layers had lower laying rate, egg mass, and higher feed-to-egg ratio (F/E), egg weight and unqualified egg rate than the younger layers (P(AGE) < 0.01) during all the experimental period. The effect of TB was found to increase egg laying rate and feed efficiency during 5 to 8 wk, 9 to 12 wk and the overall phases and decreased unqualified egg rate during 1 to 4 wk and the overall phases (P(TB) ≤ 0.05). The eggshell quality (strength, thickness), albumen quality (albumen height and Haugh unit) of eggs from older layers were decreased during overall phases (P(AGE) ≤ 0.05). TB increased eggshell strength during all phases and enhanced eggshell thickness at the end of wk 4 and 8 and increased albumen height and Haugh unit at the end of wk 8 and 12 of older layers (P(Interaction) ≤ 0.05). In addition, TB also increased egg quality of older layers after 14 d storage. A decrease in the serum concentration of progesterone, melatonin, follicle stimulating hormone, estradiol was observed in the older compared to the younger ones (P(AGE) < 0.05), while the increase in serum concentration of progesterone, melatonin, anti-Müllerian hormone (AMH) were more emphasized when older hens received TB supplemented diet (P(Interaction) < 0.05). The older layer demonstrated lower the concentration of glutathione (GSH) (P(AGE) < 0.05). And the activity of glutathione-s-transferase (GST) was significantly decreased in layers under 67-wk-old (P(AGE) <0.05). The increase in concentration of GSH and the decrease in concentration of malondialdehyde (MDA) were more pronounced when TB were supplemented in 67-wk-old layers (P(Interaction) ≤ 0.05). Layers at 67-wk-old had lower mRNA expression of Heme oxygenase 1 (HO-1) (P(AGE) < 0.01) in ovary. Dietary TB supplementation upregulated mRNA gene expression of HO-1, Nuclear factor E2 related factor 2 (Nrf2), Quinone oxidoreductase 1 (NQO1) (P(TB) < 0.01). Dietary TB upregulated mRNA expression of ovarian reproductive hormone receptor (estrogen receptor 1 [ESR1] and steroidogenic acute regulatory protein 1 [StAR1]]; P(TB) < 0.01). The results suggest feeding TB (100 mg/kg) could improve the egg production rate, egg quality, and antioxidant capacity of the ovary. Moreover, the effect of TB was more pronounced in older layers (64-wk-old vs. 47-wk-old).

Effect of benzoic acid, Enterococcus faecium, and essential oil complex on intestinal microbiota of laying hens under coccidia and Clostridium perfringens challenge.

The objective of this study was to investigate whether dietary supplementation with benzoic acid, Enterococcus faecium, and essential oil complex (BEC) could help laying hens recover from coccidia and Clostridium perfringens type A challenge. A total of 60 (35-wk-old) Lohmann-laying hens were randomly assigned to 3 experimental groups (10 replicates with 2 hens per replicate): I) control group (CON), II) challenge group (CC), and III) BEC group (2,000 mg/kg BEC). The total experimental period was 8 wk. The results shown that the challenge layers had lower egg-laying rate and average daily feed intake (ADFI) (P < 0.05), and addition of BEC after challenge increased egg-laying rate (P < 0.05). The content of propionic acid (PA) and butyric acid (BA) in short-chain fatty acid (SCFA) was significantly decreased by challenge (P < 0.05). CC and BEC groups had lower villus height to crypt depth ratio (V/C) and higher pathological scores in duodenum (P < 0.05), whereas the BEC group had lower pathological scores in jejunum when compared with the CC group (P < 0.05). The challenge increased the concentration of proinflammatory cytokines (IL-1ß and IL-6) (P < 0.05). An increase in the abundance of Bacteroidoes (genus), Bacteroidaceae (family), Bacteroidoes sp. Marseille P3166 (species), Bacteroidoes caecicola (species) was observed in the CC group, whereas the BEC group had higher abundance of Bacteroides caecigallinarum (species). The genera Faecalibacterium and Asterolplasma were positively correlated with egg-laying rate (r = 0.57, 0.60; P < 0.01); and the genera Bacteroides and Romboutsia were negatively correlated with egg-laying rate (r = -0.58, -0.74; P < 0.01). The genera Bacteroides, Lactobacillus, and Rombutzia were positively correlated with jejunal mucosa proinflammatory factor IL-1ß level (r = 0.61, 0.60, 0.59; P < 0.01), which were negatively correlated with genera Rikenbacteriaceae RC9, Faecalibacterium, and Olsenlla (r = -0.56, -0.57, -0.61; P < 0.01). There genera UCG.005 was positively correlated with proinflammatory factor IL-6 level in jejunal mucosa (r = 0.58; P < 0.01), which was negatively correlated with Rikenbacteriaceae RC9 (r = -0.62; P < 0.01). The experiment results revealed that the addition of BEC to the diet restored the production performance of the laying hens. In addition, supplementation of 2,000 mg/kg BEC modulated gut health by reducing gut damage scores and modulating microbial composition, thereby promoting recovery of laying hens after coccidia and Clostridium perfringens challenge.

Dietary 25-hydroxyvitamin D improves productive performance and intestinal health of laying hens under Escherichia coli lipopolysaccharide challenge.

The effect of 25-hydroxyvitamin D (25OHD) on the immune response of laying hens is not well elucidated. This study investigated the effects of 25OHD on egg production, egg quality, immune response, and intestinal health of laying hens challenged with Escherichia coli lipopolysaccharide (LPS). One hundred and sixty laying hens at 45 wk of age were randomly divided into 4 dietary treatments with 10 replicates of 4 birds. Hens were fed the corn-soybean based diets contained either 0 or 80 µg/kg 25OHD for 8 wks. At wk of 53 wk, birds of each dietary treatment were injected into the abdomen with 1.5 mg/kg body weight of either LPS or saline a day at 24-h intervals for continuous 7 d. LPS injection significantly decreased (PLPS < 0.05) egg laying rate, feed intake and feed efficiency; while the supplementation of 25OHD increased (PInteraction < 0.05) egg laying rate, feed efficiency and decreased (PInteraction < 0.05) the broken egg rate in layers under LPS injection. LPS challenge decreased (PLPS < 0.05) eggshell strength, eggshell thickness, albumen height and Haugh unit, while dietary 25OHD supplementation increased eggshell strength and eggshell thickness (P25OHD < 0.05). The serum proinflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6)], endotoxin and diamine oxidase (DAO) levels were higher in layers under LPS challenge (PLPS < 0.05); whereas the dietary addition of 25OHD were shown to decrease (P25OHD < 0.05) serum IL-1ß and IL-6 concentration irrespective of LPS challenge and led to a higher serum 25OHD level and a reduction in endotoxin concentration in layers under LPS challenge (PInteraction < 0.05). The layers under LPS challenge had higher crypt depth and lower villus height/crypt depth (V/C) ratio in duodenum and jejunum (PLPS < 0.05), while feeding 25OHD were shown to have decreasing effect on crypt depth and increasing effect V/C ratio in layers under LPS challenge (PInteraction < 0.05). Layers under LPS challenge had lower mRNA expression of intestinal barrier associated proteins (claudin-1 and mucin-1) (PLPS < 0.05), while the addition of 25OHD up-regulated claudin-1 and mucin-1 expression (Pinteraction < 0.05). Lower antioxidant enzymes activities, including superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), glutathione peroxidase (GPx) and higher malondialdehyde (MDA) content in jejunum were found in layers challenged with LPS (P25OHD < 0.05). The effect of 25OHD reversed the effect of LPS on SOD, T-AOC, and MDA content (PInteraction< 0.05). These results suggest that supplementing 80 µg/kg 25OHD in diets may elevate laying performance and egg quality through the improvement of intestinal barrier function, antioxidant capacity, and decreased the proinflammatory cytokines levels in laying hens with Escherichia coli LPS challenge.

Improvement in ovarian function following fecal microbiota transplantation from high-laying rate breeders.

The underlying mechanism between the gut microbiota and reproductive function is not yet well-known. This study was conducted to investigate the effect of the administration of fecal microbiota transplantation (FMT) from highly laying rate donors on the cecal microbiota, intestinal health and ovarian function in broiler breeders. A total of 60 broiler breeders (53 wk of age) were selected by their laying rate [high (HP, 90.67 ± 0.69%; n = 10) and low (LP, 70.23 ± 0.87%; n = 20)]. The LP breeders were then be transplanted with fecal microbiota from HP hens (FMTHP; n = 10) or the same dosage of PBS (FMTCON; n = 10) for 28 d. The results revealed that FMT from HP donors increased egg-laying rate and serum hormone levels [17ß-estradiol (E2), anti-Müller hormone], also decreased proinflammatory cytokine levels (interleukin-6, interleukin-8, tumor necrosis factor-α) of LP breeders (P < 0.05). The FMTHP group breeders had higher villus height, villus height/crypt depth ratio, and upregulated mRNA expression of jejunum barrier-related gene (ZO-2 and mucin-2) and estrogen, follicle-stimulating hormone (FSH) and anti-Müller hormone (AMH) receptor genes (ESR1, ESR2, FSHR, AMHR) (P < 0.05) than FMTCON group. FMT from HP donors led to higher mRNA expression of Bcl2 and sirtuin1 (SIRT1), while it downregulated the proapoptotic genes (Bax, caspase-3, caspase-8, and caspase-9) mRNA expressions in ovary compared with the FMTCON breeders (P < 0.05), and this pattern was also observed in HP donors. Also, HP breeder had higher observed_species and alpha-diversity indexes (Chao1 and ACE) than FMTCON group, while FMTHP can increase observed_species and alpha-diversity indexes (Chao1 and ACE) than FMTCON group (P < 0.05). The bacteria enrichment of Firmicutes (phylum), Bacteroidetes (phylum), Lactobacillus (genus), Enterococcus (genus), and Bacteroides (genus) were increased by FMTHP treatment. The genera Butyricicoccus, Enterococcus, and Lactobacillus were positively correlated with egg-laying rate. Therefore, cecal microbiomes of breeders with high egg-laying performance have more diverse activities, which may be related to the metabolism and health of the host; and FMT from high-yield donors can increase the hormone secretion, intestinal health, and ovarian function to improve egg-laying performance and the SIRT1-related apoptosis and cytokine signaling pathway were involved in this process.

Effects of Dietary Zinc Deficiency on Skin Breaking Strength and Fatty Acid Composition in Broiler Chickens and Pekin Ducks.

This study is aimed at investigating the effect of dietary zinc deficiency (ZnD) on skin breaking strength and skin chemical and fatty acid composition in broiler chickens and Pekin ducks. A total of 200 1-day-old male broiler chickens and 200 1-day-old male ducklings were used in a 2 × 2 factorial design and randomly allocated to 4 treatments with 10 replicated cages of 10 birds per cage. Diets containing zinc at 84.77 mg/kg and 20.42 mg/kg were regarded as the control diet and zinc-deficient diet, respectively. The results showed the following: (1) dietary ZnD decreased (P < 0.05) the breast skin weight (day 21), breast skin index (day 21), skin fat content (day 7), and skin Zn content (days 7, 14, and 21) of ducks, but only increased (P < 0.05) the skin fat content of broiler chickens at 7 days of age; ducks had a higher (P < 0.05) breast skin weight, breast skin index, and skin breaking strength as well as a lower skin fat content (days 7 and 14) than those in broiler chickens. (2) Dietary ZnD decreased the content of myristic acid (day 21) and increased the content of oleic acid (day 7) content in the skin of ducks and increased the palmitic acid (day 7) content in the skin of broiler chickens (P < 0.05) and increased the MUFA (day 7) content in the skin and the atherogenic index (day 21) in ducks. The contents of myristic acid (day 21), stearic acid (day 21), and oleic acid (days 7, 14, and 21) in the skin of broiler chickens were lower (P < 0.05) than those in ducks of the same age. In conclusion, using skin weight, skin index and skin MUFA content as criteria, the results indicated that meat ducks were more sensitive to dietary ZnD than broiler chickens. Using skin fat content as criterion, the results indicated that broiler chickens were more sensitive to dietary ZnD than meat ducks.

Determination of Time Progression and Sensitive Biomarkers of Maternal Zinc Depletion in Broiler Chickens and Pekin Ducks Fed a Zinc-Deficient Diet.

Two experiments were performed to assess maternal zinc (Zn) depletion in broiler chickens and Pekin ducks fed a Zn-deficient diet. The time of Zn depletion was assessed based on growth performance, and sensitive biomarkers were determined based on tissue Zn content via a linear regression model. A total of 200 1-day-old male broiler chickens (experiment 1) and 200 1-day-old male Pekin ducks (experiment 2) were randomly allocated to 2 diets with 10 replicate cages (10 birds/cage). The two diets were a zinc-deficient diet (ZnD, 20.42 mg Zn/kg) and a control zinc diet (CON, 84.77 mg Zn/kg). In experiment 1, compared to CON, ZnD decreased (P < 0.05) the body weight (days 7, 14, and 21), body weight gain, feed intake (days 1-7, 1-14, and 1-21), and the Zn content of plasma (days 7 and 21), pancreas (days 7, 14, and 21), and tibia in broiler chickens. The R2 of a linear model was greater at day 7 than at day 14 or day 21 for pancreatic Zn content in broiler chickens. In experiment 2, compared to CON, ZnD also decreased (P < 0.05) the body weight (days 7, 14, and 21), body weight gain (days 1-7, 1-14, and 1-21), and feed intake (days 1-14 and 1-21) and increased (P < 0.05) the feed-to-gain ratio (days 1-7 and 1-14) in ducks. Compared with CON, ZnD reduced (P < 0.05) the Zn content of the pancreas (days 7, 14, and 21), tibia (days 7, 14, and 21), and skin (days 14 and 21) and increased (P < 0.05) the Zn content of the plasma (day 21) and skin (day 7) in ducks. The R2 of a linear model was greater at day 7 than at days 14 or 21 for skin Zn content in ducks. The results indicated that the maternal Zn was depleted by 7 days of age in both birds; the sensitive biomarker for broiler chickens is pancreatic Zn content, and for ducks, it is skin Zn content.

Dietary Zinc Glycine Supplementation Improves Tibia Quality of Meat Ducks by Modulating the Intestinal Barrier and Bone Resorption.

Leg problems characterized by gait abnormity and bone structure destruction are associated with a high risk of fractures and continuous pain in poultry. Zinc (Zn) acts a pivotal part in normal bone homeostasis and has proven to be highly effective in alleviating leg problems. Therefore, the effects of graded concentration of Zn on bone quality were evaluated in this study. A total of 512 1-d-old male ducks were fed 4 basal diets added 30 mg/kg Zn, 60 mg/kg Zn, 90 mg/kg Zn, and 120 mg/kg Zn as Zn glycine for 35 d. Tibia Zn content, ash percentage, and breaking strength linearly increased with dietary elevated Zn level (P < 0.05). Broken-line analysis revealed that the recommended level of Zn from Zn glycine was 55.13 mg/kg and 64.48 mg/kg based on tibia ash and strength, respectively. To further confirm the role of dietary Zn glycine addition on bone characteristics, data from birds fed either 60 mg/kg Zn as Zn sulfate (ZnSO4), 30 mg/kg Zn, or 60 mg/kg Zn in the form of Zn glycine indicated that birds given 60 mg/kg Zn from Zn glycine diet exhibited higher tibia ash, strength, and trabecular volume compared to those fed the 30 mg/kg Zn diet (P < 0.05). Dietary 60 mg/kg Zn as Zn glycine addition decreased intestinal permeability, upregulated the mRNA expression of tight junction protein, and increased the abundance of Lactobacillus and Bifidobacterium, which was companied by declined the level of inflammatory cytokines in both the ileum and bone marrow. Regarding bone turnover, the diet with 60 mg/kg Zn from Zn glycine induced osteoprotegerin expression and thus decreased osteoclast number and serum bone resorption biomarker levels including serum tartrate-resistant acid phosphatase activity and C-terminal cross-linked telopeptide of type I collagen level when compared to 30 mg/kg Zn diet (P < 0.05). Except for the upregulation in runt-related transcription factor 2 transcription, the experimental treatments did not apparently change the bone formation biomarker contents in serum. Additionally, Zn glycine displayed a more efficient absorption rate, evidenced by higher serum Zn level, and thus had potentially greater a protective role in the intestine barrier and tibia mass as compared to ZnSO4. Collectively, the dietary supplementation of 60 mg/kg in the form of Zn glycine could suppress bone resorption mediated by osteoclast and consequently improve tibial quality of meat ducks, in which enhanced intestinal integrity and optimized gut microbiota might be involved.

Microbiome-metabolomics analysis insight into the effects of dietary resistant starch on intestinal integrity.

Resistant starch (RS) has caught much attention for its potential to exert a beneficial impact on intestine and certain members of its resident microbiota. In this study, we examined how dietary RS promotes intestinal barrier in meat ducks by microbiome-metabolomics analysis. Ducklings were fed corn-soybean basal diet or RS diet. Dietary RS improved intestinal morphology and enhanced barrier function in ileum, evidenced by lower permeability and upregulated tight junction proteins and Mucin-2 gene expression. Microbiome analysis showed that RS administration elevated the proportion of Firmicutes and butyrate-producing bacteria, and increased butyrate contents in cecum. Furthermore, significant alterations in metabolic profiles were observed, with most of these were associated with the amino acid metabolism (especially tryptophan), lipid metabolism, and intestinal inflammation. Together, diet with RS improved gut integrity and caused corresponding alterations in gut metabolome and microbiome, yielding better insights of the mechanism by RS improved the gut system of ducks.

Comparison of zinc bioavailability in zinc-glycine and zinc-methionine chelates for broilers fed with a corn-soybean meal diet.

The objective of this study was to compare the bioavailability of zinc (Zn) from zinc-glycine (Zn-Gly) and zinc-methionine (Zn-Met) as compared with zinc sulfate (ZnSO4) used as a standard in broilers. A total of 1,200 one-day-old male broilers (Cobb 500) were randomly allotted to one of 10 treatments with eight replicate cages of 15 birds each. The broilers were fed a corn-soybean meal basal diet (containing 26.46 mg Zn/kg; control) or the basal diet added with 40, 80, and 120 mg Zn/kg as Zn-Gly, Zn-Met, or ZnSO4 for 14 days. The relative bioavailability value (RBV) was calculated based on multiple linear regression slope ratios of Zn concentrations in tibia and pancreas, pancreas metallothionein (MT) concentration, and pancreas MT mRNA abundance on added Zn intake. When comparing the control with all Zn-supplemented treatments, Zn addition did not significantly affect average feed intake and bodyweight gain during days 1-14 (p > 0.10). However, Zn concentrations in the tibia, pancreas, and liver and pancreas MT concentration and MT mRNA abundance increased in all Zn-supplemented treatments compared with the control (p < 0.05), and these indices increased linearly (p < 0.001) with increasing added Zn levels on days 7 and 14. The RBV of Zn as Zn-Met was similar to that as Zn-Gly or ZnSO4 (p > 0.40) on days 7 and 14, based on tibia and pancreas Zn. In contrast, on days 7 and 14, the RBVs of Zn were in the following order: Zn-Met > Zn-Gly > ZnSO4 (p < 0.05), based on pancreas MT concentration. The bioavailable Zn from Zn-Met was 1.20 or 1.25 times that from Zn-Gly on day 7 or 14, respectively, evaluated by pancreas MT content. The RBV of Zn as Zn-Met was similar to that as Zn-Gly or ZnSO4 on day 7, whereas it was higher than that as Zn-Gly or ZnSO4 on day 14, based on pancreas MT mRNA abundance. In conclusion, Zn-Met had higher bioavailable Zn than Zn-Gly for the starter broilers fed with the corn-soybean meal diet, using pancreas MT concentration as the response criterion.

Effects of Fat Pre-Emulsification on the Growth Performance, Serum Biochemical Index, Digestive Enzyme Activities, Nutrient Utilization, and Standardized Ileal Digestibility of Amino Acids in Pekin Ducks Fed Diets with Different Fat Sources.

This experiment was conducted to evaluate the effects of fat pre-emulsification on growth performance, the serum biochemical index, intestinal digestive enzyme activities, nutrient utilization, and the standardized ileal digestibility of amino acids (SIDAA) in Pekin ducks fed diets containing different fat sources. Three hundred and twenty healthy ten-day-old Pekin male ducks (409 ± 27 g) were assigned to a 2 × 2 factorial design and given one of two types of poultry fat (duck fat or a mixed type of fat composed of chicken fat and duck fat in a 1:1 ratio) that had been pre-emulsified or not. This resulted in four treatments of eight replicates, with each replicate having ten ducks. The results showed that fat pre-emulsification (preE) significantly increased (p < 0.05) body weight and body weight gain and decreased (p < 0.05) the feed-to-gain ratio, the liver index, the activity of aspartate aminotransferase (AST) and the concentration of total cholesterol (TC) in the serum. Fat preE also tended to decrease the activity of lipase (p = 0.07) and significantly reduced (p < 0.05) the activity of trypsin in the duodenum. The utilization of dietary dry matter, ether extract (EE), energy, and total phosphorus, as well as apparent metabolizable energy (AME) and the SID of serine (p = 0.090), were improved by fat preE. Duck fat markedly increased (p < 0.05) the serum TC concentration and the utilization of dietary EE; however, it also tended to decrease the serum triglyceride (TG) concentration (p = 0.09) and markedly decreased (p < 0.05) the activity of trypsin in the jejunum and duodenum. These results indicated that fat preE contributed to the utilization of dietary nutrients, serum lipid metabolism, intestinal digestive enzyme activities, and liver health, thereby improving the growth performance of ducks. Duck fat has higher bioavailability for ducks based on dietary EE utilization.