Zhui Li

Effects of Sophy β-glucan on growth performance, carcass traits, meat composition, and immunological responses of Peking ducks.

The response of Peking ducks to supplements of Sophy β-glucan was studied. A total of 160 healthy 1-d-old mixed-sex ducklings were randomly allocated to 3 groups: Sophy β-glucan (n = 80), bacitracin zinc (n = 40), and control (n = 40), which received the same antibiotics-deficient diet supplemented with 1% β-glucan, 5% bacitracin zinc, or nothing, respectively. During 2 mo of the study, growth performance, carcass composition, and meat quality of Peking ducks were evaluated. Additionally, a separate immunological study was conducted with a total of 105 healthy male Peking ducks in 7 groups (n = 15) and immunized with different doses of β-glucan (0, 0.5, 2.5, 12.5, and 62.5 μg/duck) and BSA (200 μg/duck). Blood was taken for detection of anti-BSA-IgG antibody and peripheral blood mononuclear cells proliferation assays. Groups subjected to different dietary treatments showed almost no differences in growth performance and slaughter traits except breast muscle percentage and intestinal length. These 2 indicators were significantly higher in the bacitracin zinc group than in the control and β-glucan groups (P < 0.05). Similarly, chemical compositions, fatty acids, and amino acids of breast muscle were not significantly influenced by the diet. Ducks immunized with Sophy β-glucan did not have enhanced level of anti-BSA-IgG antibodies but had significant peripheral blood mononuclear cells proliferation compared with unchallenged ducks (P < 0.01). With an increase in the glucan concentration, the proliferative responses approximately linearly increased. These findings indicate that 1% Sophy glucan did not improve duck growth performance, carcass composition, and meat quality significantly under the conditions of the present experiment and mainly had regulatory or enhancing properties on poultry nonspecific cellular immunity.

Dietary live yeast and mannan-oligosaccharide supplementation attenuate intestinal inflammation and barrier dysfunction induced by Escherichia coli in broilers

The effects of live yeast (LY) and mannan-oligosaccharide (MOS) supplementation on intestinal disruption induced by Escherichia coli in broilers were investigated. The experimental design was a 3×2 factorial arrangement with three dietary treatments (control, 0·5 g/kg LY (Saccharomyces cerevisiae, 1·0×1010 colony-forming units/g), 0·5 g/kg MOS) and two immune treatments (with or without E. coli challenge from 7 to 11 d of age). Samples were collected at 14 d of age. The results showed that E. coli challenge impaired (P<0·05) growth performance during the grower period (1-21 d) and the overall period (1-35 d) of broilers, increased (P<0·05) serum endotoxin and diamine oxidase levels coupled with ileal myeloperoxidase and lysozyme activities, whereas reduced (P<0·05) maltase activity, and compromised the morphological structure of the ileum. Besides, it increased (P<0·05) the mRNA expressions of several inflammatory genes and reduced occludin expression in the ileum. Dietary treatment with both LY and MOS reduced (P<0·05) serum diamine oxidase and ileal myeloperoxidase levels, but elevated villus height (P<0·10) and the ratio of villus height:crypt depth (P<0·05) of the ileum. It also alleviated (P<0·05) E. coli-induced increases (P<0·05) in ileal Toll-like receptor 4, NF-κ B and IL-1 β expressions. Moreover, LY supplementation reduced (P<0·05) feed conversion ratio of birds during the grower period and enhanced (P<0·05) the community diversity (Shannon and Simpson indices) of ileal microbiota, whereas MOS addition counteracted (P<0·05) the decreased ileal IL-10 and occludin expressions in challenged birds. In conclusion, both LY and MOS supplementation could attenuate E. coli-induced intestinal disruption by alleviating intestinal inflammation and barrier dysfunction in broilers. Moreover, LY addition could improve intestinal microbial community structure and feed efficiency of broilers.

Effects of Kluyveromyces marxianus supplementation on immune responses, intestinal structure and microbiota in broiler chickens

To investigate the effects of Kluyveromyces marxianus on immune responses, intestinal structure and microbiota in broilers, 840 1-d-old broiler chicks were randomly divided into seven groups (eight replicates) and were fed basal diets without or with 0.25, 0.50, 1.0, 1.5, 2.0, and 2.5 g/kg of K. marxianus (2.0×1010 CFU/g). Serum and intestine samples were collected at 21 d of age. The results showed that increasing K. marxianus addition linearly reduced feed conversion ratio but linearly elevated relative thymus weight, as well as quadratically increased serum lysozyme and IgG levels, with the medium dose (1.0 g/kg) being the most effective. The ratio of villus height to crypt depth of jejunum and ileum, ileal villus height and sucrase activity, as well as the mRNA expression of ileal mucin-2, claudin-1 and sodium glucose cotransporter 1 linearly responded to the increasing K. marxianus addition. Supplemental K. marxianus at low (0.5 g/kg), medium (1.5 g/kg) and high (2.5 g/kg) dose all decreased the abundance of phylum Cyanobacteria, increased the abundance of phylum Firmicutes and genus Lactobacillus in ileum. The high dose of K. marxianus addition also reduced the abundance of order Rickettsiales and Pseudomonadales along with species Acinetobacter junii. Ileal bacterial communities between K. marxianus-treated and untreated groups formed distinctly different clusters. In summary, K. marxianus supplementation benefits feed efficiency and immune function, as well as intestinal structure in broilers, which might be attributed to the improved ileal microbial structure. Supplemental K. marxianus at high dose (2.5 g/kg) was more effective for feed efficiency and intestinal health of broilers, while the innate immunity was optimized at a medium dose (1.0 g/kg).

Early embryonic blood cells collect antigens and induce immunotolerance in the hatched chicken

Earlier experimental data in our laboratory showed that introduction of an exogenous protein into early chicken embryonic blood leads to immunotolerance of hatched chicken to that protein. However, the underlying mechanism is yet unknown. In the present study, we show that the blood cells collecting circulating antigen might contribute to the establishment of immunotolerance. In this experiment, most of the chicken embryo blood cells took up injected fluorescein isothiocyanate-BSA at approximately embryonic d 3. At the same stage, 1 microL of embryo blood was taken out and incubated with BSA. After being loaded with BSA in vitro and washed, these cells were injected back into the original embryo. The BSA-specific lymphocytes were depleted in chickens whose early embryo cells had been loaded with BSA, as evidenced by a significant decrease in anti-BSA antibody after challenge with BSA when the chickens were 3 wk old. In addition, by direct injection of BSA to embryonic d 3 embryo blood, the hatched chickens had decreased amounts of anti-trinitrophenol antibody after the chickens were challenged with trinitrophenol-BSA, indicating that the helper function of BSA-specific T cells was impaired. In conclusion, these observations suggest that some early embryo blood cells possibly collect and store antigen for the establishment of self-tolerance before the maturation of B and T cells.

Effects of Lactobacillus acidophilus on gut microbiota composition in broilers challenged with Clostridium perfringens

This study shows the effects of dietary supplementation with Lactobacillus acidophilus on the gut microbiota of broiler chickens challenged with Clostridium perfringens infection during a 21-day period according to pyrosequencing of the 16S ribosomal RNA gene. In a 2 × 2 factorial arrangement of treatments, 308 1-day-old male Arbor Acres broiler chicks were analyzed for the effects of the probiotic (groups without or with L. acidophilus supplementation), pathogen challenge (groups without or with C. perfringens), and the effects of interaction. The infection decreased the number of Observed species, Chao1, and ACE of ileal microbiota and increased Chao1 of cecal microbiota of broilers, whereas L. acidophilus supplementation decreased the Shannon index of the ileal microbiota. Shannon index and Simpson indices were lower in the ileal microbiota than in the cecal microbiota. In the ileal microbiota, the control group had higher relative abundance of Lachnospiraceae and Ruminococcaceae in comparison with the other groups; however, the relative abundance of Gammaproteobacteria was significantly higher in the challenge group than in the other groups. C. perfringens infection tended to increase lactate concentration and decreasedconcentrations of formate, acetate and propionate in the ileum; decreased isobutyrate concentration; and tended to decrease isovalerate concentration in the cecum. Besides, L. acidophilus supplementation increased the concentration of lactate and butyrate and decreased concentrations of formate and propionate in the ileum, and increased concentrations of lactate and valerate in the cecum. In conclusion, C. perfringens infection and/or dietary supplementation with L. acidophilus modulated the relative abundance of some bacteria taxa, and the L. acidophilus supplementation helped to restore the microbial community disrupted by C. perfringens infection.

Two Lactobacillus Species Inhibit the Growth and α-Toxin Production of Clostridium perfringens and Induced Proinflammatory Factors in Chicken Intestinal Epithelial Cells in Vitro

Clostridium perfringens is the causative pathogen of avian necrotic enteritis. Lactobacillus spp. are well-characterized probiotics with anti-microbial and immune-modulatory activities. In the present study, we investigated the effects of L. acidophilus and L. fermentum on the growth, α-toxin production and inflammatory responses of C. perfringens. In in vitro culture experiments, both lactobacilli inhibited the growth of C. perfringens (P < 0.01), accompanied with a decrease in pH (P < 0.01). Supernatants from lactobacilli cultures also suppressed the growth of C. perfringens during 24 h of incubation (P < 0.01), but this inhibitory effect disappeared after 48 h. Both lactobacilli decreased the α-toxin production of C. perfringens (P < 0.01) without influencing its biomass, and even degraded the established α-toxin (P < 0.01). Lower environmental pH reduced the α-toxin production as well (P < 0.01). Preincubation with L. acidophilus decreased the attachment of C. perfringens to cells (P < 0.01) with the cell cytotoxicity being unaffected. Both lactobacilli pretreatment reduced the up-regulation of proinflammatory factors, peptidoglycan (PGN) receptors and nuclear factor kappa B (NF-κB) p65 in C. perfringens-challenged chicken intestinal epithelial cells (P < 0.05). In conclusion, L. acidophilus and L. fermentum inhibited the pathological effects of C. perfringens in vitro conditions.

Bacillus subtilis and yeast cell wall improve the intestinal health of broilers challenged by Clostridium perfringens

ABSTRACT 1. The objective was to investigate the effects of Bacillus subtilis, yeast cell wall (YCW) and their combination on intestinal health of broilers challenged by Clostridium perfringens over a 21-d period. 2. Using a 5 × 2 factorial arrangement of treatments, 800 1-d-old male Cobb 500 broilers were used to study the effects of feed additives (without additive or with zinc bacitracin, B. subtilis, YCW, and the combination of B. subtilis and YCW), pathogen challenge (without or with Clostridium perfringens challenge), and their interactive effects. 3. C. perfringens infection increased intestinal lesions scores, damaged intestinal histomorphology, increased serum endotoxin concentration, cytokine mRNA expression and intestinal population of C. perfringens and Escherichia coli and decreased ileal bifidobacteria numbers. The 4 additives decreased serum endotoxin. Zinc bacitracin tended to decrease cytokine mRNA expression and the intestinal number of C. perfringens and E. coli. B. subtilis, YCW and their combination increased cytokine mRNA expression. B. subtilis and YCW decreased the number of C. perfringens and E. coli in the ileum, and their combination decreased pathogens numbers in the ileum and caecum. 4. In conclusion, B. subtilis, YCW and their combination improved the intestinal health of NE-infected broilers, and could be potential alternatives to antibiotics.

Absence of Donor-Derived Zona Pellucida Protein C Homolog in the Inner Perivitelline Layer of Peking Duck (Anas platyrhynchos)-Japanese Quail (Coturnix coturnix japonica) Chimeras (Duails)

Avian blastodermal cells at stage X are used to produce interspecies chimeras for heterogenous poultry reproduction. However, recipient-derived inner perivitelline layer (IPVL)-enclosed donor-derived ova may affect the efficiency of germline transmission via chimera. Among the proteins in the IPVL, zona pellucida protein C (ZPC) plays an important role in sperm-egg binding and inducing the acrosome reaction. In the present study, Peking duck blastodermal cells at stage X were transferred into subgerminal cavities of Japanese quail embryos at the same stage. Fourteen female duck-quail chimeras (duails) were hatched and raised to sexual maturity. After being screened by PCR, 3 duails were selected for examination of donor-derived ZPC. A total of 152 IPVL protein samples from the individual eggs laid by the 3 duails then underwent a preliminary examination for the presence of donor-derived ZPC by means of SDS-PAGE, periodic acid-Schiff staining, and Western blotting. A novel 35-kDa ZPC, not observed in quail but in duck, was found in the IPVL of the duails. Further analysis of peptide mass fingerprinting of Peking duck ZPC, Japanese quail ZPC, and the 35-kDa duail ZPC by matrix-assisted laser desorption-ionization reflectron time-of-flight mass spectrometry revealed that the novel ZPC was an isoform of quail ZPC. Moreover, comparison of N-terminal amino acid sequences of these 3 ZPC confirmed that the 35-kDa quail ZPC had more amino acids at the N terminus than did native quail ZPC, and none of the donor-derived ZPC was found in the duails. These findings suggest that it would be difficult to obtain donor-derived offspring by natural mating of interspecies chimeras.

Dietary l-arginine Supplementation Alleviates the Intestinal Injury and Modulates the Gut Microbiota in Broiler Chickens Challenged by Clostridium perfringens

Our previous reports suggested that Dietary l-arginine supplementation attenuated gut injury of broiler chickens infected with Clostridium perfringens by enhancing intestinal immune responses, absorption and barrier function, but its effect on the gut microbiome of broiler chickens remains unclear. This experiment aimed at evaluating the effects of Dietary l-arginine supplementation on the gut bacterial community composition and function of broiler chickens challenged with C. perfringens. In total, 105 1-day-old male Arbor Acres broiler chickens were assigned to three groups: Control (CTL), C. perfringens-challenged (CP), and C. perfringens-challenged and fed diet supplemented with 0.3% l-arginine (ARGCP) groups. The challenge led to macroscopic and histomorphological gut lesions, decreased villus height and increased the number of Observed species, Shannon, Chao1 and ACE indices of ileal microbiota, whereas l-arginine addition reversed these changes. Moreover, the three treatments harbored distinct microbial communities (ANOSIM, P < 0.05). At the genus level, 24 taxa (e.g., Nitrosomonas spp., Coxiella spp., Ruegeria spp., and Thauera spp.) were significantly more abundant in CP group than in CTL group (P < 0.05), whereas the levels of 23 genera of them were significantly decreased by l-arginine supplementation (P < 0.05). The abundances of only 3 genera were different between CTL and ARGCP groups (P < 0.05). At the species level, the challenge promoted the relative abundance of Nitrospira sp. enrichment culture clone M1-9, Bradyrhizobium elkanii, Nitrospira bacterium SG8-3, and Pseudomonas veronii, which was reversed by l-arginine supplementation (P < 0.05). Furthermore, the challenge decreased the levels of Lactobacillus gasseri (P < 0.05). Predictive functional profiling of microbial communities by PICRUSt showed that compared with CP group, ARGCP group had enriched pathways relating to membrane transport, replication and repair, translation and nucleotide metabolism and suppressed functions corresponding to amino acid and lipid metabolisms (P < 0.05). The relative abundances of KEGG pathways in l-arginine-fed broilers were almost equal to those of the controls. In conclusion, l-arginine alleviated the gut injury and normalized the ileal microbiota of C. perfringens-challenged chickens to resemble that of unchallenged controls in terms of microbial composition and functionality.

Effects of live yeast on immune responses and intestinal morphological structure in lipopolysaccharide-challenged broilers

Abstract: The effects of live yeast (LY) on immune responses and intestinal morphological structure in lipopolysaccharide (LPS) challenged broilers were investigated. A total of 480 one-day-old male broilers were divided into six groups following a 3 × 2 factorial arrangement (eight replicates) with three dietary LY dosages (0%, 0.05%, or 0.50%) and two immune treatments (injection of saline or LPS) given at an interval of 48 h between 21 and 27 d of age. Blood was sampled on days 21 and 27, while intestine were collected on day 28. Results showed that LPS damaged (P ≤ 0.05) intestinal histomorphology, and tended to reduce (P=0.071) serum anti-Newcastle disease virus (NDV) titers on day 21, but increase (P=0.089) jejunal maltase activity. Supplemental LY at 0.05% elevated (P ≤ 0.05) ileal villus width and villus surface area, and serum anti-NDV titers on day 21. The addition at 0.50% increased (P ≤ 0.05) jejunal and ileal villus height to crypt depth ratio, reduced (P ≤ 0.05) jejunal maltase activity and crypt depth, and attenuated (P ≤ 0.05) LPS-induced increase (P ≤ 0.05) in serum diamine oxidase activity on day 21. Collectively, supplemental LY at 0.05% improved antibody response and the addition at 0.50% alleviated LPS-induced intestinal damage in broilers.