This study was conducted to investigate the effects of various doses of a multistrain lactobacilli mixture (Lactobacillus salivarius, Lactobacillus reuteri, Lactobacillus crispatus, and Lactobacillus johnsonii) on the innate and adaptive immune responses in broiler chickens. At embryonic day eighteen, 200 eggs were injected with PBS, or three different doses of a multi-strain lactobacilli mixture (1 × 10 5 , 1 × 10 6 , and 1 × 10 7 CFU/egg, P1, P2, and P3 respectively) along with a group of negative control. On days 5 and 10 post-hatch, cecal tonsil, bursa of fabricius, and spleen were collected for gene expression and cellular analysis. On days 14 and 21 posthatch, birds were immunized intramuscularly with both sheep red blood cells (SRBC) and keyhole limpet hemocyanin (KLH). Serum samples were collected on days 0, 7, 14, and 21 after primary immunization. The results demonstrated that lactobacilli inoculation increased the splenic expression of cytokines, including interferon (IFN)α, IFN-β, IFN-γ, interleukin (IL)-8, and IL-12 on day 5 post-hatch compared to the control group (PBS). However, in cecal tonsils, lactobacilli treatment downregulated the expression of IL-6 on day 5 post-hatch and IL-2 and IL-8 on day 10 posthatch. No significant differences were observed in the expression of cytokine genes in the bursa except for IL-13 which was upregulated in lactobacilli-treated groups P2 and P3 on days 5 and 10 post-hatch. Flow cytometry analysis showed that the percentage of KUL01, CD4 + and CD8 + splenocytes was not affected by treatments. In addition, no significant differences were observed for antibody titers against SRBC. However, lactobacilli treatment (P1, P2, and P3) was found to increase IgM titers on day 21 post-primary immunization compared to controls. Furthermore, in ovo injection of the highest dose of probiotics (1 × 10 7 , P3) increased serum IgG titers Alizadeh et al.In ovo Application of Lactobacilli against KLH on day 7 post-primary immunization. In conclusion, this study demonstrated that that in ovo administration of lactobacilli can improve antibody-mediated immune responses and differentially modulate cytokine expression in mucosal and systemic lymphoid tissues of chickens.

Vitamins are nutritional elements which are necessary for essential activities such as development, growth, and metabolism of cells. In addition to these conventional functions, vitamins A, D, E, and C have vital roles in normal function of the immune system as their deficiency is known to impair innate and adaptive host responses. By altering transcription of multiple immune system genes and contributing to antioxidant activities, these vitamins influence the immune system in different ways including modulation of cell-mediated and antibody-mediated responses, immunoregulation, and antiinflammatory effects. Furthermore, supplementation of these vitamins to poultry may assist the immune system to combat microbial pathogens while reducing detrimental effects associated with stress and enhancing responses to vaccines. In this article, the relationship between the chicken immune system and vitamins A, D, E, and C is reviewed, and evidence from the literature pertaining to how these vitamins exert their antiinflammatory, regulatory, and antimicrobial effects is discussed.

The effect of organic trace mineral supplementation on performance, intestinal morphology, immune organ weights (bursa of Fabricius and spleen), expression of innate immune response related genes, blood heterophils/lymphocytes ratio, chemical metabolic panel, natural antibodies (IgG), and oxidative stress of broiler chickens was studied. A total of 1,080 day-old male broilers were assigned to 1 of 3 dietary treatments, which included basal diet with Monensin (control), control diet supplemented with bacitracin methylene disalicylate (BMD), and BMD diet supplemented with organic trace minerals (OTM). No difference in feed conversion ratio was observed among treatments; ileum histomorphological analysis showed a lower crypt depth, higher villi height/crypt depth ratio, and lower villi width in the OTM treatment compared to control. Furthermore, OTM treatment resulted in higher uric acid and lower plasma malondehaldehyde (MDA), indicating lower oxidative stress. Gene expression analysis showed that OTM treatment resulted in up-regulations of TLR2 bin the ileum, and TLR2b, TLR4, and IL-12p35 in the bursa of Fabricius, and down-regulation of TLR2b and TLR4 in the cecal tonsils. In the spleen, OTM treatment resulted in up-regulation of IL-10. In conclusion, OTM supplementation to broiler diets may have beneficial effects on intestinal development, immune system status, and survival by improving ileum histomorphological parameters, modulation of Toll-like receptors and anti-inflammatory cytokines, and decreasing level of MDA, which in conjunction could enhance health status.

This study evaluated the effect of yeast-derived products on growth performance, serum antibody levels, and mRNA gene expression of pattern-recognition receptors, and cytokines in broiler chickens. Two hundred and sixteen one-day-old male broiler chickens (Ross-308) were randomly assigned to six dietary treatments with six replicates (cage) of 6 birds per cage. Dietary treatments consisted of a Control diet without antibiotics (C), and diets containing 11 mg/kg of "virginiamycin", 0.25% of yeast cell wall (YCW), 0.2% of a commercial product "Maxi-Gen Plus" containing processed yeast and nucleotides, 0.05% of nucleotides, or a diet containing 8% of distiller's dried grains with solubles (DDGS). On d 21 post-hatch blood samples were collected from 6 birds per treatment and serum sample were analyzed for antibody levels. After blood sampling, birds were injected intraperitoneally with 3 mg/kg of BW of lipopolysaccharide (LPS). The unchallenged group was fed the Control diet and injected with saline solution. Spleen samples were collected to measure the gene expression of toll-like receptors (TLR)2b, TLR4, and TLR21, macrophage mannose receptor (MMR), and cytokines including interleukin (IL)-12, IL-10, IL-4, IL-6, IL-18, and interferon (IFN)-γ. No significant difference in body weight gain, feed intake, and FCR were observed among treatments. Regarding humoral immunity, the diet supplemented with YCW increased serum immunoglobulin (Ig)A level compared with the antibiotic group; however, serum concentrations of IgG and IgM were not affected by dietary treatments. Relative gene expression of TLR2 and TLR4 was not affected by dietary treatments, whereas the expression of TLR21 and MRR was upregulated in diets containing YCW and DDGS. The diet supplemented with YCW increased the expression of all cytokines, and expression of IFN-γ was upregulated in the DDGS group. However, no significant difference was observed for cytokine gene expression in the antibiotic and nucleotide diets. In conclusion, supplementation of diet with YCW stimulated the systemic innate immune responses of broiler chickens following challenge with LPS.

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