In Ovo and Oral Administration of Probiotic Lactobacilli Modulate Cell- and Antibody-Mediated Immune Responses in Newly Hatched Chicks

Abstract

There is some evidence that lactobacilli can strengthen immune system of chickens. This study evaluated the effects of in ovo and oral administration of a lactobacilli cocktail on cytokine gene expression, antibody-mediated immune response, and spleen cellularity in chickens. Lactobacilli were administered either in ovo at embryonic day 18, orally to hatched chicks at days 1, 7, 14, 21, and 28 post-hatches, or by both treatments combined. On days 5 and 10 post-hatch, spleen and bursa of Fabricius were collected for gene expression and cell composition analysis. On days 14 and 21 post-hatch, birds were injected with sheep red blood cells (SRBC) and keyhole limpet hemocyanin (KLH), and sera were collected on days 7, 14, and 21 post-primary immunization. Birds that received lactobacilli (107 CFU) via in ovo followed by weekly oral administration showed a greater immune response by enhancing antibody response, increasing the percentage of CD4+ and CD4+CD25+ T cells in the spleen, and upregulating the expression of the expression of interferon (IFN)-α, IFN-β, interleukin (IL)-12, and IL-13 in the spleen and expression of IFN-γ, IL-2, IL-6, IL-8, IL-12, and IL-18 in bursa. These findings suggest that pre-and post-hatch administration of lactobacilli modulate immune response in newly hatched chickens.

In ovo co-administration of vitamins (A and D) and probiotic lactobacilli modulates immune responses in broiler chickens

“…Macrophages are professional antigen-presenting cells that play a critical role in innate immunity through phagocytosis of pathogens and secretion of inflammatory and anti-inflammatory mediators ( Franken et al, 2016 ). In line with our previous finding ( Alizadeh et al, 2021 ), the results of our study showed that in ovo inoculation of lactobacilli alone or in combination with RA increased the number of KUL01 + cells in the spleen of treated birds. Higgins et al (2008) also showed that Lactobacillus-based probiotic supplementation increases the number of macrophages compared to the untreated control in the cecum of chickens ( Higgins et al, 2008 ).…”

“…Treatment with lactobacilli increased the absolute number of CD3 + CD4 + T cells compared to the control groups on d 10 posthatch. These results are in line with our previous finding that in ovo supplementation of lactobacilli at 10 7 CFU increased the percentage of CD3 + CD4 + T helper cells in the spleen ( Alizadeh et al, 2021 ). Other studies have also demonstrated that dietary supplementation of L. acidophilus and L. fermentum increased the number of CD3 + CD4 + T cells in peripheral blood and intestinal intraepithelial lymphocytes of chickens, respectively ( Bai et al, 2013 ; Asgari et al, 2016 ).…”

“…This study showed that the percentage and absolute number of Bu-1 + IgM + cells were not changed by in ovo inoculation of lactobacilli, RA, or VitD. A previous study also demonstrated that in ovo administration of different doses of lactobacilli may not change the percentage and absolute number of B cells in the spleen ( Alizadeh et al, 2021 ). It appears that RA has an important role in development and differentiation of B cells rather than B cell proliferation.…”

“…Mononuclear cells were prepared from the spleens of 6 birds per treatment group as previously described ( Alizadeh et al, 2021 ). Briefly, tissue samples were rinsed 3 times with PBS and crushed through a 40-μm nylon cell strainer and resuspended in 5 mL of complete RPMI (Invitrogen, Burlington, ON, Canada) medium containing 10% fetal bovine serum and 1% Penicillin-Streptomycin (Gibco, Grand Island, NY).…”

“…Detection of KLH-specific IgG (IgY) and IgM titers in serum samples was performed by ELISA as previously described ( Alizadeh et al, 2021 ). Briefly, 96-well flat-bottom Maxisorp high binding microplates were coated with 100 μL of a KLH solution (1 μg/mL KLH in 0.1 M NaHCO 3 , pH 9.6 with 30 μg/mL BSA) and incubated overnight at 4 ° C. Plates were washed (4 times) with PBST (0.05% Tween 20; P137 Sigma-Aldrich Inc., St. Louis, MO) and blocked (2 h at RT in the dark) with 200 μL blocking buffer (PBST containing 0.25% of fish skin gelatin; Sigma-Aldrich, Oakville, ON).…”