Long-Term Transcriptomic Effects of Prebiotics and Synbiotics Delivered In Ovo in Broiler Chickens

Abstract

In ovo delivery of prebiotics and synbiotics in chickens allows for the development of intestinal microflora prior to hatching, which boosts their robustness. The goal of this study was to determine the transcriptomic profile of the spleen (S), cecal tonsils (CT), and large intestine (LI) of adult chickens injected with prebiotics and synbiotics in ovo. On day 12 of embryo development, incubating eggs were injected with prebiotics: inulin alone (P1) or in combination with Lactococcus lactis subsp. lactis IBB2955 (S1), galactooligosaccharides (GOS) alone (P2) or in combination with Lactococcus lactis subsp. cremoris IBB477 (S2); control group (C) was mock injected with physiological saline. Gene expression analysis was conducted using an Affymetrix Chicken Gene 1.1 ST Array Strip. Most of the differentially expressed genes (DEG) were detected in the cecal tonsils of P2 (378 DEG), and were assigned to gene ontology categories: lymphocyte proliferation, activation and differentiation, and cytokine production. Ingenuity pathway analysis of the DEG (CT of P2) indicated the inhibition of humoral and cellular immune responses, e.g., role of NFAT in regulation of immune responses, phagocytosis, production of nitric oxide, NF-κB, IL-8, and CXCR4 signaling. The DEG with the highest up-regulation from S1 and P2 were involved in gene expression (PAPOLA, RPL27A, RPLP1, and RPS29) from P1 and P2 in transport (BEST4, SLC9A3, and SLC13A2), metabolism (OGT, ALPP, CA4, and CA7), signaling (FGG, G3BP2, UBB, G3BP2, CACNA1G, and ATP6V0A4), and immune responses (MSMB, LGALS3, CABIN1, CXCR5, PAX5, and TNFRSF14). Two DEG influencing the complement system (SERPING1 and MIR1674) were down-regulated in P2 and S1. In conclusion, GOS injected in ovo provided the most potent stimulation of the host transcriptome. This is likely due to its strong bifidogenic effect, which triggers proliferation of indigenous embryonic microflora in ovo, and indirectly influences gene expression regulation in host tissues, especially cecal tonsils.

Review: Roles of Prebiotics in Intestinal Ecosystem of Broilers

“…On the other hand, it was suggested that spore forming bacteria such as Clostridium tertium were capable of surviving the disinfection process and penetrating eggs, resulting in contamination ( 118 ). To avoid extensive pathogen infection, prebiotics were delivered in ovo , which is likely fermented by the indigenous embryonic microbiota, inhibiting pathogen proliferation and regulating gene expression of immune responses ( 119 ). Villaluenga et al ( 120 ) reported that injection of raffinose at day 12 of embryonic incubation had the highest amounts of Bifidobacteria in the ceca of 2 day-old broilers.…”

Splenic Gene Expression Signatures in Slow-Growing Chickens Stimulated in Ovo with Galactooligosaccharides and Challenged with Heat

“…On a performance level, HS causes losses in feed intake and growth rate. In broiler chickens, chronic HS has been found to significantly reduce (p < 0.01) final body weight (BW) (2.52 kg in TN vs. 3.11 kg in HS) [32,39,40]. In the current study, chronic HS also reduced final BW (p < 0.001), but the difference between TN and HS was almost two times lower than in broiler chickens (2.21 kg in TN vs. 1.94 kg in HS), even with the longer rearing period (42 days in broilers vs. 50 days in slow-growing chickens).…”

“…Galactooligosaccharides are potent prebiotics that exert beneficial effects on intestinal microbiota in chickens. Particularly interesting is the possibility of using in ovo technology for the early stimulation of the microbial communities with GOS [32,39,40]. The molecular data on broiler chickens have indicated that HS triggers systemic immune and stress responses, which are balanced by GOS that are delivered in ovo [41].…”

Metabolic Gene Expression in the Muscle and Blood Parameters of Broiler Chickens Stimulated In Ovo with Synbiotics

“…The gastrointestinal microbiota affects the host organism by regulating immune response, metabolism, digestion, and nutrient absorption [6]. A technology for reprogramming the composition of chicken microbiota at an early stage of embryonic development was developed by administering prebiotics or synbiotics using in ovo technology on day 12 of egg incubation [7,8]. The in ovo administered bioactive substances such as prebiotics like galacto-oligosaccharides (GOS) and inulin and synbiotics like Lactococcus lactis subsp.…”