Exposure to plant-oriented microbiome altered jejunal and colonic innate immune response and barrier function more strongly in suckling than in weaned piglets

Abstract

Weaning often leaves the piglet vulnerable to gut dysfunction. Little is known about the acute response of a gut mucosa primed by a milk-oriented microbiome before weaning to a plant-oriented microbiome (POM) after weaning. We evaluated the epithelial structure, secretory response and permeability in the small and large intestines of piglets receiving a milk-based (i.e., preweaning) or plant-based diet (i.e., postweaning) to POM inocula using intestinal loop perfusion assays (ILPA). The POM were prepared from jejunal and colonic digesta of four 7 week-old weaned (day 28 of life) piglets, having gut-site specific microbial and metabolite composition. Two consecutive ILPA were performed in 16 piglets pre- (days 24 to 27) and 16 piglets postweaning (days 38 to 41) in two replicate batches. Two jejunal and colonic loops per piglet were perfused with Krebs-Henseleit buffer (control) or the respective POM. The outflow fluid was analyzed for antimicrobial secretions. Jejunal and colonic loop tissue were collected after each ILPA for histomorphology and electrophysiology using Ussing chambers. ANOVA was performed using the MIXED procedure in SAS. The POM stimulated the secretory response by increasing mucin in the jejunal and colonic outflow by 99.7% and 54.1%, respectively, and jejunal IgA by 19.2%, whereas colonic lysozyme decreased 25.6% compared to the control (P < 0.05). Fittingly, the POM raised the number of goblet cells by 96.7% in jejunal and 56.9% in colonic loops compared to control loops (P < 0.05). The POM further flattened jejunal villi by 18.3% and reduced crypt depth in jejunal and colonic loops by 53.8% and 9.0% compared to the control (P < 0.05); observations typically made postweaning and indicative for mucosal recognition of 'foreign' compounds. The POM altered the jejunal and colonic net ion flux as indicated by 22.7% and 59.2% greater short-circuit current compared to control loops, respectively; the effect being stronger postweaning (P < 0.05). Colonic barrier function improved with age (P < 0.05), whereas POM perfusion compromised the mucosal barrier as suggested by 17.7% and 54.1% greater GT and mucosal-to-serosal flux of fluorescein-isothiocyanate dextran, respectively, compared to the control (P < 0.05). In conclusion, results demonstrated that the preweaning gut epithelium acutely responds to novel compounds in postweaning digesta by upregulating the first line of defense (i.e., mucin and lysozyme secretion) and impairment of the structural integrity.

Keywords: host secretory response; intestinal loop perfusion assay; intestinal permeability; microbial metabolites; neonatal piglet.

Figure 1.

Changes in species richness [Chao1(A)] and alpha diversity indices [Shannon (B) and Simpson (C)] of the jejunal and colonic plant-oriented microbiome inoculum (POM) at 0 and 90 min of incubation from the bioorthogonal noncanonical amino acid experiment using 16S rRNA gene amplicon sequencing.

Figure 2.

Changes in bacterial genera composition of the jejunal (A) and colonic (B) plant-oriented microbiome inoculum (POM) at 0 and 90 min of incubation from the bioorthogonal noncanonical amino acid tagging experiment using 16S rRNA gene amplicon sequencing.

Figure 3.

Number of goblet cells in the jejunal (A) and colonic (B) loops not perfused (NON loop), perfused with control inoculum (CON loop; Krebs-Henseleit buffer) or perfused with the respective plant-oriented microbiome inoculum (POM loop) pre- and postweaning. Values are least squares means and pooled standard error of the mean (SEM).