Figure 2

Dietary SpaC and LPS induce intestinal pyroptosis and gut microbial dysbiosis in zebrafish. (A) One‐month old zebrafish were fed with diets supplemented with 0.5 and 1.0 mg/g SpaC or diet supplemented with 0.2 mg/g LPS for three weeks. (B) Representative intestinal histology images by H&E staining. (C) Total histological score measuring the severity of the intestinal injury (Values are means. F_3,20 = 65.490; n = 6). (D) A representative western blot analysis showing Caspase‐3 activation, GSDMEa cleavage, Caspy2 activation and GSDMEb cleavage. (E) Serum LPS levels (F_3,25 = 9.826; n = 6−9, pool of three zebrafish per sample). (F) Indexes of α diversity (ACE: F_3,20 = 17.869; Chao1: F_3,20 = 20.649; Simpson: F_3,20 = 3.273; Shannon: F_3,20 = 11.611; n = 6, pool of 6 zebrafish per sample). (G) The composition of gut microbiota at phylum level (n = 6, pool of six zebrafish per sample). (H) The relative abundance of bacteria producing LPS (F_3,20 = 5.907; n = 6). (I) Total number of bacteria (F_3,20 = 2.332), the number of Proteobacteria (F_3,20 = 18.276) and the number of Entobacteriaceae (F_2,14 = 22.373) in the intestinal microbiota of one‐month old zebrafish immersed with either LGG or PB22 at 10⁷ CFUs/mL for Day 14 (n = 5−6). The Numbers of biologically independent samples are labeled on the violin plots. Box represents median ± interquartile range, whiskers represent 1.5× interquartile range. Statistics: one‐way ANOVA followed by Duncan's test. Treatments in violin or box plots labeled with different letters on top represent statistically significant results (p < 0.05).