Figure 4.

Ischemia increased opportunistic facultative anaerobes in gut microbiota. A, Feces were collected twice in the same mice, that is, before and 48 h postischemia (n=6) or sham operation (n=7), and naive mice (twice at corresponding time points; n=6). The cecum was obtained at 48 h. Quantitative polymerase chain reaction showed consistent increases in Enterobacteriaceae in feces after ischemia compared to feces before ischemia (**P=0.0015; 2-way ANOVA-repeated measures design/Sidak test), but not sham or controls. The cecum shows more Enterobacteriaceae after ischemia (**P=0.0052 vs sham; &&&P=0.0006 vs controls; 1-way ANOVA/Dunnett test). B, The Enterobacteriaceae (LOG10) change in feces after vs before ischemia in the same mice is positively correlated to the lesion volume (n=13, Pearson r=0.64, P=0.019). C, Sequencing the 16S rRNA gene from feces obtained before and after ischemia (n=8) or sham operation (n=5) using the Bray-Courtis distance shows significant changes within the ischemic (Permutational Multivariate Analysis of Variance [PERMANOVA], F value=3.5032, R²=0.20015, P<0.001), but not the sham group (PERMANOVA, F value=1.2608, r²=0.11197, P=0.26). D, Ischemia showed a trend to increase Pseudomonadota phylum. E, Sequencing data validated the ischemia-induced increase in the relative abundances of Enterobacteriaceae family and showed a small effect in sham-operated mice (2-way RM ANOVA. *P<0.05; ***P<0.0001). Data are depicted as boxplot, where the dots are individual mice and bars show median and interquartile range. F, At the Family level, ischemia not only increased Enterobacteriaceae (P<0.001, false discovery rate [FDR]<0.001), but also Clostridium family XIII (P<0.001, FDR<0.001), Enterococcaceae (P=0.001, FDR=0.007), and Porphyromonadaceae (P=0.004, FDR=0.029), whereas it decreased the highly abundant Bacteroidales_S24_7_group (P=0.003; FDR=0.024).