Fig. 4: Gut dysbiosis is associated with behavioral problems in PAE mice.

a Timeline of the experiment. b The PAE mice score lower in the learning index in the accelerated rotarod test (p = 0.0351). Student’s t-test. Control n = 12; PAE n = 13. Graph represents a box blot with 25th, median, and 75th percentiles, and whiskers extend to the lowest and highest values. Each dot represents an individual animal. c PAE mice reach lower terminal speed in the last trial (Trial 6) compared to control mice (adj.p = 0.0257). Two-way ANOVA revealed no interaction between trials and exposure. Tukey’s post hoc test was used for multiple comparisons. Control n = 12; PAE n = 13. Each dot on the graph represents a group mean ± SEM. d PAE mice spend less time in the open arm compared to the control mice (p = 0.0370). Student’s t-test. Control n = 13; PAE n = 14. c, d Graph represents mean ± SEM. Each dot represents an individual animal. e Stacked bar plots show the taxonomic composition of the microbial community at the family level in each animal used for analysis. f Microbial diversity analysis reveals that PAE mice have smaller alpha diversity compared to that of control mice (p = 0.00030926) as well as differential microbial communities (beta diversity). Alpha diversity was compared using Chao1 at the genus level. Graph represents a boxplot with 25th, median, and 75th percentiles. Whiskers represent 9 and 91 percentiles, and outside of whiskers represent outliers. Each dot represents an individual animal, and the black diamond represents the mean. Beta diversity analysis at the genus level shows that PAE and control mice have differential microbial communities. F = 3.2946, R² = 0.11246, p = 0.017 by Permutational ANOVA (PERMANOVA). Graph represents 2-D Principal Coordinate Analysis (PCoA). Each dot represents an individual animal. g Using EdgeR, 12 differentially abundant taxonomy are identified at the genus level. Graph represents log2 Fold Change (FC), and bacteria is listed in the order of the most significant (Blautia) to the least significant (Paraclostridium). Red bar increased in PAE, Blue bar decreased in PAE. h The abundance of bacterial taxa at the genus level in PAE mice correlates with levels of motor learning and anxiety. Color of heatmap indicates rho values by Spearman’s correlation analysis. i Functional profiles of the microbiome predict changes in fatty acid-related pathways in PAE mice. Graph represents the difference in mean proportions (%) with 95% confidence intervals calculated by Welch’s inverted method. *,**p < 0.05, 0.01.