Development of the gut microbiota in the first 14 years of life and its relations to internalizing and externalizing difficulties and social anxiety during puberty

Abstract

Relations between the gut microbiota and host mental health have been suggested by a growing number of case-control and cross-sectional studies, while supporting evidence is limited in large community samples followed during an extended period. Therefore, the current preregistered study ( https://osf.io/8ymav , September 7, 2022) described child gut microbiota development in the first 14 years of life and explored its relations to internalizing and externalizing difficulties and social anxiety in puberty, a period of high relevance for the development of mental health problems. Fecal microbiota composition was analysed by 16S ribosomal RNA gene amplicon sequencing in a total of 1003 samples from 193 children. Through a clustering method, four distinct microbial clusters were newly identified in puberty. Most children within three of these clusters remained in the same clusters from the age of 12 to 14 years, suggesting stability in microbial development and transition during this period. These three clusters were compositionally similar to enterotypes (i.e., a robust classification of the gut microbiota based on its composition across different populations) enriched in Bacteroides, Prevotella, and Ruminococcus, respectively. Two Prevotella 9-predominated clusters, including one reported by us earlier in middle childhood and the other one in puberty, were associated with more externalizing behavior at age 14. One Faecalibacterium-depleted pubertal cluster was related to more social anxiety at age 14. This finding was confirmed by a negative cross-sectional relation between Faecalibacterium and social anxiety in the 14-year-olds. The findings of this study continue to map gut microbiota development in a relatively large community sample followed from birth onwards, importantly extending our knowledge to puberty. Results indicate that Prevotella 9 and Faecalibacterium may be relevant microbial taxa in relation to externalizing behavior and social anxiety, respectively. These correlational findings need validations from other similar cohort studies, as well as well-designed mechanistic pre-clinical investigations before inferring cause and effect.

Keywords: Externalizing behavior; Faecalibacterium; Gut microbiota development; Prevotella 9; Puberty; Social anxiety.

Fig. 1

Microbial clusters. a Transition between microbial clusters in the first 14 years of life. Microbial clusters were determined by the DMM clustering method according to their compositional characteristics at the genus level. The clusters in infancy (i.e., ages of 1, 3, and 4 months) and middle childhood (i.e., ages of 6 and 10 years) were reported previously [8], and the clusters in puberty (i.e., ages of 12 and 14 years) were determined in the present study. Microbial clusters are presented as nodes, with the size and the number indicating how many samples belong to the corresponding cluster. The four pubertal clusters are colored in pink, grass green, lake blue, and purple, respectively. Transition rates between clusters were calculated based on complete case samples and are shown as sized lines. There are N = 130 completed cases between age 10 and age 12, and N = 116 completed cases between age 12 and age 14. The lines from ages 10 to 14 are highlighted in orange, accompanied with transition rates (> 10%) in percentages. b Phylogenetic diversity of pubertal microbial clusters. Box plots show interquartile ranges and median values. Whiskers indicate 1.5 times the interquartile ranges. Wilcoxon tests were implemented between clusters with the FDR correction (adjusted p: ns, not significant; * < 0.05; ** < 0.01). c Beta diversity between pubertal microbial clusters. It was calculated by weighted UniFrac distance based on relative abundance data of genus-level microbial taxa. Ellipses represent 95% confidence intervals for pubertal clusters assuming a multivariate normal distribution. d Differentially abundant genus-level taxa between microbial clusters in puberty. These taxa were identified through LEfSe (Linear discriminant analysis Effect Size) with FDR-adjusted p < 0.05 and LDA (Linear discriminant analysis) effect size > 4. Taxon relative abundances in individuals are shown in the heatmap on the left side. The barplot on the right side represents LDA scores, with colors indicating enriched clusters