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Updated: Jun 14, 2025

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Temperament in Early Childhood Is Associated With Gut Microbiota Composition and Diversity
Eriko Ueda1,2, Michiko Matsunaga1,2,3, Hideaki Fujihara1,2
1Graduate School of Education, Kyoto University, Kyoto, Kyoto, Japan.
Insights
Child gut bacteria composition influences temperament in preschoolers. Lower levels of beneficial bacteria and higher levels of inflammatory bacteria correlate with negative emotionality and stress, impacting early childhood development.
Area of Science:
- Microbiome research
- Developmental psychology
- Child health
Background:
- Temperament is crucial for mental health and development.
- Gut microbiome composition is established during early childhood.
- The gut microbiome may influence brain development and emotion regulation.
Purpose of the Study:
- To investigate the relationship between gut microbiota and temperament in preschool children.
- To identify specific bacterial associations with emotionality and behavioral traits.
Main Methods:
- Assessed temperament in 284 children (aged 3-4 years) using maternal reports (Children's Behavior Questionnaire).
- Analyzed gut microbiota composition and diversity from stool samples using 16S rRNA sequencing.
Main Results:
- Low abundance of anti-inflammatory bacteria (e.g., Faecalibacterium) and high abundance of pro-inflammatory bacteria (e.g., Eggerthella) linked to higher negative affectivity.
- Reduced beneficial bacteria and increased inflammatory bacteria associated with lower surgency/extraversion.
- Gut microbiota diversity correlated with impulsivity, a facet of surgency/extraversion.
Conclusions:
- Gut microbiota composition is significantly associated with preschooler temperament.
- Findings suggest potential biological mechanisms linking gut health to emotional and behavioral development.
- This research may help identify early markers for psychological and emotional risk.
Abstract:
Temperament is a key predictor of human mental health and cognitive and emotional development. Although human fear behavior is reportedly associated with gut microbiome in infancy, infant gut microbiota changes dramatically during the first 5 years, when the diversity and composition of gut microbiome are established. This period is crucial for the development of the prefrontal cortex, which is involved in emotion regulation. Therefore, this study investigated the relationship between temperament and gut microbiota in 284 preschool children aged 3-4 years. Child temperament was assessed by maternal reports of the Children's Behavior Questionnaire. Gut microbiota (alpha/beta diversity and genera abundance) was evaluated using 16S rRNA sequencing of stool samples. A low abundance of anti-inflammatory bacteria (e.g., Faecalibacterium) and a high abundance of pro-inflammatory bacteria (e.g., Eggerthella, Flavonifractor) were associated with higher negative emotionality and stress response (i.e., negative affectivity, β = -0.17, p = 0.004) and lower positive emotionality and reward-seeking (i.e., surgency/extraversion, β = 0.15, p = 0.013). Additionally, gut microbiota diversity was associated with speed of response initiation (i.e., impulsivity, a specific aspect of surgency/extraversion, β = 0.16, p = 0.008). This study provides insight into the biological mechanisms of temperament and takes important steps toward identifying predictive markers of psychological/emotional risk.
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