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Abstract
Background The gut microbiota is believed to influence the behavioral and neurodevelopmental outcomes of preterm children. This study aimed to investigate the relationships between gut microbiome composition, diversity, and mental health outcomes in preterm infants aged 3-4 years, specifically focusing on internalizing behaviors.
Methods Stool samples from 197 preterm infants aged 3-4 years were collected, and the Child Behavior Checklist (CBCL) categorized them into three groups based on Anxiety T Scores. Alpha and Beta diversity analysis assessed gut microbiota richness and evenness. Univariate and multivariable differential analyses (Mann Whitney U test and MaAsLin2) identified differences in gut microbiota composition among infants with varying levels of internalizing behaviors. Pearson correlation analysis explored network correlations between gut microbiota and internalizing behaviors. Functional prediction analysis investigated pathway and gene family abundance and correlation to uncover potential biological mechanisms influencing internalizing behaviors.
Results Significant variations in microbial diversity were observed between the Low Risk and Mid Risk groups (p=0.029, Shannon index; IDDF2024-ABS-0144 Figure 1). Beta diversity analysis showed a significant difference in microbial community structure between the ’Low Risk’ and ’Mid Risk’ groups (p=0.011; IDDF2024-ABS-0144 Figure 2). No significant differences were found in specific bacterial species among the three groups. Generalized Linear Model (GLM) analysis revealed the significance of variables such as ‘Gestation’ and ‘Delivery’ in distinguishing the Mid risk group from the Low Risk group. MaAsLin2 analysis identified significant associations, with ’Clostridium_bolteae_CAG_59’ positively correlating with the Mid Risk group (q=0.004), and ’Anaerostipes_hadrus’ negatively correlating with the Mid Risk group (q=0.037; IDDF2024-ABS-0144 Figure 3). Pathway correlations suggested that an increase in Anaerostipes hadrus might enhance certain metabolic pathways’ activity, while an increase in Clostridium bolteae CAG_59 might decrease such activity (IDDF2024-ABS-0144 Figure 4).
Conclusions The gut microbiome may play a role in the mental health outcomes of preterm infants. Variations in microbial diversity and community structure were found between infants with different levels of internalizing behaviors. Specific microbial taxa, including ’Clostridium_bolteae_CAG_59’ and ’Anaerostipes_hadrus’, were associated with internalizing behaviors. These findings provide valuable insights into the link between gut microbiota and internalizing behaviors in preterm infants aged 3-4 years.