Fecal 16S rRNA Sequencing and Metabolomics Reveal Abnormal Metabolism Activity in Preterm Infants with Different Gestational Ages

Ling Liu¹Liang Guo²Jincheng Dai²Xiangsheng Cai^3*Benqing Wu^2*

• ¹Affiliated Hospital of Guangdong Medical College Hospital, Zhanjiang, China
• ²Shenzhen Guangming District People's Hospital, Shenzhen, China
• ³Guangzhou Eleventh People's Hospital, Guangzhou, China

Objective: This study aims to conduct a comprehensive analysis of the differences in gut microbiota and metabolomics in preterm infants stratified by gestational age.Methods: Fresh fecal samples were collected from neonates within the first 3 days after birth.The gut microbiota composition and the changes in specific taxa abundance were analyzed using 16S rRNA sequencing. Metabolomic profiling was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Participants were categorized into four groups based on gestational age at birth: PreA group (34-36 weeks), PreB group (32-33 weeks), PreC group (28-31 weeks), and control group (37-42 weeks). Metabolic pathways were identified through metabolomics analysis, referencing the Kyoto Encyclopedia of Genes and Genomes (KEGG) database.Results: Notably, Principal Coordinates Analysis (PCoA) showed clear separation among samples from all groups, with significant differences noted in the PreC group when compared with the other three. We found a strong association between Escherichia-Shigella and Ureaplasma genera with infants born before 32 weeks of gestation, suggesting a higher risk of opportunistic infections for preterm infants under this gestational threshold. As gestational age increases, Megamonas and Prevotella gradually emerged, while Escherichia-Shigella and Ureaplasma progressively diminished. KEGG enrichment analysis indicated that Pyrimidine metabolism was a differentially regulated pathway between the PreA group and the control group. Interestingly, the only major differential metabolic pathway between the PreB group and the control group was Arachidonic acid metabolism. The bubble diagram revealed significant enrichment of differential metabolites in Pyrimidine and beta-Alanine metabolism pathways when comparing the PreC group with the control group.Conclusion: Significant differences were observed in the fecal microbiome and metabolome between preterm and full-term infants, particularly in those born before 32 weeks of gestation.These findings suggested that the gut microbial system in preterm infants undergoed progressive maturation, approaching a "healthy" state characteristic of full-term infants as gestational age increases.