Abstract
INTRODUCTION: Perinatal factors affect gut microbiota and infant health, but the combined impacts of preterm birth and chorionicity on gut microbiota, metabolism, and physical/neurobehavioral development in twins remain unclear. METHODS: A total of 143 twin families (12-month-old infants) were enrolled and divided into four groups by gestational age and chorionicity (dichorionic-diamniotic full-term/preterm, monochorionic-diamniotic full-term/preterm). Gut microbiota diversity and fecal metabolism were analyzed via 16S rRNA sequencing and untargeted metabolomics. Wilcoxon's rank-sum tests, generalized estimating equations, and twin-based ACE models were used for alpha diversity comparison, differential microbiota identification, and genetic/environmental contribution evaluation, respectively, with confounder adjustment in development association analysis. RESULTS: We identified 10 group-specific gut microbiota genera and 394 differential metabolites. Fifty-two microbiota taxa showed genetic variance, but none overlapped with group-specified taxa. Six genera and 18 metabolites correlated with twins' physical/neurobehavioral development. Pathway analysis highlighted three key metabolites: Morphine (isoquinoline alkaloid biosynthesis, drug metabolism, neuroactive ligand-receptor interaction), Nicotinuric acid (nicotinate/nicotinamide metabolism), and Catechin (flavonoid/phenylpropanoid biosynthesis). DISCUSSION: Preterm birth is linked to gut microbiota dysbiosis and metabolic perturbations, which may affect twin development. Notably, preterm birth exerts a stronger effect than genetic factors in shaping the gut microbiota of 12-month-old twins.