Clostridium butyricum and Bifidobacterium supplementation ameliorates gut dysbiosis and cognitive-anxiety behaviors in preterm mice

Preterm infants often exhibit gut dysbiosis and neurodevelopmental impairments, with evidence suggesting a crucial role of the microbiota-gut-brain axis in early development. To investigate whether probiotic intervention could improve these outcomes, we examined the effects of Clostridium butyricum and Bifidobacterium (LCBBCP) on gut microbiota diversity, spatial memory, and anxiety in preterm mice. We established a preterm mouse model divided into three groups: preterm experimental, preterm control, and full-term control (n=21 each). The preterm experimental group received LCBBCP oral gavage, while controls received saline. Gut microbiota analysis through 16S rRNA gene sequencing revealed higher richness and evenness at P21 than P14 across all groups, with preterm controls showing significantly lower microbial diversity at P14 compared to full-term controls. The preterm control group exhibited elevated levels of Enterobacteriaceae, Ruminococcaceae, Streptococcaceae, and Gemellaceae at P14, while Lactobacillaceae levels were higher in full-term control and preterm experimental groups at both timepoints. Behavioral tests showed that preterm experimental mice performed better than preterm controls in spatial memory tasks and displayed reduced anxiety-like behaviors, although not reaching full-term control levels. The preterm control group showed significantly reduced movement, slower speed, and increased resting time in open field tests. These findings demonstrate that LCBBCP intervention enhanced microbial diversity, improved spatial learning, and alleviated anxiety-like behaviors in preterm mice, likely through reducing Enterobacteriaceae and promoting Lactobacillaceae, suggesting a potential therapeutic strategy for preterm-related complications.