New three-stage in vitro model for infant colonic fermentation with immobilized fecal microbiota

The development and validation of a new three-stage culture system with immobilized fecal microbiota to simulate infant colonic ecosystem is described. Two continuous cultures with different fecal inocula were used to assess the validity and stability of the intestinal model. The total anaerobe populations measured in beads and effluent fermentations reached high concentrations similar to infant feces. Fluorescence in situ hybridization analyses and denaturing gradient gel electrophoresis profiles of effluent samples from the three reactors revealed complex patterns similar to that observed in the inoculum, indicating that fecal bacterial diversity was well-preserved and that dominant bacterial populations showed good stability among reactors. For both experiments, the bacterial populations and fermentation product concentrations were in the range of published data for infant feces. These results demonstrate that this new three-stage continuous culture with immobilized cells provides a useful tool for studying the infant colon ecosyste