The composition of colonic mircoflora and its changes with maturation have rarely been investigated in large samples. Methods. We used conventional microbiological testing to analyse the colonic flora (Kyberstatus, Institut forMicroecology, Herborn, Germany) of stool samples from 12 484 children with different intestinal and nonintestinal diagnoses. Stool samples were analysed for total colony forming units (CFU) (per g stool) and the abundance of Bifidobacteria, Bacteroides sp., Escherichia coli, Enterococcus sp., and Lactobacillus sp. with respect to age, gender. A subset of 1089 infants was analysed for monthly changes within the first year of life. Results. Total CFU and individual microbial species were highest during the first year of life, decreased within the first 2 years, and then stabilized for the remaining childhood. In infants, the total CFU rose until month 5, declined with weaning, and peaked at 9–10 months. Significant effects of age, but not of gender, were found in Bacteroides sp. and Lactobacilli. However Bacterioids sp. and Lactobacilli increased with age, while Enterococci and E. coli decreased, and Bifidobacteria remained stable. Conclusion. Colonic microflora show both a bacteria-specific and general pattern of maturation which is most profound within the first year.