In situ examination of microbial populations in a model drinking water distribution system

A flow cell set-up was used as a model drinking water distribution system to analyze the in situ microbial population. Biofilm growth was followed by transmission light microscopy for 81 days and showed a biofilm consisting of microcolonies separated by a monolayer of cells. Protozoans (ciliates and flagellates) were often seen attached to the microcolonies. The biofilm was hybridized with oligonucleotide probes specific for all bacteria and the α- and β-subclass of Proteobacteria and visualized with a scanning confocal laser microscope. Hybridization showed that the microcolonies primarily consisted of a mixed population of α- and β-Proteobacteria. 65 strains from the inlet water and 20 from the biofilm were isolated on R₂A agar plates and sorted into groups with amplified rDNA restriction analysis. The 16S rDNA gene was sequenced for representatives of the abundant groups. A phylogenetic analysis revealed that the majority of the isolated strains from the bulk water and biofilm were affiliated to the family of Comamonadaceae in the β-lineage of Proteobacteria. The majority of the strains from the α-lineage were affiliated to the family of Sphingomonadaceae. We were unable to detect any strains from the Pseudomonas genus and found a low abundance of bacteria affiliated to the γ-subclass of Proteobacteria where Pseudomonas and E. coli are positioned. The analysis revealed a high bacterial diversity in the water phase as well as the biofilm, but no strains were found in both environments.