Biofilms in drinking water distribution systems (DWDSs) could cause several types of problems, such as the deterioration of water quality, corrosion of pipe walls, and potential proliferation of opportunistic pathogens. In this study, ten biofilm samples from different pipe materials, including ductile cast iron pipe (DCIP), gray cast iron pipe (GCIP), galvanized steel pipe (GSP), stainless steel clad pipe (SSCP), and polyvinyl chloride (PVC), were collected from an actual DWDS to investigate the effect of pipe material on bacterial community. Real-time quantitative polymerase chain reaction (qPCR) and culture-based method were used to quantify bacteria. 454 pyrosequencing was used for bacterial community analysis. The results showed that the numbers of total bacteria and culturable heterotrophic bacteria from iron pipes were higher than that in PVC, while the numbers of Shigella and vibrios were low in biofilms from iron pipes. Bacterial community analysis showed that Hyphomicrobium or Desulfovibrio were the predominant microorganism in iron pipes, whereas Sphingomonas or Pseudomonas were dominant in other types of pipe. This study revealed differences in bacterial communities in biofilms among different pipe materials, and the results were useful for pipeline material selection in DWDSs.