Bacterial community succession related to 1,4-dioxane exposure was investigated in two different activated sludge-inoculated reactors (municipal wastewater and dye industrial wastewater sludge), with or without additional carbon source, for 7 weeks. The denaturing gradient gel electrophoresis (DGGE) analysis revealed that microbial succession varied according to the inoculum sludge sources and the presence or absence of the extra carbon source. In the reactor inoculated with the municipal sludge, bacterial species belonging to α - and γ -Proteobacteria and Nitrospira class were dominant over time. On the other hand, bacterial species showing significant homology to β -Proteobacteria (e.g., Methylibium petroleiphilum PM1) and Actinobacteria class, who have been reported to have 1,4-dioxane degradation potential, were found in the industrial sludge-inoculated reactors. The appearance of these bacteria demonstrates that the microbial community structure of the inoculum and the presence of an extra carbon source affect the microbial succession in the system exposed to 1,4-dioxane.