The potential of a native digestate microbial community for 1,4-dioxane (DX) biodegradation was evaluated under low dissolved oxygen (DO) concentrations (1-3 mg/L) under different conditions in terms of electron acceptors, co-substrates, co-contaminants and temperature. Complete DX biodegradation (detection limit of 0.01 mg/L) of initial 25 mg/L was achieved in 119 days under low DO concentrations, while complete biodegradation happened faster at 91 and 77 days, respectively in nitrate-amended and aerated conditions. In addition, conducting biodegradation at 30 ˚C showed that the time required for complete DX biodegradation in unamended flasks reduced from 119 days in ambient condition (20-25 °C) to 84 days. Oxalic acid, which is a common metabolite of DX biodegradation was identified in the flasks under different treatments including unamended, nitrate-amended and aerated conditions. Furthermore, transition of the microbial community was monitored during the DX biodegradation period. While the overall richness and diversity of the microbial community decreased, several families of known DX-degrading bacteria such as Pseudonocardiaceae, Xanthobacteraceae and Chitinophagaceae were able to maintain and grow in different electron-accepting conditions. The results suggested that DX biodegradation under low DO concentrations, where no external aeration was provided, is possible by the digestate microbial community, which can be helpful to the ongoing research for DX bioremediation and natural attenuation.