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The addition of biochar is a promising strategy to improve methane production via strengthening direct interspecies electron transfer (DIET) in anaerobic digestion (AD) field. In this study, a novel oil sludge biochar (OSBC) prepared at 500/600/700 °C was used to investigate its dosage effects on performances of sewage sludge AD. The characterization results indicated that OSBC prepared at 600 °C had highest electron accepting and donating capacity, meanwhile, it also contained moderate functional groups such as C O, which were in favor of methane production. Therefore, the highest accumulative methane yield (143.96 mL (g VS)−1) accompanying with the fast consumption of volatile fatty acids (VFAs) was achieved at dosage of 1.2 g. Additionally, the conductivity and enzyme activity (electron transport system activity and coenzyme F420) of AD with OSBC addition were also enhanced. Microbial community structures manifested that the potential syntrophic microbes including archaea Methanothrix , Methanospirillum , and bacteria Comamonas , Petrimonas and Syntrophomonas were enriched by OSBC due to its large surface area and microelement such as iron, and contributed to the improved methane production. The results suggested that OSBC exhibited excellent practical application potential in enhancing anaerobic sewage sludge treatment and biogas productivity. [Display omitted] • Oil sludge biochar prepared at 600 °C had highest electron accept/donate capacity. • Functional groups and iron of biochar were responsible for increasing CH 4 yield. • Oil sludge biochar improved the conductivity and enzyme activity of digesters. • IHT and DIET processes were promoted by oil sludge biochar from microbial perspective. [ABSTRACT FROM AUTHOR]