ABSTRACT To investigate the effects of aeration pretreatment on the anaerobic digestion (AD) of swine manure, five pretreatment groups were established with dissolved oxygen (DO) in each group set to 0.0, 0.4, 0.8, 1.4, and 2.0 mg/L, respectively. The results demonstrated that compared to the non-aeration group, methane production increased to varying degrees with different aeration pretreatments (AP), with a maximum increase of 27.98% (DO = 2.0 mg/L). AP reduced the hydrogen sulfide (H2S) content of biogas. The H2S concentration in the DO = 2.0 mg/L was only 0.209%, and this represented an increased H2S removal rate of 49.27% compared to that of the DO = 0.0 mg/L (0.412%). Simultaneously, AP increases the hydrolysis rate. When the DO concentration reached 2.0 mg/L, the hydrolysis rate reached its maximum. An increase in the hydrolysis rate further enhanced the removal rate of organic matter. The organic matter removal rate was highest (36.96%) at DO = 2.0 mg/L. AP effectively prolonged the methane generation time and shortened the lag time of methane generation. AP creates a brief micro aerobic environment, accelerates substrate hydrolysis, and promotes the production and consumption of total volatile fatty acids, particularly acetic acid. Additionally, AP promoted the symbiotic relationship between Caldicoprobacter (20.93%–34.96%) and Metanosaeta (14.73%–18.45%).