Oil pollution is still a problem that is very difficult to control. To this end, this study investigates the application effect of microbial degradation technology in oil pollution control. First, the degradation microbial strains and substrates were determined according to the type of microorganisms. Then, ultraviolet spectrophotometry was used to determine the concentration of crude oil in the culture medium. After that, the authors designed the flow chart of crude oil standard curve, extracted organic phase, determined the residual oil composition in the culture medium, and calculated the degradation rate of mixed bacteria to crude oil. By studying the degradation mechanism of aerobic bacteria, anaerobic bacteria, and fungi, the degradation products were confirmed and the treatment of oil pollution on the water surface was carried out. It can be seen from the experimental results that the microbial degradation technology is effective in solving the problem of oil pollution.