In order to study the effect of the preparation process of swine manure bio-oil modified rubber asphalt binder on the rheological properties of the binder, the matrix asphalt and asphalt rubber were used as the control group, and swine manure bio-oil modified rubber asphalt binders were obtained by three preparation processes. Then the rheological properties of five binders were evaluated by rotational viscosity, DSR, MSCR and BBR tests. The results show that the addition of swine manure bio-oil to asphalt rubber is beneficial to the workability, but cause negative effect on its high temperature performance, fatigue performance and low temperature crack resistance. The preparation method of premixing bio-oil and rubber powder can reduce the degradation of high-temperature performance of asphalt rubber caused by addition of swine manure bio-oil to a certain extent. The fatigue properties of the three bio-binders are close, and the critical temperature is less than 25 degrees C. The process of premixing bio-oil and rubber powder will increase the creep stiffness and reduce the low-temperature performance. However, the low-temperature performance of the binder obtained by this process is still better than that of the matrix asphalt and can meet the requirements of use. Therefore, it is recommended to use the preparation process of premixing bio-oil and rubber powder to prepare swine manure bio-oil modified asphalt rubber. (c) 2021 Elsevier Ltd. All rights reserved.