Veterinary medicines, including antibiotics, are utilized in large quantities in intensive livestock farming. It is evidenced that tylosin, one of the most frequently used antibiotics, is only partially metabolized in animals and not completely degraded in the manure storage stage before application to the farmland. In order to assess the mobility of tylosin in soil, a soil-column leaching study and a simple batch sorption experiment were conducted in the laboratory. Tylosin had strong sorption to various soils, with sorption distribution coefficients ranging from 42 to 65 ml/g. The range of concentrations in leachate was detected from non-detectable to 0.27 ng/mL after four simulated rainfall events in one month, and leachability of tylosin is dependent on soil properties and manure amendment. Percentage of clay, organic matter, cation exchange capacity, and manure amendment were positively correlated with sorption, and negatively correlated with mobility of tylosin in soil. The majority of tylosin was not recovered in the testing system, indicating that tylosin was most likely mineralized, or irreversibly bound to solid particles since no major degradation products were detected. Some trace level tylosin residues from manure-applied farmlands may be the major source to surface water systems through soil erosion and preferential flow processes.