This study examined the preparation, characterization, and antibacterial activity of cellulose-acetanilide. Cellulose-acetanilide ethers were prepared by homogeneous etherification of microcrystalline or kraft cellulose with 2-Chloroacetanilide in LiCl/DMAc (8%) at 60 degrees C for 24 h under mild reaction conditions. The 2-Chloroacetanilide could highly efficiently react with cellulose to synthesize cellulose-acetanilide ethers with controllable degree of substitution (DS). DS was in a range of 0.2-1.5 adjusted by the molar ratio of 2-Chloroacetanilide to anhydroglucose unit. The DS of microcrystalline cellulose is significantly higher than that of the kraft cellulose. The chemical structure and properties of the obtained products were characterized by FT-IR, NMR, and SEM. The antibacterial activity of the synthesized compounds were screened against the representative bacterium Rhodococcus sp. GK1, by the agar well diffusion method, and compared with commercials antibiotic such as Cephalothin, Ampicillin, Amoxicillin, Chloramphenicol, Kanamycin and Doxycycline. It was found that antibacterial experiment revealed excellent antibacterial activity of the cellulose-acetanilide ethers.