During the process of anaerobic digestion, hydrolysis of substrate is the key factor determining methane production efficiency. The hydrolysis efficiency are directly affected by biodegradability of substrate. In this study, three types of pre-treatment methods were tested to enhance biodegradability effectiveness. By the application of blender and the Cavitation treatment, the SCODcr increased from 24,723mg/l to 24.726~29,175mg/l and to 25,589~26,456mg/l, respectively. In case of ozone treatment, the SCODcr decreased from 24,723mg/l to 22.540~ 23,988mg/l. In batch experiment, there was a little improvement of the biochemical methane potential(BMP) by the blender and the cavitation treatment. In contrast, the BMP somewhat decreased by ozone treatment. This result shows that higher anaerobic digestion efficiency of livestock manure could be obtained through pre-treatment of substrate.