This study was carried out to determine the possibility of bottle cultivation utilizing recycled oyster mushroom culture waste as a cultivating substrate for P. ostreatus. Total nitrogen percentage was 0.76%, 1.13%, 1.16%, 1.36%, and 1.38% in the 1-, 2-, 3-, 4-, and 5-time mixed substrate, respectively; 0.95%, 1.04%, 1.34%, 1.36%, and 1.25% in the 1-, 2-, 3-, 4-, and 5-time postharvest substrate, respectively; and 0.72% and 0.68% in the 2- and 3-time nonadditive substrate, respectively. Weight of the fresh fruiting body harvest was 115 g, 120 g, 117 g, 118 g, and 114 g on 1-, 2-, 3-, 4-, and 5-time mixed substrate, respectively; and 105 g and 45 g on 2- and 3-time nonadditive substrate, respectively. The first mixed substrate (fresh) and recycled substrates generated no significant difference in the weight of fresh fruiting bodies harvested.