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To establish a protocol of estrus induction and synchronization in European mouflon, we performed artificial insemination using frozen-thawed semen and exogenous hormones. CIDR was inserted into vaginas of four mouflons for 16 days. A day before removal of CIDR, PG 600 was injected intramuscularly. PGF2α was injected when removing CIDR. Artificial insemination was cervically conducted with injecting LHRH 48 hours after CIDR withdrawal. Even though no pregnancy was confirmed, estrous signs were notified like open cervix, congestion of vaginal wall and discharge of cervical mucus. Further research in the wild sheep would be needed for development of artificial breeding methods and advancing sustainability of domestic zoos.