Abstract: Genetic difference between Cyathocotyle caecumalis sp. nov and Cyathocotyle orientalis Faust, Cyahaocotyle prussica Muhling, Cyataocotyle cornutus Rudolphi was analyzed in this study. Genic DNA of the 4 flukes was extracted, and the polymorphism and genetic distance between Cyathocotyle caecumalis sp. nov, and Cyathocotyle orientalis Faust, Cyahaocotyle prussica Muhling and Cyataocotyle cornutus Rudolphi were analyzed by RAPD (random amplified polymorphic DNA). RAPD showed: the genetic distance between Cyathocotyle caecumalis sp. nov and other flukes was respectively 0.4411, 0.4274, 0.5700. Obviously, the genetic differences between Cyathocotyle caecumalis sp. nov and other flukes were high. Based on genetics, this research discovered that Cyathocotyle caecumalis sp. nov which was a new parasite is different to Cyathocotyle orientalis Faust, Cyahaocotyle prussica Muhling and Cyataocotyle cornutus Rudolphi.