Miamiensis avidus, a causative agent of scuticociliatosis in olive flounder Paralichthys olivaceus, was previously reported to proliferate the fastest in media with an osmolarity of 300 to 500 mOsm kg(-1). This suggests that hyposaline conditions can promote the development of the disease. In the present study, olive flounder constantly showed high mortalities when they were experimentally challenged with the parasite by immersion and subsequently reared in hyposaline conditions. Furthermore, affected flounder produced by the challenge showed symptoms identical to those in naturally infected flounder. It was experimentally demonstrated that hyposaline conditions can be a key factor for the development and outbreak of scuticociliatosis in olive flounder.