Abstract
In experimental infection with Streptococcus parauberis in Japanese flounder Paralichthys olivaceus, we investigated suitable routes of inoculation for a reproduction of natural infection: intraperitoneal injection, intravascular one in tail, intramuscular one into a lateral muscle or erecting and depressing muscles of dorsal fin, and subcutaneous one at a base of dorsal fin or outside preopercular bone. Among these routes, the subcutaneous injection resulted in the challenge-dose-dependent mortality. Disease and histopathological signs of the dead fish were similar to those of spontaneously affected fish. The subcutaneous injection at a base of dorsal fin or outside preopercular bone is a recommended method at a challenge of S. parauberis.