Effects of time and duration of rearing with bottom sand on the occurrence and expansion of staining-type hypermelanosis in the Japanese flounder Paralichthys olivaceus

We previously reported that the progression of staining-type hypermelanosis spontaneously ceased at a specific time and area in Japanese flounder Paralichthys olivaceus. To examine whether time is a limiting factor in the spontaneous cessation of staining, we experimentally controlled the initiation and duration of staining by manipulating the bottom substrate condition in the fish tanks. At 151 days post hatching (DPH; 11 weeks), spontaneous cessation of staining was observed in fish reared in tanks without a sandy substrate. However, staining resumed (or was initiated) in tanks where sand was removed from 11 weeks, indicating a strong but temporary effect of bottom sand and the absence of time limitation in the staining progression by 151 DPH. Extended duration of the inhibitory period of hypermelanosis expansion (9 weeks or more) aided in only a 20 % reduction of the final staining area because of the increased rate of staining expansion. The bottom sandy substrate decreased the visibility of the staining area in individuals, but this was observed only before the completion of the staining expansion. These findings are discussed in relation to possible presence of area limitation of future staining, as well as the fundamental nature of staining.