Development of a reliable experimental set-up for Dover sole larvae Solea solea L. and exploring the possibility of implementing this housing system in a gnotobiotic model.

Due to the increasing importance of the aquaculture sector, diversification in the number of cultured species imposes itself. Dover sole Solea solea L. is put forward as an important new aquaculture candidate due to its high market value and high flesh quality. However, as for many other fish species, sole production is hampered by amongst others high susceptibility to diseases and larval mortality, rendering the need for more research in this area. In this respect, in first instance, a housing system for Dover sole larvae was pinpointed by keeping the animals individually in 24-well plates for 26days with good survival rates and initiating metamorphosis. This ensures a standardised and reliable experimental set-up in which the possible death of one larva has no effect on the other larvae, rendering experiments adopting such a system more reproducible. In addition to proving valuable in many other applications, this multi well system constitutes a firm basis to enable the gnotobiotic rearing of larvae, which hitherto is non-existing for Dover sole. In this respect, secondly, a large number of disinfection protocols were tested, making use of widely employed disinfectants as hydrogen peroxide, glutaraldehyde and/or ozone whether or not combined with a mixture of antimicrobial agents for 24h. Although none of the tested protocols was sufficient to reproducibly generate a gnotobiotic model, the combination of glutaraldehyde and hydrogen peroxide resulted in hatchable, bacteria-free eggs in some cases.