Abstract
Artificial reproduction and subsequent improved culture of the Japanese eel Anguilla japonica have not yet been achieved, but are expected in the near future. To allow the development of aquaculture activities, chromosome manipulation techniques are necessary for the genetic improvement of the eel. In the present paper suitable conditions were determined for heat shock treatment for the induction of triploidy. This was done using fertilized eggs obtained from gametes of cultured eels that were induced to mature by repeated injections of salmon pituitary extract, followed by an injection of 17α, 20β-dihydroxy-4-pregnen-3-one to induce ovulation in the female eels and by repeated injections of human chorionic gonadotropin in the male eels. Three variables (i.e. timing after fertilization, temperature, and duration of the shock) were varied to maximize the yield of triploid larvae by examining hatching and triploid rates of treated groups. Ploidy was determined by DNA content flow cytometry and/or chromosome counting. The triploid yield was the highest (70%), when a 37°C heat shock was applied for 3 min, 10 min after fertilization. Under such conditions the rate of triploidy reached 70-100% and the rate of hatching was 5.2-24.6% when the control hatching rate was 28.5% on average.
