Method of maintaining ovaries of marine fish, method of adjusting culture solution, and method of producing eggs or fertilized eggs of marine fish
Provided are a method of maintaining the ovaries of marine fish, and a method of obtaining fertilized egg. Provided is a method of maintaining ovaries in which the ovaries are removed from marine fish and the ovaries or fragmented ovaries are cultured in a culture solution. Also, provided is a method of obtaining fertilized eggs of marine fish, the method including allowing ovaries, which have been maintained after removal to ovulate and performing artificial insemination. Further, the present invention relates to a method of maintaining the fertilization capability of ovulated eggs of marine fish by using the culture solution. According to the present invention, fertilized eggs of marine fish can be easily obtained and seed can be efficiently produced.
1. A method of maintaining ovaries of marine fish, the method comprising:
removing ovaries from marine fish, wherein the ovaries comprise oocytes,
fragmenting the ovaries and conducting microplane distribution treatment immediately after the removing the ovaries from the marine fish, and
culturing the fragmented and microplane distribution treated ovaries in a culture solution,
wherein a pH of the cultured solution is adjusted to be from +0.3 to −0.3, relative to a. pH of an ovarian cavity fluid of the marine fish from which the ovaries have been removed.
2. The method according to claim 1 , wherein eggs contained in the ovaries are maintained in a fertilizable state.
3. The method according to claim 1 , wherein an osmotic pressure of the culture solution is from 100 mOsm/kg to 6000 mOsm/kg.
4. The method according to claim 1 , wherein the culture solution comprises at least one selected from the group consisting of a sodium ion, potassium ion, calcium ion, and magnesium ion.
5. The method according to claim 4 , wherein a sodium ion concentration of the culture solution is from 120 mM to 250 mM.
6. The method according to claim 4 , wherein a potassium ion concentration of the culture solution is from 5 mM to 10 mM.
7. The method according to claim 4 , wherein a calcium ion concentration of the culture solution is to 1.5 mM to 5.0 mM.
8. The method according to claim 4 , wherein a magnesium ion concentration of the culture solution is from 1.0 mM to 2.0 mM.
9. The method according to claim 1 , wherein the culture solution comprises at least one of selected from the group consisting of a chloride ion and sulfate ion.
10. The method according to claim 1 , wherein the culture solution comprises at least one selected from the group consisting of glucose, galactose, fructose, and sucrose.
11. The method according to claim 1 , wherein the culture solution comprises an antibiotic.
12. The method according to claim 1 , wherein the marine fish is a farmed fish.
13. The method according to claim 1 , wherein the marine fish is fish caught by fishing.
14. The method according to claim 1 , wherein the marine fish is a fish of the Scombridae family.
15. A method of producing eggs of marine fish in vitro, the method comprising obtaining eggs by causing ovaries cultured by the method of claim 1 to ovulate.
16. A method of producing fertilized eggs of marine fish, the method comprising cross-fertilizing eggs obtained by the method of claim 15 and sperm, thereby obtaining fertilized eggs.
17. Eggs of marine fish, the eggs being obtained by the method of claim 15 .
18. Fertilized eggs of marine fish, the eggs being obtained by the method of claim 16 .
19. A method of rearing marine fish, the method comprising rearing larvae hatched from fertilized eggs obtained by the method of claim 16 to young fry, immature fish, or adult fish.
20. A method of producing farmed marine fish, the method comprising rearing larvae hatched from fertilized eggs obtained by the method of claim 16 to young fry, immature fish, or adult fish.
21. The method according to claim 4 , wherein molar concentrations of sodium ions, potassium ions, calcium ions, and magnesium ions in the culture solution is from ¼ to 4 times of molar concentrations of sodium ions, potassium ions, calcium ions, and magnesium ions in blood serum of the marine fish from which the ovaries have been removed.
22. The method according to claim 4 , wherein a pH of the cultured solution is adjusted to be from +0.1 to −0.1, relative to a pH of an ovarian cavity fluid of the marine fish from which the ovaries have been removed.