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Method for increasing sperm of verasper variegates and preserving sperm by freezing

A cryopreservation technology for flounder and flounder, applied in the preservation of human or animal bodies, medical raw materials derived from mammals, animal husbandry, etc., can solve problems such as difficult collection, decreased fertilization rate, and asynchronous sexual maturity, etc. Achieve the effects of reducing freezing damage, promoting normal development, and reducing toxic effects

Inactive Publication Date: 2007-09-19
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
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Problems solved by technology

[0003] Star flounder is cultivated indoors, and the sexual maturity of female and male broodstock is not synchronized; star flounder is not easy to spawn naturally indoors, and the ovulation cycle is unstable under artificial breeding, and the ovarian cavity is often overripe, and the fertilization rate drops significantly; The amount of semen in male flounder is low, generally around 0.7ml for natural mature semen, which is difficult to collect and cannot reach the amount of semen required for artificial reproduction
Due to the above reasons, the improvement of the artificial propagation technology of the star flounder is seriously restricted.

Method used

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Examples

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Embodiment

[0017] (1) Select a male fish with a body weight of 1 kg and be able to squeeze out milky white semen for hormone induction. The water temperature is 12°C, and the light is 14 hours a day.

[0018] (2) According to the volume ratio of semen and antifreeze solution 1:2, put the semen into the antifreeze solution with the volume content of DMSO10%+Gly5% and the balanced salt solution Hank's as the base liquid, and equilibrate it in a refrigerator at 0-4°C for 10 Take three 2ml cryopreservation tubes from the refrigerator, absorb 2ml of antifreeze solution each, and quickly put them into the programmed cooling device to start cooling.

[0019] (3) The cooling program is: 0~-80°C, the cooling rate is 20°C / min; -80°C balance for 2 minutes; -80~-180°C, the cooling rate is 20°C / min; when it drops to -180°C, Quickly put the cryovial into the liquid nitrogen tank for storage.

[0020] (4) After 4 months of storage, take out the cryopreservation tube, quickly put it into a 43°C water t...

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Abstract

The present invention relates to method of increasing sperm and freeze preserving sperm for fish, and is especially the method of increasing sperm and freeze preserving sperm of verasper variegates as marine fish. The method includes selecting male verasper variegates capable of extruding out milk white sperm, injecting HCG in the amount of 150 IU / kg to abdominal cavity to increase sperm, adding antifreeze fluid to the obtained sperm and pre-cooling at 0-4 deg c for 9-11 min, sectionally lowering the temperature to ¿C180 deg c and maintaining in liquid nitrogen. During defrosting, the frozen sperm is first set inside warm water at 43 deg c to slight thawing and then defrosted at room temperature to obtain sperm with fertilizing capacity. The present invention is significant in breeding verasper variegates as one kind of rare marine fishes, germ plasm maintenance, genetic diversity and other aspects.

Description

technical field [0001] The invention relates to increasing the sperm count of fish and cryopreserving the sperm, in particular to a method for increasing the sperm count and freezing the sperm of the marine fish star flounder, and injecting hormones into the male star flounder to lower the temperature of the sperm. Liquid nitrogen preservation and thawing, the process of obtaining sperm with fertilization ability. Background technique [0002] Star flounder is distributed in the Yellow Sea and Bohai Sea in the north of my country, in the south of central Japan and in the coastal waters of Southwest Korea. Star flounder is larger in size, faster in growth, less in bony spurs, smaller in viscera, and more edible. It is an excellent breeding species. Due to the scarcity of natural resources, the catch of star flounder has not formed a large scale, and it is a native natural species. The sustainable use of resources and the development of its breeding have broad prospects. At ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02A61K35/12
Inventor 肖志忠丁福红李军
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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