Method for ultra-low temperature preservation and activation of sebastes schlegelii sperm of egg-viviparous fish

An ultra-low temperature preservation, ovoviviparous technology, which is applied in the field of marine organisms to achieve the effects of simple preservation and activation methods, easy operation and good stability

Active Publication Date: 2019-05-31
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, the research on sperm maturation, activation of in vivo fertilization and sperm preservation in Scorpius xu's is still blank

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Cryopreservation:

[0018] 1) In December 2017, 2 males of flounder flounder in mating period were collected in cages in Yantai, and their venous blood was obtained. The blood menstrual flow was obtained at 8000r / min, and the supernatant was obtained by centrifugation to obtain serum. At the same time, the semen of 0.5 ml, stored in a refrigerator at 4°C.

[0019] 2) Take 1 ul of the semen obtained above and mix it with 400 ul of the serum obtained above for activation, observe the motility rate of the sperm under a microscope, it is above 95%, and use it for cryopreservation experiments;

[0020] 3) Prepare the diluent, 0.7% NaCl and the serum obtained in the above step 1) are uniformly mixed in a volume ratio of 1:5 as the diluent, and then add DMSO to the diluent so that the final concentration of DMSO reaches 10% (volume ratio) , obtain antifreeze solution and store it in a refrigerator at 4°C;

[0021] 4) Mix the antifreeze solution and semen obtained above at a ...

Embodiment 2

[0025] 1) In December 2018, 4 gonad-developed males of Flounder xushii were purchased from the Qingdao market, and their venous blood was obtained. The blood menstrual flow was obtained at 8000r / min, and the supernatant was obtained by centrifugation to obtain serum, and at the same time, 0.8ml of semen during mating period was obtained. , were stored in 0-4 ℃ refrigerator.

[0026] 2) Take 1 ul of the semen obtained above and mix it with 400 ul of the serum obtained above for activation, observe the motility rate of the sperm under a microscope, it is above 95%, and use it for cryopreservation experiments;

[0027] 3) Prepare the diluent, 0.7% NaCl and the serum obtained in the above step 1) are uniformly mixed in a volume ratio of 1:5 as the diluent, and then add DMSO to the diluent so that the final concentration of DMSO reaches 12% (volume ratio) , obtain antifreeze solution and store it in a refrigerator at 4°C;

[0028] 4) Mix the antifreeze solution and semen obtained ...

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PUM

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Abstract

The invention belongs to the technical field of marine organism and particularly relates to a method for ultra-low temperature preservation and activation of sebastes schlegelii sperm of egg-viviparous fish. The method includes: evenly mixing sperm obtained in the mature period of male fish sperm of sebastes schlegelli with freezing liquid according to the proportion of 1:4-1:5, performing gradient cooling and freezing and then moving the mixture after freezing to liquid nitrogen to achieve long-term ultra-low temperature preservation of the sperm, wherein gradient cooling means that the mixture is pre-cooled for 2-5 minutes at the temperature of 0-4 DEG C, then frozen to the temperature of minus 80-100 DEG C at the speed of 10-12DEG C / min, and finally frozen to the temperature of minus 150DEG C at the speed of 15-18DEG C / min. With the method, new methods and ideals are provided for the cultivation of new marine fish species.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, in particular to a method for ultra-low temperature preservation and activation of the sperm of ovoviviparous fish flounder xushii. Background technique [0002] Sebastes schlegelii, Sebastes schlegelii (Hilgendorf, 1880), belongs to the family Scorpionidae of Scorpiomorpha, and Sebastes schlegelii is a warm-temperate demersal oviviparous fish, which is an important economically farmed fish species in northern my country. Xu's flat scorpionfish is a typical oviparous fish, fertilized in vivo, embryos develop in vivo to larvae and then produced outside the body. Xu's flat scorpionfish mates every autumn. At this time, the sperm of the male fish is mature, and the next spring is fertilized, and the embryo develops and fertilizes in vivo. Because of the special development, maturation and fertilization methods of the Scorpius spermatozoa, the sperm cryopreservation and activation methods a...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 刘清华李军王学颖徐世宏
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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