Method for cryopreservation of sperms from anoplopoma fimbria

A technology for cryopreservation and spermatozoa, which is applied in the preservation, application, animal husbandry and other directions of human or animal body, can solve problems such as difficulty in sperm preservation, and achieve the effect of convenient and effective preservation work, promotion of breeding work, and improvement of preservation quality.

Inactive Publication Date: 2020-01-17
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a method for cryopreservation of the sperm of the snail, by freezing the sperm of the scorpion, and performing activation and resuscitation treatment to obtain high-quality sperm, which provides a basis for the further implementation of the breeding work of the scorpion Technical support, solve the problem of difficult sperm preservation in the breeding process of squid, and break through the bottleneck of artificial breeding of squid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] S1, targeting mature Scorpion fish, using artificial anesthesia to obtain sperm;

[0021] S2, prepare the sperm culture solution according to the sperm osmotic pressure of the squid; the formula of the culture solution is: sodium chloride 0.8g, sodium bicarbonate 0.06g, anhydrous calcium chloride 0.064g, glucose 1.4g, Tris base 0.04g, fetal bovine serum 0.4g, glutathione 0.12g.

[0022] S3, mixing the collected semen and the culture medium at a volume ratio of 1:2 to prepare a mixed solution, and then adding dimethyl sulfoxide with a volume of 10% of the mixed solution to prepare a sperm freezing and storage mixed solution;

[0023] S4, put the mixed sample of sperm cryopreservation mixture in a 4°C refrigerator for 15 minutes, then let it stand at the mouth of the liquid nitrogen tank for 5 minutes, then move it down, and place it above the liquid nitrogen level for another 5 minutes. Place it for another 5 minutes, then immerse it in liquid nitrogen and freeze it for...

Embodiment 2

[0026] S1, targeting mature Scorpion fish, using artificial anesthesia to obtain sperm;

[0027] S2, according to the sperm osmotic pressure of the naked squid, prepare a sperm culture solution; the formula of the culture solution is: sodium chloride 1.7g, sodium bicarbonate 0.07g, anhydrous calcium chloride 0.069g, glucose 1.7g, Tris base 0.06g, fetal bovine serum 0.5g, glutathione 0.13g.

[0028] S3, mixing the collected semen and the culture medium at a volume ratio of 1:2 to prepare a mixed solution, and then adding dimethyl sulfoxide with a volume of 10% of the mixed solution to prepare a sperm freezing and storage mixed solution;

[0029] S4, put the mixed sample of sperm cryopreservation mixture in a 4°C refrigerator for 15 minutes, then let it stand at the mouth of the liquid nitrogen tank for 5 minutes, then move it down, and place it above the liquid nitrogen level for another 5 minutes. Place it for another 5 minutes, then immerse it in liquid nitrogen and freeze i...

Embodiment 3

[0032] S1, targeting mature Scorpion fish, using artificial anesthesia to obtain sperm;

[0033] S2, according to the sperm osmotic pressure of the squid, prepare the sperm culture fluid; the formula of the culture fluid is: sodium chloride 1.4g, sodium bicarbonate 0.065g, anhydrous calcium chloride 0.067g, glucose 1.5g, Tris base 0.05g, fetal bovine serum 0.45g, glutathione 0.125g.

[0034] S3, mixing the collected semen and the culture medium at a volume ratio of 1:2 to prepare a mixed solution, and then adding dimethyl sulfoxide with a volume of 10% of the mixed solution to prepare a sperm freezing and storage mixed solution;

[0035] S4, put the mixed sample of sperm cryopreservation mixture in a 4°C refrigerator for 15 minutes, then let it stand at the mouth of the liquid nitrogen tank for 5 minutes, then move it down, and place it above the liquid nitrogen level for another 5 minutes. Place it for another 5 minutes, then immerse it in liquid nitrogen and freeze it for s...

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Abstract

The invention discloses a method for cryopreservation of sperms from anoplopoma fimbria. The method comprises steps of acquisition of sperms, preparation of a culture medium, a cryopreservation process of the sperms and resuscitation and activation processes of the sperms, namely acquiring sperms from anoplopoma fimbria at sexual maturity by using an artificial anesthesia method; according to thesperm osmotic pressure of the anoplopoma fimbria, preparing a sperm culture medium; mixing the acquired sperms and the culture medium according to a volume ratio of 1:2, and further adding dimethyl sulfoxide which accounts for 10% of the volume of the mixed liquid so as to obtain a cryopreservation mixed liquid; and putting a uniformly mixed sperm cryopreservation mixed liquid sample into a refrigerator of 4 DEG C for 15 minutes, allowing the sample to stand for 5 minutes at the opening of a liquid nitrogen bottle, moving the sample downwards, further allowing the sample to stand for 5 minutesabove the liquid level of liquid nitrogen, further allowing the sample to stand for 5 minutes on the liquid level of the liquid nitrogen, and immersing the sample into the liquid nitrogen, and performing cryopreservation. By adopting the method disclosed by the invention, the anoplopoma fimbria sperm preservation operation can be convenient, efficient and effective, the anoplopoma fimbria spermscan be prevented from rapid deactivation or death in vitro, the preservation quality of the sperms can be improved, the preservation time can be prolonged, and the anoplopoma fimbria sperms can be available at any time in actual production.

Description

technical field [0001] The invention relates to the field of biological sperm preservation, in particular to a fish sperm preservation method. Background technique [0002] The scorpion belongs to the ray-finned fishes, the scorpionidae, the black scorpionidae, and the scorpion genus, and is distributed along the coast of the North Pacific Ocean. As a deep-water bottom-dwelling economic fish, the naked scorpion has a large body, white and tender meat, and rich nutrition, which has extremely high economic value. In fact there are reports of commercial exploitation of sablefish as early as 1800. At present, Russia, Japan, Canada and the United States all regard the sablefish as an important fishery resource, and gradually carry out research on its basic biology. According to reports, the body length of the adult Scorpionfish is 50-70 cm, and the life span can reach more than 100 years (the known maximum is 114 years old). Its rapid growth time is 5-7 years. At this time, abo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 王伟罗珺张赛赛李梦瑶李雪洁成智丽
Owner DALIAN OCEAN UNIV
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