Method for efficient ultralow temperature cryopreservation of turbot sperms

An ultra-low temperature freezing and turbot technology, which is applied in the field of marine biology, can solve the problems of restricting the yield of high-quality fertilized eggs and seedlings, low sperm production, short sperm production time of male turbot, etc., and achieves good repeatability and stability. , The effect of large storage capacity and easy operation

Inactive Publication Date: 2013-10-16
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology uses certain chemicals called sugar alcohol with strong antihypertensive properties when preserving DNA during cryogenic procedures such as freezing. These chemistries help keep blood vessels open without causing harmful side reactions like excessively inflamed lungs due to their ability to absorb water vapor. Additionally, this method involves adding ice particles into the vessel containing the sample before use, allowing them to dry faster over several hours compared to traditional methods where they are kept wet until ready lyophilized again. Overall, these technical improvements improve the quality control and efficiency of cryonuclear techniques involving cold temperatures.

Problems solved by technology

Technological Problem: This patented technical solution described by this patents relates to improving the efficiency at which seafoods produce their own seeds called turtles or frogs. However, current methods have limitations such as slow growing speed, poor quality control, difficulty accessing specific areas within the body, limited storage capacity, etc., making it difficult to maintain consistency across different generations without losing valuable genetic material. Therefore there remains room for improvement that could lead to more efficient use of these sources while ensuring stable yields of commercially useful products like salmon roger populations.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0020] 1) In April 2013, when the broodstock of turbot was in the reproductive period, the male broodstock was fished out from the breeding pond in the broodstock breeding workshop of Jiaonanzeng Breeding Station, Huangdao Island, and placed on a sponge. The reproductive pores were rinsed three times with distilled water, and wiped with paper towels. Clean, gently squeeze the abdomen from back to front to obtain about 30ml of fresh semen from more than ten male fish, store it in a clean centrifuge tube, keep away from light, and the viability of the microscope is higher than 90%, put it in the ice box and bring it back to the experiment room for cryopreservation;

[0021] 2) Antifreeze preparation, which is composed of diluent, antifreeze, additives and distilled water. After preparation, place it in a refrigerator at 0°C for pre-cooling and wait for use; among them, the diluent is: NaCl: 8g / L, KCl: 0.4g / L L, MgSO 4 ·7H 2 O: 0.1g / L, MgCl 2 ·6H 2 O: 0.1g / L, Na 2 HPO 4 2H ...

Embodiment 2

[0030]1) From April to May in 2012, when turbot was in the breeding season, the male fish was taken out of the breeding pond with the broodstock breeding workshop at Jiaonanzeng Breeding Station, Huangdao, and placed on a sponge. The genital pores were rinsed three times with distilled water, and wiped with paper towels. Clean, gently squeeze the abdomen from back to front to get fresh semen. A total of more than 50 tails of turbot were collected, with a total of 70ml of fresh essence, and the viability was 75-90% as detected by the microscope, and they were placed in the ice box and returned to the laboratory for cryopreservation;

[0031] 2) Antifreeze preparation, which is composed of diluent, antifreeze, additives and distilled water. After preparation, place it in a refrigerator at 0°C for pre-cooling and wait for use; among them, the diluent is: NaCl: 8g / L, KCl: 0.4g / L L, MgSO 4 ·7H 2 O: 0.1g / L, MgCl 2 ·6H 2 O: 0.1g / L, Na 2 HPO 4 2H 2 O:0.06g / L,KH 2 PO 4 :0.06g / ...

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Abstract

The invention belongs to the technical field of marine organisms, and concretely relates to a method for the efficient ultralow temperature cryopreservation of turbot sperms. The method comprises the following steps: mixing the seminal fluid of turbots with an antifreeze liquid according to a volume ratio of 3:1-4:1, carrying out two-step cooling treatment, carrying out one-step balancing, adding to liquid nitrogen (having a temperature of -196DEG C), and preserving; and the antifreeze liquid is composed of a diluent, an antifreeze agent and an additive. High-quality frozen sperms obtained through the method for the efficient ultralow temperature cryopreservation of turbot sperms are of great significance of the artificial breeding, germplasm preservation, heredity diversity and sustainable culturing of the turbots.

Description

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Claims

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Application Information

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Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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