A large-capacity, high-efficiency ultra-low temperature cryopreservation method for saddle grouper sperm

A technology of ultra-low temperature freezing of saddle grouper, which is applied in the field of marine biology, can solve the problems of restricting the development of saddle grouper breeding, unstable cultivation and production effects, immature production technology, etc., and achieve stable and repeatable protection effect The effect of good stability and accelerated storage speed

Active Publication Date: 2018-03-02
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

However, at present, the artificial breeding and seedling breeding of saddletail grouper in China is still in the experimental stage. Although some enterprises have made breakthroughs in artificial breeding technology, the production technology is not yet mature, and the production effect of seedling cultivation is unstable, which restricts the development of saddleband grouper. Development of grouper farming

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1) In August 2013, during the reproductive period of saddle grouper, in the broodstock workshop and broodstock breeding workshop of Jiaonan General Aquatic Products Co., Ltd., the male fish was removed from the breeding pond, placed on a sponge, and the genital pores were rinsed with distilled water three times. Wipe it with a paper towel, gently squeeze the abdomen from the back to the front to obtain 20ml of fresh semen from a male fish, store it in a clean centrifuge tube, keep it away from light, and check that the activity under the microscope is higher than 95%, put it in an ice box and return to the laboratory Carry out cryopreservation, place in ice box and carry out cryopreservation;

[0024] 2) Preparation of antifreeze solution, which is composed of diluent, propylene glycol, trehalose and egg yolk, pre-cooled in a refrigerator at 0°C after preparation, and ready for use; among them, the diluent is: NaCl 7.5g / L, MgSO 4 ·7H 2 O0.1g / L, CaCl 2 0.1g / L, Na 2 HP...

Embodiment 2

[0032] 1) In September 2013, the saddle grouper was in the breeding season. In the parent fish workshop of Jiaonan General Aquatic Products Co., Ltd., the male fish was fished out from the breeding pond, placed on a sponge, rinsed with distilled water for three times, wiped clean with a paper towel, and lightly rinsed with distilled water. Gently squeeze the abdomen from back to front to get fresh semen. A total of 2 saddle groupers were collected, with a total of 50ml of fresh semen, and the viability of the microscopic examination was 95%. They were placed in an ice box and returned to the laboratory for cryopreservation.

[0033] 2) Antifreeze preparation, which is composed of diluent, antifreeze, additives and distilled water. After preparation, it is pre-cooled in a refrigerator at 0°C for use; among them, the diluent is: NaCl 7.5g / L, MgSO 4 ·7H 2 O 0.1g / L, CaCl 2 0.1g / L, Na 2 HPO 4 2H 2 O0.1g / L, NaHCO 3 0.4g / L, antifreeze: 15% PG (propylene glycol); additives: treh...

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Abstract

The present invention belongs to the technical field of marine organism, and in particular relates to a giant grouper sperm large-capacity efficient ultra-low-temperature cryopreservation method. Giant grouper semen and an antifreeze liquid are mixed in the volume ratio of 2: 1-4: 1, and after one-step balancing and three-step cooling processing, the mixture is poured into liquid nitrogen (-196 DEG C), and sub-packed for storage; the antifreeze fluid comprises three parts of a diluent, an antifreeze and an additive; high-quality frozen semen obtained by the giant grouper sperm large-capacity efficient ultra-low-temperature cryopreservation method has great significance in the aspects of inter-species artificial hybridization, germplasm preservation, genetic diversity and the sustainable breeding and the like of giant grouper.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, in particular to a method for stabilizing and highly efficient cryopreservation of grouper spermatozoa. Background technique [0002] The saddle banded grouper (Epinephelus lanceolatus), also known as the grouper or gentian grouper, belongs to the order Perciformes, the family Anuridae, the grouper subfamily, the grouper genus, and is the largest fish in the grouper group , known as the giant grouper, known as the giant grouper in English. The saddle band grouper is mainly produced in Southeast Asia and Australia, and it is also distributed in the South China Sea. Saddle grouper is suitable for breeding in seawater ponds and seawater cages. It grows fast, has good meat quality and high edible value, and has a good breeding prospect. In recent years, China's Guangdong, Fujian, Hainan and other southern coastal areas have successively carried out artificial breeding of saddle grouper, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 刘清华肖志忠李军王学颖
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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