Efficient grouper sperm cryopreservation solution and use method thereof

A cryopreservation liquid and grouper technology, which is applied to the preservation, application, and animal husbandry of human or animal bodies, can solve the problems of complicated preparation and preservation methods, difficult breeding technicians, and complex ingredients, so as to improve the survival rate and fertilization rate, wide versatility, and simple ingredients

Active Publication Date: 2017-09-22
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a high-efficiency grouper sperm cryopreservation solution and its use method, which solves the problems of poor versatility, complex components and low survival rate in the grouper sperm cryopreservation solution in the prior art
[0006] The grouper sperm cryopreservation solution of the present invention has the characteristics of simple ingredients, low toxicity, easy preparation and high freezing activity, and solves the problem that the existing grouper sperm cryopreservation solution has many components, high toxicity, complicated preparation and preservation methods, and problems in breeding The problem that technical personnel are difficult to grasp; Grouper sperm cryopreservatio

Method used

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Examples

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Effect test

Embodiment 1

[0030] A method for storing sperm with a high-efficiency grouper sperm cryopreservation solution of the present invention is specifically a method for cryopreserving moire grouper sperm, comprising the following steps:

[0031] 1) Preparation of diluent: Dissolve 10 g of glucose, 8 g of sodium chloride and 0.6 g of potassium chloride in distilled water, fully stir, dissolve, and constant volume to make a diluent with a volume of 1000 mL;

[0032] 2) Preparation of preservation solution: 1,2-propanediol was dissolved in the above diluent to prepare sperm cryopreservation solution, so that the volume concentration of 1,2-propanediol in the cryopreservation solution was 10%;

[0033] 3) Pre-cooling of the preservation solution: place the above-mentioned cryopreservation solution at 4°C for pre-cooling, and the pre-cooling time is 2 hours;

[0034] 4) Dilution of semen: take the semen of moire grouper, mix the semen with the preservation solution according to the volume ratio of 1...

Embodiment 2

[0039] A method for storing sperm with a high-efficiency grouper sperm cryopreservation solution of the present invention, specifically a method for cryopreserving sperm of saddletail grouper, comprising the following steps:

[0040] 1) Preparation of diluent: Take 15.5g of glucose, 7.5g of sodium chloride and 0.70g of potassium chloride and dissolve them in distilled water, fully stir, dissolve, and constant volume to make a diluent with a volume of 1000mL;

[0041] 2) Preparation of preservation solution: 1,2-propanediol was dissolved in the above diluent to prepare sperm cryopreservation solution, so that the volume concentration of 1,2-propanediol in the cryopreservation solution was 15%;

[0042] 3) Pre-cooling of the preservation solution: place the above-mentioned cryopreservation solution in a refrigerator at 4°C for pre-cooling, and the pre-cooling time is 2 hours;

[0043] 4) Dilution of semen: take the semen of the grouper, mix the semen with the preservation soluti...

Embodiment 3

[0048] A method for using a high-efficiency grouper sperm cryopreservation liquid to preserve sperm of the present invention, specifically a method for cryopreserving brown-spotted grouper sperm, comprising the following steps:

[0049] 1) Preparation of diluent: Dissolve 15 g of glucose, 8 g of sodium chloride and 0.65 g of potassium chloride in distilled water, fully stir, dissolve, and constant volume to make a diluent with a volume of 1000 mL;

[0050] 2) Preparation of preservation solution: 1,2-propanediol was dissolved in the above diluent to prepare sperm cryopreservation solution, so that the volume concentration of 1,2-propanediol in the cryopreservation solution was 15%;

[0051] 3) Pre-cooling of the preservation solution: place the above-mentioned cryopreservation solution in a refrigerator at 4°C for pre-cooling, and the pre-cooling time is 2 hours;

[0052] 4) Dilution of semen: take the semen of brown-spotted grouper, mix the semen and preservation solution acc...

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Abstract

The invention discloses an efficient grouper sperm cryopreservation solution and a use method thereof, and belongs to the technical field of fish germplasm cryopreservation. The efficient grouper sperm cryopreservation solution is prepared from diluent and an antifreeze agent; the diluent is an aqueous solution and is prepared from, by mass, 10-20 g/L of glucose, 7.5-8.5 g/L of sodium chloride and 0.6-0.7 g/L of potassium chloride; the antifreeze agent is 1,2-propylene glycol, and the volume concentration of the antifreeze agent in the cryopreservation solution is 10-20%. The use method of the cryopreservation solution for preserving grouper sperms comprises the seven steps of diluent preparation, antifreeze agent preparation, cryopreservation solution precooling, sperm dilution, balancing and subpackaging, cryopreservation, unfreezing and fertilization. The cryopreservation solution is simple in component, low in toxicity, easy to prepare, easy to master by breeding technicians and high in freezing activity, high universality is achieved, and the grouper sperm cryopreservation survival rate is increased.

Description

technical field [0001] The invention belongs to the technical field of fish germplasm preservation, in particular to a high-efficiency cryopreservation solution of grouper sperm and its application method. Background technique [0002] Grouper (Epinephelus) is an important economic fish and excellent germplasm resource for marine aquaculture in my country. There are 36 species of grouper in China, mainly distributed in the East China Sea and South China Sea. In 2015, the grouper fishing volume reached 117,593 tons, and the breeding volume reached 100,006 tons, which played an important role in the development of blue agriculture. Because grouper is a kind of hermaphroditic and pregynous fish, the number of males in the same ecological group is quite small, and it is difficult to obtain the sperm of male fish in artificial reproduction of grouper; on the other hand, with grouper With the rapid development of the industry, as well as the problems of many diseases and high def...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 田永胜唐江陈张帆李振通成美玲
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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