Efficient grouper sperm cryopreservation solution and use method thereof

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...

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 cryopreservation liquid of the present invention has extensive versatility, can be widely used in saddle band grouper, seven band grouper, moire grouper, brown spot grouper , brown grouper, money grouper and clear water grouper and other types of grouper sperm cryopreservation, improved grouper sperm cryopreservation survival rate; sperm cryopreservation solution of the present invention uses less toxic 1,2-propanediol (PG) greatly reduces the damage to sperm cell structure and genetic material, improves the survival rate and fertilization rate of frozen sperm, and can also reduce the damage to operators and the pollution to the breeding environment, reaching It provides an efficient technical method for the establishment of grouper sperm cryopreservation