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A kind of cryopreservation method of Litopenaeus vannamei embryo

A technology of cryopreservation and vannamei, which is applied in the field of cryopreservation of Litopenaeus vannamei embryos, can solve the problems of few, efficient and easy-to-operate cryopreservation methods for shrimp embryos, and achieves improvement of extraembryonic permeability, prevention of damage, reduction of Effects of toxic effects

Active Publication Date: 2021-08-06
GUANGXI ACADEMY OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the influence of factors such as the size and structure of Litopenaeus vannamei embryos, there are few reports on cryopreservation methods for Litopenaeus vannamei embryos that are efficient and easy to operate.

Method used

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  • A kind of cryopreservation method of Litopenaeus vannamei embryo

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Effect test

Embodiment 1

[0022] A method for cryopreservation of Litopenaeus vannamei body embryos, comprising the following steps:

[0023] a. Embryo Freezing: First put the embryos of Litopenaeus vannamei hatched for 8 hours directly into the antifreeze solution for 30 seconds, then immediately perform freezing treatment, put the embryos after antifreezing treatment into a thin tube and seal it, and place it at 4°C. Then gradually lower the temperature to -7°C at a rate of 2°C / min, plant ice after 4 minutes of osmotic equilibrium, and then perform 4 minutes of osmotic equilibrium, then gradually lower the temperature to -35°C at a rate of 0.4°C / min, and then put in liquid Nitrogen preservation; the described ice planting is to use the tweezers precooled in liquid nitrogen to clamp the thin tube for 10s; Seawater is seawater obtained after being sterilized by a conventional ultraviolet sterilizer;

[0024] b. Embryo thawing: Take out the thin tube from the liquid nitrogen, quickly immerse it in wate...

Embodiment 2

[0026] A method for cryopreservation of Litopenaeus vannamei body embryos, comprising the following steps:

[0027] a. Embryo freezing: first press seawater: antifreeze is 3: 1 volume ratio evenly mixes seawater and antifreeze to obtain mixed solution B, then soaks the embryos of Litopenaeus vannamei for 8h hatching with mixed solution B for 5min, Then soak in antifreeze solution for 30 seconds, and then freeze immediately. Put the antifreeze-treated embryos into a thin tube and seal it, then place it at 4°C, and then gradually lower the temperature to -7°C at a rate of 2°C / min. Plant ice after 4 minutes of osmotic balance, and then osmotic balance for 4 minutes, then gradually lower the temperature to -35°C at a rate of 0.4°C / min, and then put it into liquid nitrogen for storage; the described ice planting is used for precooling in liquid nitrogen The tweezers clamped the thin tube for 10s; the antifreeze solution included the following components in parts by volume: 36 parts...

Embodiment 3

[0030] A method for cryopreservation of Litopenaeus vannamei body embryos, comprising the following steps:

[0031] a. Embryo freezing: put the embryos of Litopenaeus vannamei hatched for 8 hours directly into the antifreeze solution for 60 seconds, then immediately freeze them, put the embryos after the antifreeze treatment into thin tubes and seal them, and put them directly into liquid nitrogen for storage ; The antifreeze solution mainly includes the following components in parts by volume: 30 parts of glycerin, 58 parts of seawater, and 6 parts of ethylene glycol; the described antifreeze solution also includes the components of the following concentration: bovine serum albumin 3mg / mL, polysucrose 10mg / mL, sucrose 0.5mol / L; the seawater is obtained after being sterilized by a conventional ultraviolet sterilizer;

[0032] b. Embryo thawing: Take out the thin tube from the liquid nitrogen, quickly immerse it in water at 30°C, wash the embryos into the petri dish immediatel...

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Abstract

The invention discloses a cryopreservation method for embryos of Litopenaeus vannamei, which includes the steps of freezing embryos and thawing embryos; in the freezing of embryos, the embryos of Litopenaeus vannamei hatched for 8 hours are antifreeze firstly. Then the embryos are frozen by slow freezing method or quick freezing method; the thawing of the embryos is to take out the thin tube from the liquid nitrogen, quickly immerse it in water at 30°C, and elute it immediately after the thin tube becomes transparent. Embryos are rinsed into Petri dishes and placed in incubation buckets to incubate. According to the characteristics of Litopenaeus vannamei, this method prepares suitable antifreeze solution and eluent, and optimizes the process of antifreeze treatment and thawing treatment. The long-term needs of shrimp farming and research, and simple operation, easy to popularize and apply.

Description

technical field [0001] The invention belongs to the technical field of aquaculture breeding, and in particular relates to a cryopreservation method for Litopenaeus vannamei embryos. Background technique [0002] Litopenaeus vannamei, also known as Penaeus vannamei, also known as white-legged shrimp and Penaeus vannamei, is native to the Pacific coast waters of Central and South America, and is suitable for breeding in tropical, subtropical, warm temperate and temperate seas. Litopenaeus vannamei is currently one of the three top varieties with the highest aquaculture output in the world. It has strong resistance to water environment, low nutritional requirements, fast growth speed, high meat content in shrimp body, long survival time out of water and Strong disease resistance and other characteristics. Due to the significant economic benefits of Litopenaeus vannamei farming, it has become the largest and most productive prawn species in the world. [0003] In the breeding ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
Inventor 陈秀荔杨春玲彭敏曾地刚朱威霖赵永贞李强勇刘青云陈晓汉林勇杨琼王卉李旻
Owner GUANGXI ACADEMY OF FISHERY SCI
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