A cryoprotectant and freezing method for ultra-low temperature storage of copepod eggs

A cryoprotectant, freezing method technology, applied in the application, preservation of human or animal body, animal husbandry, etc., can solve the problems of unsatisfactory, effect (low hatching rate, etc., to control osmotic pressure changes, improve hatchability and Developmental potential, the effect of reducing osmotic pressure changes

Active Publication Date: 2021-06-08
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ultra-low temperature freezing methods that can be retrieved at present have not achieved satisfactory results (low hatching rate) in the application of cryopreservation of fertilized copepod eggs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A cryoprotectant for ultra-low temperature storage of copepod eggs, comprising trehalose containing L-(-)-trehalose and seawater containing eggs. The presence of L-(-)-trehalose can promote the formation of hydrogen bonds between trehalose and protein and membrane lipids at the dehydration site of egg grains during freezing, and the hydroxyl groups in L-(-)-trehalose can also interact with Phosphate radicals in the membrane phospholipids combine to displace the water around the egg cells and release it outside the cell, which speeds up the discharge rate of intracellular water and reduces the volume of ice crystals formed in the cells, thus weakening the damage of ice crystals to the eggs , In addition, L-(-)-trehalose can also play a beneficial role with ethylene glycol, glycerin, and dimethyl sulfoxide, greatly reducing the freezing point of the solution, and ensuring that the egg grains have sufficient time to decompose into the cells. Appropriate exudation of water ...

Embodiment 2

[0032] A cryoprotectant for ultra-low temperature storage of copepod eggs, specifically comprising the following components and parts by weight: 100 parts of seawater containing eggs, 3 parts of ethylene glycol, 3 parts of glycerin, and 1 part of dimethyl sulfoxide , 2 parts of trehalose.

[0033] Among them, trehalose contains 0.22% L-(-)-trehalose; the density of eggs in the seawater containing eggs is 1000 eggs / mL, and the seawater is pretreated natural seawater with a salinity of 3.0%.

[0034] The pretreatment steps of the above seawater are: filter the natural seawater through a vacuum filter pump to remove floating sand and sundries, add cysteine ​​containing D-cysteine ​​and sodium ascorbate after sterilization, and continuously inflate during the process 30h, finally adjust the salinity of the seawater, the above-mentioned cysteine ​​contains 0.55% D-cysteine; the addition amount of cysteine ​​and sodium ascorbate is 0.29mg / L and 0.11mg / L respectively .

[0035] A f...

Embodiment 3

[0042] A cryoprotectant for ultra-low temperature storage of copepod eggs, specifically comprising the following components and parts by weight: 80 parts of seawater containing eggs, 1.2 parts of ethylene glycol, 2.1 parts of glycerin, and 0.6 parts of dimethyl sulfoxide , 1.2 parts of trehalose.

[0043] Among them, trehalose contains 0.21% L-(-)-trehalose; the density of eggs in seawater containing eggs is 950 eggs / mL, and the seawater is pretreated natural seawater with a salinity of 2.7%.

[0044] The pretreatment steps of the above seawater are: filter the natural seawater through a vacuum filter pump to remove floating sand and sundries, add cysteine ​​containing D-cysteine ​​and sodium ascorbate after sterilization, and continuously inflate during the process 24h, finally adjust the salinity of seawater, the above-mentioned cysteine ​​contains 0.4% D-cysteine; the addition amount of cysteine ​​and sodium ascorbate is 0.25mg / L and 0.07mg / L respectively .

[0045] A fre...

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PUM

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Abstract

The invention discloses a cryoprotectant and a freezing method for ultra-low temperature storage of copepod eggs. The cryoprotectant comprises trehalose containing L-(-)-trehalose and seawater containing eggs, mainly comprising the following components and parts by weight: 80-110 parts of seawater containing eggs, 1.2-3.8 parts of ethylene glycol , 2.1-4.3 parts of glycerin, 0.6-1.2 parts of dimethyl sulfoxide, and 1.2-2.5 parts of trehalose. The freezing method of the present invention uses the above-mentioned cryoprotectant to carry out multi-stage freezing. The beneficial effects are: the cryoprotectant of the present invention can play a gaining role, can effectively inhibit the formation of ice crystals in egg cells, weaken the damage degree of ice crystals to eggs, improve the hatching rate of copepod eggs, and improve the efficiency of frozen-thawed eggs. developmental potential; the freezing method of the present invention can cooperate with the application of the cryoprotectant to effectively control the osmotic pressure change of the eggs, avoid the formation of intracellular ice and ice, and reduce the ice crystal damage of the copepod eggs.

Description

technical field [0001] The invention relates to the technical field of ultra-temperature storage of eggs, in particular to a cryoprotectant and a freezing method for ultra-low temperature storage of copepod eggs. [0002] technical background [0003] In marine ecosystems, changes in zooplankton populations and productivity have a very important impact on the structure, function, and replenishment of biological resources of the entire marine ecosystem. Marine Copepoda, belonging to Arthropoda, Crustacea, and Copepoda, is a small crustacean with a body length of <3mm, living in planktonic, benthic or parasitic life, distributed in In the ocean, fresh water or brackish water, there are a large number and a wide variety. It is not only the main feeding object of middle and upper layer economic fish, but also one of the important opening baits for cultured fish and shrimp larvae. important in aquaculture. The change in the number of squats is not only directly related to the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 李继姬魏可叶莹莹
Owner ZHEJIANG OCEAN UNIV
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