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Ultra-low temperature cryopreservation method of sea cucumber sperm

A technology of ultra-low temperature freezing and preservation method, which is applied in the field of ultra-low temperature freezing and preservation of sea cucumber sperm, can solve the problems of difficult application production, short sperm preservation time and high price, and achieves the effects of non-toxic cost, avoiding large loss of vitality and low cost.

Active Publication Date: 2021-09-24
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hepes buffer is not only expensive but also produces hydrogen peroxide with certain biological toxicity when exposed to visible light; the storage time of sperm is short (1 hour), and after thawing, the life span of sperm is only 1200s, which is difficult to apply in production

Method used

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Embodiment Construction

[0010] a. Male sea cucumbers cultivated by the Key Laboratory of Northern Seawater Enrichment and Aquaculture of the Department of Agriculture and Rural Affairs, Dalian Ocean University, with a body length of 20-25cm and a weight of 450-650g. During the breeding season in May 2019, take the sea cucumber out of the water and dry the surface of the sea cucumber (to avoid seawater to activate sperm), dissect the sea cucumber abdomen with sterilized scissors, avoid cutting the gonads, and pick out the complete glands with tweezers , use sterile filter paper to blot the body cavity fluid on the surface of the gonads, transfer them to a sterile petri dish, cut the gonads into pieces, draw them with a 5ml syringe, and wrap the injection port with a 300-mesh sieve (sterilized at high temperature and dried at 60°C) Carry out filtration, avoid body cavity fluid and seawater mixing during the filtration process, collect the filtered sperm into a 50ml centrifuge tube, and temporarily place...

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Abstract

The invention discloses a sea cucumber sperm ultra-low temperature cryopreservation method, which is carried out in sequence as follows: take a sea cucumber in the breeding period and dry the sea water of the sea cucumber body, dissect the abdomen, pick out the complete gland, suck the body fluid on the surface of the complete gland, and then move it to the Cut into pieces in a sterile petri dish, absorb the semen and filter it with a 300-mesh sieve; take the filtered semen and cryopreservation solution according to the volume ratio of 1:1-2 and pour them into the cryovial to mix evenly. Add 8-15g of glucose, 7-9g of sodium chloride, 0.5-0.7g of potassium chloride, 3-5g of trehalose, 0.05-0.1g of anhydrous calcium chloride and 80-120 mL of dimethyl Sulfoxide; suspend the cryovial at a distance of 10-15 cm from the surface of liquid nitrogen for 10 minutes, then suspend the cryovial on the surface of liquid nitrogen for 5 minutes, and finally immerse the cryovial in liquid nitrogen for cryopreservation.

Description

technical field [0001] The invention belongs to the technical field of sea cucumber cryobiology and ultra-low temperature cryopreservation of germplasm, and in particular relates to a method for ultra-low temperature cryopreservation of sea cucumber sperm. Background technique [0002] Sea cucumbers belong to the phylum Echinodermata, the subphylum Eleutherozea, and the class Holothuroidea. They are widely distributed in temperate and tropical oceans around the world, and are the most common marine higher invertebrates. Sea cucumber "is warm and nourishing in nature, sufficient for enemy ginseng" is rich in various nutritional elements and biologically active substances, and is a good nourishing product with the same source of medicine and food. As the market demand for sea cucumbers and the scale of the industry expand year by year, overharvesting of wild sea cucumbers is becoming more and more serious, resulting in a sharp decline in the amount of resources and germplasm q...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/021
Inventor 常亚青湛垚垚赵谭军宋坚张伟杰
Owner DALIAN OCEAN UNIV
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