Technique and process for frozen-preserving Chinese sturgeon semen under super-low temperature

An ultra-low temperature freezing, Chinese sturgeon technology, applied in the preservation of human or animal body, household refrigeration devices, lighting and heating equipment, etc., can solve the problem of not being able to meet the needs of sperm preservation, and achieve the effect of stable pH value

Inactive Publication Date: 2007-12-12
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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AI Technical Summary

Problems solved by technology

With the current technology, only 100 ml can be used for cryopreservation at most, wh

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Take the freezing 500 milliliters of Chinese sturgeon fresh essence as an example below to illustrate the specific operation process:

[0029] 1. Squeeze the abdomen of the mature male Chinese sturgeon and collect more than 500ml of fresh semen. During the collection process, the sperm should be absolutely avoided from contacting with water. The collected fresh semen should be put into the sealed container after oxygenation according to the amount of 100ml per bag. Put 5 bags in total in plastic bags, and then store them in a refrigerator with a temperature of 4°C;

[0030] 2. Microscopic examination under a microscope of 10×10 times, the computer-aided detection of fresh semen has an average liveness of 85%, the average curve velocity (VCL) of more than 160 microns / second, and the average linear velocity of more than 90 microns / second , it can be frozen when it reaches the standard;

[0031] 3. Preparation of Chinese sturgeon sperm dilution: 2000 ml dilu...

Embodiment 2

[0038] Embodiment 2: Take 1000 milliliters of frozen Chinese sturgeon as an example below, and 4000 milliliters of diluent is needed, and its configuration is as follows:

[0039] 20 ml of 1 mole of Tris-HCl, 30 ml of 1 mole of sucrose, 2 ml of 1 mole of potassium chloride, fixedly dissolved in distilled water to 1000 ml, adjusted to pH 8.5 with HCl, prepared 4 times, a total of 4000 ml was prepared, stored in a refrigerator at 4°C for later use ;

[0040] Other operating methods are the same as in Embodiment 1 above.

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PUM

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Abstract

Technology for bulk preserving spermatozoa of Chinese sturgeon through freezing at ultralow temperature comprises collecting and detecting fresh spermatozoa, diluting, balancing, adding antifreeze, subpackaging, program cooling, and preserving in liquid nitrogen container of -196deg.C. Each bulk can freeze 100 mL fresh spermatozoa. when the first bulk spermatozoa is in the procedure cooling step, after 5 minute, starting diluting the second bulk spermatozoa, and repeating above steps until all spermatozoa is preserved 8-12 h before its effective time. The diluent is Tris-HCl 20-30 millimole, sucrose 30-60 mol, potassium chloride 2-10 millimole, and distilled water 1000mL, and regulating pH to 8-8.5 with HCl. The antifreeze is methanol. The invention can efficiently carry out bulk preservation of Chinese sturgeon spermatozoa through freezing at ultralow temperature and long term ultralow temperature freezing preservation, and provide enough spermatozoa resource for artificial propagation of Chinese sturgeon.

Description

technical field [0001] The invention relates to a method for ultra-low temperature preservation of fish sperm, in particular to a method for ultra-low temperature preservation of Chinese sturgeon sperm. Background technique: [0002] With the development of fish seed artificial breeding and genetic breeding research, fish sperm cryopreservation technology has attracted people's attention. The first successful cryopreservation of fish sperm was in 1968. White eel essence, obtained a fertilization rate of 80-89%. In recent years, ultra-low temperature cryopreservation of fish sperm has been gradually extended to Cyprinidae, Perch, Catridae, Pisces and salmon trout. [0003] The Chinese sturgeon is one of the rare fish species and is a first-class protected animal in my country. It is a large freshwater fish unique to my country and is mainly distributed in the Yangtze River system and the Pearl River system. In recent years, due to the intensification of water pollution and ...

Claims

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Application Information

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IPC IPC(8): A61K35/52A01N1/00F25D3/10
Inventor 柳凌危起伟张洁明郭峰朱永久
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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