Cryopreservation method of boar semen

A technology for cryopreservation and porcine semen, which is applied in the field of livestock breeding and cryopreservation of mammalian semen, to achieve the effect of improving the integrity rate of sperm acrosome, overcoming the cooling equilibrium time, and reducing the degree of oxidative damage

Pending Publication Date: 2022-06-07
SHANGHAI ACAD OF AGRI SCI +1
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  • Abstract
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Problems solved by technology

However, during the long cooling equilibrium process, sperm respiration and metabolism will produce a large number of reactive o...

Method used

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Embodiment 1

[0021] A method of cryopreservation of pig semen consisting of the following steps:

[0022] (1) Prepare frozen base fluid TCG, add 20-25% of the yolk to the frozen base solution, stir evenly, to get the frozen dilution I. liquid, add 6% glycerol and 0.4-0.6% OEP (Orvus Es Past) to the frozen dilution I. liquid, stir well, to get the frozen dilution II. solution;

[0023] The preparation method of the frozen base liquid TCG is: weigh glucose 1.1g, citric acid 1.48g, Tris 2.42g, penicillin sodium 0.06g and streptomycin sulfate 0.1g, dissolved in double distilled water, stirred to dissolve, volume to 100mL.

[0024] (2) Preparation of dilution BTS: weigh glucose 3.7g, trisodium citrate dihydrate 0.6g, EDTA 0.125g, sodium bicarbonate 0.125g, potassium chloride 0.075g, penicillin sodium 0.06g and streptomycin sulfate 0.1g, dissolved in double distilled water, and then set the volume to 100mL, adjust the pH to 7.2, osmolality of 310mosm / L;

[0025] The dilution BTS, cryo-dilution I. li...

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Abstract

The invention relates to a freezing preservation method of boar semen, which comprises the following steps: firstly preparing a freezing dilution liquid I, a freezing dilution liquid II and a dilution liquid BTS, then carrying out hydrotreating and semen collection on the three solutions, diluting the boar semen with the hydrotreated dilution liquid BTS at the constant temperature of 1: 1, balancing at 17 DEG C for 2-4 hours, and centrifuging to remove the supernatant, thereby obtaining the boar semen. The method comprises the following steps: diluting sperm precipitates by using an isothermal hydrotreated frozen dilution liquid I, then cooling and balancing to 4 DEG C, finally diluting by using an isothermal hydrotreated frozen dilution liquid II according to a ratio of 1: 1, sub-packaging the diluted sperm by using frozen tubules, sealing, freezing in a box body of a programmed freezing instrument, and after freezing is finished, storing the frozen sperm in a refrigerator. And transferring the frozen tubule into liquid nitrogen for storage. The method is simple to operate, can effectively improve the unfreezing activity and the sperm acrosome intact rate of the frozen pig sperms, can reduce the oxidative damage degree of the sperms, and overcomes the problem that the sperms are easily subjected to oxidative damage in a long cooling balance process.

Description

Technical field [0001] The present invention belongs to the field of livestock reproduction and mammalian semen cryopreservation technology, relates to a method of cryopreservation of pig semen. Background [0002] In recent years, the cryopreservation technology of pig semen has been greatly developed. Pig frozen semen (referred to as "pig frozen semen") uses deep-site sperm infusion to achieve the effect of preserving semen at room temperature, and after cattle, it has also begun to enter the process of commercial production and popularization of applications. [0003] Compared with other animals, pig sperm is more sensitive to low temperatures, so its freeze production requires a longer cooling equilibrium time. However, in the long cooling equilibrium process, sperm respiratory metabolism will produce a large number of reactive oxygen species (ROS), and the oxidative damage caused by ROS is the main reason for the sharp decline in the quality of pig frozen sperm. Therefore, h...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/0284
Inventor 张树山张德福戴建军吴彩凤孙玲伟徐皆欢韩雪峻李步社姜红菊
Owner SHANGHAI ACAD OF AGRI SCI
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