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Buffalo seminal fluid cryopreservation method

A cryopreservation and semen technology, which is applied in the field of livestock breeding, can solve the problems of low sperm viability and achieve the effect of improving survival rate, high practicability, and simple steps

Active Publication Date: 2020-06-30
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production process of bovine frozen semen in China is implemented in accordance with the technical regulations for the production of bovine frozen semen of the People's Republic of China, but the recovery rate of frozen semen sperm is related to factors such as the quality of the original semen, the type of diluent, the freezing temperature and thawing temperature of the semen, among which , the temperature change of semen during the freezing process is one of the important factors affecting the quality of sperm freezing. At present, domestic cattle frozen semen production mostly adopts the freezing procedures provided by freezing equipment manufacturers. Basically, the same freezing procedure is used for various varieties of cattle. In the production of frozen semen, the sperm motility rate after thawing is relatively low

Method used

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  • Buffalo seminal fluid cryopreservation method
  • Buffalo seminal fluid cryopreservation method
  • Buffalo seminal fluid cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A cryopreservation method for buffalo semen, comprising the steps of:

[0021] (1) Collection of semen

[0022] Eight 2-6-year-old adult breeding male buffaloes (buffalo numbers: B18, A11, 3, E12, B12, 6, E11, Z2) were selected from Dongtan, Chongming Island, Shanghai, and the semen was collected using the false vagina method. Milky white semen, filtered through sterilized filter paper to remove jelly, and then perform quality inspection (reject sperm with more flocs, select semen with a sperm motility rate above 50%), and preheat the semen that meets the quality inspection to 35 ℃;

[0023] See Table 1 for the amount of semen collected:

[0024]

[0025] It can be seen from Table 1 that the volume of semen collected from the 8 adult bulls used in the test was 1.0-7.2 mL, and the volume changed greatly. The overall volume of semen collected on the second day was significantly better than that on the first day.

[0026] (2) Dilution of semen

[0027] Preheat the d...

Embodiment 2

[0039] The semen of the same batch of buffaloes in Example 1 was diluted and then frozen. The freezing program was changed to: start at 4°C, maintain 4°C for 30 minutes, then drop to -10°C at a rate of 5°C / min, and then start from -10°C Decrease to -140°C at a rate of 40°C / min, and finally keep at -140°C for 10 minutes to complete freezing. All the other are identical with embodiment 1.

[0040] The test results of sperm quality before and after freezing of buffalo are shown in Table 3:

[0041] Table 3 Example 2 Shanghai buffalo sperm quality before and after freezing

[0042]

[0043] Note: The same letter in the same column means no significant difference (P>0.05); different letters mean significant difference (P<0.05).

[0044] It can be seen from Table 3 that there are significant differences in the quality of frozen semen among the 8 adult buffaloes (P<0.05). The buffalo sperm numbered B18 had the highest survival rate and vitality after freezing and thawing, which ...

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Abstract

The invention relates to a buffalo seminal fluid cryopreservation method which comprises the steps of seminal fluid collection, dilution, freezing, preservation and the like, and specifically comprises the steps: firstly, collecting seminal fluid by adopting a pseudovaginal method, selecting milk white seminal fluid, filtering to remove jelly, and preheating the seminal fluid reaching the qualityinspection standard to 35 DEG C; preheating the diluent to 35 DEG C and then diluting the seminal fluid; cooling the diluted seminal fluid to 4 DEG C; and subpackaging into a 0.25 mL thin tube, and freezing after sealing, wherein the freezing procedure comprises the steps of starting from the temperature of 4 DEG C, maintaining the temperature for 30 min at 4 DEG C, then cooling to -10 DEG C at aspeed of 1-5 DEG C / min, then cooling to -140 DEG C at a speed of 30-40 DEG C / min from -10 DEG C, finally maintaining the temperature for 10 min at the temperature of -140 DEG C, completing freezing, and placing in liquid nitrogen and storing. The method is simple in step and high in practicability, and the thawing activity of the obtained frozen semen reaches up to 67.66% and greatly exceeds the collection requirements of national livestock gene banks.

Description

technical field [0001] The invention relates to a semen preservation method, in particular to a method for cryopreservation of buffalo semen, and belongs to the technical field of livestock reproduction. Background technique [0002] Shanghai buffalo are native to the suburbs of Shanghai such as Jiading, Baoshan, Fengxian and Chongming. Larger in size, cows weighing 700 to 900 kg and bulls over 1000 kg. The buffalo was originally an important meat and livestock in the southern rice region of my country. According to the "Shanghai Livestock and Poultry Breeds Record" in 1987, with the development of agricultural mechanization and rural urbanization, the value of buffalo has decreased. The number of buffalo in Shanghai has been raised since 1972. After 2000, the number of buffaloes decreased at a rate of 0.13 per year. By 1982, there were only 28,900 buffaloes left, and the number of fine-bred buffaloes dropped sharply. Shanghai buffalo was once reported to be on the verge of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0284
Inventor 孙玲伟戴建军张德福吴彩凤张树山
Owner SHANGHAI ACAD OF AGRI SCI
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