Boar semen refrigerating method and dedicated refrigerating fluid thereof
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A freezing method and pig semen technology, which is applied in the field of pig semen freezing, can solve the problems of frozen semen conception rate, low litter size, sensitive to oxidative attack, insemination and natural mating effect than fresh semen, etc., so as to reduce disease transmission , cost-saving effect
Active Publication Date: 2014-07-16
南宁壮博生物科技有限公司
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Polyunsaturated fatty acids are prone to lipid peroxidation under the action of reactive oxygen species (ROS), which reduces the number of polyunsaturated fatty acids in the plasma membrane. Since the plasma membrane of pig sperm contains a large amount of polyunsaturated fatty acids, it is extremely sensitive to oxidative attacks, which e...
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Embodiment 1
[0049] Embodiment 1, establishment of pig semen freezing method
[0050] In this example, systematic in-depth research and technical integration are carried out from various links such as pig semen collection, dilution ratio, standing cooling time after dilution, cooling equilibrium time, freezing procedure to dilution formula, in order to establish the best freezing method for pig semen.
[0051] 1. The influence of whole semen or concentrated semen and the corresponding dilution ratio on the quality of thawed sperm
[0052] 1. Tested semen from pigs
[0053] Whole semen and concentrated semen from boars with more than 90% vitality and liveness rate. Among them, all the semen ejaculated from the pig is collected, that is, the whole sperm. During the entire ejaculation process of the pig, only the part with higher sperm density in the middle section is taken, that is, the concentrated semen. The density of pig spermatozoa in concentrated semen is 6-1 billion / mL.
[0054] 2...
Embodiment 2
[0157] Practical application of the optimal freezing program for porcine semen established in embodiment 2 and embodiment 1
[0158] This embodiment utilizes the optimal freezing method for pig semen established in Example 1 (see step 4 of Example 1), and performs insemination and conception tests after thawing the frozen semen under farm conditions to detect the effectiveness of the pig semen freezing method of the present invention in production. practical application value.
[0159] The porcine semen frozen by the optimal porcine semen freezing procedure established in Example 1 was thawed according to Step 1-3 of Example 1, and 12 sows were successively deep inseminated, and the first 5 were deep insemination pre-experiments. After learning the deep insemination skills and points for attention, 7 gilts were formally inseminated. Insemination volume per sow is 80mL, sperm density: 0.82×10 8 Sperm count / mL (thawing liquid was used to adjust the sperm density, see Example 1...
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Abstract
The invention discloses a boar semen refrigerating method and dedicated refrigerating fluid thereof. The method comprises the following steps: (1) diluting dense boar semen according to a volume ratio of 1:(0.5-3), stewing at 26-30 DEG C for 25-40 minutes, and centrifuging 800g of boar semen for 10 minutes to obtain sperm precipitation; (2) adding refrigerating fluid I into the sperm precipitation according to a volume ratio of 1: (1-1.5), stewing at 17-20 DEG C for 1-2 hours, and stewing at 4-7 DEG C for 2 hours to obtain a system I; (3) adding refrigerating fluid II containing glycerin into the system I to obtain a system II with final glycerin concentration of 3%; (4) stewing the system II at 4 DEG C for 1-2 hours, and adding caffeine to obtain a system III with final caffeine concentration of 0.2g/L; and (5) stewing the system III at 4 DEG C for 0.5-2 hours, dropwise refrigerating at -120 DEG C, stewing and balancing for 5-10 minutes, and submerging the system III in liquid nitrogen for cryopreservation. After the boar semen refrigerated by the method is thawed, the sperm survival rate of the boar semen is higher than 0.45, and the in vitro survival time of the boar semen is longer than 6 hours, thereby reaching the production application level. The boar semen refrigerating method disclosed by the invention is used for providing scientific basis for the application and popularization of the refrigerated boar semen in production, and providing a technical method and standard for boar semen refrigerating.
Description
technical field [0001] The invention relates to a pig semen freezing method and a special freezing liquid thereof. Background technique [0002] With the popularization of pig artificial insemination technology, semen cryopreservation technology has an irreplaceable role in preserving high-quality germplasm resources, can prolong the service life of breeding pigs, and is not limited by factors such as the death, injury, and reproductive diseases of the boar; it can effectively Give full play to the role of high-quality boars and improve the utilization rate of high-quality breeding pigs; the introduction of high-quality boar semen can replace the introduction of live boars, reducing the cost of breeding. However, in the process of freezing pig semen, many internal and external factors will greatly affect the freezing effect. The internal factors mainly include: the internal characteristics of pig sperm, the differences between different pigs, and even different ejaculatory p...
Claims
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