Livestock semen freezing diluent as well as preparation method and application thereof

A technology for freezing diluents and semen, which is applied in applications, animal husbandry, and preservation of human or animal bodies. It can solve problems such as high prices and differences, and achieve the effects of low cost, reduced mechanical damage, and convenient sources.

Active Publication Date: 2013-04-03
YUNNAN ANIMAL SCI & VETERINARY INST +1
View PDF7 Cites 24 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of antifreeze proteins also has certain limitations
First of all, as a heterologous protein, antifreeze protein may cause certain immune responses in animals; secondly, the main source of antifreeze protein is extracted from the blood of polar marine fish, and these technolog

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The semen of Yunnan semi-fine wool sheep was collected by electrical stimulation, and then the semen quality was tested. The indicators of sheep semen used for cryopreservation must meet: the motility is greater than 70%, and the sperm density is higher than 3×10 9 / ml, the amount of semen is 0.75-2ml.

[0020] Prepare semen frozen diluent, the steps are, respectively weigh 2.71g of Tris, 1.4g of citric acid, 1.0g of glucose, 5ml of glycerin, 100,000 IU of penicillin, 100,000 IU of streptomycin, 1,3- Cyclohexanediol 1.16g, dissolve in ultrapure water, stir well, use Tris solution to adjust the pH value to 6.8-7.2, add 10ml of fresh egg yolk inactivated at 56°C for 30min before use, mix well and dilute to 100ml, Centrifuge at 15,000 rpm for 1 hour at 4°C, and filter the supernatant with a 0.45 μm Millipore filter for later use.

[0021] Mix the sheep semen and frozen diluent that meet the freezing requirements evenly in a ratio of 1:4, and then divide them into 0.25ml ...

Embodiment 2

[0024] The breeding base of Kunming Yixingheng Animal Husbandry Technology Co., Ltd. uses a fake vagina to collect the semen of Yunling black goats, and then conducts semen quality testing. The indicators of sheep semen used for cryopreservation must meet: the motility is greater than 70%, and the sperm density is higher than 3×10 9 / ml, the amount of semen is 0.75-2ml.

[0025] Prepare semen frozen diluent, the steps are, respectively weigh 2.71g of Tris, 1.4g of citric acid, 1.0g of fructose, 5ml of glycerin, 100,000 IU of penicillin, 100,000 IU of streptomycin, 1,4- Cyclohexanediol 0.697g, dissolved in ultrapure water, stirred evenly, adjusted the pH value to 6.8-7.2 with Tris solution, added 20ml of fresh egg yolk inactivated at 56°C for 30min before use, mixed evenly and adjusted to 100ml, in Centrifuge at 15,000 rpm for 1 hour at 4°C, and filter the supernatant with a 0.45 μm Millipore filter for later use.

[0026] Mix the sheep semen and frozen diluent that meet the ...

Embodiment 3

[0029] In the breeding base of Kunming Yixingheng Animal Husbandry Technology Co., Ltd., the semen of hornless Dorset rams is collected by electrical stimulation, and then the semen quality is tested immediately. The indicators of sheep semen used for cryopreservation must meet: the motility is greater than 70%, and the sperm density is higher than 3×10 9 / ml, the amount of semen is 0.75-2ml.

[0030] Prepare a frozen diluent that does not contain an osmotic protectant. The preparation method is to weigh 2.71 g of Tris, 1.4 g of citric acid, 1.0 g of glucose, 100,000 IU of penicillin, 100,000 IU of streptomycin, and 1,3,5-Cyclohexanehexanol 1.25g, dissolve in ultrapure water, stir evenly, use Tris solution to adjust the pH value to 6.8-7.2, add 15ml of fresh egg yolk inactivated at 56°C for 30min before use, mix well Dilute to 100ml, centrifuge at 15,000 rpm for 1 hour at 4°C, and filter the supernatant with a 0.45 μm Millipore filter for later use.

[0031] Prepare the froz...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a livestock semen freezing diluent as well as a preparation method and application of the livestock semen freezing diluent. The preparation method of the livestock semen freezing diluent comprises the following steps of: adding 2.71 grams of trihydroxymethyl aminomethane, 1,4 grams of citric acids, 1.0 gram of monosaccharides, 10 myriad IU (International Unit) of penicillium, 10 myriad IU of streptomycin and 0.29-6.97 grams of inositol compounds to a right amount of ultrapure water; optionally adding 5-20 milliliters of penetrability protectants; uniformly stirring; regulating a pH value to 6.8-7.2; adding 5-20 milliliters of fresh yolk subjected to inactivation treatment; uniformly mixing, and then fixing the volume to 100 milliliters; centrifugalizing at low temperature for 1 hour; and filtering supernate to prepare the livestock semen freezing diluent. Because the inositol compounds can effectively inhibit the generation of ice crystals, the livestock semen freezing diluent disclosed by the invention reduces the mechanical injury caused by the ice crystals on semens, achieves the survival rate of the defrozen semens at about 75%, the activity more than 50%, the acrosome integrality at about 60% and the plasmalemma integrality at about 50% and achieves the non-return rate more than 70% after artificial insemination; and the livestock semen freezing diluent has the advantages of no immune response generation, purity guarantee, low price, and the like by using the inositol compounds as the ice crystal inhibitors of the semen freezing diluent.

Description

technical field [0001] The invention relates to the field of animal reproductive physiology and reproductive technology, in particular to a frozen dilution of livestock semen and its preparation method and application. Background technique [0002] Although the research on cryopreservation of livestock semen has been carried out for more than 50 years, the viability of thawed semen is only about 30%-50%, and the non-return rate and litter rate after artificial insemination are significantly lower than that of fresh and low-temperature frozen semen. During the freezing process of livestock semen, the mechanical damage caused by ice crystals formed inside and outside the cells directly leads to the decline of sperm motility after freezing. How to reduce the generation of ice crystals and reduce the damage of ice crystals to livestock sperm has always been a research topic in livestock breeding technology and cryobiology. research hotspots in the field. By changing the composi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A01N1/02
Inventor 洪琼花权国波杨红远李东江吕春荣
Owner YUNNAN ANIMAL SCI & VETERINARY INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap