Additive for freezing storage protective liquid of fish spermatid

A technology of sperm cells and additives, which is applied in the field of cryopreservation protection solution, can solve the problems of unreported patent applications, etc., and achieve the effects of increasing exercise time and lifespan, reducing freezing damage, and nourishing sperm

Inactive Publication Date: 2012-07-04
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are U.S. patents (US6054491 and US6410602) disclosing a method of feeding animals with astaxanthin to increase semen volume and enhance fertility, but the research of directly adding astaxanthin to fish sperm cryopreservation liquid as a protective additive is domestic There are no reports or related patent applications abroad

Method used

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  • Additive for freezing storage protective liquid of fish spermatid
  • Additive for freezing storage protective liquid of fish spermatid
  • Additive for freezing storage protective liquid of fish spermatid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1. Extract astaxanthin: Treat 50 g of Haematococcus pluvialis algae powder with methanol / potassium hydroxide mixture (5% KOH in 30% methanol) at 50°C for 15 minutes to remove chlorophyll, wash with distilled water 2 Twice, remove the supernatant and dry at 40°C. Grinding with liquid nitrogen for 0.5 min, repeated once, to fully break the wall. Add an appropriate amount of acetone, extract by shaking, and centrifuge to take the supernatant until the algae cells turn white to obtain a clear and deep red astaxanthin extract.

[0019] 2. Dissolve the extracted astaxanthin in the basic diluent containing 10% dimethyl sulfoxide, and make the concentration of astaxanthin 1.43×10 -6 -1.43×10 -4 mol / L fish sperm cell cryopreservation protection solution.

Embodiment 2

[0021] 1. Extract astaxanthin: Take the pre-treated but unbroken Haematococcus pluvialis algae powder and put it in the extraction kettle of the device. The extraction process conditions are: extraction pressure is 35MPa, extraction temperature is 80°C; separation kettle I pressure 6MPa, temperature 50; separation tank II pressure 6MPa, temperature 45; no entrainer added; extraction time 3h, to obtain astaxanthin oil.

[0022] 2. Dissolve the extracted astaxanthin in the basic diluent containing 10% dimethyl sulfoxide, and make the concentration of astaxanthin 1.43×10 -6 -1.43×10 -4 mol / L fish sperm cell cryopreservation protection solution.

Embodiment 3

[0024] Directly commercially available synthetic astaxanthin, dissolved in the basic diluent containing 10% dimethyl sulfoxide, and the concentration of astaxanthin was 1.43×10 -6 -1.43×10 -4 mol / L fish sperm cell cryopreservation protection solution.

[0025] The impact of adding the astaxanthin-added fish sperm cell cryopreservation protection solution of the present invention on the cryopreservation performance of fish sperm cells is compared and illustrated below by specific various experiments:

[0026] 1. The effect of astaxanthin on sperm motility of large yellow croaker

[0027] In this experimental example, 10% DMSO was selected as the antifreeze agent, and other groups were added different concentrations of astaxanthin on this basis. The results showed (Table 1) that the activation rate of fresh essence was close to 100%, that is, almost all of them were activated; the exercise time was (6.02±0.10) min, and the life span was (7.71±0.29) min. Due to freezing injury...

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Abstract

The invention relates to an additive for freezing storage protective liquid of fish spermatid. The additive is characterized by comprising astaxanthin with a structural formula shown as the specification, and the addition concentration is between 1.43*10<-6> and 1.43*10<-4> mol/L. By the additive, the freezing injury of sperms can be relieved in the low-temperature storage process, the sperms are nourished, and the vigor of the sperms is prolonged, so the additive has a great significance for the artificial culture of fish and germ plasm resource protection, wherein R1 and R2 are H or any fatty acids.

Description

technical field [0001] The invention relates to a cryopreservation solution, in particular to an additive for a fish sperm cell cryopreservation solution. Background technique [0002] In recent years, due to the combined impact of humans and nature, the quantity and quality of fish species have shown a clear downward trend, and germplasm has degraded, which affects the sustainable development of the fish resource industry. Cryopreservation of fish sperm is one of the effective ways to preserve germplasm. Freezing the high-quality sperm of rare and endangered fish at ultra-low temperature and establishing a sperm bank for long-term storage is of great significance to the research of genetic breeding, production and breeding, and the protection of germplasm resources. important meaning. [0003] Sperm cells are sensitive to low temperature. Freezing and thawing make sperm structure and function extremely vulnerable. The destruction of sperm cell structure and functional comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 严小军马建陈海敏竺俊全徐善良
Owner NINGBO UNIV
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