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Urechis unicinctus sperm cryopreservation liquid and preparation method thereof

A cryopreservation solution and a technique for the single-ringed beetle, which is applied in the biological field, can solve the problems of unsuitable single-ringed beetle sperm preservation and the like, and achieve the effects of low cost, simple operation and good effect.

Active Publication Date: 2017-05-10
HUAIHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the different biological characteristics of the sperm of different species of animals, the original technology is not suitable for the preservation of the sperm of Echinopsis monocircleum

Method used

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  • Urechis unicinctus sperm cryopreservation liquid and preparation method thereof
  • Urechis unicinctus sperm cryopreservation liquid and preparation method thereof
  • Urechis unicinctus sperm cryopreservation liquid and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Material collection: C. monoringensis has two reproductive periods in a year: spring (late April to end of May) and autumn (late September to end of October), mature sperm can be collected in this stage. Take the mature male body of C. monoringensis, wipe the body surface around the kidney hole with 75% ethanol, draw the semen from the kidney hole with a straw, put it in a sterilized and dried jar, and keep it at 4°C for later use.

[0041](2.1) Sperm motility identification method: use a straw to take 1 drop of semen on a glass slide, add 1 drop of seawater to make a smear, and observe under a microscope. The sperm activation rate was calculated as the percentage of motile sperm in the total sperm count, and the sperm activation rate of the samples was counted; each experiment was repeated 3 times, and the experimental data was expressed as "mean + standard deviation"; SPSS16.0 software was used for data processing, And one-way ANOVA (One-way ANOVA), LSD (Least Sig...

Embodiment 2

[0078] With reference to the freezing liquid preparation method of Example 1, complete the following preparation process:

[0079] (1) Material collection: From the middle and late April to the end of May, or from the end of September to the end of October, take the mature male body of Echinacea monoringensis, wipe the body surface around the kidney hole with 75% ethanol, suck the semen from the kidney hole with a straw, and place Store in a sterilized and dried jar at 4°C for later use;

[0080] (2) Sperm motility identification: Use a straw to take 1 drop of semen on a glass slide, add 1 drop of seawater to make a smear, and observe under a microscope. Sperm activation rate was calculated as the percentage of motile sperm count to total sperm count. When the activation rate of the spermatozoa of the single-circle spinach reaches more than 85% under the microscope, the following freezing steps are carried out;

[0081] (3) Sperm dilution: add 20 times the sperm preservative...

experiment example

[0087] Experimental example Freezing experiment of spermatozoa of the single-ringed clamweed.

[0088] Carry out cryopreservation according to the steps described in the cryopreservation method of the single-ring echinacea sperm described in Example 2; after 15 days, open the liquid nitrogen tank, pinch the long string exposed outside the liquid nitrogen tank, and take out the cryopreservation tube, Thawing and elution were carried out according to the method described in step (8) of Example 2, and the motility test was carried out according to the method described in step (2) of Example 2, and the detected sperm activation rate was 71%.

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Abstract

The invention relates to urechis unicinctus sperm cryopreservation liquid which comprises urechis unicinctus sperms, preservatives and antifreeze agents. A preparation method of the cryopreservation liquid includes the steps: collecting sperms; identifying sperm activity; diluting the sperms; reducing temperature, and balancing; firstly pre-cooling; secondly pre-cooling; preserving; unfreezing; eluting and the like. According to the cryopreservation liquid, the urechis unicinctus sperms can be effectively preserved for a long time and are preserved in liquid nitrogen, and activation rate of unfrozen sperms can reach 70% or more. According to the preparation method, expensive instruments are omitted, and the preparation method is simple to operate, low in cost and good in effect, provides sperm source guarantee for artificial insemination operations of urechis unicinctus and can be used for other research fields such as crossbreeding, germplasm selection, gynogenesis and species protection of the urechis unicinctus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cryopreservation solution for the sperm of Echinacea monoringensis and a preparation method thereof. Background technique [0002] In aquaculture production, with the development of artificial insemination technology and the need for germplasm resources protection, the research on sperm in vitro preservation technology of aquatic economic animals has attracted more and more attention. The in vitro preservation of sperm can solve the problem of asynchronous maturation between male and female, and also reduce the cost of raising a large number of parents. It provides a technical platform for research on artificial breeding, germplasm selection, sperm metabolism and physiology. The single-ringed sea cucumber is delicious, nutritious, and affordable. It is called "naked sea cucumber", and the market demand continues to increase. In recent years, it has also been found that...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 许星鸿
Owner HUAIHAI INST OF TECH
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