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Ultra low temperature cryopreservation method for sperm of large yellow crocker and cryopreservation device

A cryopreservation method and technology of large yellow croaker, which can be applied to household refrigeration equipment, human or animal body preservation, lighting and heating equipment, etc., and can solve the problems of poor applicability of cryopreservation, poor visibility of microscopic tubes, and poor quality of sperm preservation and other problems, to achieve good antifreeze effect, reasonable mixing time and low cost

Inactive Publication Date: 2009-08-12
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is suitable for cryopreservation of freshwater fish sperm. Due to the difference in physiological characteristics between seawater fish sperm and freshwater fish, the life span of seawater fish sperm is relatively short. This method is suitable for the freezing of seawater fish sperm such as large yellow croaker. Poor preservation applicability, such as low sperm survival rate, poor sperm preservation quality, and relatively low activation rate after cryopreservation
In addition, this method is to place the straw containing semen on the hollow foam board floating on the liquid nitrogen in the foam box. Since the foam board is covered by liquid nitrogen vapor, it is difficult to see clearly when putting the straw up, and the operation is not easy. It is convenient, delays the freezing time, and is prone to misoperations where straws are placed improperly and directly dropped into liquid nitrogen, and the hollow foam board needs to be placed in a foam box, which requires a relatively large volume of the foam box. For this reason, we researched and invented A better simple freezing method and freezing device for large yellow croaker

Method used

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  • Ultra low temperature cryopreservation method for sperm of large yellow crocker and cryopreservation device
  • Ultra low temperature cryopreservation method for sperm of large yellow crocker and cryopreservation device

Examples

Experimental program
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Effect test

Embodiment 1

[0024] like figure 1 The cryopreservation device for the cryopreservation of large yellow croaker sperm includes a foam box 1 with a cover 2, liquid nitrogen 3 is built in the foam box 1, the inner length of the foam box 1 is 16 cm, the width is 12 cm, and the depth is 18 cm , the wall thickness is 2.5 centimeters, on the two long walls 12 of the foam box 1, there are grooves 4 downward from the top 11 of the foam box 1, and the depth of the groove 4 downward from the top 11 of the foam box 1 0.5 cm, the straw 5 is placed in the groove 4, and the distance between the liquid level 31 of the liquid nitrogen 3 and the groove 4 is 2.5 to 3.5 cm.

Embodiment 2

[0026] The method for ultra-low temperature freezing of large yellow croaker sperm, the steps are as follows:

[0027] 1) Antifreeze preparation: Mix the diluent and antifreeze evenly according to the volume ratio of 86.7:13.3, prepare the antifreeze and store it at 4°C for later use. The components of the diluent are as follows:

[0028] NaCl: 7.25 grams KCl: 0.38 grams CaCl 2 : 0.18g

[0029] NaHCO 3 : 1.00 g MgSO 4 ·7H 2 O: 0.23 g NaH 2 PO 4 2H 2 O: 0.41 g

[0030] Glucose: 1.00 g, dissolved in distilled water and fixed to 1000 ml; antifreeze agent is DMSO.

[0031] 2) Sperm collection: Select male large yellow croakers with mature gonads, dry the body surface around the genital opening, lightly press the belly of the fish to obtain semen, put it in a petri dish or a small beaker, and put it on an ice bath. It is more ideal to use the sperm with stronger vitality and longer activity time to be used for cryopreservation.

[0032] 3) Preparation of the mixture of an...

Embodiment 3

[0036]The activation rate of the frozen sperm of the present invention is compared with the motility time, lifespan and relative fertilization rate of fresh sperm after activation, and the data shown in Table 1 are obtained, which shows that the activation rate of the frozen sperm of the present invention is higher. The quality is better, and the relative fertilization rate is also higher.

[0037] Table 1 Comparison table of activation rate, movement time after activation, lifespan and relative fertilization rate between frozen sperm and fresh sperm:

[0038]

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Abstract

The present invention discloses a method for freezing sperm of large yellow croaker. The method comprises the steps of preparing antifreezing liquid, collecting spectrum, preparing the mixture liquid of antifreezing liquid and spectrum, separately packaging the mixture liquid, freezing the mixture liquid, etc. The method for freezing sperm of large yellow croaker according to the invention has the advantages of convenient operation, high survival rate of sperm, better freezing storing quality of sperm and relatively higher activity ratio of sperm after freezing. The invention also discloses a freezing device of the method, wherein the freezing device comprises a foam box provided with a cover. Liquid oxygen is provided in the foam box. The inner depth of foam box is 18 centimeter and the wall thickness is 2.5 centimeter. Two long walls of foam box are symmetrically provided with grooves which extend downwards from the box top of foam box. The downwards depths of grooves from the box top of foam box is 0.5 centimeter. The distance from the liquid surface of liquid nitrogen to the groove is 2.5-3.5 centimeter. The device of the invention has the advantages of more convenient operation, prevention of the phenomenon of liquid nitrogen vapor enveloping after cover opening, saving to the freezing time, prevention the phenomenon of dropping into the liquid nitrogen caused by the rolling of pulled straws, etc.

Description

technical field [0001] The invention relates to an ultra-low temperature cryopreservation method for fish sperm, in particular to an ultra-low temperature cryopreservation method and a cryopreservation device for large yellow croaker sperm. Background technique [0002] Large yellow croaker is a valuable marine economic fish in my country and an important cultured species in the southeast coast. At present, the natural resources of large yellow croaker are scarce, and the germplasm of large yellow croaker is degraded to varying degrees, which affects the healthy and sustainable development of its aquaculture industry. Cryopreservation of fish sperm is one of the effective ways of germplasm preservation. It is of great significance to protect the germplasm resources by freezing the high-quality sperm of large fish at ultra-low temperature and establishing a sperm bank for long-term preservation. The cultivation of new varieties provides excellent germplasm materials. [0003...

Claims

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Application Information

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IPC IPC(8): A01N1/02F25D3/10
Inventor 竺俊全叶霆王春琳
Owner NINGBO UNIV
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