Ultra-low temperature cryopreservation method and cryopreservation device of large yellow croaker sperm

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: 2012-02-15
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 and cryopreservation device of large yellow croaker sperm
  • Ultra-low temperature cryopreservation method and cryopreservation device of large yellow croaker sperm
  • Ultra-low temperature cryopreservation method and cryopreservation device of large yellow croaker sperm

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Experimental program
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Effect test

Embodiment 1

[0024] Such as 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: 0.41g

[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 ant...

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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PUM

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Abstract

The invention discloses a method for cryopreserving large yellow croaker sperm: through steps such as antifreeze preparation, sperm collection, preparation of a mixture of antifreeze and semen, subpackaging of the mixture and freezing of the mixture, the method is a convenient operation , the sperm survival rate is higher, the preservation quality of the sperm is better, and the activation rate of the sperm after the cryopreservation is relatively high. The large yellow croaker sperm cryopreservation method; There is liquid nitrogen built in the foam box, the depth of the foam box is 18 cm, and the wall thickness is 2.5 cm. On the two long walls of the foam box, grooves are symmetrically arranged from the top of the foam box downwards, and the grooves are formed from the foam box. The depth of the top of the box is 0.5 cm, and the distance between the liquid level of liquid nitrogen and the groove is 2.5 to 3.5 cm; the device is easy to operate, avoids the phenomenon of liquid nitrogen vapor covering after opening the cover, and saves freezing time , It also avoids the advantages of the straw falling into the liquid nitrogen due to rolling.

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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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02F25D3/10
Inventor 竺俊全叶霆王春琳
Owner NINGBO UNIV
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