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Method for ultralow temperature storage of Tilapia nilotica

A Nile tilapia cryopreservation technology, applied in the cryopreservation of Nile tilapia sperm, in the field of cryopreservation of Nile tilapia sperm, can solve the problem of different sperm membrane permeability and sperm preservation efficiency Low temperature, different temperature conduction speed and time

Inactive Publication Date: 2013-05-22
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the dilution ratio is too high, it will affect the total amount of sperm preservation, the efficiency of sperm preservation is very low, and it will also affect the later freezing and insemination
[0006] 3. Different fishes have different sperm membrane permeability
[0007] 4. Due to the different sizes and shapes of the sperm of different fishes, the conduction speed and time of temperature in the sperm are also different

Method used

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  • Method for ultralow temperature storage of Tilapia nilotica
  • Method for ultralow temperature storage of Tilapia nilotica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The following takes cryopreservation of 4ml of Nile tilapia semen as an example to illustrate the process of using the cryopreservation method of Nile tilapia sperm (-196°C) to preserve sperm. Below in conjunction with accompanying drawing, the present invention is described in further detail:

[0034] A method for cryopreservation of Nile tilapia sperm, the steps are:

[0035] 1. Preparation of sperm dilution mother solution:

[0036] (1) Prepare 1000ml of NaCl mother liquor with a concentration of 1M: Accurately weigh 58.4g of analytically pure NaCl, and distill it to 1000ml.

[0037] (2) Prepare 500ml of KCl mother liquor with a concentration of 1M: Accurately weigh 37.3g of analytically pure KCl, and dilute to 500ml with distilled water.

[0038] (3) Prepare 500ml of CaCl with a concentration of 0.5M 2 Mother liquor: Accurately weigh 55.5g of analytically pure CaCl 2 , distilled water to 500ml.

[0039] (4) Prepare 500ml of NaHCO with a concentration of 0.5M 3...

Embodiment 2

[0053] The following takes cryopreservation of 50ml of Nile tilapia semen as an example to illustrate the process of using the cryopreservation method of Nile tilapia sperm (-196°C) to preserve sperm.

[0054] A method for cryopreservation of Nile tilapia sperm, the steps are:

[0055] A. Preparation of sperm diluent mother liquor:

[0056] The preparation method of sperm diluent mother liquor is exactly the same as that of Example 1.

[0057] B. Preparation of 500ml sperm diluent:

[0058] (1) Accurately measure 63.75ml 1M NaCl mother solution; 375μl 1M KCl mother solution; 200μl 0.5M CaCl 2 Stock solution; 250 μl NaHCO 3 0.5M mother liquor in 500ml volumetric flask.

[0059] (2) After mixing the above solution, add distilled water to make the volume to 500ml.

[0060] (3) Store the prepared 500ml tilapia sperm dilution in a refrigerator at 4°C for future use.

[0061] C, cryopreservation of Nile tilapia sperm:

[0062] (1) Tilapia sperm collection: 10 to 15 Nile tila...

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Abstract

The invention discloses a method for ultralow temperature storage of Tilapia nilotica. The method comprises the following steps of: (1) collecting sperm of the Tilapia nilotica, to be specific, selecting male Tilapia nilotica with matured gonad development, extruding semen into a dry container and uniformly mixing; (2) detecting sperm activity, to be specific, carrying out microscopic examinationunder a microscope, and storing the sperm with the qualified microscopic examination into a refrigerator for later use; (3) diluting the sperm, to be the specific, precisely weighting the Tilapia nilotica semen to be frozen, adding sperm diluent, and uniformly mixing the semen and the diluent; (4) balancing the semen before freezing, to be specific, putting the diluted semen into the refrigeratorand carrying out balancing before freezing; (5) adding an anti-freezing agent and split-charging; (6) carrying out programmed cooling; and (7) feeding liquid nitrogen, to be specific, after cooling is completed, directly taking a straw with the semen from a programmed cooling instrument, putting into a liquid nitrogen tank and storing. The method is feasible and simple to operate, and the Tilapianilotica sperm is stored at the ultralow temperature, so that necessary materials and means are provided for breeding the Tilapia nilotica.

Description

technical field [0001] The invention relates to the technical fields of tilapia germplasm resource preservation and sperm cryobiology, and more specifically relates to a method for cryopreservation of Nile tilapia sperm, in particular to cryopreservation of Nile tilapia sperm. Background technique [0002] Tilapia is a small and medium-sized fish, known as one of the main sources of animal protein in the future. Originally produced in Africa, it was introduced to my country in the 1970s and has become an important aquaculture species in my country. With the selection and improvement of tilapia species in my country, many new species have been bred in addition to the originally introduced Nile tilapia (O.niloticus) and Onia tilapia (O.aureus). Such as: Oni tilapia, the hybrid offspring of Nile tilapia and Onia tilapia; Nile super male offspring selected through 3-line cross breeding; bred Gifu tilapia etc. [0003] In order to further carry out the breeding of new species ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
Inventor 柳凌邹桂伟张涛梁宏伟张洁明李忠郭峰
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI