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Method for cryopreservation of semen of siniperca scherzeri and artificial insemination

A technology of cryopreservation and semen, applied in the preservation of human or animal body, fish farming, medical science, etc., can solve the problems of small amount of mandarin fish, not paying attention to sperm-egg ratio, small body size, etc., to promote the diversity of species. The effect of synthesizing and optimizing the combination, solving the asynchrony of gonadal development, and improving the utilization rate of sperm

Active Publication Date: 2011-06-01
生生农业集团股份有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The ultra-low temperature cryopreservation method of fish semen has basically matured, but there are obvious differences among species; the storage time of spotted mandarin fish semen is very short at room temperature, and its vitality will decrease significantly over time; compared with mandarin fish, the spotted mandarin fish The amount of semen is small, and the size is small. The amount of semen that each fish can provide is usually less than 1 ml, and the stress in the reproduction process is high. How to efficiently use the semen of the mandarin mandarin fish is a key issue.
In the general reproduction process, the sperm-to-egg ratio is often not paid attention to, or the sperm-to-egg ratio cannot be flexibly adjusted according to the amount of semen or eggs, so as to improve the fertilization rate and reduce unnecessary waste of resources

Method used

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  • Method for cryopreservation of semen of siniperca scherzeri and artificial insemination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The following takes about 2 ml of semen of two-tailed mandarin mandarin fish in frozen storage as an example to illustrate the specific operation process:

[0027] (1) Semen collection: select two tails of the broodstock with good maturity; dry the abdomen of the fish body with a towel; gently squeeze the abdomen, extrude a small amount of semen, wipe it off with a dry towel or toilet paper and clean the genital opening; Gently squeeze the abdomen to squeeze out the semen without urine and feces contamination and collect it in two 2ml centrifuge tubes, numbered B-1 and B-2 respectively. The collected semen was temporarily stored on ice.

[0028] (2) Semen quality inspection: Normal semen is milky white when observed with the naked eye; put a drop of water on the glass slide, and prepare a cover glass, pick a little with a toothpick and immerse in the prepared clear water to activate, cover the slide immediately and use an ordinary microscope, Observed at 10×40 times, r...

Embodiment 2

[0033] The following is an example of thawing a tube of cryopreserved semen for insemination to illustrate the specific operation process:

[0034] (1) Semen thawing: When thawing, slowly lift a cryopreservation tube from liquid nitrogen and buffer it in liquid nitrogen vapor close to the mouth of the liquid nitrogen tank for 6 minutes, then immerse it in warm water at 40°C prepared in advance and shake it for 60 minutes. Second. The thawed semen was stored at room temperature (26°C) in the dark until use.

[0035] (2) Insemination: Select a female fish that has reached the effect period after oxytocin, artificially squeeze the eggs in a clean and dry bowl, take 10 ml of eggs and add 1 ml of frozen semen that has been thawed for 10 minutes, mix well and stir for 1 minute , add 10 ml of 0.4% Nacl solution to activate fertilization; add water and inflate to hatch, and the statistical fertilization rate in the middle gastrulation can reach 40%.

Embodiment 3

[0037] The following takes 0.5 ml of cryopreserved semen for insemination under the condition of low sperm-to-egg ratio as an example to illustrate the specific operation process:

[0038] Select a female fish that has reached the effect period after induction of labor, artificially squeeze the eggs in a clean and dry bowl, take 50 ml of eggs, add 0.5 ml of frozen semen that has been thawed for 10 minutes, mix well and stir for 1 minute, then add 50 ml 0.4% Nacl solution activates fertilization; add water and inflate to hatch, and 3000 fry can be hatched.

[0039] See attached figure 1 , the fertilization rate comparison between fresh sperm and frozen sperm under different conditions of sperm-egg ratio, artificial insemination, insemination with frozen sperm, when the sperm-egg ratio is 1,000,000:1, the fertilization rate similar to the fresh-sperm insemination ratio of 200,000:1 can be obtained . When the ratio of fresh sperm and egg is greater than 200,000:1 and the ratio ...

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Abstract

The invention belongs to the technical field of fish reproduction and relates to a method for ultralow-temperature cryopreservation of semen of siniperca scherzeri and artificial insemination in crossbreeding, which can be widely applied in culture of siniperca chuatsi in families and groups, especially large-scale culture and reproduction. The method comprises the steps of semen collection, diluent preparation, semen dilution, semen freezing, semen unfreezing, artificial insemination and the like. In the artificial insemination, as the optimum ratio of frozen sperms to fresh spawn is determined to be 1000000:1 by different experiments about sperm-spawn ratio. In the invention, the requirement for equipment is low, the operation is easy to realize, the method can be used for common fish reproduction fields, can realize storage of siniperca scherzeri resources, solves the problem of asynchronous growth of gonads of the siniperca chuatsi and the siniperca scherzeri in crossbreeding of siniperca, and increases the utilization ratio of the sperms of the siniperca scherzeri, and in the reproduction process, the sperm-spawn ratio can be flexibly adjusted according to the breeding requirements, the production need and the amount of parent fishes.

Description

Technical field [0001] The invention relates to fish breeding technology, belongs to the technical field of aquaculture, and in particular relates to a method for cryopreserving the semen of spotted mandarin fish at an ultra-low temperature, and a method for artificial insemination with frozen spotted mandarin fish semen, which can be widely used in family and collective mandarin fish farming, especially It is large-scale breeding. Background technique [0002] At present, the genetic improvement of mandarin fish is widely carried out in various places. The distribution of mandarin fish resources in my country is relatively wide, but the amount of resources is gradually decreasing, and the problem of biodiversity loss is serious; due to the wide distribution, the time difference between gonad maturation can reach two months, and the reproductive peak period is about 20 days , hybridization requires higher gonad maturity, over-maturity or under-maturity will seriously affect ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02A61D19/02A01K61/00
CPCY02A40/81
Inventor 李桂峰何建国洪锡标王小林孙际佳王海芳罗渡
Owner 生生农业集团股份有限公司
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