Method for cryopreservation of semen of siniperca scherzeri and artificial insemination
A technology of spotted mandarin fish and semen, which can be applied to the preservation of human or animal bodies, fish farming, medical science, etc., can solve the problems of small mandarin fish resources, lack of attention to the ratio of sperm to eggs, and small body size, so as to promote the variety The effects of diversifying and optimizing the combination, solving the dyssynchrony of gonad development, and improving the utilization rate of sperm
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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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