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Procypris merus sperm cryopreservation method

A technology of cryopreservation and cryopreservation of Hehua carp, which is applied in the field of cryopreservation of Hehua carp sperm. It can solve the problems of few long-term preservation of Hehua carp sperm recovery methods, low sperm concentration, and decreased sperm activity, and achieve good cryopreservation effect. , simple operation, time-saving effect

Inactive Publication Date: 2020-04-14
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the breeding of Hehua carp is seasonal, in the non-breeding season, the sperm concentration of male individuals of Hehua carp is low, and the sperm activity decreases, so it is necessary to collect and store the sperm of Hehua carp in the breeding season for a long time, so that The need for annual breeding and production needs to improve the efficiency of artificial insemination and the survival rate of seedlings in non-breeding seasons. However, there are few methods for long-term preservation and recovery of the sperm of the carp.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for cryopreservation of the sperm of the carp carp, comprising the steps of:

[0023] (1) Semen collection: During the breeding season of the carp, select robust male broodstock with mature glands. After anesthesia, dry the water and mucus around the genital pores of the fish with a dry towel, and then gently squeeze the abdomen along the midline until the genital pores flow out. Milky white semen, suck the semen with a syringe to prevent urine, blood and water from mixing, inject the collected semen into the cryopreservation tube; take 10 μl of the collected semen and mix it with an equal volume of potassium chloride with a mass fraction of 0.35%. Observe the movement of sperm under a microscope, and select semen with high sperm activity for dilution and preservation;

[0024] (2) Semen dilution: first mix and dilute the semen collected in step (1) with the diluent at a volume ratio of 1:3 to obtain solution A, and then mix solution A and antifreeze at a volum...

Embodiment 2

[0029] A method for cryopreservation of the sperm of the carp carp, comprising the steps of:

[0030] (1) Semen collection: During the breeding season of the carp, select robust male broodstock with mature glands. After anesthesia, dry the water and mucus around the genital pores of the fish with a dry towel, and then gently squeeze the abdomen along the midline until the genital pores flow out. Milky white semen, suck the semen with a syringe to prevent urine, blood and water from mixing, inject the collected semen into the cryopreservation tube; take 10 μl of the collected semen and mix it with an equal volume of potassium chloride with a mass fraction of 0.35%. Observe the movement of sperm under a microscope, and select semen with high sperm activity for dilution and preservation;

[0031] (2) Semen dilution: first mix and dilute the semen collected in step (1) with the diluent at a volume ratio of 1:3 to obtain solution A, and then mix solution A and antifreeze at a volum...

Embodiment 3

[0036] A method for cryopreservation of the sperm of the carp carp, comprising the steps of:

[0037] (1) Semen collection: During the breeding season of the carp, select robust male broodstock with mature glands. After anesthesia, dry the water and mucus around the genital pores of the fish with a dry towel, and then gently squeeze the abdomen along the midline until the genital pores flow out. Milky white semen, suck the semen with a syringe to prevent urine, blood and water from mixing, inject the collected semen into the cryopreservation tube; take 10 μl of the collected semen and mix it with an equal volume of potassium chloride with a mass fraction of 0.35%. Observe the movement of sperm under a microscope, and select semen with high sperm activity for dilution and preservation;

[0038] (2) Semen dilution: first mix and dilute the semen collected in step (1) with the diluent at a volume ratio of 1:3 to obtain solution A, and then mix solution A and antifreeze at a volum...

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PUM

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Abstract

The invention discloses a procypris merus sperm cryopreservation method. The method comprises four steps of seminal fluid collection, seminal fluid dilution, sperm cryopreservation and sperm unfreezing, procypris merus seminal fluid is collected by adopting an artificial abdominal extrusion method, then the seminal fluid is mixed with a diluent, the mixture is mixed with antifreezing liquid, and sperm preservation liquid is obtained, and the diluent is any one of D-15 diluent, Hank's liquid and Ringer's Solution for fishes, wherein the antifreeze agent is any one of methanol, dimethyl sulfoxide and glycerol; the sperm preservation liquid is injected into a cryopreservation tube, and the cryopreservation tube immersed into liquid nitrogen for long-term preservation; when the sperm preservation liquid is taken, the cryopreservation tube filled with the sperm preservation liquid is taken out of the liquid nitrogen and then immediately placed in a water bath kettle at the temperature of 27DEG C or 37 DEG C or 47 DEG C to be unfrozen, and then obtained procypris merus sperms are used for artificial insemination. The method can preserve the procypris merus sperms for a long time, and the survival rate of the thawed and recovered sperms is high.

Description

technical field [0001] The invention belongs to the technical field of biological low-temperature cryopreservation, and in particular relates to a method for cryopreservation of spermatozoa of the carp carp. Background technique [0002] Hehua carp is a small warm-water fish of Cyprinidae, also known as Hehua black carp, Hehua carp, Hehua carp and Wu carp. Located in Guilin, Guangxi, it is a native Chinese fish and is a cultured species with local characteristics. The meat of Hehua carp is delicate. Less thorns and more meat, soft bones and no fishy smell, delicious taste, high protein content, and the mixed carp in the rice field has a mixed diet, fast growth, strong reproductive ability, short breeding cycle, and has high nutritional value and economical value. value. [0003] With the increase in the scale of Hehua carp breeding year by year, the demand for Hehua carp seedlings is gradually increasing, and the quality requirements for Hehua carp seedlings are also incr...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0284
Inventor 张曼谭虹雨陈秀荔杨春玲王振光杨艳艳
Owner GUANGXI UNIV
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