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Low-temperature preserving fluid for patinopecten yessoensis sperms, preserving method and using method

A low-temperature preservation technology for Ezo scallops, which is applied to the preservation, application, and adaptation to climate change of human or animal bodies

Active Publication Date: 2021-08-20
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cryopreservation technology of scallop sperm has not been reported yet.

Method used

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  • Low-temperature preserving fluid for patinopecten yessoensis sperms, preserving method and using method
  • Low-temperature preserving fluid for patinopecten yessoensis sperms, preserving method and using method
  • Low-temperature preserving fluid for patinopecten yessoensis sperms, preserving method and using method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Screening of cryopreservation solution of Ezo scallop sperm

[0021] 1. Preparation and preservation of sperm preservation solution:

[0022] Sterile seawater was prepared. Natural seawater was filtered through a 0.2 μm microporous membrane and stored in a sterilized Erlenmeyer flask for later use.

[0023] Preparation of cryopreservation solution, weigh the cryopreservation solution reagent and place it in sterile seawater to prepare a 20 μg / ml cryopreservation solution, mix it evenly with a magnetic stirrer, filter it through a 0.2 μm microporous membrane filter, and place it in a low-temperature refrigerator for 4 Store at ℃.

[0024] Table 1 Sperm storage conditions

[0025]

[0026]

[0027] The composition and mass percentage of the sperm activator are 0.7% NaCl, 0.05% KCl, 0.2% CaCl 2 , 0.02% NaOH and 1.5% glucose, and the solvent is sterile filtered sea water.

[0028] 1.2. Observation of sperm motility

[0029] After taking a small amount ...

Embodiment 2

[0038] Example 2. Ezo scallop sperm collection method

[0039] Select Ezo scallops with full gonads during the reproductive period, take out the gonads completely and place them in a sterile petri dish, use a scalpel to chop the gonads, add a small amount of sterile sea water and stir; put the gonads on a 500-mesh sieve to filter out impurities , collect the seminal plasma in a centrifuge tube.

[0040] Scallop spermatozoa cannot be purified by centrifugation after collection, so attention should be paid to bacterial operation during sperm collection.

[0041] The sperm activity and centrifugation effect of Ezo scallop sperm at different centrifugation speeds are shown in Table 3 (centrifugation time is 5min, and the activity of fresh sperm without centrifugation is ++). The results showed that even at 1000g low-speed centrifugation, the sperm activity of Ezo scallop was significantly reduced, and the sperm could not be fully aggregated at this speed; at 2000g and 3000g, the ...

Embodiment 3

[0045] (1) Sterile seawater preparation: Take the natural seawater filtered by precipitation, filter it through a 0.2 μm microporous membrane, sterilize it in a high-temperature steam sterilizer, store it in a 1000ml Erlenmeyer flask, and seal it with tinfoil.

[0046] (2) Preparation of cryopreservation solution for Ezo scallop sperm: Use an electronic balance to accurately weigh the agent, pour it into 1000ml sterile seawater, the composition and mass percentage of the cryopreservation solution are Florfenicol 0.02g / L, compound Sulfamethoxazole 0.02g / L, sodium chloride 26.72g / L, potassium chloride 0.72g / L, calcium chloride 1.15g / L, sodium bicarbonate 0.20g / L, magnesium chloride 2.26g / L. All ingredients were uniformly mixed with seawater using a magnetic stirrer, filtered through a 0.2 μm microporous membrane, and stored in a low-temperature refrigerator at 4°C.

[0047](3) Take healthy scallops with well-developed gonads, take out their gonads, cut them into pieces, add a sm...

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Abstract

The invention relates to a low-temperature preserving fluid for patinopecten yessoensis sperms, a preserving method and a using method, and belongs to the field of aquaculture. The low-temperature preserving fluid comprises the following components by mass percent: 0.02 g / L of florfenicol, 0.02 g / L of compound sulfamethoxazole, 26.72 g / L of sodium chloride, 0.72 g / L of potassium chloride, 1.15 g / L of calcium chloride and 0.20 g / L of sodium bicarbonate, and a solvent is sterile seawater. The invention further provides a method for preserving the patinopecten yessoensis sperms by using the low-temperature preserving fluid and a using method. The patinopecten yessoensis sperms can be preserved at the low temperature of 4 DEG C, centrifugation cannot be carried out in the sperm collecting process, centrifugation can cause the motility of the patinopecten yessoensis sperms to be reduced, and the final fertilization rate is affected. The sperms preserved for 5 days by using the preserving fluid disclosed by the invention are inseminated, the sperm-egg ratio is 10000: 1, and the fertilization rate is 85% or above.

Description

technical field [0001] The invention belongs to the field of aquaculture, and in particular relates to a cryopreservation solution for the sperm of Ezo scallop and a method for preserving and using it. Background technique [0002] Patinopecten yessoensis belongs to Mollusca, Lamellibr anchia, Pteriodia, and Pectinidae. It is a typical cold-water shellfish that is naturally distributed in Japan. Hokkaido and northern Honshu, the Korean Peninsula and the Russian Far East, extending northward to Sakhalin Island and the Sea of ​​Okhotsk. Ezo scallops are delicious, nutritious, large and fast-growing. Since they were introduced to China in 1982, Ezo scallops have been one of the most important mariculture species in northern my country. At present, my country has become the world's largest scallop farming country. The area of ​​scallop cultivation in the North Yellow Sea has reached more than 700,000 hectares, with an annual output of more than 200,000 tons and an output value ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/0284Y02A40/81
Inventor 丁君常亚青丁北辰王皓宋坚王许波
Owner DALIAN OCEAN UNIV
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