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Ultralow-temperature cryopreservation and activation method of sperm of scapharca broughtonii sckrenck

A technology of ultra-low temperature freezing and sperm, which is applied in the field of marine biology, can solve the problems of complex composition of antifreeze, rough freezing procedure, large dilution ratio, etc., and achieve the effect of easy activation, good freezing effect and fast swimming speed

Active Publication Date: 2014-11-19
MARINE BIOLOGY INST OF SHANDONG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The composition of the antifreeze solution used in the existing shellfish sperm technology is relatively complex, the proportion is cumbersome, and the dilution ratio is relatively large, and the concentration of the preserved sperm is low; the antifreeze procedure usually uses a two-step cooling method, and the freezing procedure is relatively rough
Through literature search, do not see the report of ultra-low temperature cryopreservation and activation method of clam sperm of the present invention

Method used

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  • Ultralow-temperature cryopreservation and activation method of sperm of scapharca broughtonii sckrenck
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  • Ultralow-temperature cryopreservation and activation method of sperm of scapharca broughtonii sckrenck

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Experimental program
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Effect test

Embodiment 1

[0023] 1) Sperm collection: On July 23, 2014, during the reproductive period of cockles, 45 brood clams were retrieved from Rizhao Donggang Farm. After anatomical microscope examination, the male cockles with well-developed gonads and more than 85% vigor detected by microscope were selected for sperm cryopreservation test.

[0024] 2) Screening of sperm activation solution: use natural seawater, natural seawater with added fetal bovine serum and ammonium chloride, natural seawater with added fetal bovine serum and ammonium acetate, natural seawater with added fetal bovine serum and sodium hydroxide To activate the fresh semen of cockles, the microscopic observation shows that the natural seawater added with fetal bovine serum and ammonium chloride has a better effect of activating sperm. The average curve velocity of sperm is 185.1 μm / s, and the average linear velocity is 143.4 μm / s. See the table below for data comparison.

[0025]

[0026] 3) Screening of sperm antifreez...

Embodiment 2

[0038] 1) On August 7, 2014, during the reproductive period of the cockles, 20 brood clams were retrieved from the breeding workshop of Rizhao Xinhui Aquatic Products Co., Ltd. After anatomical microscope examination, the male cockles with well-developed gonads and more than 85% vigor detected by microscope were selected for sperm cryopreservation test.

[0039]2) Antifreeze configuration: The diluent is composed of base fluid, antifreeze protective agent, and additives. After preparation, it should be pre-cooled in a refrigerator at 0-4°C until use; two antifreeze solutions are prepared. One of them is marked as HS, and its base liquid is sterilized natural seawater; the antifreeze protection agent is 5% dimethyl sulfoxide (DMSO) by volume; the additive is trehalose, and the added amount is 90g / L. The other is marked as NACL, and its base liquid is a sodium chloride solution with a concentration of 0.5mol / L; the antifreeze protection agent is dimethyl sulfoxide (DMSO) with a ...

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Abstract

The invention discloses an ultralow-temperature cryopreservation and activation method of sperms of scapharca broughtonii sckrenck. The method includes following steps: mixing and diluting seminal fluid of scapharca broughtonii sckrenck and a freezing-resistant liquid, carrying out a step-by-step cooling freezing process after a pre-cooling process, and carrying out unfreeze activation and activity detection after preservation for 2 weeks. The freezing-resistant liquid consists of a base liquid, a freezing-resistant protective agent and an additive. The base liquid is sterilized natural sea water or a 0.5 mol / L sodium chloride solution. The freezing-resistant protective agent is dimethyl sulfoxide with a volume concentration being 5%. The additive is trehalose. The long-period freezing preservation method of the sperms of the scapharca broughtonii sckrenck has important significance on fields such as artificial breeding, germplasm preservation, sustainable breeding and the like of the scapharca broughtonii sckrenck.

Description

technical field [0001] The invention relates to the technical field of marine organisms, in particular to a method for cryopreserving and activating clam sperm at ultra-low temperature. Background technique [0002] Scapharca broughttonii Sckrenck, belonging to the mollusc phylum, bivalves, clam order, clam family, hair clam genus, common name red clam, large hairy clam, is a large cold-water benthic economic shellfish, mainly distributed in the Pacific Ocean The Sea of ​​Japan, the Yellow Sea, the Bohai Sea and the East China Sea in the northwest. Kui cockles are delicious and nutritious, and are very popular in domestic and foreign markets. They are excellent varieties for export and foreign exchange. In the last century, due to the over-exploitation of the clam, its wild resources have declined rapidly, coupled with the large demand for the market and exports, the scale of artificial breeding has also continued to expand, and the demand for high-quality seed has also inc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 宋爱环邹琰李翘楚王英俊辛美丽卢珺韩龙江刘洪军
Owner MARINE BIOLOGY INST OF SHANDONG PROVINCE
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