Method for collection and ultralow temperature refrigeration preservation of high-quality pacific oyster sperms

A Pacific oyster, ultra-low temperature freezing technology, which is applied in the preservation of human or animal body, medical science, climate change adaptation, etc. Exposure time, effect of good permeability

Inactive Publication Date: 2015-02-11
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Pacific oysters are mostly artificially inseminated by anatomical methods, which require high-quality ...

Method used

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Embodiment 1

[0028] 1. In mid-June, during the reproductive period of Pacific oysters, 5 offshore caged oysters were obtained from the Jiaonan breeding base, and they were temporarily raised in the shellfish culture room. Expose under the sun for 1 hour, then turn it over to the other side and put it in seawater at room temperature for another 1 hour. Take it out immediately after the male oyster ejaculate. Take out the semen by dissecting the oyster and filter it with silk to remove tissue debris. The collected vigor is higher than 80 % of sperm.

[0029] 2. Add the pre-prepared antifreeze solution to the sperm collected above with a motility higher than 80% and mix well, vigorously invert and shake the centrifuge tube 5-10 times in crushed ice (0°C), and let it stand for 2 minutes , Pour out the mixture of sperm and antifreeze in the upper layer after standing still, filter through 100 sieves of silk screen, and the filtrate is set aside; squeeze the larger tissue pieces that are filtere...

Embodiment 2

[0035]1. At the beginning of July, during the reproductive period of Pacific oysters, a male Pacific oyster was collected in the Jiaonan breeding base. The gonads were very full. It was exposed to the sun for 1 hour, then turned over to the other side for another 1 hour, and then placed in seawater at room temperature. It was observed that the male oysters were immediately taken out after ejaculation, and the semen was taken out by dissection; the tissue debris was removed by sieving, and the sperm with a motility higher than 80% was collected.

[0036] 2. Add the pre-prepared antifreeze solution to the sperm collected above with a motility higher than 80% and mix well, vigorously invert and shake the centrifuge tube 5-10 times in crushed ice (0°C), and let it stand for 2 minutes , Pour out the mixture of sperm and antifreeze in the upper layer after standing still, filter through 100 sieves of silk screen, and the filtrate is set aside; squeeze the larger tissue pieces that ar...

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Abstract

The invention belongs to the technical field of marine organisms, and concretely relates to a method for collection and ultralow temperature refrigeration preservation of high-quality pacific oyster sperms. The method comprises the following steps: carrying out illumination stimulation on male pacific oysters with the full growth sexual glands, dissecting the sexual glands, and filtering by a bolting silk net to obtain a large number of high-quality pacific oyster sperms; mixing a seminal fluid with an antifreeze liquid according to an appropriate ratio, packaging the obtained mixture in 2 ml or 5 ml cryopreserved tubes; carrying out balance incubation and programmed cooling, and preserving the obtained substances in liquid nitrogen for a long term; and carrying out water bath unfreezing, and carrying out natural water activation to obtain sperms with the frozen sperm vitality of above 70% and the fertilization rate of above 95%. The method for collection and ultralow temperature refrigeration preservation of high-quality pacific oyster sperms has great significance to artificial breeding, germplasm preservation, genetic diversity and sustainable farming.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, in particular to a method for collecting and freezing high-quality Pacific oyster sperm. Background technique [0002] Pacific oyster (Crassostrea gigas) belongs to the Oyster family, the genus Pyramid, commonly known as the true oyster, is a shellfish with a larger individual in the oyster class. Its meat is tender, delicious and nutritious, containing 45% to 57% of protein, 7% to 11% of fat, 19% to 38% of glycogen, and the content of iodine is higher than that of milk and eggs. In addition, it also contains a variety of vitamins and Trace elements, known as ocean milk. It was introduced into China from Japan, Australia and Taiwan in the 1980s. The cryopreservation of shellfish semen is of great significance in aquaculture, genetic breeding and preservation of germplasm resources. Pacific oysters are mostly artificially inseminated by anatomical methods, which require high-quality ma...

Claims

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Application Information

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IPC IPC(8): A01N1/02A61D19/02A01K61/00
CPCY02A40/81
Inventor 刘建国李军许飞张国范李莉韩龙江
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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