Utra-low temperature cryopreservation method for sperms of semilabeo obscurus

A technology of ultra-low temperature freezing and preservation method, which is applied in the field of ultra-low temperature cryopreservation of sperm of the dark lip fish, can solve the problems such as the same report of the previous invention, and achieves the effects of low cost, high vitality, high fertilization rate and hatching rate.

Inactive Publication Date: 2012-11-07
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

[0004] Through document retrieval, do not see identical report with the present invention

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Embodiment Construction

[0013] The ultra-low temperature cryopreservation method of the dark lip fish sperm of the present invention, its specific steps are as follows:

[0014] 1. The collection of gametes: the semen collection method is the method of gently pressing the abdomen, and the fine broodstock of selective maturity is mixed with 100 ppm of MS-222 (Tricaine, C 10 h 15 NO 3 S) Anesthetize for 5 minutes, dry the abdomen of the fish with a towel, then gently squeeze the abdomen, and the mature semen will naturally flow out into a 5 mL centrifuge tube;

[0015] 2. Detection of fresh semen vitality: first use a pipette to drop a drop of water on the glass slide, aim at the field of view, then use the needle tip to pick up the above semen and stir it under the microscope immediately to observe its vitality. The semen with fresh semen vitality above 85% Samples are used for sperm freezing;

[0016] 3. Freezing method: Dilute the semen and antifreeze solution according to the three-step dilution...

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Abstract

The invention relates to ultra-low temperature cryopreservation method for the sperms of the semilabeo obscurus, belonging to the technical field of cryobiology. The ultra-low temperature cryopreservation method comprises the following steps: selecting a manure high-quality protospecies or improved variety, anesthetizing for 5 minutes by 100ppm of MS-222, squeezing the dried abdomen slightly to make the manure semen flow into a 1.5ml centrifuge tube, detecting the vitality of the fresh semen, freezing the fresh semen with the vitality of more than 85%, diluting the collected high-quality semen and the antifreeze liquid (D-15 and 5% methanol) in three steps so that 1mL of the antifreeze liquid contains the number of the sperms of 2.5*108, filling into 1.8ml freezing tubes, balancing at 4 DEG C for 30 minutes, balancing in the position of 6cm above the liquid nitrogen surface for 10 minutes, balancing on the liquid nitrogen surface for 5 minutes, and putting in the liquid nitrogen to preserve. When in use, the preserved sperms are unfrozen in the water bath of 40 DEG C and are manfully fertilized. The method has low cost, is easy and convenient to operate, ensures that the sperms of the semilabeo obscurus can be preserved for a long time, the recovered sperms have high vitality and the manually propagated semilabeo obscurus has high fertilizing rate and hatching rate, can realize the large-scale production and are of great significance to the fish resource recovery and reconstruction.

Description

Technical field: [0001] The invention relates to an ultra-low temperature cryopreservation method for the sperm of the dark-colored lip fish, belonging to the technical field of cryobiology. Background technique: [0002] The dark lip fish Semilabeo obscurus Lin belongs to the genus Semilabeo of the Cyprinidae subfamily Labeoninae. Because of its fat body, tender meat and delicious taste, it has become the first choice for fishing. And because of its living habit of living in caves in groups, it has become the biggest victim of electricity, poison, and bombing by criminals. With the increase in the utilization of fish resources and the update of fishing technology, the biodiversity of fish is declining. Dark lips Fish has become a rare and endangered species in the Lixianjiang River Basin. In 1989, the dark-colored lip fish was listed as a Class II protected animal in Yunnan Province, and in 1998 it was included in the "China Red Data Book of Endangered Animals". In order...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 王晓爱潘晓赋杨君兴陈小勇
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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