Utra-low temperature cryopreservation method for sperms of semilabeo obscurus

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...

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Problems solved by technology

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