Ultra-low temperature cryopreservation and recovery methods for sperms of sinocyclocheilus grahami

Abstract

The invention relates to ultra-low temperature cryopreservation and recovery methods for the sperms of the sinocyclocheilus grahami, 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 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 semen and the antifreeze liquid (D-15 and 7.5% ethylene glycol) according to the volume ratio of 1:2, subpackaging, balancing at 4 DEG C for 60 minutes, balancing in the position of 6cm above the liquid nitrogen surface for 2 minutes, balancing on the liquid nitrogen surface for 20 seconds and putting in the liquid nitrogen to preserve. The recovery method comprises the following steps: shaking the semen rapidly in the water bath of 37 DEG C to unfreeze, and mixing the sinocyclocheilus grahami sperms squeezed out with the unfreezed semen so that the ovums are fertilized. The methods have low cost, are easy and convenient to operate, ensure that the sperms of the sinocyclocheilus grahami can be preserved for a long time, the recovered sperms have high vitality and the artificially propagated sinocyclocheilus grahami 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 and resuscitation method for the sperm of the barb in Dianchi Lake, belonging to the technical field of cryobiology. Background technique: [0002] Sinocyclocheilus grahami (Regan, 1904), also known as Dianchi golden thread fish, pineapple fish, and small cave fish, belongs to the genus Sinocyclocheilus of Cypriniformes (Cypriniformes) family Cyprinidae. Dianchi golden-thread barb is endemic to the Dianchi Lake Basin, and it is distributed in the rivers, karst caves, and underground rivers where the Dianchi Lake and its watershed are located. The golden-threaded barb in Dianchi Lake has delicious meat and is the first of the "Four Famous Fishes" in Yunnan. In 1989, it was listed as a national second-class protected animal and listed as an endangered species in the "China Red Book of Endangered Animals (Fish)". Listed as a VU rating on the Red List. Since the late 1950s, due to the in...

AI Technical Summary

Problems solved by technology

If effective rescue measures are not taken in time, it will be difficult to find the species of golden-haired barb in Dianchi Lake in nature in the future; although the artificial breeding of golden-haired barb in Dianchi Lake has been successful, due to the limitations of the cultured fish itself, in artificial breeding, It is easy to cause asynchronous development of male and female gonads; and in a long-term pond culture environment, it is easy to cause germplasm degradation