Paramisgurnus dabryanus seminal fluid ultra-low temperature cryopreservation method

Abstract

The invention discloses a paramisgurnus dabryanus seminal fluid ultra-low temperature cryopreservation method which comprises the following steps: step 1, sequentially mixing NaCl, KCl, CaCl2. 2H2O, MgCl2. 6H2O and NaHCO3 according to a concentration ratio of 80: 50: 5: 2: 50 to prepare a diluent, and storing the diluent at 4 DEG C for later use; 2, mixing the paramisgurnus dabryanus seminal fluidwith the diluent according to the volume ratio of 1: (2-4), then adding DMSO with the volume fraction of 7.5%-12.5%, performing uniform mixing to obtain seminal fluid anti-freezing mixed liquid, andsub-packaging the seminal fluid anti-freezing mixed liquid into cryopreservation tubes; and 3, balancing the cryopreservation tubes obtained in the step 2 at a position which is 6cm above liquid nitrogen surface for 10min, then balancing the cryopreservation tubes on the liquid nitrogen surface for 5min, and finally putting the cryopreservation tubes into liquid nitrogen for cryopreservation. According to the method, the sperm motility, the sperm service life and the cell membrane integrity rate are all improved, and the method becomes an effective and lasting preservation method.

Description

technical field [0001] The invention relates to the technical field of ultra-low temperature cryopreservation of semen, in particular to a method for ultra-low temperature cryopreservation of the semen of paraloach. Background technique [0002] Paramisgurnus dabryanus belongs to the genus Paramisgurnus subfamily of Cypriniformes Loachidae. It is naturally distributed in the Yangtze River, Jialing River and Minjiang River system, the middle and lower reaches of the Liao River, the Yellow River and Heilongjiang. Its meat is delicious, vitamins and unsaturated Rich in fatty acid content, Chinese medicine records have the functions of nourishing blood and Qi, strengthening yang and diuresis, and has become an important freshwater aquaculture species in my country. With the increasing scale of cultivation of Paraloach and the increasing demand for seedlings, artificial seedlings can meet the breeding needs of seedlings. However, artificial seedlings are mostly inbreeding, which ...

AI Technical Summary

Problems solved by technology

As an important economically farmed fish, both normal temperature (20-24°C) and ultra-low temperature freezing (-196°C) preservation of the semen of Paraloach grandis have been studied. Under normal temperature, the storage time is short, at most 132h; Among them, although the storage time is extended, it is impossible to determine whether there is a best solution due to the lack of discussion on various options

And the existing preservation method is all very low for the vigor of the spermatozoon, the lifespan of the sperm, and the integrity rate of the cell membrane, which is unfavorable for effective and long-lasting preservation

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