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Freezing protection solution and preservation method of Plectropomus leopardus sperms

A cryoprotectant solution, technology of leopard gillfish, used in the field of cryoprotection solution for leopard gillfish sperm, to achieve the effect of reducing adverse effects, low toxicity, and protecting sperm

Active Publication Date: 2015-01-28
SUN YAT SEN UNIV
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AI Technical Summary

Problems solved by technology

However, most osmotic antifreeze agents are inorganic chemicals, which have certain toxicity to sperm; saline solution cannot provide necessary nutrients to sperm

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0015] 1. Preparation of cryoprotectant solution: trehalose 34.23g, fetal calf serum 0.9L, ultrapure water to 1L (PH=7.45, osmotic pressure=324mOsmol / Kg).

[0016] 2. Sperm collection: Anesthetize the sexually mature male Leopard Gillfish with eugenol, dry the fish body to avoid contact with seawater, and gently press the abdomen to collect the semen in a 2ml cryopreservation tube.

[0017] 3. Freezing: mix equal volumes of semen and cryoprotectant solution in the cryopreservation tube, and then stay at 5cm above the liquid nitrogen surface for 3 minutes, then put it directly into the liquid nitrogen for storage.

[0018] 4. Thawing: Take out the cryopreservation tube with sperm stored in liquid nitrogen and put it in a water bath at 37°C for 1 minute.

[0019] 5. Sperm motility detection: The motility of leopard echinacea sperm after thawing was detected by the counting machine-assisted sperm analysis system, which was 36.33%, which was close to the fresh sperm motility of 41...

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Abstract

The invention discloses a freezing protection solution of Plectropomus leopardus sperms. The freezing protection solution comprises 34.23g of mycose, 0.9L of fetal calf serum, and water added to the total volume of 1L; and the pH value and the osmotic pressure of the freezing protection solution are 7.45 and 324mOsmol / Kg respectively. The preservation method comprises the following steps: carrying out isopyknic mixing on a seminal fluid and the freezing protection solution in 2ml of a freezing tube, allowing the freezing tube with the obtained mixed solution to stay in a position 5cm higher than a liquid nitrogen surface for 3min, and directly putting the freezing tube with the mixed solution into liquid nitrogen. The freezing tube with sperms frozen in the liquid nitrogen is taken out, and is processed in a 37DEG C water bath for 1min in order to unfreeze. Compared with freezing solutions in the prior art, the freezing protection solution has the advantages of very low toxicity to the sperms, and provision of nutrients needed by the sperms.

Description

technical field [0001] The invention relates to cryopreservation of fish sperm of the genus Echinacea, in particular to a cryoprotective liquid and a preservation method for the sperm of Echinacea genus Leopard. Background technique [0002] Prickly perch is a world-renowned rare marine fish with delicious meat, rich nutrition and high economic value. The artificial breeding of some species in the genus Acanthus genus (such as Acanthus leopardii) has been rapidly popularized and promoted in Guangdong and Hainan provinces in my country, and has become an important marine cultured species in the southern coast. [0003] Ultra-low temperature cryopreservation of semen is an effective way to protect fish germplasm. The work of cryopreservation of fish sperm in my country is mainly concentrated on freshwater fish such as the "four major family carp" and cold-water seawater fish such as flounder and large yellow croaker. The physiological characteristics of sperm of different s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 蒙子宁李波刘晓春张勇林浩然
Owner SUN YAT SEN UNIV
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