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Method for culturing myxobolus in vitro

A technology of in vitro culture and iodine ammonium, which is applied in the field of establishment and optimization of culture conditions, can solve the problems of few reports on in vitro culture and achieve easy-to-obtain results

Inactive Publication Date: 2015-12-09
FISHERIES SCI RES INST OF JILIN PROVINCE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, domestic research on the systematics of Myxospora has been carried out and some achievements have been made, but most of them focus on morphological classification and identification of new species, and there are few reports on its in vitro culture

Method used

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  • Method for culturing myxobolus in vitro
  • Method for culturing myxobolus in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, take crucian carp muscle homogenate as the screening of the optimal condition of a kind of iodophora in vitro culture of culture carrier.

[0047] Include the following test steps

[0048] (a) Take out the cysts from the diseased fish, collect the cysts with a 1.5mL centrifuge tube, wash and centrifuge repeatedly with the mother liquor of the test culture solution, and obtain pure spores.

[0049] (b) Put the pure spores into a 50 mL cell culture bottle, add 1 mL of test culture solution mother solution, 4 mL of culture solution (M199, L-15, DMEM), and 50 μL of antibiotics into the cell culture bottle.

[0050] (c) Add 100 μL of myxospores to each well of the 6-well culture plate (taken out from the culture bottle, inoculate each well with fish myxospores at a density of 20 ind / mL), 400 μL of muscle tissue homogenate supernatant culture solution , 1500 μL of culture medium (M199, L-15, DMEM), 400 μL of fetal bovine serum, continuously cultured at a temper...

Embodiment 2

[0060] Embodiment 2, the screening of the optimal condition of a kind of iodophora in vitro culture with crucian carp gill homogenate as culture carrier.

[0061] Include the following test steps

[0062] (a) Take out the cysts from the diseased fish, collect the cysts with a 1.5mL centrifuge tube, wash and centrifuge repeatedly with the mother liquor of the test culture solution, and obtain pure spores.

[0063] (b) Put the pure spores into a 50 mL cell culture bottle, add 1 mL of test culture solution mother solution, 4 mL of culture solution (M199, L-15, DMEM), and 50 μL of antibiotics into the cell culture bottle.

[0064] (c) Add 100 μL of myxospores to each well of a 6-well culture plate (taken out from the culture bottle, and inoculate each well with fish myxospores at a density of 20 ind / mL), and 400 μL of the supernatant culture solution of muscle gill homogenate , 1500 μL of culture medium (M199, L-15, DMEM), 400 μL of fetal bovine serum, continuously cultured at a ...

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Abstract

The invention provides a method for culturing myxobolus in vitro and belongs to the technical field of parasite in-vitro culture. Crucian muscle homogenate and crucian gill homogenate are adopted as culture carriers for myxobolus in-vitro culture. The method includes the following steps of firstly, preparation of antibiotics; secondly, preparation of a supernatant culture solution of the crucian gill tissue homogenate; thirdly, in-vitro culture of myxobolus; fourthly, counting and growth curve drawing. The culture conditions of myxobolus are optimized, myxobolus can propagate in a proper environment outside a host, a large number of base materials are provided for later pharmacology or vaccine research, and further research on diseases or myxosporean caused by myxobolus is promoted.

Description

technical field [0001] The invention belongs to the technical field of in vitro culture of parasites, and in particular relates to the establishment of a laboratory culture method and the optimization of culture conditions for the parasite. Background technique [0002] With the upgrading of the structure adjustment of aquaculture species and the continuous introduction of famous, special and high-quality new species, fishermen have obtained relatively rich economic benefits, but at the same time, new fish species from all over the world have also brought many diseases, among which myxospore Insect diseases are particularly prominent, and their harm is very serious. If the control is not timely, it will often cause a large number of fish deaths and cause economic losses. Myxosporum is a fish-specific parasite, and both freshwater fish and marine fish have myxospore parasites. [0003] There are more than 1020 species that have been recorded in the literature at home and abr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/10C12R1/90
Inventor 刘艳辉周井祥马悦祖岫杰楚国生杜晓燕王好李改娟王钊
Owner FISHERIES SCI RES INST OF JILIN PROVINCE
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