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A kind of in vitro culture method and application thereof

A technology of in vitro culture and Aolichun, applied in preparations for in vivo experiments, pharmaceutical formulas, compound screening/testing, etc., can solve the problem of slow progress of infectious third-stage larvae, to prevent mass reproduction and simplify collection work , to ensure the effect of no bacterial contamination

Active Publication Date: 2022-06-28
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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Problems solved by technology

However, the in vitro cultivation of infectious third-stage larvae has been progressing slowly, and there are few reports on the in vitro cultivation of pure Spring elegans infective third-stage larvae in the prior art

Method used

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  • A kind of in vitro culture method and application thereof
  • A kind of in vitro culture method and application thereof
  • A kind of in vitro culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening of the optimal developmental conditions of Orichon stipitis

[0041] The nematode egg suspension collected in the above step (6) was placed in a cell culture flask, and cultured at a constant temperature of 27°C in a constant temperature incubator for 2 weeks, and observed and recorded every day.

[0042] 1. Single factor experiment on optimization of culture conditions for larvae of Orichon nematode

[0043] (1) The effect of temperature on the hatching of eggs of S. elegans: collect the egg suspension (made by washing the eggs on the mesh screen with distilled water), adjust the pH to 7.0 and the osmotic pressure to 0, and take a 24-well culture plate , add 1 mL of worm egg suspension to each well to make about 50 worm eggs per well, and the culture temperature is set to 10°C, 17°C, 22°C, 27°C, 32°C, and 37°C, and each temperature is set to three replicates. The illumination time was the alternation of day and night during the culture period, and ...

Embodiment 2

[0058] Example 2 Establishment of the in vitro culture method of Orichonella nematode

[0059] Based on the research in Example 1, it was determined that the main influencing factor of the culture medium affecting egg hatching and larval development of Orion nematode was pH. Therefore, it was observed that in Example 1, the combination with the highest development rate in the orthogonal test , pH condition, this condition is the optimal condition, so choose 5.5. Regarding the culture temperature, it is theoretically determined according to the group with the second highest development rate, but the temperature of the second highest development rate is 32°C, which is not much different from 27°C in the single factor test, and is based on the general nematode culture temperature principle. , so choose 27°C. The osmotic pressure is theoretically determined according to the third highest growth rate group, but there are two groups with the third highest growth rate, and there is ...

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Abstract

The present invention relates to a method for in vitro cultivation of A. auritiensis and its application. The method uses distilled water with weak acidic pH value and low osmotic pressure to cultivate the eggs of A. auriscens, and supplemented by appropriate temperature and light Conditions can effectively improve the development rate of the third-stage larvae of P. auritiensis, and the development rate reaches more than 90%. The present invention provides technical support for the in vitro culture research of A. auritiensis, the research and development, screening and drug resistance detection of new veterinary drugs for the prevention and treatment of A. aureus a. It has laid a solid foundation for the study of pharmacogenes or functional genes and pathogenic mechanisms.

Description

technical field [0001] The invention relates to a method for culturing parasites in vitro, in particular to a method for culturing in vitro nematodes and application of the method. Background technique [0002] Nematodirus oiratianus belongs to Stronglata, Trichostrongylidae and Nematodirus, and is a common gastrointestinal nematode that seriously harms livestock. The nematode disease caused by this nematode is distributed worldwide. In recent years, with the abuse of some commonly used anthelmintic drugs, drug resistance of nematodes is also commonly reported. [0003] The emergence of drug resistance seriously affects the therapeutic effect of parasitic diseases and brings great challenges to the prevention and control of parasitic diseases. Therefore, timely detection and control of parasite resistance is the basis for ensuring drug effectiveness. The established methods for detecting drug resistance of gastrointestinal nematodes mainly include in vivo tests and in vit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/033A61K49/00
CPCA01K67/033A61K49/0008
Inventor 李军燕罗晓平王鹏龙耿万恒刘威高娃王凤武刘秀丽郭天龙杨晓野赵治国达来宝力格侯勇跃
Owner INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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