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A method for in vitro culture of grass carp enterocolitica and a method for preparing the culture medium.

Through the medium composed of yeast extract and other specific culture conditions, the continuous in vitro culture of bagworms has been successfully achieved, which solves the problem of lack of effective culture methods in the existing technology and provides stable experimental materials to support biological research.

Active Publication Date: 2020-10-27
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Before the present invention was made, the reports on bagworms at home and abroad mainly focused on morphology and epidemiology research, and there were no research reports on the culture medium components and culture methods of bagworms cultured in vitro at present.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A kind of culture medium of this embodiment is applied to the in vitro culture of bagworm of grass carp intestinalis, and the formulation composition and content of described culture medium are as follows: 100ml of Ren's solution, 0.5g of yeast extract, 1.0g of peptone, 3.5g of fetal bovine serum ml, horse serum 10ml, Bacillus licheniformis bacteria liquid 335μl, sterile starch 300mg;

[0041] Wherein: the formula composition and content of Ren's solution are as follows: sodium chloride (NaCl) 6.5g, potassium chloride (KCl) 0.14g, calcium chloride (CaCl) 2 ) 0.12g, sodium bicarbonate (NaHCO 3 ) 0.2g, sodium dihydrogen phosphate (NaH 2 PO 4 ) 0.01g, distilled water 1000ml.

[0042] The above-mentioned Ren's solution is prepared by the following method, comprising the steps of:

[0043](i) Dissolve 6.5g of sodium chloride, 0.14g of potassium chloride, 0.20g of sodium bicarbonate, and 0.01g of sodium dihydrogenphosphate in 999ml of distilled water in order to obtain so...

Embodiment 2

[0061] A kind of culture medium of this embodiment is applied to the culture medium of bagworm in vitro, and the formulation composition and content of described culture medium are as follows: 100ml of Ren's solution, 0.5g of yeast extract, 1.0g of peptone, 3ml of fetal bovine serum , Horse serum 6ml, Bacillus licheniformis bacteria liquid 300μl, sterile starch 200mg;

[0062] Wherein: the formula and the preparation method of described Ren's liquid are identical with embodiment 1.

[0063] The preparation method of the medium applied to the in vitro culture of bagworms of grass carp as described above in this embodiment comprises the following steps:

[0064] (a) Accurately weigh 0.5g of yeast extract and 1.0g of peptone into a 250ml Erlenmeyer flask, add 100ml of Ren's solution, adjust the pH to 7.0-7.5, seal with tinfoil and sterilize at 121°C for 20min under high temperature and high pressure. Store at 4°C after cooling;

[0065] (b) Divide the medium prepared in step (a...

Embodiment 3

[0078] A kind of culture medium of this embodiment is applied to the culture medium of bagworm in vitro, and the formulation composition and content of described culture medium are as follows: Ren's solution 100ml, yeast extract 0.5g, peptone 1.0g, fetal bovine serum 6ml , Horse serum 9ml, Bacillus licheniformis bacteria liquid 335μl, sterile starch 250mg;

[0079] Wherein: the formula and the preparation method of described Ren's liquid are identical with embodiment 1.

[0080] The preparation method of the medium applied to the in vitro culture of bagworms of grass carp as described above in this embodiment comprises the following steps:

[0081] (a) Accurately weigh 0.5g of yeast extract and 1.0g of peptone into a 250ml Erlenmeyer flask, add 100ml of Ren's solution, adjust the pH to 7.0-7.5, seal with tinfoil and sterilize at 121°C for 20min under high temperature and high pressure. Store at 4°C after cooling;

[0082] (b) Divide the medium prepared in step (a) into multi...

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Abstract

This invention relates to an in vitro culture method for *Cyclocarya paliurus* and the preparation method of the culture medium used, belonging to the field of in vitro culture technology for parasitic protozoa. The culture medium formula of this invention is: 100 ml Ringer's solution, 0.5 g yeast extract, 1.0 g peptone, 3-6 ml fetal bovine serum, 6-10 ml horse serum, 300-500 μl *Bacillus licheniformis* bacterial suspension, and 200-300 mg sterile starch. The worm culture steps of this invention are as follows: the extracted *Cyclocarya paliurus* are inoculated into the prepared culture medium, sealed after nitrogen purging, and cultured at 15°C for 48-72 hours. The worms are then transferred to another fresh culture medium, sealed after nitrogen purging, and cultured at 15°C. This cycle is repeated to allow the worms to continuously proliferate. The culture medium preparation method of this invention is simple and has good stability. The worms can achieve fission and proliferation in the culture medium, laying the foundation for physiological and experimental ecological research on *Cyclocarya paliurus*.

Description

technical field [0001] The invention belongs to the technical field of in vitro culture of parasitic protozoa, in particular, the invention relates to an in vitro culture method of bagworm and a preparation method of a culture medium used. Background technique [0002] Grass carp intestinal bagworm is an obligate intestinal parasitic ciliate of grass carp first discovered and named by Mr. Chen Qiliu (1955). Molnár and Reinhardt (1978) used histological methods to observe the intestinal tract of grass carp infected with bagworm and found that it parasitizes in the mucosal folds and contents of the hindgut, and the massive proliferation of the worms will lead to catarrh in the hindgut of grass carp enteritis and may inflame other bowel segments. Li Ming et al. (2007, 2014) observed the morphology and ultrastructure of bagworms by hematoxylin staining, scanning electron microscopy and transmission electron microscopy, and described the morphological characteristics of bagworms...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/10
CPCA01K67/30C12N5/0068C12N1/10C12N1/00
Owner INST OF AQUATIC LIFE ACAD SINICA