Culture medium, culture method and application of avian pasteurella multocida

A pasteurella and culture method technology, applied in the field of medicine and biology, can solve the problems of poor protection effect of immune vaccines and long culture period, and achieve the effects of shortening the preparation period, improving the protection effect and high protection rate

Inactive Publication Date: 2020-01-03
GUANGZHOU SOUTH CHINA BIOLOGICAL MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the traditional culture method has the disadvantages of long culture period and poor protective effect of the prepared immune vaccine.

Method used

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  • Culture medium, culture method and application of avian pasteurella multocida
  • Culture medium, culture method and application of avian pasteurella multocida
  • Culture medium, culture method and application of avian pasteurella multocida

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1 material

[0038] 1.1 Kind of virus: Pasteurella multocida CVCC44801 strain, purchased from China Veterinary Microbiological Culture Collection Management Center, batch number 19990201, namely: Pasteurella multocida C48-1, serotype A:1.

[0039] 1.2 Culture medium and feed

[0040] Table 1

[0041]

[0042] 1.2.1 Improved TSA plate: Weigh 4g of tryptone soybean agar powder, add 100mL of ultrapure water, boil to dissolve, and pressurize at 121°C for 15min, add 2mL of newborn bovine serum when it is naturally cooled to about 60°C, and mix well Divide into flat plates.

[0043] 1.2.2 50% glucose solution: weigh 2.5g of glucose, add ultrapure water to 5ml, autoclave at 121°C for 30min, and store at 4°C for later use.

[0044] 1.2.3 Improved LB medium (the medium used for the secondary seed liquid): Weigh 5g of tryptone, 2.5g of yeast powder, and 2.5g of NaCl (sodium chloride), add distilled water to make it 495mL, and press at 121°C for 30min. After the medium was ...

Embodiment 2

[0082] Example 2 Pasteurella multocida vaccine preparation

[0083] 1. Prepare several improved TSA plates according to the method 1.2.1 in the above example 1, and use the improved LB prepared by the method 1.2.3 as the dilution medium. Take 10ml of the fermented broth described in Example 1 and Comparative Examples 1-2 as a sample, and dilute the fermented broth by a 10-fold gradient. Take 100 μl of the culture dilution and add it to the modified TSA plate, spread it evenly with a sterile coating rod, incubate it upside down at 37°C for 12-16 hours, and select an appropriate dilution plate for counting. The schematic diagram of the culture plate for counting the number of living cells for the test of fermentation results is as follows: image 3 shown. The higher the dilution factor, the more the number of single colonies on the plate, and the higher the number of viable cells. The number of living cells (cfu / ml) = the number of single colony × dilution factor × 10.

[00...

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Abstract

The invention provides a culture medium, culture method and application of avian pasteurella multocida. In the culture method, the culture medium is simple and clear in composition, can be used for culturing the avian pasteurella multocida, can meet carbon sources and nitrogen sources required by microbial growth and is especially suitable for quick growth of the avian pasteurella multocida, large-scale culturing of the avian pasteurella multocida is facilitated, and the culture period is shortened. Meanwhile, adjuvants such as animal serum or whole blood do not need to be added into the culture medium, the cultured avian pasteurella multocida facilitates preparation of vaccines, and the vaccines have the characteristics that high stability is achieved, animal stress response is not proneto being caused, and the protection rate is high.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a culture medium, a culture method and an application of Pasteurella avian multocida. Background technique [0002] Pasteurella multocida is a bacterium that can cause fowl cholera disease, which is a contagious disease that infringes on the poultry breeding industry, with high morbidity and mortality. The main measures to control fowl cholera are vaccines and antibiotics. Among them, due to the shortcomings of antibacterial drugs such as high drug dependence, antibiotic residues, and easy relapse after drug withdrawal, the market gradually favors vaccine immunization methods to control the disease. Common vaccines mainly include Pasteurella multocida inactivated vaccine, propolis inactivated vaccine, oil emulsion inactivated vaccine and Pasteurella multocida live vaccine. According to part of my country's "Manufacturing and Inspection Regulations for Veterinary Biological Produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/102A61P31/04C12R1/01
CPCA61K39/102A61P31/04C12N1/20
Inventor 陈天任梁昭平许利娜王斌许芬芬李敏
Owner GUANGZHOU SOUTH CHINA BIOLOGICAL MEDICINE
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