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Method for improving living bacterium rate of brucellosis live vaccine product

A technology for brucellosis and live vaccines, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of high production cost, low viable bacteria rate, short storage period, etc., and achieve low production cost , long storage period, and high bacterial viability

Active Publication Date: 2015-04-29
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to overcome the problems of low live bacteria rate, short storage period and high production cost of the vaccine product produced in the production method of the existing brucellosis live vaccine, and provide a method to improve the live bacteria of the brucellosis live vaccine product. new method of rate

Method used

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  • Method for improving living bacterium rate of brucellosis live vaccine product
  • Method for improving living bacterium rate of brucellosis live vaccine product
  • Method for improving living bacterium rate of brucellosis live vaccine product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The preparation of embodiment 1 brucellosis live vaccine (S2 strain)

[0031] 1. Strains

[0032] The attenuated S2 strain of Brucella suis was supplied by the China Veterinary Drug Control Institute.

[0033] 2. Preparation of seeds for production

[0034] Preparation and identification of primary strains:

[0035] According to the "Manufacturing and Inspection Regulations for Live Brucellosis Vaccines" (referred to as "the Regulations"), open the ampoule of brucellosis live vaccine (S2 strain) to dissolve and dilute the freeze-dried strains, and then inoculate the enzymatic tryptone agar plate culture by streaking basal, 37°C for 72 hours.

[0036] Observe the colony shape, which should meet the standard of the procedure. Under a low-power microscope, select more than 10 qualified colonies, mix and inoculate the liver soup agar slant tube, culture at 37°C for 72 hours, and pass the inspection according to the regulations. After passing the inspection, store it at...

Embodiment 2

[0060] The preparation of embodiment 2 brucellosis live vaccines (A19 strain)

[0061] 1. Strains

[0062] Original strain:

[0063] Brucella bovis attenuated strain A19 was supplied by China Veterinary Drug Control Institute.

[0064] 2. Preparation of strains for production

[0065] Preparation and identification of primary strains:

[0066] According to the "Manufacturing and Inspection Regulations for Live Brucellosis Vaccines" (referred to as the "Regulations"), open the ampoule of brucellosis live vaccine (A19 strain) to dissolve and dilute the freeze-dried strains, and then inoculate the enzymatic tryptone agar plate culture by streaking basal, 37°C for 72 hours.

[0067] Observe the colony shape, which should meet the standard of the procedure. Under a low-power microscope, select more than 10 qualified colonies, mix and inoculate the liver soup agar slant tube, culture at 37°C for 72 hours, and pass the inspection according to the regulations. After passing the ...

Embodiment 3

[0089] The preparation of embodiment 3 brucellosis live vaccines (M5 strain)

[0090] 1. Strains

[0091] Original strain:

[0092] The attenuated M5 strain of Brucella melis was supplied by Harbin Veterinary Research Institute.

[0093] 2. Preparation of strains for production

[0094] Preparation and identification of primary strains:

[0095] According to the "Regulations for the manufacture and inspection of live brucellosis vaccines" (referred to as the "regulations"), open the ampoule of brucellosis live vaccine (M5 or M5-90 strain) after dissolving and diluting the lyophilized strain, and inoculate the enzyme trypsin in a line. Incubate on peptone agar plate medium at 37°C for 72 hours.

[0096] Observe the colony shape, which should meet the standard of the procedure. Under a low-power microscope, select more than 10 qualified colonies, mix and inoculate the liver soup agar slant tube, culture at 37°C for 72 hours, and pass the inspection according to the regulati...

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Abstract

The invention provides a method for improving living bacterium rate of a brucellosis live vaccine product. The method comprises primary strain preparation, secondary strain preparation and bacterium solution cultivation. By increasing the addition of 50% of glucose liquid, the content of ammonia nitrogen and total nitrogen in a culture medium is improved, living bacterium count is improved by controlling air filling amount, so that the method can be used for effectively improving the living bacterium rate of the brucellosis live vaccine product, keeping a relatively high bacterium activity and increasing the survival rate of bacteria during a preservation period after freeze-drying. Through the method disclosed by the invention, the living bacterium count of a cultivated bacterium liquid can achieve 180-220 billion / ml, the living bacterium count can achieve 140-170 billion / ml in 13 months of the preservation period, and the living bacterium rate is 63-77%; and the method has the advantages of being high in yield, long in preservation period, low in production cost and the like.

Description

technical field [0001] The invention relates to the field of veterinary biological products, in particular to a method for increasing the live bacteria rate of live brucellosis vaccine products. Background technique [0002] Brucellosis is a zoonotic infectious disease caused by members of the genus Brucella, which seriously threatens the life and health of humans and various animals. OIE lists it as a B-type animal disease, and our country lists it as a B-type animal disease. The disease has a wide range of epidemics and spreads all over the world. The pathogen is Brucella, which is a small, blunt-ended coccus or short bacillus, Gram-negative, without flagella, non-moving, and non-spore-forming. Sheep are the main source of infection in China, followed by cattle and pigs. Studies suggest that the use of vaccines is an effective way to prevent and control brucellosis. [0003] Vaccines commonly used in the market are mainly brucellosis live vaccine (S2 strain), brucellos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/205C12R2001/01
Inventor 罗显绎石建平雷敬宗姜晓红马海宏
Owner CHINA ANIMAL HUSBANDRY IND
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