Rhodococcus ruber fermentation method and application thereof as adjuvant in animal vaccines

A technology of Rhodococcus rhodochrous and adjuvant, applied in the field of microbiology and immunology, can solve the problems of low biological safety, edema, and poor targeting.

Active Publication Date: 2019-04-23
刘春郁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The present invention solves the technical problems of current vaccine adjuvants, such as poor immune enhancement effect, low biological safety, and poor pertinence, including the obvious adverse reactions that occur when using whole-cell complete bacterial cell adjuvants (shown as local injection of vaccines). edema, induration, and even ulceration), conventional lysate aqueous adjuvants are safe but do not significantly improve vaccine antibodies, and many specific problems faced in the prior art

Method used

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  • Rhodococcus ruber fermentation method and application thereof as adjuvant in animal vaccines
  • Rhodococcus ruber fermentation method and application thereof as adjuvant in animal vaccines
  • Rhodococcus ruber fermentation method and application thereof as adjuvant in animal vaccines

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Experimental program
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Embodiment 1

[0060] A method for fermenting Rhodococcus erythrococcus, specifically carried out according to the following steps:

[0061] 1) Preparation of eluted strains: Inoculate the Rhodococcus rubidococcus in a solid agar medium square bottle, and culture at 30°C for 4 days in aerobic or aerobic manner, until the cells in the square bottle appear light yellow or orange yellow or light red or deep red Afterwards, aseptically elute with an appropriate amount of normal saline, harvest the bacterial cells, and use them as eluted strains for later use;

[0062] 2) Preparation of the primary seed liquid: take the eluted strain prepared in step 1), inoculate the shake flask with 2% volume of the liquid culture medium, and cultivate it in a constant temperature shaker with continuous oscillation at a speed of 200 rpm, and cultivate it in aerobic or aerobic conditions at 30°C 48h, as a first-grade seed solution for subsequent use;

[0063] 3) Preparation of the secondary seed liquid: take th...

Embodiment 2

[0066] A method for fermenting Rhodococcus erythrococcus, specifically carried out according to the following steps:

[0067] 1) Preparation of eluted strains: Inoculate the Rhodococcus rubidococcus in a solid agar medium square bottle, and culture at 30°C for 4 days in aerobic or aerobic manner, until the cells in the square bottle appear light yellow or orange yellow or light red or dark red Finally, aseptically elute and harvest the thalline with an appropriate amount of normal saline, and use it as an eluting strain for subsequent use; the percentage by weight of the solid agar medium contained in the solid agar medium in the step 1) is: tryptone 1%, soybean peptone 0.5% , yeast extract powder 0.5%, NaCl 0.5%, sodium glutamate 0.5%, citric acid-citrate 0.25%, glycerin 2%, agar powder 1.5%;

[0068] 2) Preparation of the primary seed liquid: take the eluted strain prepared in step 1), inoculate the shake flask with 2% volume of the liquid culture medium, and cultivate it in...

Embodiment 3

[0072] A liquid culture medium specially used for fermenting and cultivating Rhodococcus Erythrococcus described in this application. The components and weight percentages of the liquid fermentation medium are: 1% tryptone, 1% soybean peptone, 0.1% urea, 1% yeast extract powder, NaCl 1%, sodium glutamate 1%, citric acid-citrate 0.5%, mannitol 5%, and the balance is water.

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Abstract

The invention relates to a special rhodococcus ruber (CGMCC NO. 17012) fermentation method, a preparation process of adjuvants with different dosage forms and an application as an adjuvant in animalvaccines. Rhodococcus ruber strains are separated from a farm and obtained through single colony cloning purification and identification, the invention provides a special fermentation process where the strain inactivation product is used as an animal vaccine adjuvant, and provides a preparation process of different dosage forms of adjuvants containing the strain and the product. Animal experimentsprove that when the adjuvant product is used for univalent or multivalent animal vaccines, particularly Newcastle disease inactivated vaccine, avian influenza inactivated vaccine and swine fever livevaccine, definite non-specific immune enhancement effect can be provided, specifically, the antibody peak value level induced by the animal vaccine is obviously improved, the time for producing the protective antibody level is advanced, the antibody maintenance period is prolonged, and the immune effect of the animal vaccines is enhanced.

Description

technical field [0001] The invention relates to the technical field of microbiology and immunology, in particular to a specific fermentation method of Rhodococcus ruber (preservation number: CGMCC No. 17012 Rhodococcus ruber. GD0704A) and its application as an adjuvant in the preparation of animal vaccines. Background technique [0002] In recent years, with the increasing amount of livestock and poultry breeding, animal diseases have become increasingly serious. Especially with the outbreak of highly pathogenic avian influenza, inactivated vaccines for animals, especially oil emulsion inactivated vaccines, have become the most important types of vaccines. At present, although the method of inactivated vaccines for animals is relatively mature, it is extremely susceptible to the quality of inactivated antigens. In particular, high-quality oil emulsion inactivated vaccines can only be achieved by high-power concentrated antigens, and avian influenza oil emulsions are used to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/39C12R1/01
CPCA61K39/39A61K2039/55594C12N1/20C12N1/205C12R2001/01
Inventor 付家栋
Owner 刘春郁
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