Method for preparing bacteria liquid for Riemerella anatipestifer vaccine

A technology of R. anatipestifer and vaccines, which is applied in the field of bacterial liquid for R. anatipestifer vaccines, can solve problems such as no fermentation process, and achieve good results

Active Publication Date: 2011-02-09
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Large-scale fermentation and production of Riemeria anatipestifer bacteria liquid is one of the foundations and keys for the production of R. anatipestifer inactivated vaccines. Fermentation process

Method used

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  • Method for preparing bacteria liquid for Riemerella anatipestifer vaccine
  • Method for preparing bacteria liquid for Riemerella anatipestifer vaccine
  • Method for preparing bacteria liquid for Riemerella anatipestifer vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Preparation of 1% lysed hemocyte whole blood N-J synthetic medium:

[0013] Prepare 100ml of 1% lysed blood cell whole blood N-J synthetic medium: tryptone: 1.9g; yeast extract: 0.7g; beef extract: 0.7g; hydrolyzed milk protein: 0.7g; glucose: 0.4g; NaCl: 0.5g ; Potassium dihydrogen phosphate: 0.3g; Dipotassium hydrogen phosphate: 0.3g; Na 2 HPO 4 12H 2 O: 0.9g; (NH 4 ) 2 SO 4 : 0.2g; NH 4 Cl: 0.03g; add water to 100mL; sterilize the above-mentioned components at 121°C for 15 minutes, and when the sterilized components are cooled to below 40°C, add 1ml of lysed blood cells and whole blood to the above-mentioned basal medium; After shaking well, the finished N-J synthetic medium can be obtained.

[0014] The whole blood of the added lysed hemocytes is aseptically collected from healthy bovine blood in non-epidemic areas by jugular vein blood collection or carotid artery bloodletting and aseptically defibrillated, frozen and thawed at -20°C and room temperature, an...

Embodiment 2

[0016] Preparation of duck infectious serositis inactivated vaccine vaccine preparation liquid:

[0017] 1 strain: Serum Type I Riemerella anatipestifer RA-CH-I strain.

[0018] 2 Culture medium: 1% lysed blood cell whole blood N-J synthetic medium, 10% serum nutrient agar plate, lysed whole blood and inspection medium are all produced in strict accordance with the appendix of "Chinese Veterinary Pharmacopoeia" and tested after passing the test.

[0019] 3 methods

[0020] 3.1 Propagation and identification of first-class seeds

[0021] Put the freeze-dried basal strain of serotype I Riemerella anatipestifer RA-CH-I into 5ml of nutrient broth containing 10% calf serum, and culture it at 37°C for 24 hours; then streak inoculated in 2 On the nutrient agar plate containing 10% calf serum, according to the standard [smooth surface, slightly protruding, round, creamy colonies, colony diameter 1.2 ~ 1.7mm; bacterial smears can be seen as single (occasionally in pairs) or filament...

Embodiment 3

[0035]The laboratory produced 5 batches of laboratory-made vaccines totaling 108,000 milliliters, with batch numbers 2005001, 2005002, 2005003, 2005004, and 2005005. During the vaccine production process, the semi-finished products and finished products are inspected strictly according to the inspection regulations. The results of semi-finished products and inspection reports are shown in Table 1, and the quality inspection of finished products is shown in Table 2 (the original inspection reports of the finished products of the 5 batches of laboratory trial products are detailed below).

[0036] Table 1 Production and inspection results of semi-finished duck infectious serositis inactivated vaccine

[0037]

[0038] Table 2 2005001-2005005 batches of duck infectious serositis inactivated vaccine laboratory product inspection results

[0039]

[0040]

[0041] in conclusion

[0042] Five batches of laboratory-made vaccines (2005001, 2005002, 2005003, 2005004, 2005005)...

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Abstract

The invention discloses a method for preparing bacteria liquid for Riemerella anatipestifer vaccine, which comprises the propagation and identification of the first-level seed, and the propagation and fermentation of the second-level seed; the propagation and identification of the first-level seeds comprises the following steps of: inoculating freeze-dry basic microbial strain into the nutrient broth containing 10 percent of fetal bovine serum, and culturing the nutrient broth for 24h at the temperature of 37 DEG C; inoculating the obtained product onto the nutrient agar plate containing 10 percent of fetal bovine serum by streaking, then culturing the obtained product in the environment containing 5 to 10 percent of CO2 or candle jar for 24h to 48h at the temperature of 37 DEG C; after that, selecting several typical colonies, inoculating the typical colonies onto the nutrient agar slant containing 10 percent of fetal bovine serum, culturing the product in the environment containing 5 to 10 percent of CO2 or candle jar for 24h at the temperature of 37 DEG C, and obtaining first-level seeds; the propagation of the second-level seeds comprises the following steps of: inoculating the first-level seeds into the nutrient broth containing 10 percent of fetal bovine serum; and culturing the nutrient broth for 24h at the temperature of 37 DEG C; sampling the obtained product for pureness test; and if obtained product is tested to be qualified, keeping the product at the temperature of 2 to 8 DEG C, wherein the service life thereof is not more than five days, the culture medium used in the fermentation is the N-J synthetic medium containing one percent of lysed whole blood, the fermenting microbial strain is inoculated in an amount which is 2 percent based on that of the second-level seed liquid, the fermentation manner is liquid fermentation and culture in a culture tank, and the culture condition is continuously culturing 24h at the temperature of 37 DEG C.

Description

technical field [0001] The invention relates to the technical field of veterinary biological preparations, in particular to a method for producing bacterial liquid for R. anatipestifer vaccine. Background technique [0002] Duck infectious serositis is a contagious disease that occurs in domestic ducks, geese, turkeys and a variety of poultry. Syndrome, duck septicemia and new duck disease, the pathogen is Riemerella anatipestifer (Riemerella anatipestifer, RA). The disease presents as acute sepsis or chronic infection. Clinically, the main manifestations are cough, panting, diarrhea, increased eye and nose secretions, ataxia, and head and neck tremor. Perihepatitis, air sacculitis, meningitis, and caseous salpingitis. The disease can occur all year round, and it is serious in winter, mainly through contaminated feed, drinking water, droplets, etc.; 1-8 week-old ducks are highly sensitive, especially ducklings 2-3 week old have a higher incidence High, up to 90%; the mort...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 程安春汪铭书朱德康陈孝跃
Owner SICHUAN AGRI UNIV
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