Preparation method of pig pasteurella multocida antigen and application

A Pasteurella, multocida technology, which is applied in the field of preparation of pig pasteurellosis antigen, can solve the problems such as the impact of continuous bacterial growth, and achieve the growth of the number of cultured bacteria, reduce the cost, and increase the number of cultured bacteria Effect

Inactive Publication Date: 2017-05-24
安徽东方帝维生物制品股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Cultivate according to the above-mentioned traditional fermentation process. During the culture of bacteria, nutrients such as protein and sugar in the culture medium are continuously digested, and more and more bacterial metabolites are produced, which affects the continued value-added of bacteria.

Method used

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  • Preparation method of pig pasteurella multocida antigen and application
  • Preparation method of pig pasteurella multocida antigen and application
  • Preparation method of pig pasteurella multocida antigen and application

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preparation example Construction

[0020] A method for preparing the pasteurellosis multocida antigen, characterized in that the steps of the method are as follows:

[0021] (1), preparation and disinfection of culture medium: receive porcine pneumonia dry powder culture medium, add water for injection according to the ratio of 100,000 ml / tank for each culture for preparation, and then at a temperature of 116°C, continue for 30 days in a fermenter Minutes of disinfection and sterilization, and finally cooled to 37 ° C to prepare for inoculation;

[0022] (2), bacterial inoculation: the substratum that above-mentioned step (1) is made is inoculated bacterial seed liquid according to 2% of substratum total amount, and the bacterial seed liquid 2000ml that preserves for later use is moved into aseptic room, meanwhile, cultivate by fermenter Add 0.1% whole blood of lysed hemocytes to the total amount of the base, and finally connect the various pipelines for inoculation, and use clean air to press the bacteria into...

Embodiment 1

[0025] The traditional preparation method does not add any medium during the aeration culture process, and starts from the third hour of aeration culture, samples are taken every hour to count the viable bacteria, and the growth curve of the EO630 strain is determined. The present invention adds fresh culture medium in the middle and late stages of culture, that is, when the aeration culture reaches 7-10 hours, supplements the nutrients consumed by bacterial value-added, and dilutes harmful metabolites at the same time. The middle stage of bacterial value-added logarithmic phase is the fastest time period for nutrient consumption. At this time, through the automatic control system of the fermenter, a certain amount of sterilized fresh culture medium prepared according to step 1 is added to the fermenter to make the bacterial growth reach the highest peak. The test results show that the number of viable bacteria reaches a peak in the traditional fermentation culture process afte...

Embodiment 2

[0029] In the step (3), 20% fresh medium is added to the fermenter within 7-9 hours of aerated culture. The test results show that the peak period of bacteria growth is prolonged in the fermentation culture process of one-time supplementation. Such as figure 1 , figure 2 , image 3 As shown, the average number of viable bacteria in the 7th hour of aeration culture was 8.8×10 9 CFU / mL, the average number of live bacteria in the 8th hour of aeration culture was 9.2×10 9 CFU / mL, the average number of live bacteria in the 9th hour of aeration culture was 8.8×10 9 CFU / mL, the results of the experiment all have a significant increase compared with the traditional fermentation process. Wherein, Table 2 shows the variation of the number of live bacteria in the fermenter by adding 20% ​​sterilized fresh medium to the fermenter at a time in different time periods.

[0030] Table II

[0031]

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Abstract

The invention discloses a preparation method of a pig pasteurella multocida antigen. The method is characterized by comprising the following operation steps: preparation and disinfection of a culture medium, and bacterination and bacteria culture; culturing in standing for 1 hour at 37 DEG C after the bacterination is finished, continuously performing aerobic culture, adding a certain amount of sterilized fresh culture medium in a fermentation tank in the time of aerobically culturing for 7-10 hours, and then continuously performing the aerobic culture until the viable count achieves the peak, ending the culture, and acquiring the antigen to perform various detections, saving at 2-8 DEG C for later use. The fresh culture medium is added in the middle and later period of the bacteria culture to effectively supplement nutrient ingredients consumed in bacteria fermentation in time; and meanwhile, a harmful metabolite is diluted, so that the culture bacteria count is obviously increased, the number of the viable bacteria can be greatly improved, the dosage of the antigen is lowered, the vaccine quality is improved, and the production cost is lowered.

Description

technical field [0001] The invention relates to a preparation method and application of a porcine Pasteurella multocida antigen, belonging to the technical field of veterinary biological products. Background technique [0002] Pasteurellosis multocida is an acute septic infectious disease of pigs. It mainly causes hemorrhagic sepsis or infectious pneumonia in animals. It can be transmitted between animals of the same species or different animals, and can also infect humans. According to the duration of the disease, it is divided into the most acute type, the acute type and the chronic type. Acute cases present symptoms of hemorrhagic sepsis, pharyngitis, and pneumonia; chronic cases present mainly as chronic pneumonia. The disease can occur throughout the year, mostly secondary to infectious diseases, and occurs all over the world, and sometimes it is endemic, seriously threatening the healthy development of the breeding industry and causing huge losses. The use of vaccine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/102A61P31/04
CPCA61K39/102
Inventor 肖华春王成业汪鑫鑫李玉梅王楠楠
Owner 安徽东方帝维生物制品股份有限公司
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