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Pseudomonas aeruginosa and propolis inactivated vaccine for minks and preparation process

A mink Pseudomonas aeruginosa, preparation technology, applied in the direction of antibacterial drugs, medical preparations containing active ingredients, antibody medical ingredients, etc., can solve the problems of not suitable for clinical application, complicated preparation process, acute onset, etc., and achieve good results Application prospect, the effect of simple preparation process

Inactive Publication Date: 2012-08-08
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Mink hemorrhagic pneumonia has an acute onset and rapid death, and it is often too late to treat it. Commonly used drugs such as tilmicosin, azithromycin, and kanamycin are not effective in treatment, and the abuse of antibiotics has caused Pseudomonas aeruginosa to develop drug resistance. Therefore, it is necessary to Effective vaccines are used for prevention. It has been reported abroad that bacterial cell wall extracts (JAMES E. PENNINGTON, 1979) plus toxoid component elastase are used to make immunogens, which have a certain immune effect, but the preparation process is complicated and costly, and is not suitable for large quantities. Production and use, and the number of immunizations is high, the immunization period is short, and it is not suitable for clinical application. There are many antigen types of Pseudomonas aeruginosa, and it is often difficult to achieve a good protective effect with a single-component cell extract

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Vaccine preparation: Dissolve Pseudomonas aeruginosa strains Pa1 and Pa3 purchased from the strain room of the China Veterinary Drug Control Institute in nutrient broth, inoculate them on slant medium, culture at 37°C for 24 hours, and take the cultures separately. Streak inoculation on the plate containing cetyltrimethylammonium bromide, culture at 37°C for 16-18 hours, after passing the pure inspection, as the first-class seeds, inoculate respectively in 0.3% beef extract and 2% fetal bovine Serum nutrient broth culture medium, cultivated at 37°C for 16-18 hours, after sampling and passing the pure inspection, as the second-generation seeds, inoculate the second-generation seeds in the fermenter at a ratio of 2%, and fill 70% of the total volume of the fermenter % fermentation medium and 0.01% defoaming agent, stirring speed is 150 rpm, and cultured at 37°C for 16-18 hours, which is the cultured vaccine bacteria liquid. Sampling is carried out with a suitable medium fo...

Embodiment 2

[0021] Three batches of vaccines prepared in Example 1 were injected into healthy minks in single dose, single dose repetition and overdose, and inoculated into the muscles of the hind limbs. After 14 days of observation, the body temperature and appetite of the inoculated minks were normal, there was no swelling and inflammation at the inoculation site, and no local and systemic reactions occurred: 3 animals were selected for each batch of vaccine, and the skin at the injection site was cut to observe the pathological changes of the injection site after necrosis. There was no abnormal change, and the muscle tissue at the inoculation site of the inner muscle of the hind limb was normal, only the injection site was dark brown, the size of a corn kernel, which was different from the surrounding reddish-brown muscle, and there was no inflammatory reaction at the injection site. The young minks aged 60-70 days continued to be observed until 30 days after inoculation, and the young ...

Embodiment 3

[0023] Three batches of mink Pseudomonas aeruginosa propolis inactivated vaccines were used to immunize 20 mice of 16-18g respectively, 10 mice in the experimental group and 10 mice in the control group, 0.2mL / mouse, and 21 days after the immunization, 20 mice in the control group were treated with ZC strain and RC respectively. strain challenged and observed for 7 days, the three batches of vaccine-immunized mice were 100% protected, and all the mice in the control group were 100% diseased.

[0024] Three batches of mink Pseudomonas aeruginosa propolis inactivated vaccine were used to immunize healthy minks aged 2-10 months respectively. Each batch of vaccine was inoculated with 10 minks, 1mL / mink. 21 days after immunization, together with 10 minks in the control group, Pa1 strain and Pa3 were used respectively. strain, challenged by intratracheal instillation (5 rats in each group), and observed for 7 days, 100% of the minks in the three batches of vaccine immunized groups we...

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Abstract

The invention discloses a Pseudomonas aeruginosa and propolis inactivated vaccine for minks, which is prepared from two Pseudomonas aeruginosa strains. The Pseudomonas aeruginosa and propolis divalent inactivated vaccine for the minks is prepared by the following steps of: performing aerobic amplification culturing on a Pseudomonas aeruginosa strain inoculation culture medium; inactivating harvested cultures; then mixing the inactivated cultures in the proportion of 1:1; and adding propolis. The Pseudomonas aeruginosa and propolis inactivated vaccine disclosed by the invention is good in immune effect of preventing hemorrhagic pneumonia of the minks; the protection period of at least five months can be obtained; and the protection rate is at least 80 percent. The Pseudomonas aeruginosa and propolis inactivated vaccine for the minks is good in application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a vaccine and a preparation method thereof. Background technique [0002] Mink hemorrhagic pneumonia, also known as mink Pseudomonas pneumonia, is an acute infectious disease that mainly occurs during the moulting season of mink from August to November every year. It is characterized by hemorrhagic pneumonia and acute death. Nostrils flowed out of red foamy liquid and other symptoms. Post-mortem examination showed that the entire lung lobe was enlarged with diffuse hemorrhage and sepsis. The disease is distributed all over the world, and it can cause 20%-40% mortality in farms every year. It is a traditional Chinese medicine bacterial infectious disease that endangers the mink breeding industry. [0003] The pathogen of the disease is Pseudomonas aeruginolsa (Pseudomonas aeruginolsa) in the family Pseudomonadaceae (Pseudomonas), also known as Pseudomonas aeruginosa (Bacterium pyocya...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K39/104A61P31/04
Inventor 单虎秦晓冰张传美黄娟蔡秀磊张启迪
Owner QINGDAO AGRI UNIV
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