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Mink hemorrhagic pneumonia bivalent inactivated vaccine and preparation method thereof

A technology of hemorrhagic pneumonia and mink, which is applied in the field of vaccines and its preparation, can solve the problems of insignificant therapeutic effect, unsuitability for clinical application, and complicated preparation process, and achieve the prevention of hemorrhagic pneumonia in mink, good application prospects, and simple preparation process Effect

Active Publication Date: 2011-12-21
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS +1
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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 led to strong drug resistance of Pseudomonas aeruginosa Therefore, an effective vaccine is needed to prevent it. There is no commercialized vaccine in China to prevent mink hemorrhagic pneumonia. It has been reported abroad that the bacterial cell wall extract (JAMES E.PENNINGTON, 1979) and the common protective antigen extracted from the bacteria OEP (common protective antigen) (ABEC, 1975; E.HONDA, 1976) plus toxoid component elastase to make immunogen, has a certain immune effect, but the preparation process is complicated, the cost is high, it is not suitable for mass production and use, and the immune Many times, short immunization period, not suitable for clinical application, various antigen types of Pseudomonas aeruginosa, it is often difficult to achieve a good protective effect with a single component cell extract

Method used

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  • Mink hemorrhagic pneumonia bivalent inactivated vaccine and preparation method thereof
  • Mink hemorrhagic pneumonia bivalent inactivated vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Screening of powerful virus strain

[0032] 67 strains of Pseudomonas aeruginosa were isolated from the lungs of minks suffering from hemorrhagic pneumonia in various mink farms in Shandong, Hebei, Dalian, Inner Mongolia, Jilin, etc., and were identified as Pseudomonas aeruginosa by observing biological characteristics and biochemical tests. The standard serum of Japan Seiken Co., Ltd. was used to type by slide agglutination test to determine the serotype of the isolate.

[0033] The highly virulent strains were determined by the mouse challenge experiment. The strains were inoculated in PYG liquid medium and shaken at 37°C for 12 hours. The liquid was uniformly turbid, yellow-green or white, and formed a bacterial film. After counting the bacteria, adjust the number of bacteria to 2 billion CFU / mL, dilute the bacterial solution by 10 times and 100 times respectively, that is, the bacterial volume is 20 million CFU / mL and 2 million CFU / mL respectively, and...

Embodiment 2

[0038] Embodiment 2: the preparation of bivalent inactivated vaccine of the present invention

[0039](1) Vaccine preparation: Streak inoculation of strains DL15 and JL08 on PYG agar medium and culture at 37°C for 18 hours, then pick a single colony and inoculate in 5ml PYG broth medium with shaking at 37°C for 12 hours as the first generation seed solution, and then Inoculate 3% in 300ml PYG broth and shake at 37°C for 12h as the second-generation seed liquid, inoculate 4% of the second-generation seed liquid in a 10-liter bottle of 6000mL PYG liquid and ventilate at 37°C for 12h, then it is cultivated Vaccine bacterial liquid, count the bacterial liquid by bacterial plate count, and adjust the number of bacteria of the two strains to 8 billion CFU / mL according to the counting results, and then mix the bacterial liquid according to the ratio of strain DL15 and strain JL08 at 1:1 , add formaldehyde to inactivate after mixing, inactivate according to the final concentration of ...

Embodiment 3

[0042] Embodiment 3: the safety check of bivalent inactivated vaccine of the present invention

[0043] Three batches of vaccines prepared in Example 2 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 yo...

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Abstract

The invention relates to the technical field of biology, disclosing two pseudomonas aeruginosa virulent strains and a mink hemorrhagic pneumonia divalent inactivated vaccine prepared from the two strains. The pseudomonas aeruginosa virulent strains are inoculated in a culture medium to be amplified and cultured by ventilation; after the obtained cultured object is inactivated, the products are mixed at the ratio of 1-2:1; and adjuvant is added to prepare the mink hemorrhagic pneumonia divalent inactivated vaccine. The divalent inactivated vaccine disclosed by the invention has good immunity effect on mink hemorrhagic pneumonia, at least five-month protection periods can be obtained, the protection ratio is above 80%, the mink hemorrhagic pneumonia can be prevented, and the vaccine has good 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 September to November every year. It is characterized by hemorrhagic pneumonia and acute death. Red and frothy liquid flowed out of the nostrils and other symptoms, and postmortem autopsy showed diffuse hemorrhage and sepsis changes in the entire lung lobe. The disease occurs all over the world, and it can cause 10% to 50% mortality in farms every year. It is an important bacterial infectious disease that endangers the mink breeding industry. [0003] The pathogen of the disease is Pseudomonas aeruginolsa (Pseudomonas aeruginolsa) in the Pseudomonadaceae (Pseudomonadaceae) genus (Pseud-omonas), also known as Pseudomonas aeruginosa (Bacterium pyocyaneum)...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/116A61K39/104A61P11/00A61P31/04C12R1/385
Inventor 闫喜军白雪柴秀丽罗国良赵建军张海玲邵西群张蕾高晗
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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