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A kind of Pseudomonas aeruginosa and marine mammal vaccine containing the bacteria

A technology of Pseudomonas aeruginosa and mammals, applied in the field of vaccines, to achieve the effect of effective homologous attack protection and strong immunity

Active Publication Date: 2020-11-06
BEIJING VBIOSCI INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical problem of lacking an effective bacterial pneumonia vaccine for disease protection in marine mammals at present, and provides a marine mammal vaccine obtained by inactivating Pseudomonas aeruginosa

Method used

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  • A kind of Pseudomonas aeruginosa and marine mammal vaccine containing the bacteria
  • A kind of Pseudomonas aeruginosa and marine mammal vaccine containing the bacteria
  • A kind of Pseudomonas aeruginosa and marine mammal vaccine containing the bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, acquisition and preservation of bacterial strains

[0033] 1. From a large aquarium, the stomata of marine mammals were aseptically collected in sterile plastic vials, and streaked and inoculated on hexadecanetrimethylammonium bromide agar medium (NAC). Take round, smooth, moist, and flat colonies for Gram staining and microscopic examination, select Gram-negative small bacilli for streaking and pure culture, and conduct identification according to the biochemical identification test requirements for Pseudomonas aeruginosa, and further purify and cultivate The strains were grown in pyocyanin medium, oxidase test, mannitol, nitrate reduction test, gelatin liquefaction test, 42°C growth test, etc. The results are shown in Table 1. The strain was named Pseudomonas aeruginosa DCP1 strain.

[0034] Table 1 Biochemical test results of isolated strains

[0035] Biochemical test items result Biochemical test items result Pyocyanin medium +...

Embodiment 2

[0058] Embodiment 2, the preparation of vaccine

[0059] Thaw the frozen bacterial solution obtained in step 2 of Example 1 at room temperature, and perform the following steps on the P2 generation strain:

[0060] 1. Antigen preparation

[0061] 1. Take the frozen bacterial liquid of the P2 generation DCP1 strain, thaw at room temperature, inoculate BHI liquid medium at a volume ratio of 1% to 10%, and culture at 37°C and 150rpm / min for 5 to 18 hours with shaking. The above-mentioned activated bacterial solution was inoculated into BHI medium at a ratio of 1% to 10%, cultured at 37°C and 150 rpm / min for 5 to 18 hours with shaking, and tested according to the appendix of the current Veterinary Pharmacopoeia of the People's Republic of China. The number of viable bacteria was calculated by counting viable bacteria.

[0062] 2. Add 37-40% formaldehyde solution to the above cultured bacterial solution and make the formaldehyde concentration 0.2%-0.5% (volume ratio, 37°C, 150rpm...

Embodiment 3

[0068] Embodiment 3, the inspection of vaccine

[0069] 1. Dosage Form

[0070] Get a 1ml disposable pipette, absorb a small amount of each vaccine prepared in Example 2 and drop it on the cold water surface installed on a 90mm disposable plate, except for the first drop, it does not spread.

[0071] 2. The stability of the vaccine

[0072] Take 10ml of each vaccine prepared in Example 2 and add it to a 10ml centrifuge tube, centrifuge at 3500r / min for 15 minutes, and the water phase precipitated at the bottom of the tube is not more than 0.5ml. The vaccines prepared in Example 2 were stored at 2-8°C for 24 months, at 37°C for 1 month or at 25°C for 3 months without discoloration, delamination and demulsification.

[0073] 3. Vaccine sterility test

[0074] 1. Take the above-mentioned antigens and carry out the sterility test according to the method of "The Veterinary Pharmacopoeia of the People's Republic of China", and the result is no bacterial growth.

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Abstract

The invention discloses a pseudomonas aeruginosa and a marine mammal vaccine containing the bacteria. The bacterial strain used therein is the Pseudomonas aeruginosa DCP1 strain isolated from marine mammals, and the preservation number is CGMCC NO.15418. The invention also protects a marine mammal vaccine, the active ingredient of which is inactivated marine mammal Pseudomonas aeruginosa antigen. The invention also protects a preparation method of a marine mammal vaccine, including the processes of strain isolation, identification, purification, antigen preparation and vaccine preparation. The Pseudomonas aeruginosa inactivated vaccine prepared by the invention is safe and reliable, can significantly improve the antibody level, and has wide application prospects.

Description

technical field [0001] The invention relates to the technical field of vaccines, in particular to a marine mammal vaccine prepared by inactivating Pseudomonas aeruginosa and the bacteria. Background technique [0002] With the improvement of people's living standards, the transformation of material needs to spiritual needs has promoted the rapid development of the domestic aquarium industry, and the demand for marine mammal resources has increased, especially for marine mammals with talent for performance. Such as bottlenose dolphins, beluga whales, etc., the number of breeding in various domestic aquariums continues to rise. When marine mammals are transferred from the vast wild ocean environment to the artificial breeding environment, the living space is greatly reduced, the food, water quality, ocean current, water temperature and other factors are limited, the living environment is strictly limited, and the artificial breeding conditions are very different from the natur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/104A61P31/04A61P11/00C12R1/385
CPCA61K39/104C12N1/20C12N1/205C12R2001/385
Inventor 张凌云郑杰马宁宁孙艳明张萌候野李燕李磊相伟
Owner BEIJING VBIOSCI INC
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