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Bivalent inactivated vaccine for rabbit pasteurellosis and preparation method thereof

A bivalent inactivated vaccine, Pasteurella rabbit technology, which is applied to multivalent vaccines, vaccines, veterinary vaccines, etc., can solve the problem that vaccines cannot be prevented, and achieve the effect of preventing Pasteurellosis rabbits and the preparation method is simple

Active Publication Date: 2021-05-28
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently there is only commercial inactivated vaccine against Pasteurella multocida type A in my country
At present, Pasteurella multocida type D is also very common in rabbits in my country, and the existing vaccines cannot prevent it

Method used

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  • Bivalent inactivated vaccine for rabbit pasteurellosis and preparation method thereof
  • Bivalent inactivated vaccine for rabbit pasteurellosis and preparation method thereof
  • Bivalent inactivated vaccine for rabbit pasteurellosis and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Isolation, Identification and Screening of Pasteurella multocida Type A and Pasteurella Type D in Rabbits

[0017] 1) Aseptically collect lung samples from dead rabbits with respiratory diseases, inoculate them on brain-heart infusion agar plates containing 5% (V / V) defibrinated sheep blood, incubate them upside down at 37°C for 24-48 hours, pick round, smooth, Translucent, non-hemolytic, small colonies with a diameter of less than 1.0 mm and neat edges, which were continuously purified for 3 times on a 5% (V / V) defibrated sheep blood-brain heart infusion agar plate to obtain a pure culture.

[0018] 2) Take the above-mentioned pure culture, spread it evenly on a glass slide, observe the bacterial morphology under the microscope after Gram staining, and select the isolated bacteria with Gram-negative small bacilli.

[0019] 3) Extract the sgDNA from the pure culture screened in step 2) using kmt1 , hyA and wxya Gene primers were used to amplify the isolates kmt1...

Embodiment 2

[0024]Preparation and safety testing of bivalent inactivated vaccine against pasteurellosis in rabbits

[0025] 1. Preparation of Bivalent Inactivated Vaccine against Pasteurellosis in Rabbits

[0026] Respectively, the number of viable bacteria was 2.0×10 11 CFU / mL of Pasteurella multocida type A PmA04 and Pasteurella multocida type D PmD01 were inoculated into 500 mL brain-heart infusion medium at 0.1% (V / V), and cultured at 37°C and 180rpm for 24 Hour. Formaldehyde with a final volume concentration of 0.2% was added respectively, and inactivated at 100 rpm at 37°C for 24 hours to obtain an inactivated bacterial solution. The two inactivated bacteria liquids are mixed evenly at a volume ratio of 1:1, and then the mixed inactivated bacteria liquids are mixed with the commercialized MONTANIDE of the French Sepic company. TM The GEL 02 PR adjuvant was mixed evenly at a volume ratio of 9:1 to obtain a bivalent inactivated vaccine, which was sealed in aliquots and stored at 4...

Embodiment 3

[0033] Evaluation of Immunological Efficacy of Pasteurellosis Bivalent Inactivated Vaccine in Rabbits

[0034] 1. The growth and decline of antibodies after bivalent inactivated vaccine immunization

[0035] a) Preparation of Pasteurella multocida capsular antigen for antibody detection: the number of viable bacteria was 2.0×10 11 CFU / mL of Pasteurella multocida type A PmA04 and Pasteurella multocida type D PmD01 were inoculated into 20 mL brain-heart infusion medium at 0.1% (V / V) respectively, and cultured at 37°C and 180rpm for 24 hours; centrifuge at 10000rpm at 4°C for 5 minutes, discard the supernatant, and resuspend the pellets in 5 mL of 2.5% (W / V) sodium chloride solution; incubate the resuspension in a water bath at 56°C for 2 hours, and shake well every 20 minutes in the water bath Once, centrifuge at 12,000 rpm at 4°C for 30 minutes, and take the supernatant; transfer the supernatant to dialysis bags with a molecular cut-off of 3.5 KD, and dialyze with 500 mL ster...

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Abstract

The invention relates to a bivalent inactivated vaccine for rabbit pasteurellosis and a preparation method thereof. The inactivated vaccine contains a rabbit type A pasteurella multocida PmA04 (CCTCC NO: M 2021202), and a rabbit type D pasteurella multocida PmD01 (CCTCC NO: M 2021201). The bivalent inactivated vaccine is prepared by using pasteurella multocida PmA04 and pasteurella multocida PmD01 as antigens for inactivation, and then adding a commercial MONTANIDETM GEL 02 PR adjuvant with the volume final concentration of 10% from the French Seppic Company. The bivalent inactivated vaccine can prevent infection of type A pasteurella multocida and typeD pasteurella multocida on rabbits at the same time, has the effect of preventing two diseases by one injection, is safe and reliable, and does not have the hidden danger of toxin dispersion.

Description

technical field [0001] The invention relates to a bivalent inactivated vaccine for pasteurellosis in rabbits and a preparation method thereof, belonging to the technical field of vaccine preparation for rabbits. Background technique [0002] Pasteurellosis in rabbits is an infectious disease of rabbits caused by Pasteurella multocida infection. The disease can occur all year round, and rabbits of all ages can be affected, and it is a common and frequently-occurring disease in rabbits. Clinically, pasteurellosis in rabbits is mostly characterized by respiratory symptoms, and symptoms such as otitis media, conjunctivitis, and pyometra can also be seen. Pasteurella multocida is widely prevalent in rabbits in my country, and its infection often causes serious economic losses, which is one of the important pathogens hindering the development of rabbit industry. There are many serotypes of Pasteurella multocida. According to the capsular antigens of Pasteurella multocida, it can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/102A61P31/04
CPCA61K39/102A61K39/39A61P31/04A61K2039/70A61K2039/521A61K2039/552A61K2300/00
Inventor 王锦祥谢喜平孙世坤陈岩锋陈冬金桑雷
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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