Rabbit staphylococcus aureus and application thereof in preparation of inactivated vaccine

A technology for staphylococcus and inactivated vaccine, applied in vaccines, veterinary vaccines, bacteria and other directions, can solve problems such as drug residues, and achieve the effects of simple preparation method, prevention of staphylococcus aureus infecting rabbits, and good safety.

Active Publication Date: 2021-06-11
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

At present, there is no commercialized vaccine against rabbit staphylococcus disease in my country. The prevention and treatment of the disease mainly depends on drugs. Staphylococcus aureus) and drug residues, etc.

Method used

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  • Rabbit staphylococcus aureus and application thereof in preparation of inactivated vaccine
  • Rabbit staphylococcus aureus and application thereof in preparation of inactivated vaccine
  • Rabbit staphylococcus aureus and application thereof in preparation of inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Isolation, Identification and Screening of Rabbit Staphylococcus Aureus

[0018] 1) Aseptically collect lung samples, mastitis samples, and foot dermatitis samples from dead rabbits, and inoculate each sample with 5% (V / V) defibrillated sheep blood-brain infusion (Oxoid company, article number: CM1135B) agar plate, cultivated at 37°C for 24 hours, pick a round colony with a slight bulge in the center, light yellow, complete hemolysis, and neat edges, on a 5% (V / V) defibrillated sheep blood brain heart infusion agar plate Purified 3 times in succession to obtain a purified culture.

[0019] 2) Take the above-mentioned pure culture, spread it evenly on a glass slide to prepare a smear, and observe the bacterial morphology with a microscope after Gram staining. Select single, paired or clustered Gram-positive cocci for further identification.

[0020] 3) Extract the genomic DNA of the strain screened in step 2) using 16S rRNA with nuc Gene primers were used to amplif...

Embodiment 2

[0024] Preparation and Safety Test of Rabbit Staphylococcus Aureus Inactivated Vaccine

[0025] 1. Preparation of Rabbit Staphylococcus aureus Inactivated Vaccine

[0026] Rabbit Staphylococcus aureus SA472 was inoculated into 500 mL of brain heart infusion medium at a volume ratio of 0.1%, and cultured at 180 rpm at 37°C for 24 hours; added formaldehyde to a final volume concentration of 0.3%, and inactivated at 100 rpm at 37°C for 24 hours to obtain inactivated Live bacteria liquid; add the commercialized MONTANIDE of the French Sepic company to the inactivated bacteria liquid TM GEL 02 PR adjuvant to a final volume concentration of 10%, mixed evenly to obtain an inactivated vaccine, sealed after aliquoting, and stored at 4°C.

[0027] 2. Safety testing of rabbit Staphylococcus aureus inactivated vaccine

[0028] 1) Sterility test: Take 0.2 mL of the inactivated vaccine prepared in step 1, spread evenly on the brain-heart infusion agar plate containing 5% (V / V) defibrinat...

Embodiment 3

[0033] Evaluation of Immunity Efficacy of Inactivated Vaccines

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

[0035] A total of 40 healthy rabbits negative for Staphylococcus aureus and negative for anti-Staphylococcus aureus antibodies were selected and divided into two groups on average. Immune group: 20 experimental rabbits, 10 each of male and female, 1 mL of the vaccine of the present invention was injected subcutaneously at the back of the neck at the age of 35 days, and 1 mL of the vaccine of the present invention was immunized for the second time at the age of 50 days, about 230 days of age (female) Rabbits were immunized with 1 mL of the vaccine of the present invention for the third time; control group: 20 experimental rabbits, 10 male and female, subcutaneously injected 1 mL of sterilized brain-heart infusion medium at the back of the neck at the age of 35 days, and 50 days 1 mL of sterilized brain-heart infusion medium wa...

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PUM

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Abstract

The invention discloses rabbit staphylococcus aureus and application thereof in preparation of an inactivated vaccine. A strain of staphylococcus aureus SA472 with strong pathogenicity to rabbits is screened from suppurative pneumonia lung samples, mastitis samples and foot dermatitis samples of multiple dead rabbits through bacterial isolation and identification and animal regression tests, and the preservation number of the staphylococcus aureus SA472 is CCTCC NO: M 2021200. The invention also discloses an application of the staphylococcus aureus SA472 in preparation of an inactivated vaccine, which comprises the following steps of: carrying out enlarged culture on the strain to obtain a bacterial solution, inactivating the bacterial solution to obtain an inactivated bacterial solution, adding an adjuvant, and uniformly mixing to obtain an inactivated vaccine. The inactivated vaccine disclosed by the invention is high in safety, the titer of an antibody generated after immunization is high, and the occurrence of staphylococcus aureus infected rabbits can be effectively prevented.

Description

technical field [0001] The invention relates to the technical field of vaccine preparation, in particular to a strain of rabbit Staphylococcus aureus and its application in the preparation of inactivated vaccines. Background technique [0002] Rabbit staphylococcal disease is an infectious disease caused by Staphylococcus aureus infection in rabbits. The disease occurs throughout the year, especially in high temperature and high humidity seasons. The disease is particularly harmful to breeding male rabbits and breeding female rabbits. Clinically, it is mainly manifested as mastitis, foot dermatitis and respiratory symptoms. Mastitis, foot dermatitis and respiratory symptoms caused by Staphylococcus aureus infection not only seriously affect the production performance of male rabbits and female rabbits, but also lead to the elimination or death of male rabbits and female rabbits, causing huge losses to rabbit farms. Economic losses. Research investigations found that Staph...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/085A61P31/04C12R1/445
CPCA61K39/085A61P31/04A61K2039/521A61K2039/552
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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