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Triple inactivated vaccine for haemophilus parasuis diseases, streptococcosis suis diseases and pasteurella multocida diseases of pigs and preparation method of triple inactivated vaccine

A technology for Haemophilus suis disease and Haemophilus suis, applied in the field of veterinary biological products, can solve the problems of increasing the morbidity and mortality of respiratory diseases in pigs, affecting the safety of animal body and human consumption, and damaging the antigenicity of vaccines. , to achieve the effect of improving antigen utilization, convenient and fast immunization, and improving targeted delivery

Inactive Publication Date: 2020-02-21
广东君睿生物技术研究有限公司
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Problems solved by technology

According to the capsular polysaccharide of bacteria, Pasteurella multocida can be divided into types A, B, D, E, and F. Pasteurellosis in swine is also called swine pneumonia, which is mainly caused by type A and B infections. Sexual Pasteurella infection can cause acute or sporadic infectious diseases, and the pathogen can also be co-infected with a variety of pathogens, thereby increasing the morbidity and mortality of respiratory diseases in pigs and causing serious economic losses
[0005] Most of the vaccines on the market for the prevention and control of Streptococcus suis, Haemophilus parasuis, and Pasteurella multocida are single vaccines and dual vaccines for Streptococcus suis and Haemophilus parasuis , Pasteurellosis multocida co-infection inactivated vaccines are still blank in China, and commercial vaccines can no longer meet clinical needs, and the abuse of antibiotics has resulted in the emergence of drug-resistant strains, making multivalent vaccines for co-infection even more difficult. popular
[0006] Most veterinary vaccines are inactivated with formaldehyde. Formaldehyde inactivation is easily affected by temperature, pH and concentration. Excessive formaldehyde inactivation will not only damage the antigenicity of the vaccine, but also cause a large amount of residue, which is harmful to animals and humans. Eating safety is affected
Aluminum adjuvant is one of the recognized vaccine adjuvants. The application of conventional aluminum adjuvants is limited due to shortcomings such as large immune stress. Therefore, research and development of a simple preparation method can effectively prevent Streptococcus suis Haemophilus disease, Pasteurellosis multocida, and small side effects, safe and efficient inactivated vaccines are imperative

Method used

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  • Triple inactivated vaccine for haemophilus parasuis diseases, streptococcosis suis diseases and pasteurella multocida diseases of pigs and preparation method of triple inactivated vaccine
  • Triple inactivated vaccine for haemophilus parasuis diseases, streptococcosis suis diseases and pasteurella multocida diseases of pigs and preparation method of triple inactivated vaccine
  • Triple inactivated vaccine for haemophilus parasuis diseases, streptococcosis suis diseases and pasteurella multocida diseases of pigs and preparation method of triple inactivated vaccine

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Experimental program
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Effect test

Embodiment 1

[0032] The acquisition of embodiment 1 bacterial strain antigen solution

[0033] 1. Isolation, identification and virulence test of Haemophilus parasuis H23 and H24 strains

[0034] 1.1 Isolation and identification of Haemophilus parasuis strains H23 and H24

[0035] Animals: From 2016 to 2018, fresh diseased pigs with clinical manifestations such as panting, pleurisy, peritonitis, pericarditis, arthritis or encephalitis were collected from South China. They were nursery piglets 1-2 weeks after weaning.

[0036] Method: Aseptically collect the lungs, heart blood, and joint fluid of diseased pigs, inoculate them in TSA medium (containing 5% bovine serum, 0.002% NAD, and 100 μg / mL lincomycin) respectively, and culture them in a 37°C incubator for 24-48 After hours, observe the shape of bacterial colonies, select suspicious colonies for pure culture, and carry out 16S rRNA identification after expanding the culture of suspicious strains. The primer sequences are as follows:

...

Embodiment 2

[0090] Embodiment 2 Preparation of a triple inactivated vaccine of Haemophilus parasuis, Streptococcus suis and Pasteurellosis multocida

[0091] 1. Seed Preparation for Production

[0092] Primary seed propagation: Freeze-dried Haemophilus parasuis serotype 4 H23 strain (GDMCC No: 60722, storage place: Guangdong Microbial Culture Collection Center), Haemophilus parasuis serotype 5 H24 strain (GDMCC No: 60723, preservation place: Guangdong Microbial Culture Collection Center) inoculated in TSA containing 5% calf serum and 0.002% NAD, freeze-dried Streptococcus suis serotype 2 S23 strain (GDMCC No: 60724, preservation place: Guangdong Provincial Microbial Culture Collection Center) inoculated in TSA containing 5% calf serum, and Pasteurella multocida capsular type A P13 strain (GDMCC No: 60720, preservation place: Guangdong Provincial Microbiological Culture Collection Center ), Pasteurella multocida capsular type B P11 strain (GDMCC No: 60721, storage place: Guangdong Microbi...

Embodiment 3 3

[0108] The sterility test and safety test of embodiment 3 triple inactivated vaccines

[0109] Materials: Vaccine 1 and Vaccine 2. Vaccine 2 was prepared by inactivating each bacterial solution according to two inactivation methods respectively, and prepared according to the components of vaccine 2, and the preparation method was the same as that in Example 2.

[0110] The first inactivation method uses β-propiolactone solution as an inactivating agent to inactivate each bacterial liquid. The specific method is: slowly add β-propiolactone solution according to 0.1%-0.15% of the total bacterial liquid, and heat it at 4°C. Inactivate for 72 hours, stir several times during the period, and then hydrolyze the inactivator in a water bath at 37°C for 2 hours;

[0111] The second inactivation method is to use formaldehyde solution as an inactivator to inactivate each bacterial solution. The specific method is: slowly add formaldehyde solution according to 0.4% of the total bacterial...

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Abstract

The invention belongs to the field of veterinary biological products and in particular relates to a triple inactivated vaccine for haemophilus parasuis diseases, streptococcosis suis diseases and pasteurella multocida diseases of pigs and a preparation method of the triple inactivated vaccine. The triple inactivated vaccine of haemophilus parasuis, streptococcosis suis and pasteurella multocida inpigs, which is provided by the invention, comprises an antigen concentrated solution and a nano aluminum adjuvant, wherein the antigen concentrated solution comprises a deactivated haemophilus parasuis serum type-4 H24 strain, a deactivated haemophilus parasuis serum type-5 H24 strain, a streptococcosis suis type-2 serum S23 strain, a pasteurella multocida type-A P13 strain and a pasteurella multocida type-B P11 strain. The triple inactivated vaccine is capable of effectively preventing haemophilus parasuis diseases, streptococcosis suis diseases and pasteurella multocida diseases of pigs, the purposes that multiple effects are achieved with one injection and stress is reduced are achieved, and the epidemic prevention cost is reduced.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to a triple inactivated vaccine of Haemophilus parasuis, Streptococcus suis and Pasteurella multocida and a preparation method thereof. Background technique [0002] Haemophilus parasuis (HPS) is an opportunistic pathogen in the upper respiratory tract of pigs. Under certain conditions, it invades the body and causes fibrinous polyserositis, arthritis, meningitis and high mortality. A systemic infectious disease. At present, Haemophilus parasuis disease is one of the major bacterial diseases prevalent in my country. The bacterium can affect piglets and fattening pigs aged 2-25 weeks. For pigs, the morbidity rate is 10%-90%, and the mortality rate is as high as 50-90% in severe cases. The infection fatality rate of adult pigs is about 20%, and the fatality rate of sows can be as high as 30%-40%. huge economic losses. According to the KRG serotype, it can be ...

Claims

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Application Information

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IPC IPC(8): A61K39/09A61K39/102A61K39/39A61P31/04
CPCA61K39/092A61K39/102A61K39/39A61K2039/552A61K2039/55505A61K2039/70A61P31/04A61K2300/00
Inventor 郭鑫黄剑华陈奕梅敏敏
Owner 广东君睿生物技术研究有限公司
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