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Haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae triple inactivated vaccine and application thereof

A technology of Haemophilus suis and Streptococcus suis, which is applied in the field of veterinary biological products, can solve the problems of lack of immune cross-protection, and achieve good immunogenicity, good safety, and long-term immunity

Pending Publication Date: 2019-08-02
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Aiming at the current mixed infection of Haemophilus parasuis, Streptococcus suis, and Actinobacillus pleuropneumoniae, and the lack of immune cross-protection between each serotype, the invention provides a serum of Haemophilus parasuis, Streptococcus suis and pleuropneumoniae Actinobacillus triple inactivated vaccine

Method used

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  • Haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae triple inactivated vaccine and application thereof
  • Haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae triple inactivated vaccine and application thereof
  • Haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae triple inactivated vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Isolation and identification of strain HNHPS1

[0046] 1.1 Collection of disease data

[0047] The disease material came from a 2-month-old nursery pig that died of severe pneumonia, dyspnea and neurological symptoms in a large pig farm in Ruyang County, Henan Province in November 2016.

[0048] 1.2 Preparation of medium

[0049] TSB liquid medium: Dissolve 30 g of TSB broth powder (Tryptic Soy broth, tryptone soybean broth) in 1000 mL of ultrapure water, sterilize at 115 °C for 15 min, add 50 mL of fetal bovine serum, sterile 1% NAD (Nicotinamide adeninedinucleotide, nicotinamide adenine dinucleotide, coenzyme Ⅰ) 1 mL;

[0050] TSA solid medium: Dissolve 40g of TSA agar powder (Tryptic Soy Agar, tryptone soybean agar) in 1000mL ultrapure water, sterilize at 115°C for 15min, add 50mL of fetal bovine serum, 1mL of sterile 1% NAD, in 4 Store at ℃ for later use.

[0051] 1.3 Isolation and culture of bacteria

[0052] Aseptically collect the brain tissue of ...

Embodiment 2

[0072] Example 2: Isolation and Identification of Bacterial Strain HN1570

[0073] 2.1 Culture medium preparation

[0074] TSB liquid medium and TSA solid medium were prepared according to the method in Example 1.

[0075] 2.2 Isolation and cultivation of strain HN1570

[0076] In 2017, the applicant collected the heart blood of nursery pigs with typical polyserositis under aseptic conditions, inoculated them on TSA solid medium containing NAD, cultured them in a constant temperature incubator at 37°C for 36 hours, and picked suspected colonies for passage Purify and culture for another 24-48 hours to observe the colony morphology of Haemophilus parasuis, and grow consistent needle-shaped, colorless, transparent, smooth, moist colonies with a diameter of 1-2 mm (see Figure 4 ); while purified and inoculated on NAD-free TSA solid medium, it cannot grow on NAD-free TSA medium;

[0077] Pick a single purified colony for Gram staining and microscopic examination, and observe t...

Embodiment 3

[0091] Example 3: Isolation and identification of strain HNSS1

[0092] 3.1 Culture medium preparation

[0093] TSB liquid medium and TSA solid medium were prepared according to the method in Example 1.

[0094] 3.2 Isolation and cultivation of strain HNSS1

[0095] The disease material came from a 2-month-old nursery pig that suffered from severe pneumonia, dyspnea and neurological symptoms and died in a large pig farm in Jun County, Henan Province in November 2016. The brain tissue of the diseased pig was aseptically collected and inoculated on TSA solid culture containing NAD On the base, cultured in a constant temperature incubator at 37°C for 24 hours, a uniform needle-like size, a single colony of 0.5-1 mm on the blood plate, off-white translucent, round, smooth, and neatly edged α-hemolyzed colonies (see Figure 7 ). After purification and inoculation, carry out Gram staining. The bacteria are Gram-positive bacteria, round or oval, arranged in chains or in pairs, and...

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Abstract

The invention relates to the technical field of animal biological products and specifically relates to a haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae triple inactivatedvaccine. The inactivated vaccine includes an inactivated haemophilus parasuis HNHPS1 serotype antigen, an inactivated haemophilus parasuis strain HN1570 serotype antigen, an inactivated streptococcussuis strain HNSS1 serotype antigen and an inactivated actinobacillus pleuropneumoniae strain HNAPP1 antigen; the four strains all have excellent immunogenicity; when the four strain inactivated antigens are used for preparing the vaccine, the infection of clinic haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae can be effectively prevented; the vaccine is characterizedby simple preparation process, high safety, excellent immune effect and long immunity period and meanwhile is capable of achieving the purposes of preventing various diseases with one injection, reducing cost and reducing stress.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of veterinary biological products, and in particular relates to the preparation and application of a triple inactivated vaccine of Haemophilus parasuis serum types 4 and 5, Streptococcus suis type 2 and Actinobacillus pleuropneumoniae type 1. [0003] Background technique: [0004] Haemophilus parasuis ( Haemophilus parasuis , HPS) is a Gram-negative polymorphic small bacterium of Pasteurellaceae, which is the pathogen of Haemophilus parasuis disease, mainly causing polyserositis, arthritis and meningitis in pigs. This disease exists widely all over the world. It was first reported by German scholar Glsser in 1910. It is also known as Glsser's disease. Haemophilus parasuis can infect pigs of various ages. It mainly occurs after weaning and in the nursery stage. It mainly infects pigs aged 2 weeks to 4 months. The incidence rate is generally 10% to 15%. The disease has become one of the main causes o...

Claims

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Application Information

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IPC IPC(8): A61K39/102A61K39/09A61K39/02A61P31/04C12N1/20C12R1/21C12R1/46C12R1/01
CPCA61K39/102A61K39/09A61K39/0208A61P31/04C12N1/20A61K2039/70A61K2039/552A61K2039/521C12R2001/21C12N1/205C12R2001/46C12R2001/01
Inventor 王治方徐引弟张青娴朱文豪许峰焦文强李海利郎利敏张立宪游一王克领
Owner INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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