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Quadruple inactivated vaccine of mycoplasma pneumoniae and haemophilus parasuis, streptococcus suis, and actinobacillus pleuropneumoniae and application thereof

A technology of Mycoplasma hyopneumoniae and Haemophilus suis, applied in vaccines, multivalent vaccines, veterinary vaccines, etc., can solve the problems of lack of immune cross-protection, achieve good immunogenicity, reduce costs, and reduce stress effects

Active Publication Date: 2019-08-16
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

[0012]Aiming at the current mixed infection of Mycoplasma hyopneumoniae, Haemophilus parasuis, Streptococcus suis, and Actinobacillus pleuropneumoniae, there is a lack of immune cross-protection between each serotype , the present invention provides a quadruple inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis serotypes 4 and 5, Streptococcus suis type 2 and Actinobacillus pleuropneumoniae type 1

Method used

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  • Quadruple inactivated vaccine of mycoplasma pneumoniae and haemophilus parasuis, streptococcus suis, and actinobacillus pleuropneumoniae and application thereof
  • Quadruple inactivated vaccine of mycoplasma pneumoniae and haemophilus parasuis, streptococcus suis, and actinobacillus pleuropneumoniae and application thereof
  • Quadruple inactivated vaccine of mycoplasma pneumoniae and haemophilus parasuis, streptococcus suis, and actinobacillus pleuropneumoniae and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1, the separation and identification of Mycoplasma hyopneumoniae

[0052] 1.1 Collection of disease data

[0053] The disease material came from the lungs of a 2-month-old nursery pig that had severe pneumonia and dyspnea in a large pig farm in Puyang City, Henan Province in April 2017.

[0054] 1.2 Preparation of medium

[0055] PPLO liquid medium: 10.5 g of PPLO broth powder, 2.5 g of glucose, and 2.5 g of yeast powder, dissolved in 440 mL of ultrapure water, sterilized at 115°C for 15 min, added with 5 mL of MEM medium, 50 mL of horse serum, penicillin 80,000 units, 10 mL of sterile 10% arginine and 500 μL of 1% (w / v) phenol red. After the culture was sterilized at 115°C for 15 minutes, it was stored at 4°C for later use;

[0056] PPLO solid medium: PPLO liquid medium with 1.5% (w / v) agar powder added. After the culture was sterilized at 115°C for 15 minutes, it was stored at 4°C for later use.

[0057] 1.3 Isolation and culture of mycoplasma

[0058...

Embodiment 2

[0070] Example 2: Isolation and identification of strain HNHPS1

[0071] 2.1 Collection of disease data

[0072] 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.

[0073] 2.2 Preparation of medium

[0074] 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;

[0075] 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.

[0076] 2.3 Isolation and culture of bacteria

[0077] Aseptically collect the brain tissue of ...

Embodiment 3

[0097] Example 3: Isolation and Identification of Bacterial Strain HN1570

[0098] 3.1 Culture medium preparation

[0099] TSB liquid medium and TSA solid medium were prepared according to the method in Example 2.

[0100] 3.2 Isolation and cultivation of strain HN1570

[0101] 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 7 ); while purified and inoculated on NAD-free TSA solid medium, it cannot grow on NAD-free TSA medium;

[0102] Pick a single purified colony for Gram staining and microscopic examination, and observe t...

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Abstract

The invention relates to the technical field of veterinary biological products, in particular to a quadruple inactivated vaccine of mycoplasma pneumoniae and haemophilus parasuis, streptococcus suis,and actinobacillus pleuropneumoniae. The quadruple inactivated vaccine comprises inactivated mycoplasma hyopneumoniae strain HNMhy1 serotype antigen, inactivated haemophilus parasuis strain HNHPS1 serotype antigen, inactivated haemophilus parasuis strain HN1570 serotype antigen, inactivated streptococcus suis strain HNSS1 serotype antigen, inactivated actinobacillus pleuropneumoniae strain HNAPP1serotype antigen, and an immune adjuvant, the five strains have good immunogenicity and can prevent the infection of clinical mycoplasma pneumoniae and haemophilus parasuis, streptococcus suis, and actinobacillus pleuropneumoniae, the quadruple inactivated vaccine has good safety, good immune effect and long immunization period, can achieve a purpose of preventing various diseases by one-time injection, and reduces costs.

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 quadruple inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis serotypes 4 and 5, Streptococcus suis type 2 and Actinobacillus pleuropneumoniae type 1 . [0003] Background technique: [0004] Mycoplasma hyopneumoniae (Mhp) is the main pathogen of Mycoplasma pneumonia of Swine (MPS). Mycoplasma swine pneumonia is a chronic, contagious infectious disease with high morbidity and low fatality rate. It mainly manifests as loss of appetite, fever, cough, wheezing, dyspnea and other symptoms. The diseased pigs grow slowly and the feed conversion rate Decreased, often induce the infection of other pathogens, especially immunosuppressive pathogens such as porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome, PRRSV), porcine circovirus type 2 (P...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02A61K39/102A61K39/09A61P31/04C12N1/20C12R1/35C12R1/21C12R1/46C12R1/01
CPCA61K39/0241A61K39/102A61K39/092A61K39/0208A61P31/04C12N1/20A61K2039/70A61K2039/521A61K2039/552
Inventor 徐引弟王治方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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