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A kind of dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis and its application

A technology of Mycoplasma hyopneumoniae and double inactivated vaccine, which is applied in the direction of antibacterial drugs, antibody medical components, bacterial antigen components, etc., can solve the problem of lack of immune cross-protection, etc., and achieve good immunogenicity, long immune period, and feed remuneration. reduced effect

Active Publication Date: 2022-01-28
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

[0006] In view of the current mixed infection of Mycoplasma hyopneumoniae and Haemophilus parasuis, the lack of immune cross-protection among the serotypes of Haemophilus parasuis, and the problem that the inactivated Haemophilus parasuis vaccine only protects against strains of the same serotype, The invention provides a dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis

Method used

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  • A kind of dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis and its application
  • A kind of dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis and its application
  • A kind of dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Isolation and identification of Mycoplasma hyopneumoniae

[0038] The mycoplasma hyopneumoniae disease material of the present invention comes 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.

[0039] 1.1 Preparation of medium

[0040] PPLO liquid medium: PPLO broth powder 10.5g, glucose 2.5g, yeast powder 2.5g, dissolved in 440mL ultrapure water, sterilized at 115°C for 15min, added MEM medium 5mL, horse serum 50mL, penicillin 80,000 units, Sterile 10% arginine 10 mL and 1% (w / v) phenol red 500 μL. After the culture was sterilized at 115°C for 15 minutes, it was stored at 4°C for later use.

[0041] PPLO solid medium: PPLO liquid medium added with 1.5% (w / v) agar powder. After the culture was sterilized at 115°C for 15 minutes, it was stored at 4°C for later use.

[0042] 1.2 Isolation and culture of mycoplasma

[0043] Take an appropriate amount of dis...

Embodiment 2

[0057] Example 2: Isolation and identification of bacterial strain HN1553

[0058] 2.1 Culture medium preparation

[0059]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 adenine dinucleotide, nicotinamide adenine dinucleotide, coenzyme Ⅰ) 1mL.

[0060] TSA solid medium: Dissolve 40 g of TSA agar powder (Tryptic Soy Agar, tryptone soybean agar) in 1000 mL of ultrapure water, sterilize at 115 ° C for 15 min, add 50 mL of fetal bovine serum, sterile 1% NAD (add as needed ) 1 mL; store at 4°C for future use.

[0061] 2.2 Isolation and cultivation of strain HN1553

[0062] In 2018, the applicant collected the 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...

Embodiment 3

[0079] Example 3: Isolation and identification of bacterial strain HN1570

[0080] 3.1 Culture medium preparation

[0081] TSB liquid medium and TSA solid medium were prepared according to the method in Example 2.

[0082] 3.2 Isolation and cultivation of strain HN1570

[0083] 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 and purification to cultivate.

[0084] 3.3 Identification of strain HN1570

[0085] 3.3.1 Morphological observation and biochemical identification

[0086] Observe the colony morphology of Haemophilus parasuis after subculture of suspected colonies for 24 to 48 hours, and grow consistent needle-shaped, colorless, transparent, smooth, moist colonies with a diameter of 1 to 2 mm (see Figure 7 ); Purify and inoculate on ...

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Abstract

The invention belongs to the technical field of veterinary biological products, in particular to a dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis serotypes 2, 5 and 7. The vaccine is made of Mycoplasma hyopneumoniae antigen, three Haemophilus parasuis serotype antigens and immune adjuvants, wherein the Mycoplasma hyopneumoniae antigen is the inactivated Mycoplasma hyopneumoniae HNMhy1 strain antigen, and the Mycoplasma hyopneumoniae HNMhy1 strain , the preservation number is CGMCC NO: 13858, and the preservation date is May 26, 2017; the three serotype antigens of Haemophilus parasuis are respectively inactivated Haemophilus parasuis type 2 HN1553 strain and type 5 HN1570 strain and type 7 HN1565 strain antigen, wherein the Haemophilus parasuis type 2 HN1553 strain has a preservation number of CGMCC NO: 16801; the Haemophilus parasuis serotype 5 HN1570 strain has a preservation number of CGMCC NO: 16802; The Haemophilus parasuis serotype 7 HN1565 strain has a preservation number of CGMCC NO: 16803; the immune adjuvant is aluminum hydroxide gel adjuvant.

Description

Technical field: [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to the preparation and application of a dual inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis serotypes 2, 5 and 7. Background technique: [0002] Mycoplasma hyopneumoniae (Mhp) is the main pathogen of Mycoplasma hyopneumoniae (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 (Porcine cirovirus 2, PCV-2), causing Induction of immunosuppression, often resulting in death ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/116A61K39/02A61K39/102A61K39/39A61P31/04
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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