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Mycoplasma hyopneumoniae and haemophilus parasuis combined inactivated vaccine and application thereof

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

Active Publication Date: 2019-05-14
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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  • 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 serotypes 2, 5 and 7

Method used

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  • Mycoplasma hyopneumoniae and haemophilus parasuis combined inactivated vaccine and application thereof
  • Mycoplasma hyopneumoniae and haemophilus parasuis combined inactivated vaccine and application thereof
  • Mycoplasma hyopneumoniae and haemophilus parasuis combined inactivated vaccine and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: Isolation and identification of Mycoplasma hyopneumoniae

[0035] 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.

[0036] 1.1 Preparation of medium

[0037] 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.

[0038] 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.

[0039] 1.2 Isolation and culture of mycoplasma

...

Embodiment 2

[0053] Example 2: Isolation and identification of bacterial strain HN1553

[0054] 2.1 Culture medium preparation

[0055] 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, coenzyme I) 1 mL.

[0056] 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 ) 1mL; store at 4°C for later use.

[0057] 2.2 Isolation and cultivation of strain HN1553

[0058] In 2018, 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...

Embodiment 3

[0075] Example 3: Isolation and identification of bacterial strain HN1570

[0076] 3.1 Culture medium preparation

[0077] TSB liquid medium and TSA solid medium were prepared according to the method in Example 2.

[0078] 3.2 Isolation and cultivation of strain HN1570

[0079] 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 Purification culture.

[0080] 3.3 Identification of strain HN1570

[0081] 3.3.1 Morphological observation and biochemical identification

[0082] Observe the colony morphology of Haemophilus parasuis after 24 to 48 hours of subculture of suspected colonies, and grow consistent needle-shaped, colorless, transparent, smooth, moist colonies with a diameter of 1 to 2 mm (see Figure 7 ); at the same time of purification a...

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Abstract

The invention belongs to the technical field of animal biological products, and particularly relates to a mycoplasma hyopneumoniae and haemophilus parasuis serotype-2, serotype-5, serotype-7 combinedinactivated vaccine. The vaccine is prepared from a mycoplasma hyopneumoniae antigen, three haemophilus parasuis serotype antigens and an immunologic adjuvant, wherein the mycoplasma hyopneumoniae antigen is the inactivated mycoplasma hyopneumoniae HNMhy1 antigen, the preservation number of the mycoplasma hyopneumoniae HNMhy1 antigen is CGMCC NO:13858, and the preservation date of the mycoplasma hyopneumoniae HNMhy1 antigen is May 26 th, 2017. The three haemophilus parasuis serotype antigens are inactivated haemophilus parasuis type-2 HN1553, type-5 HN1570 and type-7 HN1565 antigens respectively, wherein the preservation number of the haemophilus parasuis type-2 HN1553 strain is CGMCC NO:16801, the preservation number of the haemophilus parasuis serotype-5 HN1570 strain is CGMCC NO:16802,and the preservation number of haemophilus parasuis serotype-7 HN1565 strain is CGMCC NO:16803. The immunologic adjuvant is an 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 f...

Claims

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Application Information

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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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