Mycoplasma hyopneumoniae and haemophilus parasuis bivalent pentavalent inactivated vaccine and application thereof

A technology of Mycoplasma hyopneumoniae and Haemophilus suis applied in the direction of multivalent vaccines, vaccines, veterinary vaccines, etc., can solve the problems affecting the healthy development of the pig industry, economic losses of the pig industry, and lack of immune cross-protection ability. Good immunogenicity, good immune effect, good cross-protection effect

Pending Publication Date: 2021-06-25
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

The disease is widespread all over the world and can infect pigs of various ages. It mainly occurs after weaning and nursery. It mainly infects pigs aged 2 weeks to 4 months. The incidence rate is generally 10% to 15%. In severe cases, the mortality rate can reach 80%. The disease has become one of the main reasons for the death of weaned and nursery piglets in recent years. It is one of the main pathogens of clinical mixed infection. Healthy Development of Pig Industry
[0004] There are many clinical serotypes of Haemophilus parasuis, and there are 15 serotypes in the standard typing, and there are more than 20% clinically non-typing strains, and the serotypes of the popular strains in different places are different, and there is a lack of immune cross-protection ability between the serotypes

Method used

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  • Mycoplasma hyopneumoniae and haemophilus parasuis bivalent pentavalent inactivated vaccine and application thereof
  • Mycoplasma hyopneumoniae and haemophilus parasuis bivalent pentavalent inactivated vaccine and application thereof
  • Mycoplasma hyopneumoniae and haemophilus parasuis bivalent pentavalent inactivated vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Isolation and identification of Mycoplasma hyopneumoniae

[0027] 1.1 Collection of disease data

[0028] The disease material of Mycoplasma hyopneumoniae 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.

[0029] 1.2 Culture medium preparation

[0030]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.

[0031] 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.

[0032] 1.3 Isolation and cul...

Embodiment 2

[0047] Embodiment 2: Isolation and identification of Haemophilus parasuis

[0048] 2.1 Sources of disease materials

[0049] From January 2016 to December 2019, a total of lung, heart blood, lymph node, Spleen, brain, joint fluid and other disease materials, the diseased pigs often manifested as fever, dyspnea, joint swelling, lameness, cyanosis of the skin and mucous membranes, difficulty in standing or even paralysis, stiff pigs or death.

[0050] 2.2 Culture medium preparation

[0051] TSB liquid medium: 30 g of TSB broth powder (Tryptic Soy broth, tryptone soybean broth, BD Company) was dissolved in 1000 mL of ultrapure water, sterilized at 115 °C for 15 min, and 50 mL of fetal bovine serum (Hychone Company), sterile 1% NAD (Nicotinamide adenine dinucleotide, coenzyme Ⅰ, Roche Company) 1mL.

[0052] TSA solid medium: Dissolve 40 g of TSA agar powder (Tryptic Soy Agar, Tryptone Soy Agar, BD Company) in 1000 mL of ultrapure water, sterilize at 115 ° C for 15 min, add 50 m...

Embodiment 3

[0072] Embodiment 3: Preparation, safety and efficacy test of vaccine

[0073] 3.1 Preparation of the vaccine

[0074] 3.1.1 Breeding of strains and strains

[0075] Primary Seed Propagation

[0076] The Mycoplasma hyopneumoniae HNMhy1 strain was inoculated on PPLO solid medium at 37°C, 5% CO 2 Cultured in an incubator, harvested after 3 days.

[0077] The freeze-dried strains of Haemophilus parasuis type 4 HNHPS1 strain, type 5 HN1570 strain, type 12 HN486 strain and type 13 HN1601 strain were respectively inoculated in the culture medium containing newborn bovine serum (final concentration 5%) and NAD (final concentration 0.01%). On the TSA plate, cultivate at 37°C for 18-24h, pick typical colonies that meet the requirements, subculture and inoculate the TSA plate containing newborn bovine serum (final concentration 5%) and NAD (final concentration 0.01%), and culture at 37°C for 24h. After passing the inspection, it will be regarded as the first-class seed.

[0078] se...

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Abstract

The invention belongs to the technical field of veterinary biological products, and particularly relates to preparation and application of a mycoplasma hyopneumoniae and haemophilus parasuis serum type 4, type 5, type 12 and type 13 bivalent pentavalent inactivated vaccine. The inactivated vaccine is prepared from an inactivated mycoplasma hyopneumoniae HNMhy1 strain antigen liquid, inactivated haemophilus parasuis type 4 HNHPS1 strain antigen liquid, inactivated haemophilus parasuis type 5 HN1570 strain antigen liquid, inactivated haemophilus parasuis type 12 HN486 strain antigen liquid and inactivated haemophilus parasuis type 13 HN1601 strain antigen liquid, and then immunologic adjuvants for livestock and poultry are added. The vaccine not only can well prevent infection of mycoplasma hyopneumoniae and haemophilus parasuis serotypes 4, 5, 12 and 13, but also has good cross protection force on other serotypes and non-typing haemophilus parasuis.

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 pentavalent inactivated vaccine of Mycoplasma hyopneumoniae and Haemophilus parasuis serotypes 4, 5, 12 and 13. Background technique [0002] Mycoplasma hyopneumoniae ( 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 (Porcine cirovirus 2, PCV-2), causing Inductio...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61K39/102A61P31/04
CPCA61K39/0241A61K39/102A61P31/04A61K2039/70A61K2039/552A61K2039/521
Inventor 徐引弟焦文强王治方张青娴朱文豪许峰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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