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Combined strain for preparing haemophilus parasuis vaccine, octavalent inactivated vaccine and application thereof

A technology of Haemophilus suis and inactivated vaccines, applied in the field of veterinary biological products, can solve the problems of poor vaccine effect and lack of effective cross-protection, and achieve the effect of no toxic side effects, good cross-protection and good immune effect

Active 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

[0004] In view of the defects and problems of the different circulating serotypes of Haemophilus parasuis in various places, and the lack of effective cross-protection among the serotypes, which leads to the defects and problems of poor vaccine effect, the present invention provides a kind of Haemophilus parasuis serotype 1, 2, 4 Type, Type 5, Type 7, Type 12, Type 13, Type 14 Octavalent Inactivated Vaccine

Method used

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  • Combined strain for preparing haemophilus parasuis vaccine, octavalent inactivated vaccine and application thereof
  • Combined strain for preparing haemophilus parasuis vaccine, octavalent inactivated vaccine and application thereof
  • Combined strain for preparing haemophilus parasuis vaccine, octavalent inactivated vaccine and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: Isolation and identification of Haemophilus parasuis

[0020] 1.1 Source of disease material

[0021] 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.

[0022] 1.2 Culture medium preparation

[0023] 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.

[0024] 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 mL ...

Embodiment 2

[0033] Embodiment 2: toxicity test

[0034] A total of 45 weaned and healthy nursery pigs aged 9-10 weeks were selected as experimental animals. The test animals were randomly divided into 9 groups, 1 control group and 8 test groups, 5 animals in each group. The Haemophilus parasuis strains HN1397, HN1553, HNHPS1, HN1570, HN1565, HN486, HN1601 and HN1513 were respectively inoculated in TSB liquid medium, cultured on a shaker at 37°C for 24 hours, and then the bacterial concentration was measured , continuous 10-fold dilution coating solid plate, count viable bacteria, calculate the bacterial concentration of the stock solution, adjust to 10 9 CFU / mL; 8 groups of animals were inoculated intraperitoneally with 3 mL of liquid culture of Haemophilus parasuis strains HN1397, HN1553, HNHPS1, HN1570, HN1565, HN486, HN1601 and HN1513, and the control group Animals were inoculated intraperitoneally with 3 mL of sterilized TSB liquid medium. Continuously observe for 2 weeks after inj...

Embodiment 3

[0039] Embodiment 3: Preparation, safety and efficacy test of vaccine

[0040] 3.1 Preparation of the vaccine

[0041] 3.1.1 Strain and strain propagation

[0042] Primary Seed Propagation

[0043] The freeze-dried strains of Haemophilus parasuis type 1 HN1397, type 2 HN1553, type 4 HNHPS1, type 5 HN1570, type 7 HN1565, type 12 HN486, type 13 HN1601 and type 14 HN1513 were respectively Inoculate on TSA plates containing newborn bovine serum (final concentration 5%) and NAD (final concentration 0.01%), culture at 37°C for 18-24 hours, pick typical colonies that meet the requirements, and subculture and inoculate them with fetal bovine serum (final concentration 5%) and NAD (final concentration 0.01%) on TSA plates, cultivated at 37°C for 24 hours, and after passing the inspection, they were used as primary seeds.

[0044] secondary seed propagation

[0045] The first-grade seeds of each strain were inoculated in TSB liquid medium containing newborn bovine serum (final conce...

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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 haemophilus parasuis serum type 1, type 2, type 4, type 5, type 7, type 12, type 13 and type 14 octavalent inactivated vaccine. The vaccine has 100% protection rate on infection of 1 type, 2 type, 4 type, 5 type, 7 type, 12 type, 13 type and 14 type strains, has good cross protection force on infection of 3 type, 6 type, 8 type, 9 type, 11 type, 15 type and haemophilus parasuis incapable of being classified, and is simple in preparation process, free of toxic and side effects, good in safety, long in immune period and good in immune effect.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to an eight-valent inactivated vaccine of Haemophilus parasuis serotypes 1, 2, 4, 5, 7, 12, 13 and 14 and its application . Background technique [0002] Haemophilus parasuis ( Haemophilus parasuis , HPS) is the pathogenic bacterium of Haemophilus parasuis, which mainly causes polyserositis, arthritis and meningitis in pigs. The disease is widespread worldwide. 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 of death of weaned and nursery piglets in recent years, and is one of the main pathogens of clinical mixed infection, which has caused large economic losses to the pig industry and seriously affected the healthy development of the pig industry. . [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/116A61K39/102A61K39/39A61P31/04C12N1/20C12R1/21
CPCA61K39/102A61K39/39A61P31/04C12N1/20A61K2039/70A61K2039/55566A61K2039/521Y02A50/30
Inventor 徐引弟王治方冯丽丽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