Serum 14-type haemophilus parasuis with cross protection force and application thereof

A technology of Haemophilus suis and bacterial liquid, which is applied in the directions of medical preparations, bacteria, and antibacterial drugs containing active ingredients, can solve the problems of lack of 14 type Haemophilus parasuis vaccine, different serotypes, etc., and achieve good results. The effect of immunogenicity, good safety and no toxic side effects

Active Publication Date: 2021-05-25
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] Aiming at the defects and problems that the current serotypes of Haemophilus parasuis are different in various places and there are many vaccines but still lacking the vaccine of Haemophilus parasuis type 14, the present invention provides a virulent strain of Haemophilus parasuis serotype 14 and its application

Method used

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  • Serum 14-type haemophilus parasuis with cross protection force and application thereof
  • Serum 14-type haemophilus parasuis with cross protection force and application thereof
  • Serum 14-type haemophilus parasuis with cross protection force and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Isolation and identification of Haemophilus parasuis

[0021] 1.1 Source of disease material

[0022] In August 2018, the lungs, heart blood, lymph nodes, spleen, brain, joint fluid and other disease materials of pigs with suspected clinical symptoms of Haemophilus parasuis were collected from a large-scale pig farm in Henan. The affected pigs showed fever and dyspnea , joint swelling, lameness, cyanosis of the skin and mucous membranes, difficulty standing or even paralysis, stiffness or death.

[0023] 1.2 Culture medium preparation

[0024] 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.

[0025] TSA solid medium: Dissolve 40 g of TSA agar powder (Tryptic Soy Agar, Tryptone Soy Agar, BD Company...

Embodiment 2

[0036] Embodiment 2: Toxicity test of Haemophilus parasuis

[0037] A total of 20 weaned and healthy nursery pigs aged 9-10 weeks were selected as experimental animals. The test animals were randomly divided into two groups, one control group and one test group, with 10 animals in each group. Inoculate Haemophilus parasuis strain HN1513 into TSB liquid medium respectively, and culture it on a shaker at 37°C for 24 hours, measure the bacterial concentration, serially dilute 10 times to coat a solid plate, count viable bacteria, calculate the bacterial concentration of the original solution, and adjust to 10 9 CFU / mL. Animals in the test group were inoculated intraperitoneally with 3 mL liquid culture of Haemophilus parasuis strain HN1513, and animals in the control group were inoculated with 3 mL sterilized TSB liquid culture medium. Continuously observe for 2 weeks after injection, observe and record the number of cases and deaths.

[0038] It was observed that the HN1513...

Embodiment 3

[0041] Embodiment 3: Preparation, safety and efficacy test of vaccine

[0042] 3.1 Preparation of the vaccine

[0043] 3.1.1 Strains and strain propagation

[0044] Primary Seed Propagation

[0045] Inoculate the freeze-dried strains of Haemophilus parasuis serotype 14 strain HN1513 on TSA plates containing neonatal 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 are subcultured and inoculated on TSA plates containing fetal bovine serum (final concentration 5%) and NAD (final concentration 0.01%), cultivated at 37°C for 24 hours, and are used as first-class seeds after passing the inspection.

[0046] secondary seed propagation

[0047] The first-grade seeds of each strain were inoculated in TSB liquid medium containing neonatal bovine serum (final concentration 5%) and NAD (final concentration 0.01%), cultured on a shaker at 37°C for 18-24 hours, and after the pure ...

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Abstract

The invention belongs to the technical field of veterinary biological products, and particularly relates to a serum 14-type haemophilus parasuis virulent strain and application thereof. The 14-type haemophilus parasuis HN1513 is preserved in the China General Microbiological Culture Collection Center on November 9, 2018, and the preservation number of the 14-type haemophilus parasuis HN1513 is CGMCC (China General Microbiological Culture Collection Center) NO: 16807. The strain has relatively strong pathogenicity to nursery piglets and can cause morbidity and death of nursery pigs, and an inactivated vaccine prepared by taking the strain as an antigen has 100% protection rate to infection of type 14 and has relatively good cross protection force to infection of type 1-13, type 15 and haemophilus parasuis which cannot be classified.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a virulent strain of Haemophilus parasuis serotype 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. Haemophilus parasuis is widespread all over the world and has become one of the main causes of death of weaned and nursery piglets in recent years. Seriously affect the healthy development of the pig industry. [0003] 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 . Vaccination is the best way to p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/102A61P31/04C12R1/21
CPCA61K39/102A61P31/04A61K2039/521A61K2039/552Y02A50/30
Inventor 徐引弟王治方张青娴焦文强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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