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Serum 4 type haemophilus parasuis and application thereof

A technology of Haemophilus suis and Haemophilus suis disease, applied in the field of biological immunity, can solve the problems of farmers increasing the dosage of antibiotics, pig products exceeding the standard of antibiotics, and poor treatment effect, etc., and achieve stable biological performance and strong pathogenicity Sexuality and the effect of reducing economic losses

Inactive Publication Date: 2015-01-28
CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, a large number of antibiotics are used for treatment. The result is: on the one hand, the antibiotics in pig products exceed the standard, which is harmful to human health; It is very prone to drug resistance, resulting in poor treatment effect, so farmers increase the amount of antibiotics, resulting in a vicious circle

Method used

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  • Serum 4 type haemophilus parasuis and application thereof
  • Serum 4 type haemophilus parasuis and application thereof
  • Serum 4 type haemophilus parasuis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation and identification of serotype 4 Haemophilus parasuis

[0035] 1. Isolation of pathogenic bacteria

[0036] Use a sterile inoculation loop to take samples from fresh lungs, pericardial effusion, joint fluid and other disease materials of suspected sick pigs, inoculate them by streaking on TSA medium (containing final concentration of 5% serum and 0.001% NAD), and culture at 37°C After 24h to 48h, the growth was observed, and Gram staining was performed on suspected colonies. Pick a single colony, adopt the method of partitioning and marking on the TSA solid medium, and carry out 3 to 5 times of single colony purification and culture until there is no miscellaneous bacteria.

[0037] 2. Identification of pathogenic bacteria

[0038] 2.1 NAD dependence test

[0039] In a Class II biological safety cabinet, use an inoculation loop to pick up a single colony of the above-mentioned suspicious bacteria, and streak horizontally on a NAD-free TSA plate. Aft...

Embodiment 2

[0058] Embodiment 2 white oil adjuvant inactivated vaccine preparation

[0059] 1. Antigen preparation

[0060] Inoculate Haemophilus parasuis HN1009 strain on TSA solid medium for resuscitation, pass three generations continuously for rejuvenation, inoculate the rejuvenated Haemophilus parasuis in TSB liquid medium, and culture in a shaker at 37°C for about 12h to 14h , as the seed liquid, then inoculate the seed liquid into TSB liquid medium preheated to 37°C according to 1% of the total volume of the medium, 37°C, 180r / min shaking and expanding culture for 14h-16h, and measure the bacterial liquid with a spectrophotometer OD 600 Value, according to the number of viable bacteria and OD 600 Value correlation standard curve formula to calculate the total bacterial content. Then add formaldehyde with a final concentration of 0.2% to the bacterial solution to inactivate the bacteria, place in a 37°C incubator to inactivate for 14h-16h, and shake once every 4h-5h during the in...

Embodiment 3

[0063] Embodiment 3 preparation of aluminum gel adjuvant inactivated vaccine

[0064] Antigen preparation steps are as in Example 2.

[0065] Vaccine preparation:

[0066] Evenly mix the bacteria solution that has passed the inactivation test with 20% sterile aluminum hydroxide saline dilution solution, let it stand for precipitation at room temperature for 24 hours, suck out the supernatant according to 1 / 2 of the total volume, and then add Thimerosal solution with a final content of 1 / 10,000. The content of bacteria in the finished vaccine is 2.5×10 9 CFU / mL, stored at 4°C.

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Abstract

The invention provides a serum 4 type haemophilus parasuis strain HN1009 which is preserved in CCTCC (China Center For Type Culture Collection) with the preservation number of NO: M20-14126. The strain is stable in biological performance, has relatively strong pathogenicity and is used for grafting piglets after being activated, so that the strain has good immunogenicity. The strain which is used as a monovalent vaccine prepared from a vaccine candidate strain is good in safety, can be used for generating relatively high antibodies to the piglets, is long in duration, can be used for resisting attack of homotype wild strains and has a good immune efficacy. In clinical use, the deactivated vaccine is used for effectively reducing occurrence of haemophilus parasuis so as to reduce the economical loss of a pig farm, so that the immune effect reaches or is superior to existing marketed commercialized vaccines.

Description

technical field [0001] The invention relates to the field of biological immunity, in particular to a strain of serotype 4 Haemophilus parasuis and its application. Background technique [0002] Haemophilus parasuis disease, also known as Glaser's disease, is caused by Haemophilus parasuis (Hps). The disease was first reported by German scholar Glasser in 1910, and now it is widely distributed all over the world. At present, Haemophilus parasuis infection is one of the main causes of high mortality of piglets in pig farms. In the United States, the disease has become an important infectious disease that endangers the health of nursery pigs. It ranks first with PRRS and circovirus (PCV), and before influenza, streptococcal and colibacillosis. The disease is currently also a great threat to Chinese pigs. With the prevalence of porcine reproductive and respiratory syndrome (PRRS, blue ear disease), Haemophilus parasuis has caused more serious losses to pigs than before. And o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/11A61K39/102A61P31/04C12R1/21
CPCA61K39/102A61K2039/52A61K2039/521A61K2039/555A61K2039/55505A61K2039/55511A61K2039/55566C12N1/205C12R2001/21
Inventor 李晓成邵卫星吴发兴张志刘爽董雅琴李鹏王佳于美芳
Owner CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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