Blood serum 5 type haemophilus parasuis and application thereof

A technology of Haemophilus suis and Haemophilus suis disease, which is applied in the direction of bacteria, antibacterial drugs, and medical raw materials derived from bacteria, can solve the problems of farmers increasing the amount of antibiotics, pig products exceeding the standard of antibiotics, and endangering health. Achieve stable biological performance, strong pathogenicity, and reduce economic losses

Inactive Publication Date: 2015-03-04
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

Method used

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  • Blood serum 5 type haemophilus parasuis and application thereof
  • Blood serum 5 type haemophilus parasuis and application thereof
  • Blood serum 5 type haemophilus parasuis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0034] Example 1 Isolation and identification of serotype 5 Haemophilus parasuis

[0035] 1. Isolation of pathogenic bacteria

[0036] Use a sterile inoculation loop to sample fresh lungs, pericardial effusion, joint fluid and other disease materials of suspected pigs, streak inoculation on TSA medium (containing final concentration of 5% serum and 0.001% NAD), culture at 37°C Observe the growth after 24h~48h, and perform Gram staining microscopy on the suspected colonies. Pick a single colony, and use the partition streaking method on the TSA solid medium to perform 3 to 5 single colony purification cultures until there are no other 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 single colonies of the above suspicious bacteria, horizontally streak it on a TSA plate without NAD, pick out Staphylococcus aureus and streak it vertically, and incubate at 37°C ...

Example Embodiment

[0058] Example 2 Preparation of white oil adjuvant inactivated vaccine

[0059] 1. Antigen preparation

[0060] Inoculate Haemophilus parasuis JX1002 strain on TSA solid medium for resuscitation, pass 3 consecutive generations to rejuvenate, inoculate the rejuvenated Haemophilus parasuis in TSB liquid medium, and cultivate in a shaker at 37°C for 12h-14h. As the seed solution, inoculate the seed solution in TSB liquid medium preheated to 37℃ by 1% of the total volume of the culture medium. Then, at 37℃, 180r / min shaking and expanding for 14h-16h, and measuring the bacterial solution by spectrophotometer OD 600 Value, based on 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, and place it in a 37°C incubator to inactivate the bacteria for 14 to 16 hours. During the inactivation period, shake once eve...

Example Embodiment

[0063] Example 3 Preparation of aluminum gel adjuvant inactivated vaccine

[0064] The antigen preparation steps are as in Example 2.

[0065] Vaccine preparation:

[0066] Mix the qualified bacterial solution in inactivation test with the same amount of 20% sterile aluminum hydroxide saline diluent, let stand at room temperature for 24 hours, aspirate the supernatant at 1 / 2 of the total volume, and then add A solution of thimerosal with a final content of one part in 10,000. The content of bacteria in the finished vaccine is 2.0×10 9 CFU / mL, store at 4°C.

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Abstract

The invention provides a blood serum 5 type haemophilus parasuis strain JX1002. The blood serum 5 type haemophilus parasuis strain JX1002 is collected in China Center for Type Culture Collection (CCTCC), and the collection number is CCTCC M 2014127. The strain provided by the invention is stable in biological performance, has very strong pathogenicity to piglets, and has very good immunogenicity when the strain is inoculated to the piglets after inactivation. A monovalent vaccine prepared by using the strain as 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 the attack of wild strains with the same type, and has very good immune efficiency. An inactivated vaccine can be used for effectively reducing the occurrence of diseases and reducing the economic loss of a pig farm in clinical use, and the immune effect of the inactivated vaccine can reach or exceed that of existing commercial vaccines in markets.

Description

technical field [0001] The invention relates to the field of biological immunity, in particular to a strain of Haemophilus parasuis serotype 5 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): C12N1/20A61K35/74A61K39/102A61P31/04C12N15/11C12R1/21
CPCA61K39/102A61K2039/55505A61K2039/55566C12N1/205C12R2001/21
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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