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Infectious actinobacillus pleuropneumoniae and application thereof

A technology of pleuropneumonia and actinobacillus, which is applied in the direction of bacteria, antibacterial drugs, veterinary vaccines, etc., can solve the problems of lack of cross-immune protection and achieve good protection effect, strong pathogenicity, and good immunogenicity

Active Publication Date: 2017-09-26
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are as many as 15 serotypes of Actinobacillus pleuropneumoniae, and there are still many clinically isolated strains that cannot be typed. There is a lack of effective cross-immune protection among the serotypes, and the dominant serotypes that are popular in different countries, regions and pig farms are different. Therefore, the isolation, identification and typing of APP are particularly important in the prevention and treatment of this disease.

Method used

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  • Infectious actinobacillus pleuropneumoniae and application thereof
  • Infectious actinobacillus pleuropneumoniae and application thereof
  • Infectious actinobacillus pleuropneumoniae and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1, Isolation and identification of infectious Actinobacillus pleuropneumoniae

[0022] 1.1 Disease data collection

[0023] The disease material came from a 5-month-old fattening pig that died of severe pneumonia and dyspnea in a large pig farm in Ruzhou City, Henan Province in November 2016.

[0024] 1.2 Preparation of medium

[0025] TSB liquid medium: Dissolve 30 g of TSB broth powder (Tryptic Soy broth, tryptone soybean broth) in 1000 mL of ultrapure water, sterilize at 115 °C for 15 min, add 50 mL of fetal bovine serum, sterile 1% NAD (Nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide, coenzyme Ⅰ) 1mL.

[0026] TSA solid medium: Dissolve 40 g of TSA agar powder (Tryptic Soy Agar, tryptone soybean agar) in 1000 mL of ultrapure water, sterilize at 115 ° C for 15 min, add 50 mL of fetal bovine serum, sterile 1% NAD (add as needed ) 1 mL; store at 4°C for future use.

[0027] 1.3 Isolation and culture of bacteria

[0028] Aseptically col...

Embodiment 2

[0050] Embodiment 2, the preparation of vaccine, safety and efficacy test

[0051] 2.1 Preparation of vaccine

[0052] Inoculate the infectious Actinobacillus pleuropneumoniae strain HNAPP1 into the TSB liquid medium, culture it in a shaker at 37°C, collect the culture after 18 hours, measure the concentration, and adjust the number of bacteria in the culture to 2×10 9 CFU / mL, add formaldehyde to a final concentration of 0.2%, inactivate at 37°C for 12h. The inactivated culture is added with aluminum hydroxide adjuvant at a volume ratio of 1:1, and mixed to prepare an inactivated vaccine of infectious Actinobacillus pleuropneumoniae.

[0053] 2.2 Vaccine safety test:

[0054] Ten healthy nursery pigs aged 2 months were selected and divided into 2 groups with 5 pigs in each group. Vaccine group: intramuscular injection of 4 mL of this vaccine in the neck; control group: intramuscular injection of 4 mL of sterilized normal saline in the neck. The body temperature of the anim...

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Abstract

The invention discloses an infectious actinobacillus pleuropneumoniae and application thereof. The infectious actinobacillus pleuropneumoniae is an actinobacillus pleuropneumoniae HNAPP1 with a preservation number of CGMCC NO:13333 and preserved on November 22, 2016 in the China General Microbiological Culture Collection Center (3#, 1# Yard, West Beichen road, Chaoyang District, Beijing). The strain HNAPP1 provided by the invention is separated from a heart of an acute-death fattening pig with typical dyspnea, no other bacteria grow but only the infectious actinobacillus pleuropneumoniae grows during separation on a TSA solid culture medium, and the proliferation titer in a TSB liquid culture medium is high and reaches 109 CFU / mL or above. The strain HNAPP1 in the invention has strong pathogenicity for nursery pigs to cause diseases and death to the nursery pigs, and has high immunogenicity.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a strain of infectious actinobacillus pleuropneumoniae and its application. Background technique [0002] Actinobacillus Pleuropneumoniae (APP) is the pathogen that causes porcine infectious pleuropneumonia (Porcine infection Pleuropneumonia), which is a highly contagious infectious respiratory disease of pigs. , hemorrhagic and necrotizing pneumonia, the incidence rate is between 8.5% and 100%, and the mortality rate is between 0.4% and 100%. It is one of the main reasons for the death of fattening pigs and breeding pigs. Therefore, the control of infectious Actinobacillus pleuropneumoniae is of great significance to the healthy development of pig industry. [0003] There are as many as 15 serotypes of Actinobacillus pleuropneumoniae, and there are still many clinically isolated strains that cannot be typed. There is a lack of effective cross-immune protection among t...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/102A61P31/04C12R1/01
CPCA61K39/102A61K2039/521A61K2039/552C12N1/205C12R2001/01
Inventor 徐引弟李海利王治方张青娴郎利敏朱文豪焦文强王克领张立宪游一
Owner INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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