Sturgeon-source aeromonas hydrophila and application thereof

A technology of Aeromonas hydrophila and sturgeon, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problem of unsatisfactory immune prevention effect of heterogeneous serotype strains, and achieve the prevention of bacterial sepsis in sturgeon , the effect of good marketing prospects

Active Publication Date: 2017-03-15
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, antibiotics have been mainly used to treat sturgeon bacterial septicemia. However, with the emergence of drug resistance of pathogenic bacteria, the quality and safety of aquatic products have become increasingly apparent. The search for new prevention methods has become a hot spot in scientific research.
At present, immunoprophylaxis is an important means to control the outbreak of Aeromonas hydrophila infection in fish. Scholars at home and abroad have done a lot of work in the research and development of the vaccine. The immune prevention effect of serotype strains is not ideal, and it is difficult to promote the use on a large scale. The antigenic diversity of Aeromonas hydrophila has become a major obstacle to the development of effective vaccines

Method used

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  • Sturgeon-source aeromonas hydrophila and application thereof
  • Sturgeon-source aeromonas hydrophila and application thereof
  • Sturgeon-source aeromonas hydrophila and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Isolation and identification of Aeromonas hydrophila HZ8:

[0032] The present invention adopts the method of molecular biology to conduct experimental research on pathogenicity, phenotype analysis, molecular identification and virulence gene detection of eight strains of Sturgeon-derived Aeromonas hydrophila pathogenic bacteria. A phylogenetic tree was constructed based on the gyrB gene sequences of all strains, and the eight strains were all clustered with Aeromonas hydrophila; the distribution pattern of nine virulence genes in the strains was detected by PCR. Only the HZ8 strain contains all tested virulence genes: aerolysin (aer), flagellar gene (fla), heat-stable cell excitatory enterotoxin (ast), heat-unstable cell excitatory enterotoxin (alt), hemolysin (hly), elastase (ahyB), transverse flagellin (laf), enterotoxin (act) and lipase (lip). In addition, taking Sturgeon's sturgeon as the infection object, the half-lethal dose of HZ8 strain was the lowest, and the...

Embodiment 2

[0049] Preparation of immunogen and phagogen

[0050] Preparation of the immunogen:

[0051] Pick a single colony of Aeromonas hydrophila HZ8 in LB medium, culture at 37°C, 200 rpm for 12-16 hours.

[0052] Formalin is added to the cultured Aeromonas hydrophila bacteria solution to a final concentration of 0.3%, and inactivated in a constant temperature incubator at 37° C. for 48 hours, which is the inactivated vaccine of Aeromonas hydrophila. Centrifuge the inactivated vaccine at 5000r / min for 10min to precipitate the bacteria, suspend the bacteria in PBS and then centrifuge to precipitate the bacteria, repeat 3 times. Adjust the bacterial concentration to 1.0×10 8 CFU / ml, which is the immunogen, was stored in a refrigerator at 4°C for use in the following examples.

[0053] In the same way as above, the prepared inactivated Staphylococcus aureus was used as a phagogen, and it was stored in a refrigerator at 4°C for use in the following examples.

Embodiment 3

[0055] Safety detection of Aeromonas hydrophila HZ8 from sturgeon

[0056] (1) Plane sterility testing

[0057] Take 100 μL of the prepared immunogen, spread it on LB solid medium in a sterile operating bench, and incubate it in a constant temperature incubator at 37°C for 72 hours. Observation results show that there is no bacterial growth.

[0058] (2) Test fish safety testing

[0059] Take 30 healthy test fish (body length 15-20 cm), and immunize with the prepared immunogen by intraperitoneal injection, and the immunization dose is 1 ml / tail. Another 30 healthy test fish were intraperitoneally injected with the same dose of PBS as the control group. The test fish were fed normally and observed continuously for 14 days. There was no fish body death or typical symptoms after being infected by Aeromonas hydrophila.

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Abstract

The invention discloses a sturgeon-source aeromonas hydrophila and an application thereof. The collection number of the sturgeon-source aeromonas hydrophila is CCTCC NO:M 2016555. Inactivated vaccine prepared with the sturgeon-source aeromonas hydrophila can prevent sturgeon bacterial septicemia. The quantity of the aeromonas hydrophila HZ8 (Aeromonas HydrophilaHZ8) in the inactivated vaccine is larger than or equal to 1.0*10<8> CFU/ml. The screened sturgeon-source aeromonas hydrophila has the high pathogenicity, the inactivated vaccine prepared with the sturgeon-source aeromonas hydrophila can prevent sturgeon bacterial septicemia, and the sturgeon-source aeromonas hydrophila has good market popularization prospects.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a sturgeon-derived Aeromonas hydrophila and an application thereof. Background technique [0002] Sturgeon is one of the important freshwater aquaculture species in China, and plays an important role in my country's freshwater aquaculture industry. my country's sturgeon farming is mainly concentrated in Shandong, Hubei, Yunnan, Sichuan, Zhejiang, Hunan, Hebei, Guangdong, Guizhou, Fujian, Beijing and other provinces (municipalities). According to statistics, the total national output of sturgeon in 2014 was 76,000 tons, an increase of 17.43% over the output in 2013, ranking third in the value-added output of major freshwater aquaculture fish. Amur sturgeon (Acipenserschrenckii Brandt, 1869) is the main species of sturgeon cultured in northern my country, and has a large wild output. With the continuous expansion of the breeding scale of Strycke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
CPCA61K39/0208A61K2039/521A61K2039/55C12N1/205C12R2001/01
Inventor 周勇曾令兵范玉顶史玉恒
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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