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A kind of highly pathogenic Aeromonas hydrophila and its application

A technology of Aeromonas hydrophila and ZYAH72, applied in bacteria, antibacterial drugs, veterinary vaccines, etc., to reduce economic losses, enhance immunity, and prevent bacterial sepsis

Active Publication Date: 2018-05-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, several Aeromonas hydrophila inactivated vaccines have been prepared and proved to have partial immune protection against specific strains, but because Aeromonas hydrophila strains are divided into pathogenic and nonpathogenic strains, the virus There are many combinations of virulence factors, so it is generally believed that only by screening highly virulent strains with multiple virulence factors for the preparation of fire-fighting vaccines can better protect aquatic organisms against infection by different pathogenic strains of Aeromonas hydrophila

Method used

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  • A kind of highly pathogenic Aeromonas hydrophila and its application
  • A kind of highly pathogenic Aeromonas hydrophila and its application
  • A kind of highly pathogenic Aeromonas hydrophila and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Obtainment of a highly pathogenic Aeromonas hydrophila:

[0032] (1) Screening of highly pathogenic Aeromonas hydrophila with multiple virulence factors

[0033] A. Dissect diseased or dying fish in the ultra-clean bench, streak the diseased part onto TSB solid medium, cultivate at 28°C for 18h-24h, pick a single colony on the plate and transfer it to pure culture on fresh medium ; The typical colony on TSB solid medium is round, light yellow, translucent, smooth surface, moist colony with neat edges.

[0034] B. Take the pure cultured single colony on the above plate, carry out biochemical identification with the API bacterial identification reagent strip of Mérieux, France (see Table 1), and carry out Gram staining to observe the colony morphology.

[0035] Table 1: Physiological and biochemical identification results of some isolates

[0036]

[0037] C. The 16S rDNA of the candidate strain was amplified and sequenced, and the homology comparison with the existi...

Embodiment 2

[0054] Recovery and growth curve of Aeromonas hydrophila ZYAH72:

[0055] Cultivation of Aeromonas hydrophila ZYAH72: Take out the strain stored in glycerin from the -80 degree refrigerator, streak it on the TSB solid medium, cultivate it in a 28°C incubator for about 24 hours, and then pick a single colony on the plate Streak culture on fresh TSB solid medium, after subculture and recovery twice, pick a single colony from TSB solid medium and inoculate it in TSB liquid medium, then culture at 28°C, 180r / min shaker, every 1 Hourly measure its OD 600nm Absorbance value, the same operation is applied to the pathogenic standard strain, the results show that both ZYAH72 and the pathogenic standard strain enter the logarithmic phase at about 2h, enter the plateau phase at 4h-5h, and contain about 10 8 ~10 9 CFU / mL, the absorbance value of ZYAH72 will be higher than that of the pathogenic standard strain after entering the stationary phase, indicating that the growth rate of the i...

Embodiment 3

[0057] Preparation of Aeromonas hydrophila ZYAH72 inactivated vaccine:

[0058] Selection of optimal formaldehyde concentration:

[0059] Add formaldehyde solutions with a final concentration of 0.1%-0.8% (volume fraction) to the cultured bacteria solution to inactivate the highly pathogenic Aeromonas hydrophila, and continue culturing in a shaker at 28°C for 12 hours , different inactivated vaccines were coated on TSB solid medium, and the results showed that complete inactivated vaccines could be prepared when the formaldehyde concentration was 0.3%-0.8%. %, the antigen loss of the inactivated vaccine is the smallest, and the inactivated vaccine prepared with this formaldehyde concentration is used as the immune vaccine.

[0060] Preparation of inactivated vaccine:

[0061] The isolated highly pathogenic Aeromonas hydrophila ZYAH72 was rejuvenated for 2 generations on the Columbia blood plate, and a single colony on the blood plate was picked and inoculated in TSB medium, ...

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Abstract

The invention discloses a highly pathogenic Aeromonas hydrophila and its application. The highly pathogenic Aeromonas hydrophila is Aeromonas hydrophila (Aeromonas hydrophila) ZYAH72, preservation number: CCTCC NO :M2015797. The Aeromonas hydrophila is prepared into an inactivated vaccine, and the obtained vaccine is safe and non-toxic, and can obtain better immune effects through soaking and injection, and has good broad-spectrum. The preparation method of the vaccine is fast and simple, and it is easy to prepare in large quantities. The invention provides a practical method for the prevention and treatment of aquatic bacterial sepsis.

Description

technical field [0001] The invention relates to the technical field of animal vaccines, in particular to a highly pathogenic Aeromonas hydrophila and its application. Background technique [0002] Aeromonas hydrophila is a gram-negative bacterium belonging to the family Aeromonas, belonging to the genus Aeromonas. The bacteria can infect fish, amphibians, mollusks, and even birds, reptiles, mammals and other animals, and the infection or bacterial septicemia caused by it can lead to a large number of animal deaths. Aeromonas hydrophila is an important acute infectious disease with the largest number of fish species, the largest fish age range, the widest distribution area and the longest epidemic season in the history of fish farming in my country. At the same time, the infection caused by this pathogen is widely distributed around the world, bringing economic losses to the major aquaculture countries. Studies have found that once the fish is infected with Aeromonas hydrop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
CPCA61K39/0208A61K2039/521A61K2039/552C12N1/205C12R2001/01
Inventor 周洋李锦铨林蠡赵丽娟王小红李智喻蔚
Owner HUAZHONG AGRI UNIV
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