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Aeromonas salmonicida from cold water rainbow trout

A technology for killing Aeromonas salmonella and vaccines, applied in the directions of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of ineffectiveness and reduced immune effect, and achieve the effect of perfect immune protection

Active Publication Date: 2019-07-26
MARINE BIOLOGY INST OF SHANDONG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutant strains have a certain chance to reduce the immune effect of existing vaccines or have no effect at all

Method used

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  • Aeromonas salmonicida from cold water rainbow trout

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Isolation and toxicity analysis of Aeromonas salmonicida strains

[0013] 1) The diseased rainbow trout was obtained from Shandong Rizhao Fish Farm, but the diseased rainbow trout had been immunized with the monocystus vaccine before. The average body weight of the diseased fish was (100±5) g, and the average body length was (17±2) cm. The main symptoms were body surface ulceration, redness at the base of the pelvic fin, protruding anus, and intestinal congestion.

[0014] 2) Take dying diseased fish with typical symptoms, pick some tissues from lesions such as the base of fins and intestinal tract under aseptic conditions, streak them on the MH agar plate, culture them at 28°C for 24 hours, and pick the dominant colonies with the same shape to carry out the test. Purify and cultivate until pure culture bacteria are obtained.

[0015] 3) The isolated pure cultures of each bacterial strain were expanded on the MH agar plate, and after culturing at 28°C for 24...

Embodiment 2

[0022] Example 2: Preparation of Inactivated Vaccine Using Aeromonas salmonicida

[0023] Centrifuge the Aeromonas salmonicida RB0506 strain overnight cultured in MH broth at 6000rpm for 10min at room temperature, resuspend the bacterial pellet with normal saline (0.9% NaCl solution), then collect the precipitate by centrifugation, repeat twice, and finally resuspend with normal saline And adjust the bacterial concentration to 1.5×10 9 Individuals / mL, distributed in 50mL centrifuge tubes, 30mL per tube, using 0.1% formaldehyde solution to inactivate Aeromonas salmonicida for 5 hours at 28°C to prepare an inactivated vaccine antigen suspension.

[0024] Take 19ml of the above-mentioned inactivated vaccine antigen suspension, add 1ml of sterilized Tween-80 and mix well to prepare the aqueous phase of the emulsified vaccine; then use 38.4ml of vaccine white oil, 1.6ml of Span-80, 0.6g of aluminum stearate Mix the phases, heat to completely dissolve the aluminum stearate, and ste...

Embodiment 3

[0025] Embodiment 3: the effect detection of vaccine

[0026] According to the steps of preparing the vaccine in step 2, the Aeromonas salmonicida CICC 23564 strain and the Aeromonas salmonicida E11I4 strain (CGMCC1.16015) were used as antigens to prepare a vaccine,

[0027] Adjust the 3 inactivated vaccine concentrations to OD 600 0.1±0.01, healthy rainbow trout were immunized by intraperitoneal injection, 100 μL per fish, and the control group was injected with the same volume of sterile saline, 20 fish in each group, with 4 repetitions in each group. Breeding water temperature was 17±1°C, inflated throughout the day, and 50% of the water was changed every day; 40 days after immunization, three strains of monocytocidal bacteria were used to challenge the virus, and the intraperitoneal injection concentration of each tail was 5.0×10 8 CFU / mL bacterial solution was 100 μL; the number of dead fish in each experimental group was regularly observed and recorded every day, and th...

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Abstract

The invention provides aeromonas salmonicida from cold water rainbow trout. The preservation number of the aeromonas salmonicida is CCTCC NO: M2018849. The aeromonas salmonicida is used for preparinga vaccine. The aeromonas salmonicida is obtained by screening pathogenic cold-water rainbow trout, and compared with existing aeromonas salmonicida, the strain has variation and is used for preparinga vaccine, and accordingly more perfect immune protection is provided for rainbow trout.

Description

technical field [0001] The invention belongs to the technical field of screening pathogenic microorganisms, and in particular relates to Aeromonas salmonicida derived from cold-water rainbow trout. Background technique [0002] Rainbow trout is a cold-water fish belonging to the family Salmonidae. It is one of the most widely farmed freshwater fishes in the world. Because rainbow trout is most suitable for artificial intensive farming, it has the characteristics of high yield, high efficiency, and a broad market. Therefore, rainbow trout farming is developing rapidly, and the scope of farming area and farming output are increasing year by year. [0003] However, with the expansion of the breeding scale and the gradual increase of the breeding density, many disease problems have appeared in the breeding process of cold-water rainbow trout. In recent years, bacterial diseases have occurred frequently in the breeding process of rainbow trout, which has caused a lot of problems ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
CPCA61K39/0208A61K2039/521A61K2039/552A61P31/04C12N1/205C12R2001/01
Inventor 刁菁李乐于晓清王晓璐许拉樊英盖春蕾王淑娴叶海斌
Owner MARINE BIOLOGY INST OF SHANDONG PROVINCE
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