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Aeromonas salmonicida and edwardsiella tarda bivalent vaccine and application thereof

A technology for killing Aeromonas salmonicum and Edwardsiella tarda, which is applied in the field of Aeromonas salmonicum killing and Edwardsiella tarda dual vaccine, can solve the problems of waste of manpower and material resources, no turbot scabies and ascites diseases, etc. , to reduce environmental hazards, eliminate residual hidden dangers, and enhance immune response.

Active Publication Date: 2021-10-08
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the fish diseases caused by Aeromonas salmonicida and Edwardsiella tarda are vaccines with a single antigen, and there is no vaccine against turbot scabies and scabies caused by Aeromonas salmonicida and Edwardsiella tarda. Effective Vaccines for Ascites Disease
In addition, in order to achieve the prevention and control of diseases caused by multiple pathogenic bacteria, the existing single-antigen vaccines also need repeated immunization against diseases caused by two pathogenic bacteria. The secondary and mixed infection of the pathogens of Monascus and Edwardsiella tarda urgently requires the development of a new vaccine to solve the defects in the prior art that a single antigen is used to prevent and treat fish diseases caused by the above two pathogens

Method used

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  • Aeromonas salmonicida and edwardsiella tarda bivalent vaccine and application thereof
  • Aeromonas salmonicida and edwardsiella tarda bivalent vaccine and application thereof
  • Aeromonas salmonicida and edwardsiella tarda bivalent vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and identification of Aeromonas salmonicida and E. tarda embodiment

[0030] 1 strain

[0031] Aeromonas salmonicida and tarda, from a farm in Shandong diseased fish separate.

[0032] 2, method

[0033] 2.1 Screening of vaccine strains

[0034] The prevalence of a turbot farm from Yantai, Shandong, diseased fish mainly for the abdominal part of the red area, body surface ulcers, liver local redness. With saline diseased turbot washed three times, the removal of kidney tissue of diseased fish in aseptic technique. Normal saline tissue block is placed in a sterile 2.0ml centrifuge tube, cut into pieces and a small amount streaked on TSB plates, 28 deg.] C incubation 24h. After colonies grow, the color of the same size were picked single colonies were identified. After delivery company sequenced, and the resulting sequence homology in the NCBI site sequence alignment analysis.

[0035] 2.2 Identification of results

[0036] 1, the results of morphological ...

Embodiment 2 2

[0044] Example 2 Preparation of two vaccines

[0045] After the above-mentioned murderous mono cyclasmia is activated, the scribe is inoculated on the LB solid petriper, 28 ° C culture 24 h, and picks up a single bacterius to the LB liquid medium culture for 24 h, and send the company to sequencing and identifying. After the monohydrococcus, the bacterium in the medium was collected from centrifugation at 4 ° C, and the precipitate was cleaned by sterile PBS (pH 7.3), and finally containing 0.5% formaldehyde (V / V). The PBS was resuspended and placed at 4 ° C for 24 h. The next day, a small portion of the solution was washed three times with a sterile PBS. After the coating plate was determined, the precipitate was collected by centrifugation (4 ° C) at 5000 r / min (4 ° C), removed, and washed 3 times with Sterilization. , And heavy suspension, the concentration is about 1 × 10 9 CFU / mL, the resulting solution is a mild splita vaccine. The same method was used to get slowed Ed...

Embodiment 3

[0047] Example 3 Immune animal experiment

[0048] 1, immunization method

[0049] Choose 300 tail healthy tailings, divided into vaccine groups and control groups, each group of 150 tail fish. Immunization in the abdominal injection. The vaccine group is injected with a concentration of 0.1 ml of a concentration of 1 × 10, respectively. 9 The inactivated vaccine of CFU / mL was injected with 0.1 ml of sterile PBS in the control group.

[0050] 2. Determination of acid phosphatase (ACP) and lysozyme (LZM) vitality during immunized serum

[0051] The blood samples were performed on Daxue (1, 2, 3, 4w) before immunization (1, 2, 3, 4w), each of which were randomly taking 9 tail fish each time. After standing at room temperature for 2 h, 4 ° C, 3000r / min centrifuge 15min, take the supernatant, according to the corresponding step of the kit (Nanjing Implementation Biological Engineering Research Institute), using the kit to measure the acid phosphatase in serum Activity to lysozyme....

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Abstract

The invention discloses an aeromonas salmonicida and edwardsiella tarda bivalent vaccine and application thereof, and belongs to the technical field of biological products. A vaccine strain comprises aeromonas salmonicida and edwardsiella tarda, the aeromonas salmonicida and the edwardsiella tarda are preserved in the China Center for Type Culture Collection on April 19, 2021 and April 21, 2021 respectively, the preservation numbers are CCTCC NO: M 2021397 and CCTCC NO: M2021422 respectively, and the preservation addresses are Wuhan University. The antigens of the bivalent vaccine are the aeromonas salmonicida and the edwardsiella tarda. The antibody titer of the bivalent vaccine is 1:210, and the bivalent vaccine can effectively prevent and treat turbot diseases caused by aeromonas salmonicida and edwardsiella tarda, and provides reference for development of fish vaccines and prevention and control of various diseases in aquaculture.

Description

Technical field [0001] The present invention relates to the technical field of biological products, to Aeromonas salmonicida and E. tarda bivalent vaccine and its application. Background technique [0002] Turbot (Scophthalmus maximus) is native to the cold water benthic marine fish in Europe, introduced into my country in 1992. Because of its nutrient-rich, high economic value, in northern my country has gradually become an important economic farmed fish. With the development of intensive farming, large-scale, turbot frequent disease, a serious impact on the sustainable development of the industry. [0003] Boils disease and ascites disease is more common in aquaculture bacterial diseases, which kill salmon Aeromonas is one of the main pathogens boils disease, and kill salmon Aeromonas (Aeromonas salmonicida) was first proposed by Emmerich and Weible in 1890 isolated and described from trout. Bacteria can cause salmon trout fish boils disease, the prevalence of red fish body sur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02A61K39/39A61P31/04A61P37/04
CPCA61K39/0208A61K39/025A61K39/39A61P31/04A61P37/04A61K2039/70A61K2039/552A61K2039/521A61K2039/54A61K2039/55566Y02A40/81
Inventor 周顺修云吉郭慧敏李影苏琳
Owner QINGDAO AGRI UNIV
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