Nocardia seriolae induced low virulent strain and application thereof

A technology of Nocardia amberjack and Nocardia, which is applied in the directions of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problem that the development process of fish Nocardia vaccine is not ideal and the development route of inactivated vaccine Loss of feasibility and inability to provide immune protection, etc., to eliminate the possibility of spreading a large number of virulent pathogens, achieve significant immune effects, and achieve good control effects

Active Publication Date: 2014-12-24
SHENZHEN INST OF GUANGDONG OCEAN UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Worldwide, however, the development of vaccines against Nocardia spp. for fish has been suboptimal because, unlike other fish pathogens, it has been found that multiple inactivated vaccines against Nocardia spp. effect, which makes the development route of inactivated vaccines unfeasible

Method used

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  • Nocardia seriolae induced low virulent strain and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation of embodiment 1 attenuated live vaccine

[0022] (1) Mutagenesis, screening and identification of a mutagenic attenuated strain of Nocardia japonica

[0023] 1. Preparation of Nocardia wild strain ZJ0503 bacterial suspension from yellowtail

[0024]In March 2005, our laboratory isolated a wild yellowtail Nocardia strain ZJ0503 (Huang Yucong et al. , Isolation and Identification of the Pathogen of Sarcoidosis in Pompano ovata, Journal of Guangdong Ocean University, 2008, 28(4): 49-53). The amberjack Nocardia ZJ0503 strain stored in a -80°C refrigerator was streaked on BHI solid medium, cultured upside down at a constant temperature of 28°C for 7 days, a single colony was picked and placed in a sterile phosphate buffer, and mixed to prepare bacterial suspension.

[0025] 2. UV mutagenesis

[0026] Pipette 0.1ml of the ZJ0503 bacterial suspension and spread it on the modified solid medium plate of Nocardia amberii containing 0.3%~0.5% lithium chloride (...

Embodiment 2

[0052] Example 2 Detection of the pathogenicity of the mutagenized attenuated strain NSX1 of Nocardia japonica

[0053] Under laboratory conditions, zebrafish was used as a model animal to detect the toxicity of the attenuated strain NSX1 of Nocardia amberii obtained through mutagenesis screening. The experimental zebrafish (average body length 2.5-3cm, body weight about 0.2g) was first placed in a clean water tank to adapt to breeding for 1 week to eliminate abnormal individuals, and the mortality rate should be less than 2%. Before the infection test, the test fish were randomly divided into two water tanks in each group, and 50 fish were cultured in each tank (20L). The test water tank is replaced with 1 / 2 volume of aquaculture water with tap water that has been oxygenated for more than 24 hours every day, and the water temperature is 28±2°C. In the infection test, each group of test fish was treated with a certain gradient dose (10 7 ~10 10 CFU / ml) of Nocardia japonica ...

Embodiment 3

[0057] Example 3 Evaluation of the immune protection rate of the mutagenized attenuated strain NSX1 of the yellowtail Nocardia

[0058] Zebrafish were used as experimental animals for the immune protection test. The zebrafish were randomly divided into 3 groups, each group had 3 parallel water tanks, 16 fish / box. The attenuated strain NHX1 was used as a live vaccine to immunize zebrafish by intraperitoneal injection, and the immunization dose was 10 7 CFU / tail. The control group was injected with sterile phosphate buffer. After 14 days of immunization, the zebrafish in each group were artificially infected and challenged with the wild strain ZJ0503 of Nocardia japonica, and the challenge dose was 10 6 CFU / tail. The number of deaths in the control group and the immune group was observed and counted within 14 days, and the immune protection rate of each group was calculated (see Table 4).

[0059] Among them, the immune protection rate is calculated according to the followi...

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Abstract

The invention relates to a nocardia seriolae induced low virulent strain and an application thereof, and belongs to the technical field of bacterial strain induced mutation. Particularly, the nocardia seriolae induced low virulent strain, named NSX1 (Nocardia seriolae NSX1), of which the culture preservation number is CGMCCNO.9437. An immune compound comprises the nocardia seriolae induced low virulent strain used as an immune an immunogenicity component, wherein the concentration of the nocardia seriolae induced low virulent strain is 10<3>-10<9>CFU / ml; the nocardia seriolae induced low virulent strain is used for preparing drugs for resisting fish nocardiosis and has significant low toxicity effect; meanwhile, the fish can be effectively protected from the invasion of nocardia seriolae caused by nocardia seriolae pathogenic strains or virulence strains; therefore, the nocardia seriolae induced low virulent strain has significant immune effect and good nocardia seriolae disease control effect.

Description

technical field [0001] The invention relates to a mutagenesis of fish Nocardia and its application, in particular to a mutagenic attenuated strain of Nocardia amberjack and its application, and belongs to the technical field of strain mutagenesis. Background technique [0002] Fish nocardiosis is one of the common diseases in pond culture, floating raft cage culture and deep water cage culture mode. It is a chronic systemic granulomatous disease. Nocardia seriolae ) is the main pathogenic bacteria. The disease can form a large number of nodules in the body and surface of the fish, and can also cause bleeding and ulcers on the surface of the fish, which seriously affects the commercial value of adult fish. Its infection rate and mortality are high, and the natural incidence rate can reach 15~ 30%, up to 50-60% in severe cases, the average mortality rate is about 20%, and the mortality rate of artificial infection can be as high as 90%-100%. In recent years, fish nocardiosis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
Inventor 夏立群陈建林鲁义善简纪常蔡佳张红莲
Owner SHENZHEN INST OF GUANGDONG OCEAN UNIV
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