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Vibrio harveyi inactivated vaccine, and preparation method and application thereof

A technology of vibrio harveyi and inactivated vaccines, which is applied in the field of aquatic vaccinology to achieve the effect of simple preparation, no safety hazard, and significant immune protection effect

Inactive Publication Date: 2018-12-18
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prevention and treatment of Vibrio harveyi is still based on antibiotics and chemical drugs, and there is no effective vaccine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Preparation of Vibrio harveyi inactivated vaccine.

[0018] step 1)

[0019] The Vibrio harveyi T4 is preserved in CGMCC, General Microorganism Center of China Microbiological Culture Collection Management Committee. The address of the preservation unit is No. 13, North Zhongguancun, Haidian District, Beijing. The preservation number is: CGMCC No.1985, and the preservation date is March 22, 2007. Classification named Vibrio harveyi (Vibrio harveyi).

[0020] Step 1) Proliferation and cultivation of Vibrio harveyi. Vibrio harveyi was resuscitated and inoculated on LB plates, and a single colony was picked and passed on for 2 times. The single colony was picked for Gram staining and microscopic examination to observe its morphology and stainability to initially verify the strain, and the 16S rDNA of the bacteria was further analyzed by colony PCR. Sequencing identification.

[0021] Inoculate the identified correct strains into 6ml liquid LB medium and culture at 28°C ...

Embodiment 2

[0027] Immunization application of inactivated Vibrio harveyi vaccine.

[0028] Step 1) Vaccine injection immunization. 60 turbots (each weighing about 15g) were randomly divided into 2 groups, 30 in each group. These 2 groups are named as Group A and Group B, respectively. Each fish in Group A was injected intraperitoneally with 100ul of the Vibrio harveyi vaccine mixture in step 3) of Example 1 above, and each fish in Group B was injected intraperitoneally with 100ul of the control solution in Step 3) of Example 1 above.

[0029] Step 2) Preparation of Vibrio harveyi suspension for challenge. Pick a single colony of Vibrio harveyi and inoculate it in LB liquid medium, cultivate to OD 600 0.8-1.0, and then centrifuged (5000g, 4°C) for 5min. Collect the cells and suspend them in PBS to a final concentration of 5x10 7 cfu / ml.

[0030] Step 3) Detection of vaccine immune protection effect. On the 30th day after the immunization injection in step 1), the 2 groups of fish i...

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PUM

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Abstract

The invention relates to the field of aquatic product vaccinology, and concretely relates to a Vibrio harveyi inactivated vaccine, and a preparation method and an application thereof. The preparationmethod comprises the following steps: inactivating a cultured Vibrio harveyi strain T4 by formaldehyde for 24 h, re-suspending the inactivated strain in PBS until the final concentration is 8.0 * 10<9> cfu / ml, and mixing and emulsifying the obtained mixture with an adjuvant to obtain the Vibrio harveyi inactivated vaccine. The immunoprotection efficiency of the obtained inactivated vaccine againstVibrio harveyi infection can reach 85% or above. The vaccine obtained in the invention has the advantages of simple preparation process, no extraction or purification, no hidden safety hazards, and achieving high-efficiency and long-term immunoprotective effect by one-time immunization.

Description

technical field [0001] The invention relates to the field of aquatic vaccinology, in particular to an inactivated vaccine of Vibrio harveyi and its preparation and application. Background technique [0002] After decades of continuous growth, China has become the world's largest aquaculture country. However, with the expansion of factory farming scale, aquatic diseases occur frequently, seriously restricting the development of aquaculture industry. At present, the main means of prevention and treatment of bacterial diseases in China is the use of antibiotics, but the extensive use of antibiotics has caused drug residues in aquatic products and bacterial resistance, disrupting the balance of the ecological environment and posing potential hazards to human health. As a means of preventing and controlling aquatic animal diseases, vaccines are superior to antibiotics and other drug treatments in terms of specificity, safety and protective effect, and are widely accepted and use...

Claims

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Application Information

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IPC IPC(8): A61K39/106A61P31/04C12N1/20C12R1/63
CPCA61K39/107A61K2039/521A61K2039/55A61P31/04C12N1/20C12N1/205C12R2001/63
Inventor 张健孙黎
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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