A vibrio anguillarum divalent vaccine, a preparing method thereof and a using method of the vaccine

A technology of bivalent inactivated vaccine and Vibrio anguillarum, which is applied in the direction of antibacterial drugs, pharmaceutical formulations, bacterial antigen components, etc., can solve the problems of low protection and inability to provide complete protection for aquaculture animals, and achieve good safety, The effect of widening the scope of protection and low production cost

Active Publication Date: 2014-11-12
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a bivalent vaccine for Vibrio anguillaris and its preparation and use method to overcome the shortcomings of existing Vibrio anguillarum vaccines that cannot provide complete protection and low protection for aquaculture animals

Method used

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  • A vibrio anguillarum divalent vaccine, a preparing method thereof and a using method of the vaccine
  • A vibrio anguillarum divalent vaccine, a preparing method thereof and a using method of the vaccine
  • A vibrio anguillarum divalent vaccine, a preparing method thereof and a using method of the vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Pathogenicity of Vibrio anguillaris to flounder (Paralichthys olivaceus)

[0024] 1) The median lethal dose (LD) of flounder infected by Vibrio anguillarum injection 50 )

[0025] Healthy test fish with a body weight of 10-12g are temporarily raised in a 1000L aquarium, equipped with a flowing circulating water treatment and purification system, and the temperature of the breeding water is maintained at 18-20°C. After 4 weeks of temporary breeding, they were randomly divided into 40L experimental aquariums, with 20 fish in each group, and continued to be temporarily raised for 2 weeks. In the test aquarium, the water was changed once a day in the morning and afternoon, the amount of water change was 1 / 2, and the water temperature was maintained at 18-20°C.

[0026] In the artificial infection test, live Vibrio anguillarum CGMCC No.6942 or CGMCC No.6944 cells were diluted with sterile PBS buffer solution to a serial concentration of 10 5 -10 9 Bacterial su...

Embodiment 2

[0036] Example 2: Pathogenicity of Vibrio anguillaris to turbot (Scophthalmus maximus)

[0037] 1) The median lethal dose (LD) of turbot infected by Vibrio anguillarum injection 50 )

[0038] Healthy test fish with a body weight of 9-12g are temporarily raised in a 1000L aquarium, equipped with a flowing circulating water treatment and purification system, and the temperature of the breeding water is maintained at 16-18°C. After 4 weeks of temporary raising, they were randomly divided into 30L experimental aquariums, with 10 fish in each aquarium, and continued to be temporarily raised for 2 weeks. In the test aquarium, the water was changed once a day in the morning and afternoon, the amount of water changed was 1 / 2, and the water temperature was maintained at 16-18°C.

[0039] In the artificial infection test, live Vibrio anguillarum CGMCC No.6942 or CGMCC No.6944 cells were diluted with sterile PBS buffer solution to a serial concentration of 10 5 -10 9 cfu / ml bacterial...

Embodiment 3

[0049] Embodiment 3: Preparation of Vibrio anguillarum inactivated vaccine

[0050] A. Take Vibrio anguillarum CGMCC No.6942 and CGMCC No.6944 and inoculate them into common broth medium containing 1.5% NaCl (w / v), respectively, and culture them in shake flasks at 25-30°C for 24-36 hours ; Add formaldehyde with a final concentration of 0.3-0.5% to the obtained cultures, and place them in shake flasks at 25-30° C. for 24-36 hours;

[0051] B. Centrifuge the cultures obtained above at a speed of 4000-5000rpm / min for 10-20min, respectively collect the precipitated bacteria, and then use PBS phosphate buffer solution to suspend the sediment respectively, and adjust the bacterial cell concentration to ~10 9 cells / ml, respectively Vibrio anguillarum strain CGMCC No.6942 inactivated vaccine and Vibrio anguillarum CGMCC No.6944 inactivated vaccine.

[0052] C. Mix the inactivated cells of Vibrio anguillarum CGMCC No.6942 and CGMCC No.6944 in a sterile phosphate-buffered saline soluti...

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Abstract

The invention relates to aquaculture animal disease control technologies, and particularly relates to a vibrio anguillarum divalent vaccine, a preparing method thereof and a using method of the vaccine. The vaccine is a bacterial suspension mixture of a vibrio anguillarum 01 serotype and a 02 serotype. The vibrio anguillarum 01 serotype and the 02 serotype are deposited in the general microbiology center of the China Committee for Culture Collection of Microorganisms on December 7, 2012 and respectively have an accession number namely CGMCC No.6942 and CGMCC No.6944. The divalent inactivated vaccine can protect susceptible fishes from being infected by pathogenic vibrio anguillarum. The divalent vaccine broadens the vaccine protection range, effectively irritates cultured fishes to generate specific immune response, and effectively provides immune protection for diseases. Preparation of the divalent vaccine is low in cost of used materials and safe to animals and the environment.

Description

technical field [0001] The invention relates to the disease prevention technology of aquaculture animals, in particular to a bivalent vaccine of Vibrio anguillarum and its preparation and use method. Background technique [0002] Vibrio anguillarum is one of the important pathogens causing sepsis in aquaculture animals. The vibriosis caused by it is the main disease of aquaculture fish in my country. The infected farmed fish involve turbot, flounder, half smooth tongue sole, perch, American Important cultured species such as redfish have caused significant economic losses to the cultured fish industry. The main methods of prevention and control of vibriosis in eels are drug therapy and vaccine immunization. Drug treatment involves the use of antibiotics and chemical drugs, which is likely to cause drug residues in aquatic products and bacterial resistance, disrupt the balance of the ecological environment, and pose potential hazards to human health. Due to the harm caused b...

Claims

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Application Information

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IPC IPC(8): A61K39/116A61P31/04A61K39/106
Inventor 莫照兰李贵阳李杰
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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