Combined vaccine for bacterial septicemia and red skin disease of grass carps, and preparation method of combined vaccine

A technology of bacterial sepsis and septicemia, applied in the field of grass carp bacterial septicemia and red skin disease combined vaccine and its preparation, can solve problems such as unsatisfactory results, achieve control of grass carp bacterial septicemia and red skin disease, reduce antibiotics and chemical The use of drugs, the effect of reducing damage

Active Publication Date: 2019-04-26
广州普麟生物制品有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For the prevention and control of grass carp bacterial diseases, tissue plasma vaccines or...

Method used

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  • Combined vaccine for bacterial septicemia and red skin disease of grass carps, and preparation method of combined vaccine
  • Combined vaccine for bacterial septicemia and red skin disease of grass carps, and preparation method of combined vaccine
  • Combined vaccine for bacterial septicemia and red skin disease of grass carps, and preparation method of combined vaccine

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Experimental program
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Effect test

Embodiment 1

[0023] Embodiment 1 vaccine preparation

[0024] (1) Inoculate the nutrient agar medium with Aeromonas hydrophila (GA201 strain) and Pseudomonas aeruginosa (JP802 strain) stored at -70°C respectively, culture at a constant temperature of 28°C for 24 hours, and pick single colonies to inoculate Cultivate overnight (18h) at 28°C in nutrient broth.

[0025] (2) Draw 100 μL each of Aeromonas hydrophila and Pseudomonas aeruginosa cultured overnight in step (1), and re-inoculate them in nutrient broth medium respectively, at 28°C, 180 rpm, shaker Shake for 18 to 20 hours for expanded culture, and the concentration of Aeromonas hydrophila was 7.5×10 8 CFU mL -1 , Pseudomonas aeruginosa was 4.2×10 8 CFU mL -1 .

[0026] (3) Add β-propiolactone with a final concentration of 0.1% (V / V) to the enlarged cultured Aeromonas hydrophila liquid and Pseudomonas aeruginosa liquid respectively, inactivate at 4°C for 72h, and then inactivate at 37°C Water bath for 2h to hydrolyze the inactiv...

Embodiment 2

[0029] Embodiment 2 vaccine preparation

[0030] (1) Inoculate the nutrient agar medium with Aeromonas hydrophila (GA201 strain) and Pseudomonas aeruginosa (JP802 strain) stored at -70°C respectively, culture at a constant temperature of 28°C for 24 hours, and pick single colonies to inoculate Cultivate overnight (24h) at 28°C in nutrient broth.

[0031] (2) Draw 90 μL each of Aeromonas hydrophila and Pseudomonas aeruginosa cultured overnight in step (1), and re-inoculate them in nutrient broth medium respectively, at 28°C, 180 rpm, shaker Shake for 18 to 20 hours to expand the culture, and the concentration of Aeromonas hydrophila was determined to be 9×10 8 CFU mL -1 , Pseudomonas aeruginosa is 6×10 8 CFU mL -1 .

[0032] (3) Add β-propiolactone with a final concentration of 0.5% (V / V) to the expanded cultured Aeromonas hydrophila liquid and Pseudomonas aeruginosa liquid respectively, inactivate at 5°C for 80h, then inactivate at 40°C Water bath for 3h to hydrolyze the...

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Abstract

The invention discloses a combined vaccine for bacterial septicemia and red skin disease of grass carps, and a preparation method of the combined vaccine. The safe and efficient two-tier white oil inactivated vaccine against aeromonas hydrophila septicaemia and pseudomonas aeruginosa red skin disease is developed by using beta-propiolactone as an inactivator, and can effectively control the occurrence of the bacterial septicemia and red skin disease of the grass carps. After the grass carps are immunized with the vaccine, the vaccine can stimulate the expression of IgM, IL-1beta, IFN, Mx, C3 and TLR3 immune-related genes in fish spleen and head kidney tissues. The vaccine can produce up to 80-90% relative immune protection rate for the grass carps; the relative protection rate can still bemaintained at 50% after immunization is performed for one year. The combined vaccine is simple to operate, is safe and effective, can prevent two diseases after being used for immunizing at a time, and not only reduces the use of antibiotics and chemical drugs, but also reduces the pollution and destruction of the external environments such as breeding environment and ecological environment, thusguaranteeing the food safety and water ecological security of the grass carps.

Description

technical field [0001] The invention belongs to the field of fish farming, and in particular relates to a combined vaccine of grass carp bacterial sepsis and red skin disease and a preparation method thereof. Background technique [0002] Grass carp is one of the main breeding species in Guangdong Province, with an annual output of more than 4 million tons, but the annual loss due to disease is as high as more than 2 billion yuan. Grass carp septicemia and red skin disease are extremely serious bacterial diseases that harm grass carp, with a wide range of prevalence and high mortality. Grass carp bacterial sepsis is caused by the infection of Aeromonas hydrophila. It is a high-incidence and high-risk disease of grass carp in recent years. The onset season is long and the epidemic range is wide, and the disease occurs to varying degrees in the main breeding areas of grass carp across the country. Grass carp red skin disease has plagued grass carp farmers in my country for a...

Claims

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Application Information

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IPC IPC(8): A61K39/106A61K39/104A61P31/04
CPCA61K39/104A61K39/107A61P31/04A61K2039/70A61K2039/55511A61K2039/55A61K2039/521
Inventor 罗霞黄志斌江小燕刘春花陈总会赵长臣孙承文巩华陶家发
Owner 广州普麟生物制品有限公司
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