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Turbot-derived Edwardsiella piscicida and application thereof

A technology of turbot and source, applied in the field of piscicidal Edwardsiella, can solve the problems of high frequency of bacterial diseases, large economic losses, serious problems, etc., and achieve the effect of improving the immune protection rate

Active Publication Date: 2020-12-04
烟台市海洋经济研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the continuous development of scale and the improvement of intensive farming, turbot diseases frequently occur during the breeding period, causing huge economic losses. Among the reported turbot diseases, bacterial diseases have the highest frequency of occurrence and the highest losses. more serious

Method used

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  • Turbot-derived Edwardsiella piscicida and application thereof
  • Turbot-derived Edwardsiella piscicida and application thereof
  • Turbot-derived Edwardsiella piscicida and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] This example relates to the isolation and identification of Edwardsiella piscicida. The Edwardsiella piscicida derived from turbot is Edwardsiella piscicida H4-S18 strain, and its gene sequence is shown in SEQ ID No.1.

[0016] Specific steps are as follows:

[0017] The samples of diseased turbot were collected from the turbot farm in Yantai, Shandong Province. The individual weight ranged from 205 g to 518 g, the average weight was 436±5 g, and the average body length was 29±2 cm. The diseased turbot showed weakened feeding ability, sluggish movement, swollen abdomen, red and swollen anus, congested and red snout, fin rays and base. Autopsy revealed pale yellow ascites in the abdominal cavity, intestinal enlargement, thin intestinal wall with pale yellow liquid, and liver congestion.

[0018] Take dying individuals with typical symptoms, aseptically dissect internal organs, take liver, kidney, spleen, heart and other tissues, rinse with sterile water, grind, streak o...

Embodiment 2

[0025] This embodiment involves the use of Edwardsiella piscicida to prepare inactivated vaccines. The Edwardsiella piscicida derived from turbot is Edwardsiella piscicida H4-S18 strain, the gene sequence of which is shown in SEQ ID No.1.

[0026] Specific steps are as follows:

[0027] Inoculate the Edwardsiella piscicidae H4-S18 strain on a nutrient agar medium (NA) plate, and incubate at 37°C for 24-48h for recovery; pick the above pathogenic bacteria into NB medium, culture with shaking at 37°C for 24h, and collect the culture solution Centrifuge at 5000rpm for 10min at 4°C, wash the centrifuged bacterial cells with PBS buffer solution with pH=7.2 for 3 times, and then use sterile normal saline (containing 2% NaCl) to make the concentration of 10 9 cfu / mL bacterial suspension stock solution; add 37% formalin solution until the final concentration of formaldehyde is 0.2%, inactivate at 37°C and 140r / min, take the inactivated 12h and 24h bacterial solution respectively Dete...

Embodiment 3

[0029] This embodiment relates to the detection of vaccine effects, and the Edwardsiella piscicida derived from turbot is Edwardsiella piscicidae H4-S18 strain.

[0030] Its gene sequence is shown in SEQ ID No.1.

[0031] According to the steps of preparing the vaccine in step 2, Edwardsiella piscicida (ET883) was used as the antigen to prepare the vaccine.

[0032] Adjust the 2 inactivated vaccine concentrations to OD 600 0.1±0.01, intraperitoneal injection of immune turbot, each injection of 0.1mL, the control group injection of the same dose of sterile saline. 20 tails were immunized in each group, with 3 replicates and 1 control. Breeding water temperature 18±1℃, inflated throughout the day, daily water change 30%; 30 days after immunization, it was challenged with Edwardsiella piscicida, and the concentration of each tail was 8.9×10 10 CFU / mL of bacterial solution 0.1mL; observe and record the number of deaths in each experimental group every day, and calculate the im...

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Abstract

The invention provides turbot-derived Edwardsiella piscicida and an application thereof. The turbot-derived Edwardsiella piscicida is an Edwardsiella piscicida H4-S18 strain, the gene sequences of theEdwardsiella piscicida H4-S18 strain are shown as SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3, and the Edwardsiella piscicida H4-S18 strain is preserved with the preservation number of CCTCC NO: M 2020450. The invention further relates to an application of the turbot-derived Edwardsiella piscicida in preparation of vaccines. The turbot-derived Edwardsiella piscicida provided by the invention is obtained by screening from diseased turbots, is the turbot-derived Edwardsiella piscicida carrying drug resistance genes for the first time in China, the turbot-derived Edwardsiella piscicida carrying thedrug resistance genes is applied for the first time to prepare the vaccines, and therefore, the immune protection rate of the turbots to the drug resistance Edwardsiella piscicida is effectively improved.

Description

technical field [0001] The invention belongs to the field of pathogenic microorganism screening, and in particular relates to Edwardsiella piscicida derived from turbot and application thereof. Background technique [0002] Turbot is rich in nutrition and high in economic value. It is a cold-water bottom-dwelling marine fish. It is native to Europe. It was introduced to my country in 1992 and has become an important economically farmed fish in northern my country. However, with the continuous development of scale and the improvement of intensive farming, turbot diseases frequently occur during the breeding period, causing huge economic losses. Among the reported turbot diseases, bacterial diseases have the highest frequency of occurrence and the highest losses. more serious. The infection rate and fatality rate of turbot to Edwardsiella are high, and Edwardsiella is the main disease that endangers the healthy breeding of turbot. Edwardsiella piscicida of the genus Edward...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
CPCA61K39/02A61K2039/521A61K2039/552A61P31/04C12N1/205C12R2001/01
Inventor 黄华王鹤张建柏王力勇徐晓莹张岚王晓飞史文凯
Owner 烟台市海洋经济研究院
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