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A kind of streptococcus parauberis derived from turbot and its application

A technology of parauterine and streptococci, applied to turbot-derived streptococcus parauterine and its application field

Active Publication Date: 2021-01-22
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

There is no report of fish streptococcosis caused by Streptococcus parauberis in my country

Method used

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  • A kind of streptococcus parauberis derived from turbot and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] This embodiment relates to the isolation and identification of Streptococcus parauberis. The Streptococcus parauberis is Streptococcus parauberis BH2-4 strain, and its gene sequence is shown in SEQ ID No.1.

[0017] Specific steps are as follows:

[0018] The samples of the diseased turbot were collected from the turbot farm in Yantai, Shandong Province. The individual weight ranged from 195 g to 535 g, and the average weight was 458 g. The diseased turbot showed loss of appetite, black body color, unbalanced swimming state, spinning, exophthalmos, hyperemia at the base of the mandible and fin rays, and individual abdominal swelling and body surface ulcers. Autopsy revealed mottled congestion in the liver, splenomegaly, dark color, and a few cases of peritoneal ascites.

[0019] Take a dying individual with typical symptoms, aseptically dissect the viscera, take liver, kidney, spleen, brain, eye and other tissues, rinse with sterile water, grind, streak on a BHI agar p...

Embodiment 2

[0026] This example relates to the preparation of inactivated vaccines using Streptococcus parauberis.

[0027] Specific steps are as follows:

[0028] Inoculate the BH2-4 strain of Streptococcus parauberis on a nutrient agar medium (NA) plate, and incubate at 37°C for 24-48h for recovery; pick the above-mentioned pathogenic bacteria into NB medium, culture with shaking at 37°C for 24h, and collect the culture solution in 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 Detection of inactivation and observation of inactivation. Centrifuge the inactivated bacterial solution at 5000rpm for 10m...

Embodiment 3

[0030] This embodiment relates to the detection of the effect of the vaccine.

[0031] Specific steps are as follows:

[0032] According to the step 2 for preparing the vaccine, the Streptococcus parauberis strain BH2-4 is used as the antigen to prepare the vaccine.

[0033] Adjust inactivated vaccine concentration 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 all day, daily water change 30%; 30 days after immunization, it was challenged with Streptococcus parauberis, and the concentration of each tail was 9.2×10 10 CFU / mL of bacterial solution 0.1mL; observe and record the number of deaths in each experimental group every day, and calculate the immune protection rate of the inactivated vaccine against Streptococcus parauberis.

[0034] The results showe...

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Abstract

The present invention provides a Streptococcus parauberis derived from turbot and its application; the Streptococcus parauberis is Streptococcus parauberis BH2‑4 (Streptococcus parauberis BH2‑4), the gene sequence of which is shown in SEQ ID No.1 Show. The strain has been submitted for culture deposit, and the deposit number is CCTCC NO:M 2020451. The present invention also relates to the application of said turbot-derived Streptococcus parauberis in preparing vaccines. The Streptococcus parauberis provided by the invention is obtained by screening from diseased turbot, and is used to prepare a vaccine, so as to effectively prevent and treat the turbot streptococcosis.

Description

technical field [0001] The invention belongs to the field of pathogenic microorganism screening, and in particular relates to a turbot-derived Streptococcus parauberis and application thereof. Background technique [0002] The breeding of flounder and flounder has successfully promoted the formation of the fourth wave of industrialization of mariculture in my country, and is an important industry in northern mariculture. Turbot is the most important species of flounder and flounder in northern my country. Since the breakthrough in seed breeding technology, Shandong Province's aquaculture output and aquaculture area are among the top in the country. [0003] In recent years, with the expansion of aquaculture scale and the degradation of germplasm resources, turbot diseases have occurred frequently, seriously restricting the green and healthy development of mariculture. Among turbot diseases, bacterial diseases occur more frequently, causing huge economic losses to farmers. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/09A61P31/04C12R1/46
CPCA61K39/092A61K2039/521A61K2039/552A61P31/04C12N1/205C12R2001/46
Inventor 王鹤黄华李战军张建柏王力勇王文豪贺加贝曹亚男
Owner 烟台市海洋经济研究院
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