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Marine Bacillus amyloliquefaciens and culturing method and application thereof

A technology of amylolytic spores and marine bacteria, applied in the field of microorganisms

Active Publication Date: 2014-09-24
连云港忠华农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of marine Bacillus amyloliquefaciens or its fermentation products or extracts in the prevention and control of aquatic diseases, especially in the prevention and control of Vibrio eel diseases and drug development, has not seen relevant patents and literature reports at home and abroad.

Method used

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  • Marine Bacillus amyloliquefaciens and culturing method and application thereof
  • Marine Bacillus amyloliquefaciens and culturing method and application thereof
  • Marine Bacillus amyloliquefaciens and culturing method and application thereof

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Embodiment 1

[0052] The separation method of embodiment 1 marine bacillus amyloliquefaciens B5:

[0053] ⑴Sample collection and enrichment Use sterile sampling bags and sterile sampling tools to collect samples of seawater, sea mud, kelp, enteromorpha, wakame, asparagus, and seaweed in Lianyungang sea area. Indicate the name of the sample, date of collection, and location of collection.

[0054] (2) The sample is processed as follows: the seawater collected from the seawater sample is mixed and 10mL is added to a 500ml Erlenmeyer flask containing 100mL of sterile seawater TSB liquid enrichment medium, and the shaker enrichment culture is carried out at 28°C and 180r / min for 2 days. . Sea mud sample collection The sea mud mixed with the five-point method is used to sample and mix again. Take 10g of sea mud and put it into a 500ml Erlenmeyer flask with 100ml of seawater TSB liquid enrichment medium sterilized in advance at 28°C, 180r / The shaker enrichment culture was carried out under the...

Embodiment 2

[0058] The identification of embodiment 2 marine bacillus amyloliquefaciens B5 and the research of aspects such as phenotype:

[0059] The biological form of the above-mentioned marine bacterium B5 is described as follows: On the seawater TSB medium, the colony form: round protrusions, irregular edges, white, dry and dull, the strain’s body form: rod-shaped, with blunt round ends, Gram-positive, spores are ellipsoid, with periosteum flagella. The size is (0.5~0.8)×(1.5.~4.5)μm. Such as figure 1 , figure 2 , image 3 shown.

[0060] The physiological and biochemical indicators of the above-mentioned marine bacteria B5: the physiological and biochemical test results show that the physiological and biochemical test results of the bacterial strain B5 are consistent with the characteristics of the relevant marine bacillus amyloliquefaciens in the literature, and can utilize glucose, sucrose, galactose, mannose, mannitol, Does not utilize fructose, lactose, rhamnose, arabinose...

Embodiment 3

[0072] The fermentation method of embodiment 3 marine bacillus amyloliquefaciens B5:

[0073] Strain activation: Marine Bacillus amyloliquefaciens B5 was inoculated on the seawater TSB solid slant medium by streaking method, cultured at 28°C for 18 hours, and the slant culture in logarithmic growth phase was obtained.

[0074] The configuration method of the seawater TSB solid slant medium is as follows: Weigh 17g of tryptone, 3g of soybean peptone, 2.5g of dipotassium hydrogen phosphate, 2.5g of glucose, and 5g of agar powder, and use aged seawater to heat and stir to dissolve, then add aged seawater to constant volume. 1000ml, use NaOH to adjust the pH=7.2-7.5, and then sterilize at 121°C for 15min to obtain seawater TSB solid slant medium.

[0075] Seed liquid preparation process: Under sterile conditions, pick 2-3 loops of the solid slant culture of Bacillus amyloliquefaciens B5 and inoculate it into a 500ml Erlenmeyer flask filled with 100mL sterile seawater TSB liquid me...

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Abstract

The invention discloses a marine Bacillus amyloliquefaciens and a culturing method and application thereof. Marine Bacillus amyloliquefaciens B5 (Bacillus amyloliquefaciens strain chenj-5) is isolated from sea mud, kelp, Enteromorpha, undaria pinnatifida, asparagus and laver in Lianyungang sea area. The strain is preserved in China Center for Type Culture Collection on May 27, 2014, and has a preservation number of CCTCC NO:M 2014229. The gene sequence of the strain has been registered in NCBI on January 8, 2012, with registration number of JN859130.1. The strain has strong antagonism against aquatic disease bacteria Vibrio anguillarum. The fermentation products, fermentation broth extract and thalline extract have significant antimicrobial properties against Vibrio anguillarum. With the antagonistic properties to marine bacteria Vibrio anguillarum, the marine bacteria can be prepared into aquaculture probiotics and biological feed additives. With property of Vibrio anguillarum inhibition, the fermentation product of the bacteria and fermentation product extract can be used for the preparation of medicaments for controlling Vibrio anguillarum disease. The fermentation product of the bacteria has high heat tolerance.

Description

technical field [0001] The present invention relates to the technical field of microorganisms. The present invention relates to a method for isolating a new strain of Vibrio anguillarum antagonistic bacteria, in particular to a method for isolating marine Bacillus amyloliquefaciens B5. The present invention also relates to marine Bacillus amyloliquefaciens B5 obtained by the aforementioned separation method and the application of its fermentation products. Background technique [0002] Vibrio is the most important group of pathogens causing bacterial diseases in aquaculture fish. The diseases caused by Vibrio have a wide epidemic area and a high incidence rate, which has caused great harm to the aquaculture industry and has been a hot spot in the field of aquatic research for a long time. [0003] Vibrio anguillaris can infect most fish, and it is the most harmful Vibrio to aquaculture, and it is also the most studied and reported Vibrio. As early as 1718, there were writt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/74A61P31/04C12R1/07
Inventor 朱强陈静夏艳秋魏贤勇宗志敏史大华詹永成许福泉张晓君阎斌伦
Owner 连云港忠华农业科技有限公司
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