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A strain of bacillus subtilis dcu and its use

A technology of Bacillus subtilis and bacteria powder, which is applied in the direction of bacteria, applications, and microorganisms, can solve the problems of ineffective effects, achieve good anti-pathogenic activity, broad application prospects, and inhibit the growth of pathogenic bacteria

Active Publication Date: 2018-07-06
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Probiotics have been applied to prawns, shellfish and other marine aquaculture animals, but the probiotics suitable for shrimp and shellfish do not necessarily work or the effect is not obvious in mud crab culture

Method used

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  • A strain of bacillus subtilis dcu and its use
  • A strain of bacillus subtilis dcu and its use
  • A strain of bacillus subtilis dcu and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Separation and purification of Bacillus subtilis DCU

[0024] 1) Isolation and purification of bacterial strains: blue crabs were collected from Niutianyang blue crab farm in Shantou, Guangdong Province (blue crabs are disease-resistant crabs that survived "milk disease"), brought back to the laboratory, rinsed with distilled water and other debris on the surface, alcohol After spraying, they were dissected under the ultra-clean bench and the intestinal contents were taken to complete the collection of samples of the intestinal contents of blue crabs. The strain was isolated from the intestinal tract of cultured mud crabs, diluted with LB, 2216E, TCBS medium, plated, and cultured overnight at 28°C. One of the strains had a suspected inhibitory effect on other surrounding strains, and it was re-plated for separation and purification.

[0025] 2) Identification of strains: preliminary identification was carried out through plate colony morphology and scanni...

Embodiment 2

[0030] Embodiment 2: the antibacterial test of bacillus subtilis DCU

[0031] Bacillus subtilis DCU was cultured, and the antibacterial effect on Vibrio parahaemolyticus, a common pathogenic bacteria in the cultivation of mud crabs, was detected by the disc method. The antibacterial effect was observed according to the size of the inhibition zone.

[0032] Specific steps: Inoculate the pathogenic bacteria isolated from the intestinal tract of blue crabs in the corresponding liquid medium and culture overnight at 28°C with 180rpm shaking, draw 100 μL of the culture solution and spread it on a 90mm petri dish, spread it evenly, and place it on a flat surface Dry upwards. A filter paper piece with a diameter of about 6mm that was cut in advance and sterilized by high pressure steam and dried in an oven was immersed in the activated and cultured bacterial solution for about 24 hours for 20 minutes. Use sterilized tweezers to clamp out the filter paper, stick it on the wall of th...

Embodiment 3

[0034] Embodiment 3: the cultivation method of bacillus subtilis DCU

[0035] The inventors of the present application have found that the Bacillus subtilis DCU can be cultivated using 2216E (Zobell) medium: yeast extract 1g, peptone 5g, NaCl 34g, Fe 3 PO 4 0.1g, add 800mL of distilled water, adjust the natural pH to 1000mL. Add 1.5% agar to the solid plate. The culture conditions are: rotating speed 180rpm, temperature 28°C, culture for 12h.

[0036] The inventors of the present application also used nutrient agar medium (NA) to cultivate the Bacillus subtilis DCU. The NA medium consisted of: peptone 10g, beef extract 3g, sodium chloride 5g, natural pH, and distilled water to 1000mL. The cultivation conditions were the same as above. The results showed that the strain growth rate and final concentration in NA medium were better than those in 2216E medium.

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Abstract

The invention provides a strain of Bacillus subtilis DCU, the preservation number of which is CCTCC NO.2014118. The Bacillus subtilis not only has good anti-pathogenic activity, but also can well inhibit the growth and reproduction of pathogenic bacteria, and can also improve the body's Immunity has a wide application prospect. It can be applied to probiotic preparations, or it can be made into bacterial powder and added to feed as a feed additive to inhibit the growth of pathogenic bacteria in animals and reduce animal mortality.

Description

technical field [0001] The invention relates to a strain of Bacillus subtilis, in particular to a strain of Bacillus subtilis DCU capable of inhibiting the growth of Vibrio parahaemolyticus and its application. Background technique [0002] Scylla paramamosain, referred to as blue crab, belongs to wide-salt and wide-temperature seawater economic crustaceans, mainly distributed in the southeastern coastal areas of my country, and is the main marine culture species in eastern Guangdong, especially Shantou. It has fast growth, individual Large, adaptable and disease-resistant, resistant to dry dew, easy to transport, delicious and unique meat, rich in nutrition and other characteristics, it is favored by domestic and foreign producers and consumers. In recent years, blue crab diseases, especially viral diseases (such as blue crab baculovirus disease, reovirus disease), bacterial venereal diseases and parasitic diseases have caused serious economic losses to the blue crab breedin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18C12R1/125
CPCC12N1/20C12N1/205C12R2001/125
Inventor 李升康吴慧娟孙令斌刘文华胡忠温小波
Owner SHANTOU UNIV
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