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Bacillus amyloliquefaciens B7 with immune growth promoting effect and application method of bacillus amyloliquefaciens B7

A technology of decomposing starch spores and promoting growth, which is applied in the field of microorganisms to achieve the effects of promoting growth and development, reducing usage, green environmental protection, efficient and healthy breeding

Active Publication Date: 2017-05-24
天津市水生动物疫病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reported research results show that this technology has the advantages of green environmental protection, practicality and high efficiency, but it has not yet formed a technology and product that can be applied in aquaculture production, especially a technical product that has immune growth-promoting effect on the marine treasure fish Semi-smooth tongue sole There are no related reports

Method used

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  • Bacillus amyloliquefaciens B7 with immune growth promoting effect and application method of bacillus amyloliquefaciens B7
  • Bacillus amyloliquefaciens B7 with immune growth promoting effect and application method of bacillus amyloliquefaciens B7
  • Bacillus amyloliquefaciens B7 with immune growth promoting effect and application method of bacillus amyloliquefaciens B7

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Experimental program
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Effect test

Embodiment 1

[0013] Obtaining of Bacillus amyloliquefaciens B7:

[0014] (1) Separation and purification: Take a healthy semi-smooth tongue sole with a weight of about 15 g, dissect the intestinal tract under aseptic conditions, remove the intestinal contents, wash it with sterile saline for 3 times, weigh it and place it in a pre-cooled sterile glass Homogenize in a homogenizer, bathe the homogenate in water at 100°C for 10 minutes, kill non-spore cells, and then dilute to 10 -2 , take the homogenate stock solution, 10 -1 、10 -2 100 μL of three gradient dilutions were spread on nutrient agar plates, and cultured in a constant temperature incubator at 30°C for 48 hours. Observe the number, shape, color and size of the colonies on each plate, pick a single colony for purification and culture, and check the purified strains with simple fuchsin staining under the microscope, remove non-bacillus, and perform Gram staining with carbolate fuchsin staining Stained with spores to obtain 14 stra...

Embodiment 2

[0026] Features of Bacillus amyloliquefaciens B7:

[0027] (1) Colony characteristics: Inoculated on nutrient agar plate and cultured at 30°C for 24 hours, the colonies are white, rough, flat, with irregular edges; Gram-positive, the bacteria are rod-shaped, arranged singly, in pairs or in chains.

[0028] (2) Hemolyticity: Inoculate the isolated and purified Bacillus amyloliquefaciens B7 in sterile nutrient broth, culture with shaking at 150 rpm at 30°C for 24 hours, take 10 μl of spot seed on a blood agar plate, and culture in a constant temperature incubator at 30°C for 24 hours , It was observed that no transparent circle was formed around the colony, indicating that Bacillus amyloliquefaciens B7 did not produce hemolysin.

[0029] (3) Amylase activity: Inoculate Bacillus amyloliquefaciens B7 into sterile nutrient broth, shake and culture at 30°C at 150 rpm for 24 hours, centrifuge at 4500 rpm for 20 minutes, take 100 μL of the supernatant and add it to an Oxford cup place...

Embodiment 3

[0043] Bacillus amyloliquefaciens B7 fermentation broth preparation method:

[0044] The laboratory preparation method is:

[0045] (1) Seed liquid culture: activate Bacillus amyloliquefaciens B7, inoculate it into sterile nutrient broth, and culture it with shaking at 30°C and 150 rpm for 24 hours to obtain the seed liquid;

[0046] (2) Fermentation: put 400mL of sterile nutrient broth in a 1L conical flask, inoculate the seed liquid into the conical flask with 10% inoculum, 30°C, 150rpm constant temperature shaking culture for 48h, the number of bacteria reached 8 × 10 8 CFU / mL~1×10 9 CFU / mL;

[0047] (3) Harvesting: transfer the cultured fermentation broth to a 500 mL sterile Erlenmeyer flask and store at 4° C. in the dark.

[0048] The large-scale preparation method is:

[0049] (1) Seed solution cultivation: Prepare 4L of seed solution according to the above-mentioned laboratory preparation method;

[0050] (2) Fermentation medium: 36L medium in a 50L fermentation ta...

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Abstract

The invention discloses bacillus amyloliquefaciens B7 with an immune growth promoting effect and application method of the bacillus amyloliquefaciens B7. The bacterium is named as bacillus amyloliquefaciens B7 according to the classification, and is collected in the China Center for Type Culture Collection, with the collection number of CCTCC NO: M 2016128. An experiment shows that the bacterium disclosed by the invention can improve the immune function of cynoglossus semilaevis and effectively resist invasion of pathogenic microorganisms; the disease morbidity is reduced to 4 percent or below, and the aquaculture survival rate is increased to 96 percent or above; the digestive enzyme activity and the feed utilization rate of the cynoglossus semilaevis are improved; the food coefficient is reduced by 12 percent or above, and the specific growth rate is increased by 4 percent or above; the use amounts of antibiotic and chemical medicines are effectively reduced, the environmental pollution is relieved, the quality safety of an aquaculture product is guaranteed, and the bacillus amyloliquefaciens B7 has a good ecological benefit. The key technical problem in disease damage prevention and treatment, which restricts healthy development of cynoglossus semilaevis aquaculture industry is solved, and the bacillus amyloliquefaciens B7 realizes environmentally friendly, efficient and healthy aquaculture.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to bacillus amyloliquefaciens B7 with the effect of promoting immune growth and its application method. Background technique [0002] Semi-smooth tongue sole is an important economically valuable fish in my country. It has delicious meat, wide temperature, wide salt and adapts to changing environments. It is one of the ideal breeding species. With the development of industrial aquaculture in seawater, tongue sole has become one of the main species. Long-term high-density intensive farming has led to more and more serious diseases. The main diseases are: fester disease, ascites disease, spleen and kidney necrosis, etc. The mortality rate of diseased fish is as high as 70%, and the economic loss is huge, which seriously restricts the culture of tongue sole healthy development of the industry. The traditional treatment method is to use large doses of antibiotics or chemi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K10/18A23K50/80A61K35/742A61P37/04C12R1/07
CPCA61K2035/115C12N1/205C12R2001/07
Inventor 孙金生王雪惠耿绪云董学旺薛淑霞魏俊利李晶晶孙妍
Owner 天津市水生动物疫病预防控制中心
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