Bacillus pumilus BP and application thereof
A technology of bacillus pumilus, which is applied in the direction of application, bacteria, antibacterial drugs, etc., can solve the problem of ineffective effect, achieve the effect of inhibiting the growth and reproduction of pathogenic bacteria, broad application prospects, and inhibit the growth of pathogenic bacteria
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Embodiment 1
[0023] Embodiment 1: Isolation and purification of Bacillus pumilus BP
[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 scanning ...
Embodiment 2
[0030] Embodiment 2: the antibacterial test of bacillus pumilus BP
[0031] Bacillus pumilus BP was cultured, and the antibacterial effect on Vibrio parahaemolyticus, a common pathogenic bacteria in the breeding 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. The filter paper pieces with a diameter of about 6mm that were cut in advance and sterilized by high pressure steam and then dried in an oven were immersed in the activated and cultured bacteria solution for about 24 hours for 20 minutes. Use sterilized tweezers to take out the filter paper, stick it on the bottle wa...
Embodiment 3
[0034] Embodiment 3: the cultivation method of bacillus pumilus BP
[0035]The inventors of the present application have found that the Bacillus pumilus BP can be cultivated using 2216E (Zobell) medium, 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 pumilus BP. The NA medium consisted of: 10 g of peptone, 3 g of beef extract, 5 g of sodium chloride, natural pH, and distilled water to 1000 mL. 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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