Bacillus megaterium P5-2 and separation method and application thereof
A technique for the isolation of Bacillus megaterium, applied in the field of Bacillus megaterium P5-2 and its isolation, can solve the problems of weak environmental adaptability, slow growth of strains, and low denitrification efficiency, and achieve strong environmental adaptability and growth Fast speed and high nitrogen removal efficiency
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Embodiment 1
[0030] Isolation of Bacillus megaterium P5-2
[0031] S1, sampling
[0032] In the tilapia industrialized recirculating aquaculture workshop of the Panyu Fine Breed Farm in Guangzhou City, water samples of the aquaculture water and biological fillers were taken and stored in a refrigerator at 4°C for enrichment and separation;
[0033] S2, primary screening
[0034] Add 120mL of No. 5 enrichment and separation medium to a 250ml Erlenmeyer flask, use a 100mL graduated cylinder to take 20mL of water sample, add it to a 50mL centrifuge tube and shake for about 30s, then put it into No. 5 enrichment and separation solid medium (containing 15 biological Filler), placed in a shaker at 32°C and 180r / min for constant temperature cultivation for 5-7 days, repeated the above operation twice, and took the third enriched culture solution for gradient dilution and diluted it to 10 -3 、10 -4 、10 -5 , take 150 μL each of the dilutions, spread them on No. 5 enrichment and separation solid...
Embodiment 2
[0046] The isolated and purified heterotrophic Bacillus megaterium (Bacillus megaterium) P5-2 with high denitrification efficiency, the colony morphology on LB solid medium is as follows: figure 1 As shown, the Gram stain was positive ( figure 2 ); hydrolyzable starch ( Figure 3-1 ) and casein ( Figure 3-2 );
Embodiment 3
[0048] The safety of strain P5-2 to tilapia was tested by intraperitoneal injection and soaking. The results of the injection experiment showed that the concentrations of 1.5×10 7 CFU / ml, 1.5×10 8 CFU / ml and 1.5×10 9 Under the condition of CFU / mL bacterial liquid, there was no death or other abnormal conditions; the results of immersion test showed that the concentration of P5-2 in water was 1.5×10 6 CFU / mL and 1.5×10 7 In the case of CFU / mL, tilapia with body weight of (1.0±0.5) g did not appear premature death and other abnormal phenomena. The above results show that P5-2 is a strain with good safety performance.
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