A strain of Bacillus megaterium p5-2 and its isolation method and application
A technology of Bacillus megaterium and microbial strains, applied in the field of Bacillus megaterium P5-2 and its separation, can solve the problems of slow growth rate of strains, weak environmental adaptability, low denitrification efficiency, etc. The effect of strong adaptability and high denitrification 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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