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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[0029] Example 1
[0030] Isolation of Bacillus megaterium P5-2
[0031] S1, sampling
[0032] In the Tilapia Factory Recirculating Aquaculture Workshop of Panyu Fine Breeding Farm in Guangzhou City, aquaculture water samples and biological fillers are taken and stored in a refrigerator at 4°C for enrichment and separation;
[0033] S2, preliminary screening
[0034] Add 120mL of No. 5 enrichment and separation medium in a 250ml Erlenmeyer flask, use a 100mL graduated cylinder to take 20mL water sample and add it to a 50mL centrifuge tube, shake for about 30s, and 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 5-7 days, repeat the above operation twice, take the third enriched culture solution and dilute it to 10 -3 , 10 -4 , 10 -5 , Take 150μL each of the dilutions, apply to No. 5 enriched and separate solid medium, make 2 dishes for each dilution concentration, and place them in a constant temper...
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[0045] Example 2
[0046] The isolated and purified Bacillus megaterium (Bacillus megaterium) P5-2 with high efficient denitrification, the colony morphology on LB solid medium is like figure 1 As shown, the Gram stain is positive ( figure 2 ); Hydrolyzable starch ( Figure 3-1 ) And casein ( Figure 3-2 );
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[0047] Example 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 concentration of 100μL injection of tilapia with a body weight of (6.5±1.5)g was 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 immersion experiment results showed that the concentration of P5-2 in the water body was 1.5×10 6 CFU / mL and 1.5×10 7 In the case of CFU / mL, tilapia weighing (1.0±0.5)g did not die first and other abnormal phenomena. The above results indicate that P5-2 is a strain with good safety performance.
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