Bacillius firmus GY-49, screening method and applications thereof
A technology of Bacillus firmus and GY-49, which is applied in the direction of chemical instruments and methods, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of high cost of oxygenation, waste of resources, low treatment efficiency, etc., and achieve detoxification Stable nitrogen effect, lower COD, and high treatment efficiency
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Embodiment 1
[0025] 1. Material preparation
[0026] (1). Bacterial source
[0027] Bacterial source: the soil at a depth of 10-15 cm below the surface of the vegetable field in Wanshou Village, Qixia District, Nanjing City, Jiangsu Province.
[0028] (2). Medium
[0029] (1) Beef extract peptone medium: 5.0 g beef extract, 10.0 g peptone, 5.0 g KCl, MgSO 4 ·7H 2 O 2.5 g, dilute to 1 L with water, and adjust the pH to 7.0~7.2.
[0030] (2) LB medium: 10.0 g peptone, 5.0 g yeast powder, 10.0 g NaCl, dilute to 1 L with water, and adjust the pH to 7.0~7.2.
[0031] (3) Denitrification medium: NaNO 3 0.6 g, trisodium citrate 4.1 g, MgSO 4 ·7H 2 O 0.1 g, KH 2 PO 4 1.5 g, 1 mL of trace element solution, dilute to 1 L with water, adjust pH to 7.0-7.5.
[0032] Trace element solution: Na 2 EDTA 63.7 g, ZnSO 4 2.2 g, CaCl 2 5.5 g, MnCl 2 4H 2 O 5.1 g, FeSO 4 ·7H 2 O 5.0 g, Na 2 MoO 4 4H 2 O 1.1 g, CuSO 4 ·5H 2 O 1.6 g, CoCl 2 ·6H 2 O 1.6 g, dilute to 1 L with water, adju...
Embodiment 2
[0042] Example 2: Identification of strains.
[0043] 1. Morphological, physiological and biochemical characteristics of Bacillus firmus GY-49.
[0044] (1) Bacillus firmus Bacillus firmus. Morphological characteristics of GY-49 colonies ( figure 1 )
[0045] Strain GY-49 can be seen after being cultured on LB medium for 2 days, and the colony shape is round, white, with neat edges, rough surface, dry, and protrusions.
[0046] (2) Bacillus firmus Bacillus firmus. Physiological and biochemical characteristics of GY-49
[0047]
[0048] Note: + means positive, - means negative.
[0049] 2. Determination of the 16S rDNA sequence of strain GY-49
[0050] (1) Extraction of bacterial total DNA
[0051] Pick a single colony and activate it on an LB plate first, then transfer it to 5 mL LB liquid medium, culture it to the logarithmic phase overnight with shaking, centrifuge at 8000 rpm for 2 min, collect the bacterial liquid, and wash it with STE for 2 to 3 times; use 0....
Embodiment 3
[0099] Embodiment 3: bacterial strain of the present invention adds KNO with beef extract peptone medium 3 To simulate the anaerobic denitrification and COD removal of high-nitrogen and high-COD aquaculture wastewater, the steps are as follows:
[0100] 1. The Bacillus firmus ( Bacillus firmus. ) GY-49 was inoculated onto the slant of sterilized LB medium, and activated at 30°C for 24-36h; the activated strain was inoculated in beef extract peptone liquid medium, and 100ml of medium was filled in a 250ml Erlenmeyer flask. The medium is beef extract 5.0 g, peptone 10.0 g, NaCl 10.0 g, KCl 5.0 g, MgSO 4 ·7H 2 O 2.5 g, distilled water 1 L, pH7.0-7.5, 30°C, 120rpm shaking culture for 24-36h;
[0101] 2. Centrifuge the bacterial solution obtained in step 1 at 5000rpm for 3 minutes to collect the bacterial cells, and transfer to the KNO-containing 3 Cultured in the beef extract peptone medium for 48 hours, the bacterial agent must be applied; containing KNO 3 The formula of be...
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