Application of Providencia sp. 2D in Remediation of Dibutyl Phthalate Contaminated Soil
A Providencia and contaminated soil technology is applied in the application field of Providencia 2D in the remediation of dibutyl phthalate contaminated soil to achieve the effect of efficient remediation
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Embodiment 1
[0026] Example 1 Isolation and Identification of Dibutyl Phthalate Degrading Bacteria
[0027] 1. Culture medium formula
[0028] Inorganic salt culture medium (MSM, g / L): K 2 HPO 4 : 5.8; KH 2 PO 4 : 4.5; (NH 4 ) 2 SO 4 : 2.0; MgCl 2 : 0.16; CaCl 2 : 0.02; Na 2 MoO 4 2H 2 O: 0.0024; FeCl 3 : 0.0018; MnCl 2 2H 2 O: 0.0015; adjust the final pH of the inorganic salt culture solution to 7.5. Add DBP so that its concentration in the inorganic salt culture solution is 50 mg / L, and DBP is used as the sole carbon source.
[0029] Beef extract peptone medium (LB): Yeast powder 5.0g, peptone 10.0g, sodium chloride 10.0g, add ultrapure water to 1L, adjust pH=7.0, sterilize at 121°C for 20 minutes. Add 1.5% (W / V) agar powder to the solid plate.
[0030] 2. Separation and identification
[0031]The separation method adopts the sample suspension shaking flask method: Weigh 5g of compost samples (taken from farm animal compost) in a 150mL Erlenmeyer flask containing 50mL o...
Embodiment 2
[0037] Example 2Providencia sp.2D degradation experiment of DBP and its intermediate product phthalic acid
[0038] 1. Preparation of bacterial suspension
[0039] Put the purified strain Providencia sp.2D into 10 mL of LB liquid medium for overnight activation to the logarithmic phase, collect the bacteria by centrifugation at 5000 rpm for 10 min, wash the bacteria with PBS for 3 times and resuspend to adjust the OD 600nm =0.8 as the strain suspension.
[0040] 2. Degradation ability determination
[0041] To the MSM culture fluid (inorganic salt culture fluid g / L) containing different concentrations of DBP (50, 100, 200, 500, 1000mg / L) and phthalic acid (25, 50, 100, 200, 500mg / L) respectively: K 2 HPO 4 : 5.8; KH 2 PO 4 : 4.5; (NH 4 ) 2 SO 4 : 2.0; MgCl 2 : 0.16; CaCl 2 : 0.02; Na 2 MoO 4 2H 2 O: 0.0024; FeCl 3 : 0.0018; MnCl 2 2H 2 O: 0.0015) was inoculated with 1 mL of bacteria, and no bacteria was used as a control, and the pH was adjusted to 8.0, and ea...
Embodiment 3
[0045] Embodiment 3Providencia sp.2D is to the degradation research of DBP in polluted soil
[0046] 1. Preparation of test soil
[0047] The soil is the paddy soil of the farm of South China Agricultural University, the pH is 6.7, the C / N is 5.98, and it is passed through a 60-mesh sieve after air-drying. Animal manure compost was collected from the farm with a pH of 8.8 and a C / N of 17.49. Add compost to the paddy soil according to the common addition ratio on the farm (compost: soil = 1:15, w / w), and mix well to obtain an improved soil with pH = 7.54 and C / N of 10.8. DBP was added to the above-mentioned non-composted soil (conventional soil) and improved soil respectively, so that the concentration reached 100 mg / kg.
[0048] 1. Analysis of DBP degradation effect in soil
[0049] The following treatments were set up in the experiment: conventional soil and improved soil inoculated with bacteria as the experimental group; conventional soil and improved soil without bacter...
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