Dicofol degrading bacterium and soil restoration application
A technology of dicofol and degrading bacteria, applied in the field of biological treatment of environmental pollutants, can solve the problems of entering the soil, harm, pollution, etc., and achieve the effect of slow application effect, good general performance and fast degradation rate
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Embodiment 1
[0031] Example 1: Pseudomonas putida Separation of DCF-7 and its ability to degrade dicofol
[0032] The soil near the sewage outlet of Tonglu Pesticide Factory in Zhejiang was collected on site, packed in a 150 mL triangular flask, and quickly brought back to the laboratory. The retrieved samples were placed in dicofol-selective basic inorganic salt liquid medium for acclimation culture. The medium composition is (g / L): MgCl 2 0.2, NH 4 NO 3 1, KH 2 PO 4 2,K 2 HPO 4 7.5, NaCl 1, dicofol 0.15 (pre-dissolved in acetone), pH 6.8, 1 L of deionized water, sterilized by moist heat at 115°C for 30 minutes before use. The culture conditions are: 25°C, 120 r / min shaking culture, and the liquid volume in the container is 1 / 3 of the container volume. Thereafter, the shake flask was kept still for a while every 5 days, the supernatant was discarded, and then an equal amount of freshly prepared selective liquid medium was added and the shaking enrichment culture was continue...
Embodiment 2
[0035] Example 2: Pseudomonas putida Drug resistance experiment of DCF-7
[0036] Tetracycline, gentamicin and neomycin were selected for testing by the filter paper method, and the diameter of the inhibition zone of each antibiotic filter paper (30 μg / piece) on the cultured degrading bacteria DCF-7 plate was used as the sensitivity or resistance basis for judgment. The test results showed that the diameters of the inhibition zone were 24, 28 and 32 mm in sequence, showing moderate, highly and extremely sensitive reactions respectively (the sensitive range is: the moderately sensitive range of tetracycline is 23-25 mm; the highly sensitive range of gentamicin is 26 mm; -30 mm; neomycin extremely sensitive range >30 mm).
[0037] This example shows that when the bacteria are released into the natural environment, super bacteria will not be produced due to the problem of drug resistance (resistance), which provides a safety guarantee for its future practical application.
Embodiment 3
[0038] Example 3: Pseudomonas putida Heavy Metal Minimal Inhibitory Concentration (MIC) Test of DCF-7
[0039] The design of the MIC experiment was carried out with reference to the research of Filali et al. (Filali B.K., Taoufik J., Zeroual Y., Dzairi F.Z., Talbi M., Blaghen M. Waste water bacterial isolates resistant to heavy metals and antibiotics. Current Microbiology, 2000, 41: 151-156), pick the same size DCF-7 single colony after activation treatment, put it in the culture test tube, 30 ℃, 150 r / min shaking culture for 48 hours. The MIC value was determined according to the growth of the bacteria, and three replicates were set up in each group to test the tolerance of the bacteria to 13 metal ions. The results are shown in Table 2, the strain has multiple tolerance and strong tolerance.
[0040]
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