Complex microbial inoculant and remediation of petroleum-chlorophenol combined pollution environment by using complex microbial inoculant
A compound bacterial agent and compound pollution technology, applied in water pollutants, bacteria, biological water/sewage treatment, etc., can solve the problems of inability to degrade pollutants and weak degradation ability, and achieve obvious advantages in biodegradation and comprehensive functions. , easy to prepare
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Embodiment 1
[0029] Example 1. Strain screening
[0030] Materials and methods
[0031] Soil samples: Dongying Dongxin Oil Production Plant has long been contaminated by oil.
[0032] Sludge sample: activated sludge in the secondary sedimentation tank of Qingdao Sewage Treatment Plant.
[0033] Inorganic salt medium: (NH 4 ) 2 SO 4 1.0 g, K 2 HPO 4 1.0 g, KH 2 PO 4 1.0 g, anhydrous CaCl 2 0.01 g, MgSO 4 ·7H 2 O 0.2g, NaNO 3 5.0 g, FeCl 3 Trace amount, distilled water 1000mL, adjust pH to 7.2~7.5, sterilize at 121℃ for 20min before use.
[0034] Petroleum hydrocarbon degrading bacteria screening medium: add a certain amount of petroleum on the basis of inorganic salt medium.
[0035] Chlorophenol-degrading bacteria screening medium: add a certain amount of chlorophenol on the basis of inorganic salt medium.
[0036] Screening of Petroleum Hydrocarbon Degrading Strain
[0037] Weigh 10g of petroleum-contaminated soil and add it to a sterilized Erlenmeyer flask containing 90mL...
Embodiment 2
[0050] Example 2. Strain compounding
[0051] Materials and Methods
[0052] Strains: After screening, two petroleum hydrocarbon-degrading bacteria C4 and C5, and two chlorophenol-degrading bacteria L3 and L4 were obtained.
[0053] culture medium
[0054] Beef extract liquid medium, petroleum-chlorophenol inorganic salt medium
[0055] Assay method: prepare petroleum-chlorophenol inorganic salt medium (the concentration of chlorophenol in the medium is 50mg / L; the content of petroleum is 2g / L), sterilized for later use;
[0056] Two petroleum hydrocarbon-degrading bacteria Alcaligenes faecails Y5 C4 and Bacillus sp.7614 C5; two chlorophenol-degrading bacteria Bacillus marisflavi strain BCB4-1 L3, Bacillus sp. aquimaris T 172) L4, according to the inoculation amount of compound bacteria is 10%, the bacterial liquid is added in the inorganic salt liquid medium with chlorophenol concentration of 50mg / L, oil concentration of 2g / L, pH=7.5, and different V C4 :V C5 :V L3 :V ...
Embodiment 3
[0064] Example 3. Remediation of petroleum-chlorophenol compound polluted environment by compound bacterial agent
[0065] Experimental setup:
[0066] The petroleum inorganic salt medium was prepared, and after sterilization, an appropriate amount of chlorophenol compound was added for use.
[0067] The mixed fermentation broth of 10% (V:V) compound bacteria was added to the prepared medium containing petroleum-chlorophenol inorganic salt.
[0068] The inorganic salt medium without the inoculated strain was used as the blank control group.
[0069] All treatments were incubated at 35°C for 7 days at 160 r / min.
[0070] The organic phase of the cultured medium was extracted with n-hexane, dried over anhydrous sodium sulfate, the operation was repeated three times, and concentrated to 2 mL with a nitrogen blower, and relevant assays were carried out.
[0071] The infrared analysis of the remaining oil was determined by Fourier transform infrared spectroscopy (FTIR), and the ...
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