Preparation method and application method of mushroom bran immobilized microbial agent
An application method, the technology of bacteria chaff, applied in the field of environmental restoration, to achieve the effect of improving the degradation rate of restoration, maintaining the activity for a long time, and increasing the number of viable bacteria
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Embodiment 1
[0033] Taking the aging contaminated soil of crude oil in Shengli Oilfield as the source of bacteria, take 5g of soil samples, add 100ml of sterile water, and shake in a shaker for 30min to make a soil bacteria suspension, and use the dilution coating method to obtain different forms of strains. Take a single bacterium and enrich it in the beef extract peptone liquid medium for about 18 hours, then absorb 2ml and put it in the crude oil medium, and adopt the oil concentration gradient method (5000mg / L, 10000mg / L, 15000mg / L) to acclimatize and cultivate for 7 days ; Take 1ml of the bacterial solution in the 15000 mg / L medium for dilution and coating, pick the bacteria, enrich and spread until a single colony is formed, put the selected single colony in the crude oil medium for cultivation, and use it after 7 days Ultrasonic liquid separation extraction-ultraviolet spectrophotometry is used to measure the remaining petroleum hydrocarbon content at a wavelength of 225nm, so as to ...
Embodiment 2
[0038] A kind of preparation of bacterium chaff immobilized bacterial agent, comprises the following steps:
[0039] 1) Preparation of bacterial suspension: two strains of homopallidum strains screened in Example 1 ( Ochrobactrum sp.) Q1 and Microbacteria ( Microbacterium sp.) Q5, cultured in beef extract peptone liquid medium to the end of the logarithmic phase, adjust the OD600 of each bacterial solution between 0.4-0.8, then mix the two evenly at a volume ratio of 2:3, and then Centrifuge the high-efficiency petroleum hydrocarbon-degrading bacteria liquid, discard the supernatant, rinse with sterile normal saline, and then add normal saline to prepare a high-efficiency petroleum hydrocarbon-degrading bacterial suspension. The control of the bacterial suspension: measure its wavelength at 600nm with an ultraviolet spectrophotometer The lower absorbance value is recorded as OD600, which is determined to be 0.4-0.8.
[0040] 2) Preparation of the carrier: clean the fungu...
Embodiment 3
[0046] The bacterial chaff-immobilized bacterial agent prepared in Example 2 was applied to the remediation of petroleum hydrocarbon-contaminated soil.
[0047] The oil-contaminated soil of Shengli Oilfield was used as the experimental soil. The soil sample was crushed and turned until loose, air-dried and sieved, and the soil between 80 mesh and 40 mesh was taken, that is, after removing large impurities and fine soil with too small diameter for use In the flowerpot restoration experiment, the ratio of soil C, N, and P was adjusted to 100:10:1. During the restoration process, the water content in each flowerpot was adjusted by weight reduction method to keep it at 15-30%.
[0048] Natural restoration (CK) was used as the blank control group, and the four components in petroleum hydrocarbons were used as the target pollutants. The free bacteria group (JK1, 10% free bacteria), the bacterial chaff group (JK2, 5% bacterial chaff), and the bacterial chaff group were respectively ad...
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