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Solid microbial preparation for petroleum pollutant and oil product degradation, preparation and use

A technology for microbial inoculants and petroleum products, which is applied in the field of bioremediation of oil-contaminated soil and emergency treatment of oil product leakage accidents, can solve the problem that the production of surfactants affects the restoration process, the effect is not ideal, and the growth of strains cannot be achieved. Ideal state and other problems to avoid poor synergy effect, improve bioremediation effect, and easy operation of preparation process

Inactive Publication Date: 2009-07-22
于洋
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effect obtained in the actual application process is not ideal, the main reason is that the ideal state of the growth of the strains in the laboratory cannot be achieved in the harsh environment, and the growth of the surfactant-producing strains is restricted, which will affect the entire restoration process.

Method used

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  • Solid microbial preparation for petroleum pollutant and oil product degradation, preparation and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: the preparation of bacterial fermentation liquid

[0026] The bacterium used was Pseudomonas aeruginosa. Bacterial fermentation broth is obtained by culturing on inclined planes, activating culture in shake flasks, and fermentation in shake flasks respectively. The specific operation steps are as follows:

[0027] (1) Slant culture medium: Prepare beef extract peptone solid medium by conventional methods. The specific steps are: 3g beef extract, 5g peptone, 5g NaCl, 1000ml distilled water, pH7.2~7.4, 1.5%~2% agar, stir well , at 1.05kg / cm 2 , Sterilize at 121.3°C for 20-30 minutes. Make slopes after sterilization, and inoculate the preserved strains onto the slopes of test tubes on ultra-clean workbench respectively, culture the strains at 25-30°C for 16-32 hours, and wait for the bacteria lawn to cover the slopes for later use;

[0028] (2) Preparation of seed liquid: the liquid medium is still beef extract peptone medium, and the prepared liquid mediu...

Embodiment 2

[0030] Embodiment 2: Bacteria is to the degradation ability test of crude oil

[0031] Pick a ring of bacterial lawn from the slant of the activated bacteria, activate it in beef extract peptone liquid medium for 16-32 hours, make a certain concentration of bacterial suspension, and then inoculate according to 2-10% Transfer to crude oil medium, place on a shaker at 20-30°C for 5-12 days, and cultivate at a speed of 150-250rpm. The culture medium without bacteria was used as blank control. At the end of the cultivation, add dichloromethane (DCM) to the culture medium, centrifuge to break the oil-in-water emulsion, filter and dehydrate the organic phase with granular anhydrous sodium sulfate, place it in a pre-constant weight beaker, blow it off with nitrogen at room temperature until constant Weight, the residual oil content in the culture medium was determined by gravimetric method. Adopt the following formula to calculate the biodegradation rate (η%) of petroleum hydrocarb...

Embodiment 3

[0037] Embodiment 3: the preparation method of solid bacterial agent, concrete operating steps are:

[0038] (1) Prepare Pseudomonas aeruginosa bacterial fermentation broth according to the method of Example 1.

[0039] (2) Weigh 100g of peat soil and 20g of bran, put them into a 500mL beaker, wrap them and sterilize at 121.3°C for 30min-2h, and set aside after sterilization.

[0040] (3) Mix the bacterial fermentation liquid in step (1) with the peat and bran in step (2) at a ratio of 0.5:1:0.2, mix thoroughly on a clean bench, and store at room temperature. Two days later, measure the effective viable count of bacteria in the made solid microbial agent, the effective viable count of Pseudomonas aeruginosa is 10 7 ~10 9 pieces / ml.

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Abstract

The invention discloses a solid microbial inoculum that is used for decomposing petroleum pollutants and petroleum products, and a preparation method and applications of the solid microbial inoculum; and the invention belongs to the technical field of biological reparation in the emergency processing of accidents including petroleum pollution of soils and petroleum product leakage. The solid microbial inoculum is prepared by mixing a pseudomonas aeruginosa bacteria fermented fluid, turfy soil and bran with proportion by weight: 0.1-1:1:0.2-0.5, and the effective living bacteria number of pseudomonas aeruginosa bacteria inside the pseudomonas aeruginosa bacteria fermented fluid is 10 to 10 in each milliliter. The solid microbial inoculum has the advantages of high liveness in soil and quick growth, does not cause secondary environmental pollution, and can be used in the biological reparation on the spot of accidents including petroleum pollution of soils and petroleum product leakage.

Description

technical field [0001] The invention relates to a solid microbial bacterial agent for degradation of petroleum pollutants, a preparation method and application, and belongs to the technical field of bioremediation for emergency treatment of petroleum-contaminated soil and petroleum product leakage accidents. Background technique [0002] Bioremediation, also known as biological remediation, biological restoration, ecological restoration or ecological restoration, refers to the process of using the organisms in the treatment system, mainly the metabolic activities of microorganisms, to reduce the concentration of pollutants on the polluted site or make them harmless. Since bioremediation degrades pollutants by dosing or stimulating organisms in the soil, it does not destroy the soil environment required for plant growth. The final product is CO 2 、H 2 O and other substances, so it is environmentally friendly and does not produce secondary pollution. In addition, compared w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/385
Inventor 于洋
Owner 于洋
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