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

Pending Publication Date: 2018-11-16
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the use of bacteria chaff immobilized microorganisms to remediate petroleum hydrocarbons and other organic pollutants, so it is particularly important to explore the preparation and application of bacteria chaff immobilized bacteria agents

Method used

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  • Preparation method and application method of mushroom bran immobilized microbial agent
  • Preparation method and application method of mushroom bran immobilized microbial agent
  • Preparation method and application method of mushroom bran immobilized microbial agent

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a preparation method of a mushroom bran immobilized microbial agent. The preparation method comprises steps as follows: (1) preparation of a bacterial suspension: hydrocarbon degradation bacteria are added to normal saline to be prepared into the bacterial suspension; (2) preparation of a carrier: air drying is performed after mushroom bran is washed clean, and a product issieved by a 20-80-mesh sieve for later use; (3) adsorption: the mushroom bran in the step (2) and the bacterial suspension in the step (1) are mixed in the proportion of 1:(15-25) (v/v), the mixtureis put in a shaking table at the temperature of 30-37 DEG C at the speed of 120-160 rpm to be adsorbed for 16-24 h, and an upper solution is discarded; (4) centrifugation: the adsorbed mushroom bran is transferred to a centrifugal tube with normal saline, and a liquid supernatant is discarded through centrifugation; the centrifugation operation is repeated multiple times, and precipitates obtainedafter centrifugation are the immobilized microbial agent. The prepared mushroom bran immobilized microbial agent can be applied to remediation of petroleum-polluted soil, has high removal efficiencyfor petroleum hydrocarbon, causes no secondary pollution and has the remediation and degradation rate substantially increased as compared with independent mushroom bran and microorganisms.

Description

technical field [0001] The invention relates to the technical field of environmental restoration, in particular to the preparation and application of a bacterial chaff-immobilized bacterial agent capable of repairing petroleum-contaminated soil. Background technique [0002] With the development of the petroleum industry, petroleum pollution has gradually intensified. Organic pollutants remain in the environment for a long time, have a stable structure and are not easy to degrade, posing a serious threat to the environment and human body. Due to the advantages of low cost, no secondary pollution and good treatment effect, microbial remediation technology has been widely used in organic pollution remediation in recent years. Immobilized microorganism technology has received more and more attention and application because of its advantages such as high bacterial density, shielding from harsh environments, and improving degradation efficiency. [0003] Mushroom chaff is the re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02B09C1/10C12R1/01
CPCB09C1/10C12N11/02
Inventor 张秀霞张博凡熊鑫张钊徐文斐贾宏宇辛瑞刘会娥顾莹莹刘春爽
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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