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Composite functional petroleum hydrocarbon degradation microbial agent, as well as preparation method and application thereof

A technology of petroleum hydrocarbon degrading bacteria and petroleum degrading bacteria, which is applied in the field of microorganisms, can solve the problems that the functions of bacteria cannot be completely degraded, and cannot solve the problems of petroleum degradation, etc., and achieve outstanding effects, improve salt resistance, and increase speed.

Active Publication Date: 2019-02-12
ECOLOGY INST SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It cannot solve the problem of petroleum degradation, mainly because the function of bacteria cannot achieve the purpose of complete degradation

Method used

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  • Composite functional petroleum hydrocarbon degradation microbial agent, as well as preparation method and application thereof
  • Composite functional petroleum hydrocarbon degradation microbial agent, as well as preparation method and application thereof
  • Composite functional petroleum hydrocarbon degradation microbial agent, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] A preparation method of sludge petroleum degrading bacterial agent, the steps are as follows:

[0068] (1) Activate and culture Pseudomonas fluorescens ECO, Azotobacter chroococcum and Bacillus paramycoides 3 at 35°C for 2 days to prepare activated fluorescent pseudomonas. Monascus, activated Azotobacter rotundum and activated petroleum hydrocarbon degrading bacteria;

[0069] The components of Pseudomonas fluorescens activation medium are as follows:

[0070] Glucose 10g, MgCl 2 5g, MgSO 4 ·7H 2 O 0.25g, KCl 0.2g, (NH 4 ) 2 SO 4 0.1g, Ca 3 (PO 4 ) 2 5g, agar 15g / L, distilled water 1L, pH 7.0;

[0071] The components of the activation medium for Azotobacter rotundum are as follows:

[0072] K H 2 PO 4 0.2g, K 2 HPO 4 0.8g, MgSO 4 ·7H 2 O 0.2g, CaSO 4 2H 2 O 0.1g, FeCl 3 0.01g, Na 2 MoO 4 2H 2 O 0.01g, yeast extract 0.5g, mannitol 20g, agar 15g, distilled water 1L, pH 7.2;

[0073] The components of the activation medium for petroleum hydroca...

Embodiment 2

[0093] The composite functional petroleum hydrocarbon degrading bacterial agent as described in Example 1, the difference is that Azotobacter vinelandii is replaced by Azotobacter vinelandii, and the preservation number is CGMCC No.1.1005.

[0094] The Vineland nitrogen-fixing bacteria culture medium and the culture method are consistent with those of the nitrogen-fixing bacteria rotundum in Example 1.

[0095] The cultivation method of Pseudomonas fluorescens ECO and Bacillus paramycoides 3 is the same as that in Example 1.

[0096] The Bacillus subtilis bacterium liquid obtained in the present embodiment is mixed with the Pseudomonas fluorescens bacterium liquid and the Vineland azotobacter bacterium liquid prepared in Example 1, and the mass ratio is:

[0097] 1 part of Pseudomonas fluorescens, 1 part of Azotobacter Vinelande, and 1 part of Bacillus subtilis were mixed with peat soil at a mass ratio of 1:5 to prepare a sludge oil-degrading bacterial agent.

[0098] Applica...

experiment example 1

[0127] Take the bacterial agents prepared in Example 1-2 and Comparative Example 1-2, respectively numbered D3-1, D3-2, D3-3, D3-4, and carry out the site restoration experiment under the same conditions:

[0128] The petroleum hydrocarbon content of the polluted soil on the site is 5.17%, and the salt content is 2.04%;

[0129] D3-1, D3-2, D3-3, and D3-4 are all 2% of the quality of oil-contaminated soil, spread evenly on the site to be repaired, plow the plowing layer (30cm), and keep moisture for 15 -20%, measure the content of remaining petroleum hydrocarbons after 120d.

[0130] Petroleum hydrocarbon determination method:

[0131] (1) Extraction of petroleum in soil

[0132] Take 3 g of freeze-dried soil samples for extraction.

[0133] Put the sample into a 50 mL screw-cap centrifuge tube, add 25 mL of dichloromethane, vortex and mix for 1 min, sonicate for 15 min, centrifuge at 6000 rpm for 10 min, and transfer the supernatant to a dry Erlenmeyer flask with a stopper...

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Abstract

The invention relates to a composite functional petroleum hydrocarbon degradation microbial agent, as well as a preparation method and application thereof. The sludge petroleum degradation microbial agent is prepared from the following effective living bacteria components: phosphate dissolving bacteria, nitrogen fixing bacteria and petroleum hydrocarbon degrading bacteria; the phosphate dissolvingbacteria are pseudomonas fluorescens ECO, and the strain storage number thereof is CGMCC No. 16104; the petroleum hydrocarbon degrading bacteria are bacillus paramycoides 3, and the strain storage number thereof is CGMCC No. 16238. The prepared microbial agent has the advantages that the degradation rate of petroleum in sludge is detected according to a gravimetric method, and the degradation rate of the petroleum-contaminated soil or sludge with the petroleum content of 3-10% in a high salinity environment can reach 21-50% within 30 days; the products obtained after degradation by the microbial agent are CO2 and H2O, and secondary pollution is avoided.

Description

technical field [0001] The invention relates to a composite functional petroleum hydrocarbon degrading microbial bacterial agent and its preparation method and application, belonging to the technical field of microorganisms. Background technique [0002] The long-term exploitation of oil has caused serious soil pollution in the oilfield area, and there is a serious conflict between oil exploitation and environmental protection. For the ecological health and ecological safety of the oilfield area, a large amount of oil-contaminated soil formed after oil exploitation must be safely restored. Bioremediation is an effective means of oil-contaminated soil remediation. In the process of microbial remediation of oil-contaminated soil, the growth, reproduction and metabolism of oil-degrading microorganisms require a fixed ratio of carbon, nitrogen, and phosphorus to ensure the rapid reproduction of microorganisms and the protection of petroleum hydrocarbons. Research results at home...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F11/02C12R1/39C12R1/07C12R1/065C02F101/32
CPCC02F11/02C02F2101/32C12N1/20C12N1/205C12R2001/065C12R2001/07C12R2001/39
Inventor 张强季蕾王加宁傅晓文陈贯虹
Owner ECOLOGY INST SHANDONG ACAD OF SCI
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