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Efficient marine petroleum degrading composite microbial agent and preparation method and application thereof

A compound bacterial agent, offshore oil technology, applied in the field of marine environment oil pollution control, can solve the problems of secondary pollution, difficult to completely remove oil, unable to effectively control and other problems, achieve good tolerance, improve survivability, and degrade Ability-enhancing effect

Active Publication Date: 2019-09-10
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although physical restoration and chemical restoration can quickly remove most of the leaked oil, it is difficult to completely remove the oil film on the ocean surface and the dissolved oil in the water by physical restoration; The above is to add chemical pollutants to the ocean, causing secondary pollution
For seabed oil spills or oil pollution that settles into seabed sediments with the movement of water bodies, current physical and chemical remediation methods cannot effectively control

Method used

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  • Efficient marine petroleum degrading composite microbial agent and preparation method and application thereof
  • Efficient marine petroleum degrading composite microbial agent and preparation method and application thereof
  • Efficient marine petroleum degrading composite microbial agent and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] In the screening method of marine oil-degrading bacteria, the screening medium and degradation medium used in the method are all based on petroleum as the only carbon source and energy source. The specific steps are as follows:

[0038] 1) Source of marine oil-degrading bacteria

[0039] (1) The samples of marine oil-degrading bacteria were taken from the seabed sediments of the Penglai 19-3 oil spill platform.

[0040] (2) Oil is obtained from heavy oil in Shengli Oilfield, Dongying City, Shandong Province, with a viscosity of 4896 mPa·s.

[0041] 2) Screening of marine oil-degrading bacteria

[0042] (1) Preliminary screening of marine oil-degrading bacteria

[0043] Weigh 10g of the above-mentioned sediment sample and put it into 100mL BHB inorganic salt medium after high-temperature sterilization (121°C, 20min), in which 0.5g of the above-mentioned petroleum is added as the only carbon source and energy source. 30°C, 180rpm shaker for 7 days.

[0044] (2) Domes...

Embodiment 2

[0075] Preparation method of high-efficiency marine oil-degrading compound bacterial agent:

[0076] (1) Primary seed cultivation

[0077] The above three petroleum hydrocarbon degrading bacteria Pseudoxanthomonas sp.S1-2, Acinetobacter sp.HC8-3S and Dietzia sp.CN-3 were activated and cultivated respectively. The specific operation is as follows: under sterile conditions, take the petroleum-degrading bacteria in the slant culture medium of the test tube and culture it in 10mL 2216E medium at 30°C in a shaker at 180rpm as the primary seed solution.

[0078] (2) Secondary seed cultivation

[0079] Under sterile conditions, 5 mL of the primary seed liquid of Pseudoxanthomonas sp.S1-2, Acinetobacter sp.HC8-3S and Dietzia sp.CN-3 were inserted into 100 mL of 2216E medium, the inoculum size was 5%, 30°C, Cultivate in a 180rpm shaker as a secondary seed solution.

[0080] (3) Mixed fermentation

[0081] Adjusting the OD of the secondary seed solution of Pseudoxanthomonas sp.S1-2,...

Embodiment 3

[0088] Degradation of oil by high-efficiency marine oil-degrading bacterial strains and compound bacterial agents

[0089] The above-mentioned three kinds of marine oil-degrading bacteria Pseudoxanthomonas sp.S1-2, Acinetobactersp.HC8-3S and Dietzia sp.CN-3 and the marine oil-degrading composite bacterial agent were respectively activated and cultivated in 2216E medium, and then used BHB medium Wash 3 times and resuspend bacteria to OD 600 1.0, according to 10% inoculum size, insert in the 100mL BHB medium containing petroleum concentration 0.5% (w / v); For the control group, each group has three parallels; then cultured on a shaker at 30°C and 180rpm for 7 days, and the culture solution was used for the determination of the oil degradation effect.

[0090] After culturing for 7 days, the culture solution was poured into a separatory funnel, and the oil was extracted three times with 100 mL of normal hexane. After filtering and concentrating, the petroleum sample was fixed to...

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Abstract

The invention relates to the technical field of marine environment petroleum pollution control, in particular to an efficient screening method of marine petroleum degrading bacteria and preparation and application of a composite microbial agent. The composite microbial agent comprises mixed strains of Pseudoxanthomonas sp. S1-2, Acinetobacter sp. HC8-3S and Dietzia sp. CN-3. The marine petroleum degrading composite microbial agent has a better degradation effect on petroleum in a neutral alkaline environment, can resist to a high-salt environment, is particularly adaptive to the marine environment, and expands the application range and prospect of the petroleum degrading composite microbial agent in the marine petroleum pollution control.

Description

technical field [0001] The invention relates to the technical field of marine environment oil pollution control, in particular to a screening method for efficient marine oil-degrading bacteria and the preparation and application of a composite bacterial agent. Background technique [0002] At present, for the treatment of marine oil pollution, especially sudden oil spill accidents, emergency treatment methods include mechanical seals, oil spill recovery, and the use of chemical dispersants. The physical restoration method is to use different types of oil booms, oil suction vessels, and natural or synthetic adsorption materials to recover oil. The chemical repair method is mainly to add oil dispersants, oil coagulants, etc. to seawater to treat oil. Although physical restoration and chemical restoration can quickly remove most of the leaked oil, it is difficult to completely remove the oil film on the ocean surface and the dissolved oil in the water by physical restoration; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C02F103/08C02F101/32C12R1/01
CPCC12N1/20C12N1/02C02F3/34C02F2103/08C02F2101/32C12R2001/01C12N1/205Y02A20/204
Inventor 胡晓珂陈卫卫李俊德
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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