Complex microbial agent for restoring coastline with heavy oil pollution, as well as preparation method and applications thereof

A compound inoculant, coastline technology, applied in the directions of microorganism-based methods, restoration of polluted soil, biochemical equipment and methods, etc., can solve the problems of no large-scale practice, late start, etc., and achieve outstanding competitive advantages and salt tolerance. The effect of good performance and efficient repair effect

Active Publication Date: 2015-07-01
BC P INC CHINA NAT PETROLEUM CORP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The above achievements once again fully affirmed the role of bioremediation technology in oil pollution control. However, due to the late start of this field in China, this type of technology has not yet been practiced on a large scale in the environmental treatment of oil spill pollution in my country's coastlines.

Method used

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  • Complex microbial agent for restoring coastline with heavy oil pollution, as well as preparation method and applications thereof
  • Complex microbial agent for restoring coastline with heavy oil pollution, as well as preparation method and applications thereof
  • Complex microbial agent for restoring coastline with heavy oil pollution, as well as preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Enrichment and Screening of Example 1 Petroleum Degrading Strains

[0060] The bacterial strain involved in the present invention was isolated from the coastline of the Yellow Sea, which was severely polluted by oil spills during the explosion accident of the Dalian Xingang oil pipeline in 2010.

[0061]The specific separation operation is carried out as follows: Add 2 g of coastline and beach samples and a small amount of trace element solution to 100 mL of sterilized broad-spectrum culture solution, and enrich and cultivate on a shaking table at 25 ° C ~ 30 ° C, 140 rpm, and avoid light. 5d; After two consecutive enrichment treatments, transfer 1mL of the enrichment solution into the ASM medium containing 1g / L Venezuelan heavy oil, add a small amount of trace element solution, and culture for 7d at 25°C-30°C, 140rpm, and dark conditions , and then the culture medium was continuously transferred and cultured three times in the same fresh ASM medium containing Venezuela...

Embodiment 2

[0072] The degradability of embodiment 2 single bacterium and mixed flora to Venezuelan heavy oil

[0073] Taking Venezuelan heavy oil as pollutant, the petroleum degradation performance of the above single and mixed strains was investigated under simulated seawater salinity. Follow the steps below: (1) Pipette 200uL of cryopreserved strains into sterilized liquid LB medium with a pipette gun, vibrate and activate at 25°C-30°C, 140rpm for 48h, and transfer to fresh LB culture repeatedly Base 1 time. At this point, take the turbid solution in the logarithmic phase of growth and centrifuge at room temperature at 5000rpm for 10min, collect the thalline and wash it with sterile phosphate buffered saline, after repeating this at least 3 times, dilute it to a specific strain density with phosphate buffered saline ( OD 600 ), prepared as a single strain suspension. The above phosphate buffer contains 21.759g K per liter 2 HPO 4 ·H 2 O, 33.48 g Na 2 HPO 4 12H 2 O, 8.79 g KH ...

Embodiment 3

[0075]Embodiment 3 relates to the salt tolerance of strain

[0076] Affected by seawater tides and evaporation, the salinity of the oil spilled coastline environment is relatively high and presents complex changes, which is an important factor affecting the growth, metabolism and competition of microorganisms. This embodiment is carried out according to the following steps: take the suspensions of Sphingobacterium mizutaii XG-3, Achromobacter xylosoxidans XG-14, and Achromobacter piechaudii XG-11 with the same strain concentration as Inoculate at 5% volume ratio in LB medium with NaCl mass concentration gradients of 0, 1%, 3%, 5%, 7%, and 10%, and culture for 24 hours under shaking at 28°C, 140rpm, and dark conditions , measure the OD of each mixture 600 , according to the test results to evaluate the saline-alkali tolerance performance of each dominant strain. If the OD of the bacterial solution 600 >0.25, it is considered that the strain grows well in the salinity environ...

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Abstract

The invention relates to a complex microbial agent for restoring coastline with heavy oil pollution, as well as a preparation method and applications thereof. The complex microbial agent comprises Sphingobacterium mizutaii, Achromobacter xylosoxidans and Achromobacter piechaudii, furthermore, the collection number of the Sphingobacterium mizutaii is CGMCC No.800; the collection number of the Achromobacter xylosoxidans is CGMCC No.8022; and the collection number of Achromobacter piechaudii is CGMCC No.8021. The complex microbial agent is good in salt resistance property, prominent in competitive advantages, and has a high-efficiency restoration effect on the crude oil pollution of coastline, especially refractory heavy oil contamination through the synergistic effect of the three bacteria, and can also be applied to restoration of saline alkali soil with overflow oil contamination.

Description

technical field [0001] The invention belongs to the technical field of microbial technology and oil-polluted coastline bioremediation, and in particular relates to a composite bacterial agent capable of specifically degrading heavy oil pollutants for repairing heavy oil-polluted coastlines, and a preparation method and application thereof. Background technique [0002] The ocean is not only a treasure house of resources for human survival and development, but also an important transportation environment. With the increasing demand for oil, the rapid development of offshore oil industry and transportation, and the continuous expansion of offshore oil exploration, exploitation and transportation, marine oil spills and pollution accidents occur frequently, and oil and its products have become second only to cyanobacteria. , the second largest marine pollution source of red tide. And when the oil spill accident occurs in the sea, due to the action of tides and waves, the threat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10A62D3/02C12R1/025C12R1/01A62D101/20
CPCA62D3/02A62D2101/20B09C1/10C12N1/20
Inventor 宋佳宇蔡斌刘光全陈春茂岳勇阎光绪陈梅梅代小丽刘思敏贾伟超
Owner BC P INC CHINA NAT PETROLEUM CORP
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