Preparation method of petroleum degrading bacteria solid inoculant and method for restoring petroleum-polluted soil by using prepared solid inoculant

A technology of petroleum-degrading bacteria and solid bacterial agent, which is applied in the field of microbial remediation of petroleum-contaminated soil, and can solve problems such as unguaranteed actual effects, unrealized industrialized production, suboptimal viability, and unsatisfactory degradation efficiency.

Active Publication Date: 2015-03-25
QINGDAO AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The large-scale promotion of microbial remediation of oil-contaminated soil technology requires a large number of microbial agents. Domestic research on oil-degrading agents has only carried out small-scale laboratory cultivation, and has not achieved industrial production.
[0007] There have been reports on methods for screening petroleum-degrading bacteria using petroleum hydrocarbons as the only carbon source, but most of them use a single petroleum hydrocarbon or several petroleum hydrocarbons and their intermediates as carbon sources for screening. There are many kinds of actual conditions, so the viabil

Method used

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  • Preparation method of petroleum degrading bacteria solid inoculant and method for restoring petroleum-polluted soil by using prepared solid inoculant
  • Preparation method of petroleum degrading bacteria solid inoculant and method for restoring petroleum-polluted soil by using prepared solid inoculant
  • Preparation method of petroleum degrading bacteria solid inoculant and method for restoring petroleum-polluted soil by using prepared solid inoculant

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0067] Example 1

[0068] Screening of high-efficiency oil-degrading bacteria strains

[0069] 1. Collection of the source of indigenous bacteria from petroleum degradation

[0070] The oil-polluted soil near the oil storage tank of the Shengli Oilfield Refinery in Shandong Province was used as the source of the indigenous bacteria for oil degradation. Sampling method: Use a small shovel to take a soil sample with a thickness of 10 cm, and mix it evenly after sampling at multiple points. Take 3 kg of the soil sample and bring it back to the laboratory for later use.

[0071] 2. Preparation of petroleum polysorbate-80 solubilizing solution

[0072] After the surfactant reaches the critical micelle concentration (CMC) in the aqueous solution, the solubility of some water-insoluble or slightly soluble substances in the micellar solution can be significantly increased to form a transparent colloidal solution. This effect is called solubilization (solubilization). Polysorbate-80 (polyoxye...

Example Embodiment

[0119] Example 2

[0120] Liquid fermentation production of petroleum degrading bacteria

[0121] 1. Primary seed cultivation

[0122] Under aseptic conditions, the purified oil-degrading bacteria mixed strain 2 of Example 1 was inoculated into the test tube slant medium. Culture medium composition per 1000ml sterile water: Petroleum polysorbate-80 solubilization solution containing 0.125g equivalent of petroleum (the mass concentration of petroleum is 2%, the mass concentration of polysorbate-80 is 4%, and the rest is water.), beef Paste 3g, peptone 10g, NaCl 5g, agar powder 20g, pH 7, 121°C, sterilize for 20 minutes. The cultivation conditions are: 30°C, 24 hours cultivation, as the first-level seed.

[0123] 2. Secondary seed culture

[0124] Under aseptic conditions, take two rings of first-grade seeds and put them into a 500ml Erlenmeyer flask containing 200ml culture medium. Shake flask culture medium composition per 1000ml sterile water: petroleum polysorbate-80 solubilizatio...

Example Embodiment

[0131] Example 3

[0132] Petroleum-degrading solid composite microbial agent and plant joint remediation of oil-contaminated soil

[0133] 1. Selection and processing of experimental bases

[0134] This experiment selects the land near the oil extractor of an oil extraction plant in Dongying City. This land is located near the oil extractor and is polluted by petroleum. The total petroleum hydrocarbon content is 4g / Kg. The basic physical and chemical properties of the soil are shown in Table 5. In the present invention, alfalfa-microbes are used to repair the oil-contaminated soil, and the planting density is about 8000 plants / mu. The prepared oil-degrading bacteria solid inoculum is added to the oil-contaminated soil at 10-100 kg / mu of land, and plow 0-30cm topsoil and mix well. When planting alfalfa, cover the soil with a film to maintain moisture, and apply controlled-release fertilizer to ensure the nutrients required for the normal growth of plants. In the seedling stage of ...

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Abstract

The invention relates to a preparation method of a petroleum degrading bacteria solid inoculant. The preparation method comprises the following steps: A, screening and domestication of the petroleum degrading bacteria; B, preparation of a seed culture solution; C, fermentation of the solid inoculant; and D, drying, pulverization, measurement and packaging of composted products. The petroleum degrading bacteria solid inoculant has the beneficial effect of improving the petroleum dissolution effect by taking a nonionic surfactantpolysorbate-80 (tween 80) as a solubilizer. The preparation method has the advantages that solid fermentation raw materials are easy to get and the process is relatively simple; and the solid inoculant contains a large amount of carbon and nutrient elements so as to provide more proper matrix for the growth of the bacteria, has strong affinity for the microorganisms, has immobilization efficiency, is capable of improving the competitiveness and degrading efficiency of the added microorganisms and indigenous microorganisms, and is convenient to transport and operate agriculturally and suitable for the large-scale in-situ biological remediation of the petroleum-polluted soil.

Description

technical field [0001] The invention relates to the technical field of microorganism restoration of petroleum-contaminated soil, in particular to a method for preparing a solid bacterial agent of petroleum-degrading bacteria and a method for repairing oil-contaminated soil by the solid bacterial agent prepared therefrom. Background technique [0002] With the rapid development of economy, soil environmental problems caused by humans are becoming more and more serious. As a non-renewable resource for human beings, soil is the basis for human survival. The toxic and harmful substances discharged by industry and agriculture are increasing day by day, and the organic pollution caused by the soil is on the rise. Among them, the oil discharged in the process of oil development, transportation, processing, storage and transportation is particularly harmful to the soil environment. Petroleum is a mixture of various chemical substances with different chemical properties, mainly inc...

Claims

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Application Information

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IPC IPC(8): C12N1/36B09C1/10
CPCB09C1/10C12N1/36
Inventor 郑庆柱田侠
Owner QINGDAO AGRI UNIV
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