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Offshore oil degrading microbial inoculum and preparation method thereof

A technology of oil-degrading bacteria and offshore oil, applied in the direction of fixing on or in inorganic carriers, fixing on/in organic carriers, biological water/sewage treatment, etc., can solve the problem of low concentration of effective degradation bacteria and environmental conditions Sensitivity, poor resistance to poisonous damage, etc.

Inactive Publication Date: 2012-07-25
NAVY MEDICINE RES INST OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, petroleum hydrocarbon degrading bacteria often cannot be directly applied in the field. The reasons may be due to the low concentration of effective degrading bacteria per unit volume, slow reaction initiation, inferior competition with indigenous bacteria, poor resistance to toxicity, and sensitivity to environmental conditions.

Method used

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  • Offshore oil degrading microbial inoculum and preparation method thereof
  • Offshore oil degrading microbial inoculum and preparation method thereof
  • Offshore oil degrading microbial inoculum and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1. Enrichment and cultivation of crude oil degrading bacterial flora

[0035] Collect surface water samples from the seawater area polluted by crude oil, take 2L of surface seawater in a sterilized 4L brown glass bottle and seal it for preservation, and carry out microbial enrichment and culture within 24 hours.

[0036] 1 mL of the collected water sample was inoculated into 100 mL of sterilized artificial seawater medium (MMC) containing 0.5% (v / v) crude oil. The formula of MMC is (content per liter): NaCl 24g; MgSO 4 ·7H 2 O 0.7g; NH 4 NO 3 1g; KCl 0.7g; KH 2 PO 4 2g; Na 2 HPO 4 12H 2 O 3g; pH7.5, add appropriate amount of trace elements to mix after sterilization. Trace elements are filtered and sterilized by 0.22μm filter membrane, and its composition (content per liter) is as follows: CaCl 2 2 mg; FeCl 3 ·6H 2 O 50mg; CuSO 4 0.5 mg; MnCl 2 4H 2 O 0.5mg; ZnSO 4 ·7H 2 O 10mg / L.

[0037] After cultivating on a shaker at 30°C for 7 days...

Embodiment 2

[0038] Example 2. Preparation of floating and degradable biological immobilization carrier

[0039] Prepare 1% CaCl separately 2 solution, 1% sodium alginate solution and 1% chitosan and 1% CaCl 2 mixture.

[0040] Weigh an appropriate amount of puffed small yellow rice, add 1% CaCl 2 solution, stir and infiltrate for 5-10 minutes; from CaCl 2 Take out the infiltrated puffed millet from the solution, add it to 1% sodium alginate solution, stir for 5-10 minutes, and carry out film-forming reaction; It is composed of a calcium alginate film, and the semi-finished carrier is added with 1% chitosan and 1% CaCl 2 In the mixed solution, stir for 5-10 minutes to carry out re-coagulation, so that the calcium alginate film of the outer layer and chitosan are re-coagulated into a composite film; the carrier is taken out from the mixed solution and set aside, such as figure 1 shown.

Embodiment 3

[0041] Embodiment 3. Activation, cultivation of petroleum degrading flora

[0042] The oil-degrading bacterial population stored at -70°C was inoculated into 5ml LB medium at a ratio of 1%, and cultured on a shaker at 180rpm at 30°C for 8 hours for activation. The composition of the LB medium is as follows (content per liter): 10 g of NaCl, 10 g of peptone, and 5 g of yeast extract. The activated culture was then inoculated into 500ml LB medium at a ratio of 1%. 180rpm, shaker at 30°C for 3-6 hours, OD 600 The value is between 0.6 and 1.0, and it is used as a degrading bacteria culture solution for preparing bacterial agents.

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Abstract

The invention relates to the technical field of marine environment management, in particular to an offshore oil degrading microbial inoculum and a preparation method thereof. The invention discloses an offshore oil degrading microbial inoculum, which comprises a petroleum degrading flora and a biological fixation carrier, and is characterized in that the microbial inoculum is prepared by attaching the petroleum degrading flora on the floating degradable biological fixation carrier. According to the invention, the biological fixation carrier takes an expanding degradable material as a core, and an outer layer is coated with a degradable composite membrane with positive electricity; the biological fixation carrier has floating property through the expanding degradable material; the degradable composite membrane with positive electricity and a thallus with negative electricity can attract mutually, so that the bacterium are firmly attached on the carrier, and the composite membrane is a porous loose structure, so that the degrading flora can be attached on the wall of the porous structure of the composite membrane; in comparison with the simple petroleum degrading flora, the degrading efficiency of the degrading microbial inoculum is remarkably improved. The offshore oil degrading microbial inoculum, provided by the invention, is expected to be used for bio-remediation of marine oil pollution.

Description

1 technical field [0001] The invention relates to the technical field of marine environment treatment, in particular to a marine oil-degrading bacterial agent and a preparation method thereof. 2 background technology [0002] With the world's increasing demand for oil and its products, oil spill accidents in the process of offshore exploitation, transportation, loading and unloading, and utilization of oil are also increasing. Oil spills not only cause serious environmental pollution, but also lead to a serious decline in the environmental quality and biodiversity index of coastal waters due to the potential toxicity and bioaccumulation effects of petroleum hydrocarbon pollutants, destroying the function of the ecosystem, and affecting the aquatic industry and tourism. It will also cause huge economic losses. In addition, polycyclic aromatic hydrocarbons in petroleum hydrocarbons have strong teratogenic, carcinogenic and mutagenic effects, and may enter the human body throu...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/12C12N11/10C12N11/02C02F3/34
Inventor 侯登勇沈先荣何颖王庆蓉蒋定文刘玉明刘琼陈伟李珂娴莫琳芳钱甜甜
Owner NAVY MEDICINE RES INST OF PLA
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