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Bacterial agent for bioremediation of oil-polluted water area and preparation method

A technology of water organisms and inoculants used in the field of oil pollution environmental control can solve the problems of limiting the effectiveness of bio-enhanced restoration and easy loss of survival time of oil-degrading bacteria, and achieve the effects of degradable buoyancy and superior buoyancy.

Inactive Publication Date: 2014-01-22
NAVY MEDICINE RES INST OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Under the natural conditions of oil pollution, the inoculated oil-degrading bacteria face competition with indigenous microorganisms, and need to adapt to the new growth environment and withstand environmental pressures such as the toxicity of environmental pollutants, resulting in the easy loss and survival time of the added oil-degrading bacteria Short, this is the main issue limiting the effectiveness of bioaugmented remediation

Method used

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  • Bacterial agent for bioremediation of oil-polluted water area and preparation method
  • Bacterial agent for bioremediation of oil-polluted water area and preparation method
  • Bacterial agent for bioremediation of oil-polluted water area and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The enrichment and separation of embodiment 1 petroleum degrading bacteria

[0026] Inoculate 1 mL of collected oil-contaminated seawater into 100 mL of sterilized MMC with a volume fraction of 0.5% (v / v) of diesel. After cultivating on a shaker at 30°C and 200r / min for 5-6 days, take 1 mL from the culture medium and transfer it to 100 mL of fresh medium, and increase the oil concentration of the medium to 1% (v / v). Cultivate; the oil concentration is increased according to the gradient (0.5%, 1%, 1.5%, 2%, 2.5%), and enriched for five cycles. Use a pipette gun (the tip of which has been sterilized) to draw 1 mL of enriched water sample and place it in 9 mL of sterile water as 10 -1 Diluent; after mixing, draw 1mL of the liquid in the test tube and place it in 9mL sterile water as 10 -2 Dilutions, and so on, were made with serial dilutions of 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 Diluent: draw 0.2mL samples of different dilutions and drop them into LB solid m...

Embodiment 2

[0027] The construction of embodiment 2 petroleum degrading mixed flora

[0028] Inoculate the 4 strains of petroleum-degrading bacteria (Acinetobacter Y9, W3, F9 and Gordonella X1) isolated in Example 1 into LB liquid medium, and after culturing overnight, take equal amounts of bacterial liquids and mix them respectively. Form a mixed flora, collect the bacteria by centrifugation, inoculate into an inorganic salt medium containing 1% diesel oil, cultivate at 180rpm, 30°C for a certain period of time, measure the remaining diesel concentration in the medium, and calculate the degradation of diesel oil rate, such as figure 1 As shown, the degradation rate of diesel oil by bacteria group was significantly higher than that of single strain.

Embodiment 3

[0029] Embodiment 3 takes calcium alginate and chitosan as the preparation of the bacterial agent of material

[0030] Weigh 3g chitosan, dissolve in 90ml of 2% acetic acid solution to make solution A; weigh 5g urea and 1.2gNaH 2 PO 4 , dissolved in 10ml of pure water to prepare solution B; mix and dissolve solutions A and B, drop them into 1mol / L NaOH solution, solidify for 5min, and rinse the formed solidified pellets with clean water until they are neutral. Add it to 3% sodium alginate solution, stir well, drop into the solution containing 2% CaCl 2And in 0.5% chitosan solution, solidify 30min, the wet carrier that makes is added in the LB bacterial liquid of the mixed bacterial liquid among the embodiment 2, at 140rpm, cultivate 3h under 30 ℃ of conditions, take out the carrier that is fixed with degrading bacteria, Add protective agent, pre-freeze at -70°C, and vacuum freeze-dry in a lyophilizer to prepare the bacterial agent. Store at 4°C. The appearance of the bacteri...

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Abstract

The invention relates to the technical field of petroleum-polluted environment management, and particularly relates to a bacterial agent for bioremediation of an oil-polluted water area and a preparation method. The invention discloses a bacterial agent for petroleum bioremediation, which comprises a petroleum degradation bacterial colony and a biological immobilization carrier and is characterized in that the bacterial colony is fixed in a floatable and degradable biological carrier capable of slowly releasing nutrient substances such as nitrogen, phosphorus and the like. In the biological immobilization carrier, the slow-release fertilizer capable of releasing nutrient substances such as nitrogen, phosphorus and the like serves as a core, and the periphery is enveloped by the biological carrier with the immobilized petroleum degradation bacterial colony. The interior of the biological carrier is of a porous hollow structure so as to keep the floatability, the slow-release fertilizer can continuously provide nutrient substances such as nitrogen, phosphorus and the like for petroleum degradation bacteria, and additional nutrient substances are not required. The petroleum degradation bacterial agent provided by the invention can be applied to the bioremediation of a petroleum-polluted water body.

Description

1 technical field [0001] The invention relates to the technical field of oil-polluted environment treatment, in particular to a bacterial agent for bioremediation of oil-polluted waters and a preparation method thereof. 2 background technology [0002] With the rapid development of the petroleum industry, the damage of petroleum to the environment has attracted more and more attention. Especially for the marine environment, various oil spill accidents have caused serious harm to the marine ecosystem and restricted the sustainable development of human society and the environment. The bioremediation technology of oil pollution is considered to be one of the most effective ways to control oil pollution. It has the advantages of high efficiency, economy and no secondary pollution. The process of bioremediation generally includes two ways of biopromoting and bioaugmentation. Bioaugmentation refers to the addition of microorganisms that can degrade petroleum; biopromotion refers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C02F3/34C12N11/10C02F101/30
Inventor 罗群沈先荣侯登勇何颖刘琼王庆蓉张建国
Owner NAVY MEDICINE RES INST OF PLA
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