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Composite bacteria liquid for cooperative degradation of petroleum and preparation method thereof

A compound bacterial solution and synergistic degradation technology, applied in the field of bioremediation of polluted marine environment, can solve the problems of low oil degradation rate and poor restoration effect, and achieve the effects of fast degradation rate, sufficient degradation and high oil degradation rate

Active Publication Date: 2010-12-15
THE FIRST INST OF OCEANOGRAPHY SOA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of a single type of obligate hydrocarbon-decomposing bacteria often has the problems of low oil degradation rate and poor repair effect.

Method used

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  • Composite bacteria liquid for cooperative degradation of petroleum and preparation method thereof
  • Composite bacteria liquid for cooperative degradation of petroleum and preparation method thereof
  • Composite bacteria liquid for cooperative degradation of petroleum and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: the preparation steps of the composite bacteria liquid of synergistically degrading petroleum:

[0027] (1) Preparation of seed solution: 10 L of M8 medium was prepared for the cultivation of strains 97CO-5 and 97CO-6, and 10 L of sodium acetate medium was prepared for the cultivation of strain PY97S. The cryopreserved strains were inoculated into the above two media according to the inoculation amount of 2%, that is, the strains 97CO-5 and 97CO-6 were inoculated into 10LM8 medium respectively, and the strain PY97S was inoculated into 10L sodium acetate medium. Under the conditions of 25° C., 150 rpm, and protected from light, culture until late logarithmic growth, and prepare seed liquid.

[0028] (2) Bacterial liquid fermentation culture

[0029] Prepare 200L of M8 medium for the fermenter culture of strains 97CO-5 and 97CO-6, prepare 200L of sodium acetate medium for the fermenter culture of strain PY97S; inoculate the seed solution into M8 according to ...

Embodiment 2

[0032] Embodiment 2 The test method of the oil degradation effect of the present invention is:

[0033] (1) Prepare the oil degradation effect test medium containing 22.79g NaCl and 11.18g MgCl per liter 2 ·6H 2 O, 3.98 g Na 2 SO 4 , 1.46gCaCl 2 2H 2 O, 1.30g TAPSO, 0.72g KCl, 0.27g NH 4 Cl, 89.00 mg Na 2 HPO 4 ·7H 2 O, 83.00 mg NaBr, 31.00 mg NaHCO 3 , 27.00 mg H 3 BO 3 , 24.00mg SrCl 2 ·6H 2 O, 2.60mg NaF, 2.00mg FeCl 2 4H 2 O, 10g crude oil, pH7.6.

[0034] (2) Petroleum degradation experiment: the bacteria solution and composite bacteria solution of single degrading bacteria PY97S, 97CO-5, 97CO-6 were received in 100mL petroleum medium according to the ratio of 2%; the treatment without inoculation of microorganisms was used as a negative control ; There were 3 repetitions for each of the above treatments. Cultivate for 2 weeks at 150 rpm, 20°C, and protected from light. Gravimetric method and GC-MS were used to analyze and compare the degradation rate of...

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Abstract

The invention relates to a composite bacteria liquid for cooperative degradation of petroleum and a preparation method thereof. The composite bacteria liquid is formed by mixing fermentation liquids of two strains of alcanivorax 97CO-5 and 97CO-6 and a strain of marinobacter PY97S wherein cell concentration is 107 to 109 CFU / mL. The preparation method comprises the steps of: inoculating seed liquids of the alcanivorax and the marinobacter respectively by 5% of inoculation quantity into an M8 culture medium or sodium acetate culture medium; fermenting and culturing under the conditions of 25 DEG C, the ventilation volume of 0.25m<3> / h, the stirring speed of 150rpm and defoaming by a fed defoamer; mixing all the fermentation liquids to prepare the composite bacteria liquid; centrifuging to prepare 5 times concentrate; and finally storing at 4 DEG C. The composite bacteria liquid is suitable for biodegradation of petroleum pollution at lower environmental temperature and thoroughly degrading alkane components and aromatic components in the petroleum, has higher petroleum degradation ratio than bacteria liquid with single degradation bacteria, has obvious cooperative degradation effect, and is applied to biological repair of coastlines or beaches polluted by ocean spilled petroleum.

Description

technical field [0001] The invention belongs to the technical field of bioremediation of polluted marine environments, and in particular relates to a composite bacterial solution based on the synergistic petroleum degradation effect of marine "obligate hydrocarbonoclastic bacteria" (OHCB) and a preparation method thereof. Background technique [0002] The frequent occurrence of marine oil spills not only causes huge economic losses to the marine aquaculture and tourism industries, but also has long-term negative effects on the marine ecological environment and human health. Therefore, how to eliminate marine oil spill pollution and restore damaged marine ecological environment has become one of the important contents of marine environmental protection. Due to the advantages of low cost, thoroughness and no secondary pollution, bioremediation technology has gradually become an effective way to deal with oil spills. [0003] Marine obligate hydrocarbon-decomposing bacteria ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N1/20A62D3/02C12R1/01A62D101/20
Inventor 郑立崔志松陈宇高伟王绍良李倩吴亮朱生凤王东王胜周文俊
Owner THE FIRST INST OF OCEANOGRAPHY SOA
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