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Ecological microbial inoculum for remedying oil-polluted soils and preparation method thereof

A technology for oil pollution and bacterial agent, which is applied in the field of bacterial preparation and its preparation for restoring the ecology of oil-polluted soil, and can solve the problems of high density and high activity that have never been seen before.

Inactive Publication Date: 2010-12-15
NANKAI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been some studies at home and abroad on the restoration of oil-contaminated soil ecology, but there have been no reports of high-density, high-activity bacterial preparations that can be used in situ to degrade oil and restore oil-contaminated soil ecology.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Follow these steps:

[0021] 1) Activate the slant planes of the four strains stored at 4°C and culture them in shake flasks respectively. Cultured in soluble starch medium, the specific formula is as follows: soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.01g, distilled water 1000mL, pH7.2, 0.1Mpa, autoclave at 121℃ for 30min. Incubate at 28°C for 24h.

[0022] 2) Secondary liquid expansion culture (6L), replace the soluble starch in the above medium with cornmeal, add 0.1% humic acid (mass fraction), inoculum size 5%, culture at 28°C for 24h. .

[0023] 3) Tertiary liquid expansion culture (60 L), the medium is the same as above; the inoculum size is 10%, and cultured at 28° C. for 24 hours.

[0024] 4) Using humic acid as a carrier to fix and adsorb the bacterial solution on it. Sealed in a polyethylene bag and stored at room temperature.

Embodiment 2

[0026] Follow these steps:

[0027] 1) Activate the slant planes of the four strains stored at 4°C and culture them in shake flasks respectively. Expand the culture with soluble starch medium, the specific formula is as follows: soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.01g, distilled water 1000mL, pH7.0, 0.1Mpa, autoclave at 121℃ for 30min. Incubate at 28°C for 12h.

[0028] 2) Secondary liquid expansion culture (10L), replace the soluble starch in the above medium with cornmeal, add 0.1% humic acid (mass fraction), inoculum size 5%, culture at 28°C for 12h. .

[0029] 3) Tertiary liquid expansion culture (100 L), the medium is the same as above, the inoculum size is 10%, and cultured at 28° C. for 12 hours.

[0030] 4) Using humic acid as a carrier to fix and adsorb the bacterial solution on the humic acid. Sealed in a polyethylene bag and stored at room temperature.

Embodiment 3

[0032] Follow these steps:

[0033] 1) Activate the slant planes of the three strains stored at 4°C, and culture them in shake flasks respectively. Expand the culture with soluble starch medium, the specific formula is as follows: soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.01g, distilled water 1000mL, pH7.2, 0.1Mpa, autoclave at 121℃ for 30min. Incubate at 28°C for 12-24 hours.

[0034] 2) Secondary liquid expansion culture (30L), replace the soluble starch in the above medium with cornmeal, add 0.1% humic acid (mass fraction), inoculum size 5%, culture at 28°C for 24h.

[0035] 3) Tertiary liquid expansion culture (300 L), the medium is the same as above, the inoculum size is 10%, and cultured at 28° C. for 12 to 24 hours.

[0036] 4) Using humic acid as a carrier to fix and adsorb the bacterial solution on the humic acid. Sealed in a polyethylene bag and stored at room temperature.

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Abstract

The invention relates to an ecological microbial inoculum for remedying oil-polluted soils and a preparation method thereof. The ecological microbial inoculum for remedying oil-polluted soils is a mixed microflora of bacillus subtilis, pseudomonas fluorescens, berea trichosporon asahii and candida tropicalis, which are optimally mixed in a ratio of 1:1:1:1 basically. The invention has the advantages that: after the microbial inoculum is applied to the oil-polluted soils, a micro-ecological dominant microflora can be formed rapidly; and in case that the microbial inoculum in an amount which is 2 to 5 percent based on the weight of surface soils (cultivation layer: about 15 to 20cm) is applied to the oil-polluted soils, the oil removing rate is more than or equal to 75 percent (by using the methylene chloride refluxing extraction / gravimetric method) within three months; and the ecological microbial inoculum has a high effective colony count, a strong adaptability, and an obvious effect on remedying the ecological environments of oil-polluted soils.

Description

technical field [0001] In the field of biological technology, the present invention relates to a bacterial preparation for restoring the ecology of petroleum-contaminated soil and a preparation method thereof. Background technique [0002] The pollution of the surrounding environment will inevitably be caused in the oil extraction, transportation and use, especially the pollution of the soil ecosystem is becoming more and more serious. There have been some studies at home and abroad on the restoration of oil-contaminated soil ecology. However, there have been no reports of high-density, high-activity bacterial preparations that can be used on a large scale in situ to degrade oil and restore oil-contaminated soil ecology. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a high-density, high-activity microbial preparation for degrading petroleum and restoring the ecology of petroleum-contaminated soil and a preparat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N1/16C12N1/20B09C1/10C12R1/125C12R1/39C12R1/645C12R1/74
Inventor 唐景春孙正贵王如刚祝威乔俊桂召龙秦晓董健张清敏王田丽
Owner NANKAI UNIV
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