CN112694984A - Microbial compound inoculant for restoring mining area soil and use method thereof - Google Patents
Microbial compound inoculant for restoring mining area soil and use method thereof Download PDFInfo
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- CN112694984A CN112694984A CN201911011934.4A CN201911011934A CN112694984A CN 112694984 A CN112694984 A CN 112694984A CN 201911011934 A CN201911011934 A CN 201911011934A CN 112694984 A CN112694984 A CN 112694984A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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Abstract
The invention relates to a microbial composite inoculant for restoring mine soil and a using method thereof. The composite microbial repairing microbial inoculum is prepared by mixing Azotobacter chroococcum, Bacillus coagulans and Pseudomonas stutzeri. The compound microbial agent provided by the invention has the advantages of easiness in obtaining, simplicity in preparation and no pollution, and can be used for efficiently improving the soil fertility.
Description
Technical Field
The invention belongs to the field of environmental remediation, and relates to a method for remedying the soil in a mining area by using soil microbial flora so as to achieve the purpose of remedying the ecological environment in the mining area.
Background
Mineral products are basic resources for economic development and make great contribution to the high-speed economic development of China. However, unreasonable mining activities bring serious ecological environmental problems to the periphery of mines, such as soil degradation, natural landscape destruction, geological disasters, water pollution, reduction in biodiversity, and the like. These environmental problems exacerbate the loss of mine ecosystem function, ultimately leading to the loss of the original environmental value of the mine.
The ecological restoration of the mining area is based on the restoration of soil, and the soil restoration method comprises a physical method, a chemical method and a biological method. The physical method needs a large amount of soil covering, the thickness of the soil covering is at least 30cm, and the repairing mode has high cost; the biggest problem of the chemical method is that the addition of chemical agents has the consequences of changing soil properties, reducing yield of plants and the like, and is not beneficial to the sustainable development of the ecology in a mining area. The microbial remediation has the advantages of low cost, good remediation effect, small damage to the environment and soil structure, contribution to ecological restoration and the like, is widely applied to ecological remediation of mines, and is a more effective soil improvement mode. Although the microbial remediation technology has attracted increasing attention, most of the reported microbial remediation methods have insignificant effect, slow effect and unstable remediation effect due to the difficulty in finding suitable strains, so that the popularization and application of the technology are limited.
Disclosure of Invention
The invention provides a compound microbial agent for restoring soil in a mining area and a using method thereof, so that soil damaged by mine development can be effectively restored.
The composite microbial repairing microbial inoculum provided by the invention comprises a mixture of Azotobacter chroococcum, Bacillus coagulans and Pseudomonas stutzeri.
Wherein the microbial agent combination comprises:
20-30 wt% of Pseudomonas stutzeri (China microbial species Inquiry Net number: bio-52779);
30-60 wt% of azotobacter chroococcum (Chinese microbial strain inquiry net number: bio-05484);
15-50 wt% of bacillus coagulans (Chinese microbial strain inquiry net number: bio-53285).
The sum of the weight percentages of all the microbial components is 100 percent.
The formula of the culture medium for enriching the compound microbial agent is as follows: 10 parts by weight of glucose, 5 parts by weight of calcium phosphate, 0.3 part by weight of sodium chloride, 0.03 part by weight of ferrous sulfate, 1 part by weight of yeast extract and 1000 parts by weight of water, and the pH value is controlled to be 7.0-7.5.
The composite microbial repairing microbial inoculum is used according to the following steps: carrying out enrichment culture on the compound microbial agent by using a liquid culture medium, wherein the culture temperature and time are as follows: above 30 ℃ (e.g. 30-35 ℃), 48-96 h; or 20-30 ℃ for 72-120 h. After the enrichment culture is finished, the concentration of the microbial inoculum is 0.7-2g/L, and the obtained flora is uniformly sprayed to the area to be repaired, wherein the inoculation amount is every m2The area is 1-10L; repeating the above steps for 3-5 times at intervals of 5-7 days. After the compound microbial agent is inoculated, the soil humidity needs to be kept at 35% +/-5%.
The compound microbial agent provided by the invention has the advantages of easiness in obtaining, simple preparation structure and no pollution, and can be used for efficiently improving the soil fertility.
Detailed Description
For a further understanding of the invention, reference will now be made to the preferred embodiments of the invention by way of example, and it is to be understood that the description is intended to further illustrate features and advantages of the invention, and not to limit the scope of the claims.
The strains Pseudomonas stutzeri (China microbial species Inquiry Net number: bio-52779), azotobacter chroococcum (China microbial species Inquiry Net number: bio-05484) and Bacillus coagulans (China microbial species Inquiry Net number: bio-53285) used in the following examples were purchased from China microbial species Inquiry Net (www.biobw.org).
The soil sample required by the experiment is from an open coal mine refuse dump, and the standard soil sample for evaluating the quality of the remediation microbial inoculum is from a natural grassland soil sample in the same area.
Example 1
(1) Preparing a composite microbial repairing microbial inoculum: 20 wt% of pseudomonas stutzeri, 60 wt% of azotobacter chroococcum and 15 wt% of bacillus coagulans, wherein the sum of the weight percentages of all the microbial components is 100%.
(2) Carrying out enrichment culture on the composite microbial remediation microbial inoculum, wherein the formula of a liquid culture medium is as follows: 10 parts by weight of glucose, 5 parts by weight of calcium phosphate, 0.3 part by weight of sodium chloride, 0.03 part by weight of ferrous sulfate, 1 part by weight of yeast extract and 1000 parts by weight of water, and the pH value is controlled to be 7.0-7.5.
(3) Carrying out enrichment culture on the composite microbial remediation microbial inoculum by using a liquid culture medium, wherein the culture temperature and time are as follows: 20 ℃ and 120 h. After the enrichment culture is finished, the concentration of the microbial inoculum is 2g/L, the microbial inoculum is uniformly sprayed to soil of a certain mining area to be repaired in the north, and the area of the repair area is about 200m2The inoculation amount is per m2The area is 10L; this was repeated 3 times with 5 days intervals.
(4) After each inoculation, water should be sprayed to keep the moisture content of the soil to 35% + -5%. The physical and chemical indexes of the soil can be close to the natural grassland state through detection.
Example 2
(1) Preparing a composite microbial repairing microbial inoculum: 30 wt% of pseudomonas stutzeri, 50 wt% of azotobacter chroococcum and 20 wt% of bacillus coagulans, wherein the sum of the weight percentages of all the microbial components is 100%.
(2) Carrying out enrichment culture on the composite microbial remediation microbial inoculum, wherein the formula of a liquid culture medium is as follows: 10 parts by weight of glucose, 5 parts by weight of calcium phosphate, 0.3 part by weight of sodium chloride, 0.03 part by weight of ferrous sulfate, 1 part by weight of yeast extract and 1000 parts by weight of water, and the pH value is controlled to be 7.0-7.5.
(3) Carrying out enrichment culture on the composite microbial remediation microbial inoculum by using a liquid culture medium, wherein the culture time is as follows: 35 ℃ and 48 h. After the enrichment culture is finished, the concentration of the microbial inoculum is 0.7g/L, the microbial inoculum is uniformly sprayed to soil of a certain mining area to be repaired in the north, and the area of the repair area is about 200m2The inoculation amount is per m2The area is 5L; this was repeated 4 times with 6 days intervals.
(4) After each inoculation, water should be sprayed to keep the moisture content of the soil to 35% + -5%. The physical and chemical indexes of the soil can be close to the natural grassland state through detection.
Example 3
(1) Preparing a composite microbial repairing microbial inoculum: 30 wt% of pseudomonas stutzeri, 40 wt% of azotobacter chroococcum and 30 wt% of bacillus coagulans, wherein the sum of the weight percentages of all the microbial components is 100%.
(2) Carrying out enrichment culture on the composite microbial remediation microbial inoculum, wherein the formula of a liquid culture medium is as follows: 10 parts by weight of glucose, 5 parts by weight of calcium phosphate, 0.3 part by weight of sodium chloride, 0.03 part by weight of ferrous sulfate, 1 part by weight of yeast extract and 1000 parts by weight of water, and the pH value is controlled to be 7.0-7.5.
(3) Carrying out enrichment culture on the composite microbial remediation microbial inoculum by using a liquid culture medium, wherein the culture time is as follows: 25 ℃ and 72 h. After the enrichment culture is finished, the concentration of the microbial inoculum is 1g/L, the microbial inoculum is uniformly sprayed to soil of a certain mining area to be repaired in the north, and the area of the repair area is about 200m2The inoculation amount is per m2The area is 1L; this was repeated 5 times with 7 days intervals.
(4) After each inoculation, water should be sprayed to keep the moisture content of the soil to 35% + -5%. The physical and chemical indexes of the soil can be close to the natural grassland state through detection.
Example 4
(1) Preparing a composite microbial repairing microbial inoculum: 25 wt% of pseudomonas stutzeri, 50 wt% of azotobacter chroococcum and 25 wt% of bacillus coagulans, wherein the sum of the weight percentages of all the microbial components is 100%.
(2) Carrying out enrichment culture on the composite microbial remediation microbial inoculum, wherein the formula of a liquid culture medium is as follows: 10 parts by weight of glucose, 5 parts by weight of calcium phosphate, 0.3 part by weight of sodium chloride, 0.03 part by weight of ferrous sulfate, 1 part by weight of yeast extract and 1000 parts by weight of water, and the pH value is controlled to be 7.0-7.5.
(3) Carrying out enrichment culture on the composite microbial remediation microbial inoculum by using a liquid culture medium, wherein the culture time is as follows: 30 ℃ and 96 h. After the enrichment culture is finished, the concentration of the microbial inoculum is 0.7g/L, the microbial inoculum is uniformly sprayed to soil of a certain mining area to be repaired in the north, and the area of the repair area is about 200m2The inoculation amount is per m2The area is 8L; this was repeated 5 times with 5 days intervals.
(4) After each inoculation, water should be sprayed to keep the moisture content of the soil to 35% + -5%. The physical and chemical indexes of the soil can be close to the natural grassland state through detection.
The soil and the control group (soil without spraying bacteria and natural grassland soil samples) repaired in the examples 1 to 4 were sent to Chongqing Sooo monitoring Co., Ltd for soil nutrient test, and the experimental results are shown in Table 1.
TABLE 1
The soil nutrient is characterized by an index of the carbon-nitrogen ratio of the soil. Soil carbon to nitrogen ratios between 15 and 25 generally indicate good soil nutrient status, with higher indicating more nutrients. In addition, the increase of the proportion content of the clay particles indicates that the water retention capacity of the soil is strengthened again, and the soil condition is gradually improved.
Claims (9)
1. A composite microbial repairing microbial inoculum comprises Azotobacter chroococcum, Bacillus coagulans and Pseudomonas stutzeri.
2. The composite microbial remediation agent of claim 1, comprising:
20-30 wt% pseudomonas stutzeri;
30-60 wt% of azotobacter chroococcum;
15-50 wt% of Bacillus coagulans,
wherein the sum of the weight percentages of all microorganism components is 100 percent.
3. A liquid medium for enrichment culture of the composite microbial remediation agent of claim 1, comprising 10 parts by weight of glucose, 5 parts by weight of calcium phosphate, 0.3 part by weight of sodium chloride, 0.03 part by weight of ferrous sulfate, 1 part by weight of yeast extract, 1000 parts by weight of water, and having a pH of 7.0 to 7.5.
4. The method for restoring the soil of the mining area is characterized in that the composite microbial restoration microbial inoculum of claim 1 or 2 is subjected to enrichment culture by using the liquid culture medium of claim 3, and then the obtained microbial inoculum is uniformly sprayed to the area to be restored.
5. The method of claim 4, wherein the enrichment culture is performed at 30 ℃ or higher for 48-96 hours, or at 20-30 ℃ for 72-120 hours.
6. The method according to claim 4, wherein the concentration of the composite microbial remediation agent after the enrichment culture is finished is 0.7-2 g/L.
7. A method according to claim 4, wherein the uniform spraying of the resulting flora on the area to be repaired is characterized by an inoculum size of inoculum per m2The area is 1-10L.
8. The method of claim 4, wherein the spraying is repeated 3-5 times with 5-7 days intervals.
9. The method according to claim 4, wherein the soil humidity needs to be kept at 35% ± 5% after inoculation of the composite microbial remediation agent.
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Citations (7)
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| CN104673319A (en) * | 2013-11-29 | 2015-06-03 | 大连三科生物工程有限公司 | Microbial soil conditioner for lithified soil |
| CN105838644A (en) * | 2016-05-05 | 2016-08-10 | 陈五岭 | Compound microbial agents, bacterial manure, preparation methods thereof and application thereof to restoration of saline alkali soil |
| CN108239611A (en) * | 2016-12-26 | 2018-07-03 | 北京有色金属研究总院 | One plant of brevibacterium and its method for heavy metals in farmland pollution in-situ immobilization |
| CN108707558A (en) * | 2018-04-25 | 2018-10-26 | 江苏世邦生物工程科技有限公司 | Microorganism formulation and its preparation method and application for restoration of soil polluted by heavy metal |
| CN109926445A (en) * | 2017-12-19 | 2019-06-25 | 国家电投集团远达环保工程有限公司重庆科技分公司 | A kind of soil remediation method |
| CN110026430A (en) * | 2019-04-12 | 2019-07-19 | 云南农业大学 | A kind of ecological recovery method in land reclamation in mining area |
-
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- 2019-10-23 CN CN201911011934.4A patent/CN112694984A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104673319A (en) * | 2013-11-29 | 2015-06-03 | 大连三科生物工程有限公司 | Microbial soil conditioner for lithified soil |
| CN104492805A (en) * | 2014-11-28 | 2015-04-08 | 云南圣清环保科技有限公司 | Soil remediation method |
| CN105838644A (en) * | 2016-05-05 | 2016-08-10 | 陈五岭 | Compound microbial agents, bacterial manure, preparation methods thereof and application thereof to restoration of saline alkali soil |
| CN108239611A (en) * | 2016-12-26 | 2018-07-03 | 北京有色金属研究总院 | One plant of brevibacterium and its method for heavy metals in farmland pollution in-situ immobilization |
| CN109926445A (en) * | 2017-12-19 | 2019-06-25 | 国家电投集团远达环保工程有限公司重庆科技分公司 | A kind of soil remediation method |
| CN108707558A (en) * | 2018-04-25 | 2018-10-26 | 江苏世邦生物工程科技有限公司 | Microorganism formulation and its preparation method and application for restoration of soil polluted by heavy metal |
| CN110026430A (en) * | 2019-04-12 | 2019-07-19 | 云南农业大学 | A kind of ecological recovery method in land reclamation in mining area |
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