CN115055515A - Microbial remediation method for micro-plastics in garbage-polluted soil - Google Patents
Microbial remediation method for micro-plastics in garbage-polluted soil Download PDFInfo
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- CN115055515A CN115055515A CN202210923383.4A CN202210923383A CN115055515A CN 115055515 A CN115055515 A CN 115055515A CN 202210923383 A CN202210923383 A CN 202210923383A CN 115055515 A CN115055515 A CN 115055515A
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- 238000005067 remediation Methods 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 34
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- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 12
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- 238000004132 cross linking Methods 0.000 claims abstract description 12
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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
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/08—Reclamation of contaminated soil chemically
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/50—Reuse, recycling or recovery technologies
- Y02W30/62—Plastics recycling; Rubber recycling
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- Biotechnology (AREA)
- Biomedical Technology (AREA)
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Abstract
The invention discloses a microbial remediation method for micro-plastics in garbage-polluted soil, and belongs to the technical field of in-situ remediation of garbage-polluted soil. The method comprises the following steps: (1) expanding and culturing microorganisms on the surface of the plastic in the garbage polluted site by using a liquid culture medium, transferring the obtained bacterial liquid into a selective inorganic salt culture medium for enrichment culture, and enriching to obtain functional floras with plastic degradation capability; (2) mixing the bacterial suspension and the biomass charcoal for adsorption, freezing and drying to obtain microorganism immobilized biomass charcoal; (3) adding the microorganism immobilized biochar into a mixed solution of carboxymethyl cellulose and sodium alginate, uniformly stirring, and then dropwise adding the mixture into a calcium chloride solution for cross-linking reaction to obtain biochar-based microorganism immobilized particles; (4) and adding the biochar-based microorganism immobilized particles into the garbage-polluted soil, and carrying out in-situ remediation on the garbage-polluted soil. The invention solves the problems of difficult degradation of plastics in the garbage polluted soil and migration removal of micro plastics.
Description
Technical Field
The invention relates to the technical field of in-situ remediation of garbage-polluted soil, in particular to a microbial remediation method of micro-plastics in the garbage-polluted soil.
Background
With the rapid development of economy, the wide use of plastics and products thereof leads to an increase in the proportion of plastic waste. The plastic has the characteristics of difficult decomposition, low recovery rate and the like, so that the problem of serious environmental pollution is caused, particularly in garbage polluted soil. The natural decomposition of plastics and improper treatment mode can generate a new pollutant, namely micro plastics, which is widely distributed in various water and soil environments at present. The micro plastic generally refers to plastic particles with the particle diameter of less than 5mm, has hydrophobicity, durability and difficult degradability, and is difficult to recover and obtain from soil. At present, most of plastic wastes are treated by landfill, incineration, reprocessing and the like, but the treatment cost is high and secondary pollution is generated to the environment. In contrast, the key core of the technology for microbial degradation treatment is the selection of microbial degrading strains, which is safe and environmentally friendly.
The method for removing or repairing the micro-plastic is shallow, more micro-plastic is concentrated in a water environment and mainly comprises biodegradation, physical flocculation and the like, but the problems of long treatment period, low adsorption efficiency and the like exist. The micro plastic has mobility, can flow with water, is enriched in soil, can be ingested by passive plants, and seriously threatens the ecological environment and human health. The content of the micro-plastic in the soil can be increased by simply applying the plastic degradation flora, and the micro-plastic pollution is aggravated. Therefore, it is of great significance to explore an in-situ soil remediation technology with both plastic degradation and micro-plastic removal.
Disclosure of Invention
The invention aims to provide a microbial remediation method for micro-plastics in garbage-polluted soil, which aims to solve the problems of difficult degradation of the plastics in the garbage-polluted soil and migration and removal of the micro-plastics.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a microbial remediation method of micro-plastics in garbage contaminated soil, which comprises the following steps:
(1) culturing microorganisms on the surface of plastic in a garbage polluted site by using a liquid LB culture medium, transferring the obtained bacterial liquid into a selective inorganic salt culture medium according to the volume ratio of (1-3) to (10), and transferring for 3-5 times to obtain a functional flora with plastic degradation capability;
(2) preparing the functional flora into a bacterial suspension, wherein the bacterial suspension and the biomass charcoal are mixed according to the mass ratio of (10-20): 1, mixing, adsorbing, freezing and drying to obtain microorganism immobilized biochar;
(3) adding the microorganism immobilized biochar into a mixed solution of carboxymethyl cellulose and sodium alginate, stirring uniformly, and then slowly dropwise adding the mixture into a calcium chloride solution for crosslinking reaction to obtain biochar-based microorganism immobilized particles;
(4) adding the biochar-based microorganism immobilized particles into garbage contaminated soil according to the mass percent of 0.5-5%, and carrying out in-situ remediation on the garbage contaminated soil.
Preferably, in the step (1), the selective inorganic salt medium comprises the following components in parts by weight: carbon source 5.0g, (NH) 4 )NO 3 1g、Na 2 HPO 4 1g、KCl 0.5g、CaCl 2 0.3g、FeSO 4 0.2g、MgSO 4 0.1g、ZnSO 4 0.1g, 0.08g of vitamin complex and Na 2 SeO 3 0.03g、H 3 BO 3 0.02g, 1L of water; wherein the carbon source is at least one of PE, PP and PVC. Preferably, in the step (2), the biomass charcoal is obtained by crushing the waste biomass, calcining the crushed waste biomass at a high temperature of 300-600 ℃ for 1-3h in a nitrogen environment, and grinding and sieving the crushed waste biomass.
Preferably, the waste biomass includes at least one of corn stover, rice stover, wood chips, fruit shells, and shells.
Preferably, the conditions of the mixed adsorption are as follows: oscillating for 6-12h at 15-20 ℃ and 150 r/min.
Preferably, the particle size of the microorganism immobilized biomass charcoal is less than 100 μm.
Preferably, the OD of the bacterial suspension 600 Is 0.8.
Preferably, in the step (3), the mass ratio of the mixed solution to the microorganism immobilized biomass charcoal is 100 (0.5-2).
Preferably, in the mixed solution, the mass fraction of the carboxymethyl cellulose is 1-3%, and the mass fraction of the sodium alginate is 1-3%; the mass fraction of the calcium chloride solution is 1-4%.
The invention discloses the following technical effects:
according to the invention, biomass charcoal is used as a carrier, microbial flora with strong plastic degradation capability obtained by screening from the surface of plastic garbage is fixed on the biomass charcoal through adsorption, biomass charcoal-based microorganism immobilized particles are obtained through embedding and crosslinking, and the biomass charcoal-based microorganism immobilized particles are applied to in-situ remediation of garbage-polluted soil. The invention can improve the degradation of plastics in the garbage polluted soil by functional flora in the immobilized particles, and in addition, both biomass charcoal and cellulose components can adsorb the micro plastics in the polluted soil so as to prevent and control the migration and diffusion of the micro plastics in the soil.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required in the embodiments will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a technical route chart of the microbial remediation method of the micro-plastics in the garbage-contaminated soil according to the present invention.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
According to the invention, biomass carbon is used as a carrier, microbial flora with strong plastic degradation capability obtained by screening from the surface of plastic garbage is fixed on the biomass carbon through adsorption, biomass carbon-based microbial immobilized particles are obtained through embedding and crosslinking, and the immobilized particles are applied to in-situ remediation of garbage-polluted soil, and the technical scheme is shown in figure 1.
Example 1
(1) Collecting a plurality of plastic wastes made of PE, PP, PVC and the like from a plurality of waste pollution sites, slightly washing off attachments on the surfaces by using sterile water, shearing the plastic wastes to the size of 1cm, putting the plastic wastes into a liquid LB culture medium according to the mass ratio of 1:25, and oscillating the plastic wastes at the conditions of 150rpm and 25 ℃ for 48 hours to enrich microorganisms on the surfaces of the plastics;
transferring LB culture solution into selective inorganic salt culture medium (flake PE 2.5g, flake PP2.5g, NH) according to volume ratio of 1:10 4 NO 3 1g、Na 2 HPO 4 1g、KCl 0.5g、CaCl 2 0.3g、FeSO 4 0.2g、MgSO 4 0.1g、ZnSO 4 0.1g, 0.08g of vitamin complex and Na 2 SeO 3 0.03g、H 3 BO 3 0.02g, 1L of water), and carrying out shake culture at 150rpm and 25 ℃ for 7 d; and transferring the obtained culture solution into a selective inorganic salt culture medium again for culture, and repeating the transfer for 3 times to reduce the mass of the flaky plastics (PE and PP) in the culture medium by 12 percent, namely: enriching to obtain functional flora with plastic degradation capacity; the functional flora with plastic degradation capability takes at least one of PE, PP and PVC as a unique carbon source, and the weight of the degraded plastic is reduced by more than 10%.
Centrifuging the microorganism liquid at 5000rpm for 10min, and resuspending the thallus with sterile deionized water to obtain bacterial suspension (OD) 600 0.8).
(2) Crushing waste biomass wood chips, calcining at 500 ℃ for 2h in a nitrogen environment, and grinding and sieving to obtain wood chip biomass charcoal with the particle size of less than 100 microns.
(3) And mixing the wood chip biomass charcoal and the bacterial suspension according to the mass ratio of 1:10, oscillating for 8 hours at 15 ℃ at 150r/min for mixed adsorption, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microbial immobilized biomass charcoal (1%, w/w) into a mixed solution containing 1.5% (w/w) of carboxymethyl cellulose and 1.5% (w/w) of sodium alginate, uniformly stirring, slowly dropwise adding the mixture into a 2% calcium chloride solution, and crosslinking for 16h to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
(4) The experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 1.5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect. The results are shown in Table 1.
Example 2
(1) The same as in (1) of example 1.
(3) Mixing the wood chip biomass charcoal and the bacterial suspension according to a ratio (w/w) of 1:10, oscillating for 8h at 15 ℃ and 150r/min for mixing and adsorbing, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microbial immobilized biochar (1 percent, w/w) into a mixed solution containing 3 percent (w/w) of carboxymethyl cellulose and 1.5 percent (w/w) of sodium alginate, uniformly stirring, slowly dripping the mixture into a 2 percent calcium chloride solution, and crosslinking for 16 hours to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
The rest steps are the same. The results are shown in Table 1.
Example 3
(1) The same as in (1) of example 1.
(2) Crushing waste biomass wood chips, calcining at 300 ℃ for 1h in a nitrogen environment, and grinding and sieving to obtain wood chip biomass charcoal with the particle size of less than 100 microns.
(3) Mixing the wood chip biomass charcoal and the bacterial suspension according to a ratio (w/w) of 1:10, oscillating for 6h at 18 ℃ and 150r/min for mixing and adsorbing, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microbial immobilized biochar (0.5 percent, w/w) into a mixed solution containing 1 percent (w/w) of carboxymethyl cellulose and 1 percent (w/w) of sodium alginate, uniformly stirring, slowly dripping the mixture into a 1 percent calcium chloride solution, and crosslinking for 20 hours to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
(4) The experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 0.5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect. The results are shown in Table 1.
Example 4
(1) The same as in (1) of example 1.
(2) Crushing waste biomass wood chips, calcining at 600 ℃ for 3h in a nitrogen environment, and grinding and sieving to obtain wood chip biomass charcoal with the particle size of less than 100 microns.
(3) Mixing the wood chip biomass charcoal and the bacterial suspension according to a ratio (w/w) of 1:10, oscillating for 12h at 20 ℃ and 150r/min for mixed adsorption, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microorganism immobilized biochar (2 percent, w/v) into a mixed solution containing 3 percent (w/v) of carboxymethyl cellulose and 3 percent (w/v) of sodium alginate, uniformly stirring, slowly dripping the mixture into a 4 percent calcium chloride solution, and crosslinking for 24 hours to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
(4) The experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass carbon-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation and analysis to judge the treatment effect. The results are shown in Table 1.
Example 5
(1) The same as in (1) of example 1;
(2) crushing waste biomass corn straws, calcining at the high temperature of 500 ℃ for 2h in a nitrogen environment, and grinding and sieving to obtain corn straw biomass charcoal with the particle size of less than 100 mu m;
(3) same as in (3) of example 1;
(4) the same as in (4) of example 1.
The results are shown in Table 1.
Example 6
The difference from the example 5 is that:
(1) collecting a plurality of plastic wastes made of PE, PP, PVC and the like from a plurality of waste pollution sites, slightly washing off attachments on the surfaces of the plastic wastes by using sterile water, shearing the plastic wastes to be 1cm in size, putting the plastic wastes into a liquid LB culture medium according to the mass ratio of 1:25, and oscillating the plastic wastes for 48 hours at the conditions of 150rpm and 30 ℃ to enrich microorganisms on the surfaces of the plastic wastes;
transferring LB culture solution into selective inorganic salt culture medium (PE 5g, NH) according to the volume ratio of 1:10 4 NO 3 1g、Na 2 HPO 4 1g、KCl 0.5g、CaCl 2 0.3g、FeSO 4 0.2g、MgSO 4 0.1g、ZnSO 4 0.1g, 0.08g of vitamin complex and Na 2 SeO 3 0.03g、H 3 BO 3 0.02g, 1L of water), and carrying out shake culture at 150rpm and 25 ℃ for 7 d; transferring the obtained culture solution into a selective inorganic salt culture medium again for culture, repeating the transfer for 3 times, and enriching to obtain a functional flora with plastic degradation capacity (the weight of PE plastic is reduced by 10%);
(2) crushing waste biomass corn straws, calcining at 300 ℃ for 2h in a nitrogen environment, and grinding and sieving to obtain the corn straw biomass charcoal with the particle size less than 100 mu m.
The rest steps are the same. The results are shown in Table 1.
Example 7
The difference from the example 6 lies in: crushing waste biomass corn straws, calcining at 600 ℃ for 3h in a nitrogen environment, and grinding and sieving to obtain the corn straw biomass charcoal with the particle size less than 100 mu m.
The rest steps are the same. The results are shown in Table 1.
Example 8
The difference from the example 6 lies in: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 0.5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation and analysis to judge the treatment effect.
The rest steps are the same. The results are shown in Table 1.
Example 9
The difference from the example 6 lies in: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 4% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
The rest steps are the same. The results are shown in Table 1.
Example 10
The difference from the example 6 lies in: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
The rest steps are the same. The results are shown in Table 1.
Example 11
The difference from the example 6 lies in: (3) mixing the corn straw biomass charcoal and the bacterial suspension according to a ratio (w/w) of 1:10, oscillating for 8 hours at 15 ℃ at 150r/min for mixed adsorption, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microorganism immobilized biochar (1 percent, w/w) into a mixed solution containing 3 percent (w/w) of carboxymethyl cellulose and 1.5 percent (w/w) of sodium alginate, uniformly stirring, slowly dripping the mixture into a 2 percent calcium chloride solution, and crosslinking for 16 hours to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
The rest steps are the same. The results are shown in Table 1.
Example 12
(1) The same as in (1) of example 6;
(2) crushing waste biomass shells, calcining at the high temperature of 500 ℃ for 2h in a nitrogen environment, and grinding and sieving to obtain shell biomass charcoal with the particle size of less than 100 mu m;
(3) same as in (3) of example 6;
(4) the same as in (4) of example 6.
The results are shown in Table 1.
Example 13
The difference from example 12 is that:
(1) collecting a plurality of plastic wastes made of PE, PP, PVC and the like from a plurality of waste pollution sites, slightly washing off attachments on the surfaces by using sterile water, shearing the plastic wastes to the size of 1cm, putting the plastic wastes into a liquid LB culture medium according to the mass ratio of 1:25, and oscillating the plastic wastes at the conditions of 150rpm and 25 ℃ for 48 hours to enrich microorganisms on the surfaces of the plastics;
transferring LB culture solution into selective inorganic salt culture medium (PP 2.5g, PVC 2.5g, NH) according to volume ratio of 1:10 4 NO 3 1g、Na 2 HPO 4 1g、KCl 0.5g、CaCl 2 0.3g、FeSO 4 0.2g、MgSO 4 0.1g、ZnSO 4 0.1g, 0.08g of vitamin complex and Na 2 SeO 3 0.03g、H 3 BO 3 0.02g), and carrying out shaking culture at 150rpm and 25 ℃ for 7 d; transferring the obtained culture solution into a selective inorganic salt culture medium again for culture, repeating the transfer for 4 times, and enriching to obtain a functional flora with plastic degradation capacity (the total weight of PP and PVC plastic is reduced by 13%);
(2) crushing waste biomass shells, calcining at 300 ℃ for 2h in a nitrogen environment, and grinding and sieving to obtain shell biomass charcoal with the particle size of less than 100 mu m.
The rest steps are the same. The results are shown in Table 1.
Example 14
The difference from example 13 is that: crushing waste biomass shells, calcining at 600 ℃ for 3h in a nitrogen environment, and grinding and sieving to obtain shell biomass charcoal with the particle size of less than 100 mu m.
The rest steps are the same. The results are shown in Table 1.
Example 15
The difference from example 13 is that: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 0.5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
Example 16
The difference from example 13 is that: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 4% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
Example 17
The difference from example 13 is that: (4) the garbage contaminated soil remediation effect experiment: uniformly adding the biomass carbon-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
Example 18
The difference from example 13 is that: (3) mixing the shell biomass charcoal and the bacterial suspension according to a ratio (w/w) of 1:10, oscillating for 8h at 15 ℃ and 150r/min for mixed adsorption, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microbial immobilized biochar (1 percent, w/w) into a mixed solution containing 3 percent (w/w) of carboxymethyl cellulose and 1.5 percent (w/w) of sodium alginate, uniformly stirring, slowly dripping the mixture into a 2 percent calcium chloride solution, and crosslinking for 16 hours to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
The rest steps are the same. The results are shown in Table 1.
Example 19
(1) The same as in (1) of example 13;
(2) crushing the wood chips, the corn straws and the shells, mixing according to the mass ratio of 1:1:1, and calcining at the high temperature of 500 ℃ for 2 hours in a nitrogen environment to obtain the mixed biomass charcoal.
(3) The same as in (3) of example 13.
(4) The same as in (4) of example 13.
The results are shown in Table 1.
Example 20
The difference from example 19 is:
(1) collecting a plurality of plastic wastes made of PE, PP, PVC and the like from a plurality of waste pollution sites, slightly washing off attachments on the surfaces by using sterile water, shearing the plastic wastes to the size of 1cm, putting the plastic wastes into a liquid LB culture medium according to the mass ratio of 1:25, and oscillating the plastic wastes at the conditions of 150rpm and 25 ℃ for 48 hours to enrich microorganisms on the surfaces of the plastics;
transferring LB culture solution into selective inorganic salt culture medium (PE 2g, PP 1.5g, PVC 1.5g, NH) according to volume ratio of 1:10 4 NO 3 1g、Na 2 HPO 4 1g、KCl 0.5g、CaCl 2 0.3g、FeSO 4 0.2g、MgSO 4 0.1g、ZnSO 4 0.1g, 0.08g of vitamin complex and Na 2 SeO 3 0.03g、H 3 BO 3 0.02g), and carrying out shaking culture at 150rpm and 25 ℃ for 7 d; transferring the obtained culture solution into a selective inorganic salt culture medium again for culture, repeating the transfer for 5 times, and enriching to obtain a functional flora with strong plastic degradation capacity (the total weight of PP, PE and PVC plastic is reduced by 15%);
(2) crushing the wood chips, the corn straws and the shells, mixing according to the mass ratio of 1:1:1, and calcining at the high temperature of 300 ℃ for 2 hours in a nitrogen environment to obtain the mixed biomass charcoal.
The rest steps are the same. The results are shown in Table 1.
Example 21
The difference from example 20 is: (2) crushing the wood chips, the corn straws and the shells, mixing according to the mass ratio of 1:1:1, and calcining at 600 ℃ for 3 hours in a nitrogen environment to obtain the mixed biomass charcoal.
The rest steps are the same. The results are shown in Table 1.
Example 22
The difference from example 20 is: (4) the garbage contaminated soil remediation effect experiment: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 0.5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
The rest steps are the same. The results are shown in Table 1.
Example 23
The difference from example 20 is: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass carbon-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 5% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
The rest steps are the same. The results are shown in Table 1.
Example 24
The difference from example 20 is: (4) the experiment on the remediation effect of the garbage-polluted soil comprises the following steps: uniformly adding the biomass charcoal-based microorganism immobilized particles into garbage contaminated soil according to the proportion of 4% (w/w), fully and uniformly mixing by using soil turning equipment, and standing for 6 months. And the abundance ratio of the micro-plastics with different particle sizes in the soil is detected by a density flotation method and microscopic observation analysis to judge the treatment effect.
The rest steps are the same. The results are shown in Table 1.
Example 25
The difference from example 20 is: (3) mixing the mixed biomass charcoal and the bacterial suspension according to a ratio (w/w) of 1:10, oscillating for 8h at 15 ℃ and 150r/min for mixed adsorption, freezing, drying and grinding to obtain the microorganism immobilized biomass charcoal (the particle size is less than 100 mu m).
Adding the microorganism immobilized biochar (1 percent, w/w) into a mixed solution containing 3 percent (w/w) of carboxymethyl cellulose and 1.5 percent (w/w) of sodium alginate, uniformly stirring, slowly dripping the mixture into a 2 percent calcium chloride solution, and crosslinking for 16 hours to obtain the immobilized particles.
And washing the immobilized particles with deionized water for 3 times, and drying at low temperature to obtain the biomass carbon-based microorganism immobilized particles.
The rest steps are the same. The results are shown in Table 1.
TABLE 1 effect of microorganism-biomass charcoal immobilized particles on remediation of garbage contaminated soil
The results in Table 1 show that the application of different biomass charcoal-based immobilized microorganism particles can improve the degradation of plastics, 0.1-1cm 2 The proportion of plastic debris in size increased from 32.15% to 36.05% -40.98% before treatment (blank control) to 1-5cm 2 And 5-10cm 2 The proportion of plastic fragments is reduced, which shows that the biomass charcoal-based immobilized microorganism particles can realize the decomposition of plastics in the garbage-polluted soil. In addition, the quantity of the micro-plastics in the soil is obviously reduced from 2654 per kilogram to 1642-2219 per kilogram after the biomass charcoal-based immobilized microorganism particles are applied, which shows that the invention can realize the adsorption of the micro-plastics in the soil polluted by garbage.
The invention enriches the microbial flora with the capacity of degrading plastics from the surface of the plastic waste, can avoid the loss of some strains which are difficult to separate and culture, more comprehensively obtains the functional flora and realizes the high-efficiency degradation of the plastics. Specifically, the adsorption method is higher in feasibility for micro-plastics in soil, the biomass material is rich in active groups such as hydroxyl and carboxyl, the structure of the biomass material can be improved through high-temperature calcination, the adsorption efficiency is improved, and the resource utilization of solid wastes can be realized by selecting waste biomass (crop straws, sawdust, shells and the like) as a material source. Therefore, the invention can fix the high-efficiency functional bacteria on the adsorbing material by utilizing the immobilized microorganism technology, and can effectively maintain the quantity and the activity of the microorganisms by embedding and fixing, and the service cycle is long.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (9)
1. A microbial remediation method for micro-plastics in garbage contaminated soil is characterized by comprising the following steps:
(1) culturing microorganisms on the surface of plastic in a garbage polluted site by using a liquid LB culture medium, transferring the obtained bacterial liquid into a selective inorganic salt culture medium according to the volume ratio of (1-3) to (10), and transferring for 3-5 times to obtain a functional flora with plastic degradation capability;
(2) preparing the functional flora into a bacterial suspension, wherein the bacterial suspension and the biomass charcoal are mixed according to the mass ratio of (10-20): 1, mixing, adsorbing, freezing and drying to obtain microorganism immobilized biochar;
(3) adding the microbial immobilized biochar into a mixed solution of carboxymethyl cellulose and sodium alginate, uniformly stirring, and then slowly dropwise adding the mixture into a calcium chloride solution for crosslinking reaction to obtain biochar-based microbial immobilized particles;
(4) adding the biochar-based microorganism immobilized particles into garbage contaminated soil according to the mass percent of 0.5-5%, and carrying out in-situ remediation on the garbage contaminated soil.
2. The method for microbial remediation of a micro-plastic in soil contaminated with garbage of claim 1, wherein in step (1),
the selective inorganicThe salt culture medium comprises the following components in parts by weight: carbon source 5.0g, (NH) 4 )NO 3 1g、Na 2 HPO 4 1g、KCl 0.5g、CaCl 2 0.3g、FeSO 4 0.2g、MgSO 4 0.1g、ZnSO 4 0.1g, 0.08g of vitamin complex and Na 2 SeO 3 0.03g、H 3 BO 3 0.02g, 1L of water; wherein the carbon source is at least one of PE, PP and PVC.
3. The microbial remediation method of micro-plastics in garbage-polluted soil as claimed in claim 1, wherein in the step (2), the biomass charcoal is obtained by crushing the waste biomass, calcining the crushed waste biomass at a high temperature of 300-600 ℃ in a nitrogen environment for 1-3h, grinding and sieving the crushed waste biomass.
4. The method of claim 3, wherein the waste biomass comprises at least one of corn stover, rice straw, wood chips, fruit shells, and shells.
5. The method for microbial remediation of a micro-plastic in soil contaminated with garbage of claim 1 wherein the conditions of said mixed adsorption are: oscillating for 6-12h at 15-20 ℃ and 150 r/min.
6. The method for microbial remediation of a micro-plastic in soil contaminated with debris of claim 1 wherein the microbial immobilized biomass charcoal has a particle size of < 100 μm.
7. The method of claim 1, wherein the OD of the bacterial suspension is greater than the OD of the soil contaminated with the waste 600 Is 0.8.
8. The method for microbial remediation of a micro-plastic in a soil contaminated by garbage according to claim 1, wherein in the step (3), the mass ratio of the mixed solution to the microbial immobilized biomass charcoal is 100 (0.5-2).
9. The microbial remediation method of claim 8 of a micro-plastic in soil contaminated with waste, wherein in the mixed solution, the mass fraction of the carboxymethyl cellulose is 1% to 3%, and the mass fraction of the sodium alginate is 1% to 3%; the mass fraction of the calcium chloride solution is 1-4%.
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Application publication date: 20220916 Assignee: Guizhou Jiamu Environmental Protection Technology Co.,Ltd. Assignor: GUIZHOU INSTITUTE OF BIOLOGY Contract record no.: X2024980009186 Denomination of invention: A microbial remediation method for microplastics in garbage contaminated soil Granted publication date: 20230526 License type: Common License Record date: 20240709 |