CN112090954A - Activated and degraded coagulated beads, preparation method thereof and degradation method of polycyclic aromatic hydrocarbon-polluted soil - Google Patents
Activated and degraded coagulated beads, preparation method thereof and degradation method of polycyclic aromatic hydrocarbon-polluted soil Download PDFInfo
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Abstract
The invention discloses an activated degradation coagulated bead, preparation and application thereof and a degradation method of polycyclic aromatic hydrocarbon polluted soil. The activated and degraded bead consists of two parts, namely a bead outer wall and a bead core, wherein the bead core consists of a bead membrane and a bead inclusion compound; the condensed bead inclusion compound is a degrading bacterial liquid, and the outer wall of the condensed bead is formed by mixing crop straw powder, rhamnolipid and citric acid. Wherein the crop straw powder is preferably corn straw; the degrading bacterial liquid is preferably bacillus subtilis. The activation degradation coagulated bead is an activation-degradation integrated material which is simple to operate and environment-friendly, and can activate polycyclic aromatic hydrocarbon in soil, repair polluted soil and improve the microenvironment of the soil. The method can sequentially release effective components according to requirements and time sequence while maintaining good degradation performance of the degradation strain in the soil, is environment-friendly, can be biodegraded, cannot cause secondary pollution, does not damage the soil structure, is simple and convenient to use and operate, and avoids recovery.
Description
Technical Field
The invention belongs to the field of soil pollution control and restoration, and particularly relates to an activated degradation bead, a preparation method and application thereof, and a degradation method of polycyclic aromatic hydrocarbon polluted soil.
Background
The volatile organic polycyclic aromatic hydrocarbon has 'three-cause effect', biological accumulation and the like, and can cause great threat to the ecological environment and human health. Polycyclic aromatic hydrocarbons are possibly generated in the processes of processing, abandoning and burning of organic matters, wherein the problem of polycyclic aromatic hydrocarbon pollution in a coking site is prominent, so that rapid and efficient treatment measures are urgently needed to reduce the environmental risk and the ecological toxicity of the organic matters.
The traditional polycyclic aromatic hydrocarbon polluted soil remediation technology is a chemical method. For example, patent CN110589951A discloses a method for degrading polycyclic aromatic hydrocarbons by using zero-valent iron to activate persulfate. The method utilizes a chemical oxidation method to degrade the polycyclic aromatic hydrocarbon, has high cost although the operation is simple, and is easy to destroy the ecological environment, cause secondary pollution and the like.
At present, microbial remediation is considered to be one of the main approaches for removing polycyclic aromatic hydrocarbons in soil environment, but the degradation effect of the microbes on the polycyclic aromatic hydrocarbons in the remediation process is limited by a plurality of factors, such as the concentration of the polycyclic aromatic hydrocarbons, particularly the concentration and the range of the polycyclic aromatic hydrocarbons in coking fields and surrounding soil are extremely high, so that the microbes are not suitable for directly degrading. Patent CN108543808A discloses a method for degrading polycyclic aromatic hydrocarbon pollutants in soil by using a chemical oxidation-anaerobic microorganism combination. The PAHs are degraded through chemical oxidation at high concentration, the concentration of the polycyclic aromatic hydrocarbon is reduced to a lower concentration level, and then the PAHs are degraded through microorganisms, but the use of an oxidant can reduce the soil environment quality to a certain extent and influence the activity of the microorganisms, and the application is suitable for the soil polluted by the high-concentration polycyclic aromatic hydrocarbon and is not suitable for the soil polluted by medium-light polycyclic aromatic hydrocarbon.
The surfactant and microorganism combined remediation of the polycyclic aromatic hydrocarbon contaminated soil is also one of common methods, and can promote the migration process of polycyclic aromatic hydrocarbon from soil to a water phase, enhance the mobility of hydrophobic pollutants, and reduce the content of polycyclic aromatic hydrocarbon and other organic pollutants in the contaminated soil. Patent CN105013815A discloses a bioremediation method of polycyclic aromatic hydrocarbon heavy metal combined pollution. The method comprises the steps of mixing white rot fungi, tween 80, deionized water and polycyclic aromatic hydrocarbon-heavy metal composite contaminated soil, then carrying out solid state fermentation, and carrying out ultrasonic treatment and oscillation treatment on the fermented contaminated soil mixture to finish remediation of the composite contaminated soil. However, the surfactant Tween 80 is a chemical surfactant, is difficult to biodegrade, can obviously reduce the activity of indigenous microorganisms and white rot fungi, adopts ectopic repair, has complex operation and high repair cost, is easy to damage an ecosystem, and is not suitable for large-scale engineering treatment.
Disclosure of Invention
One of the purposes of the invention is to provide an activation degradation bead which is an environment-friendly activation-degradation integrated material, can be suitable for novel activation degradation of polycyclic aromatic hydrocarbon contaminated soil, and solves the problems of secondary pollution treatment and high cost of contaminated soil in the prior art.
In order to achieve the purpose, the invention provides the following technical scheme: the activation degradation condensation bead consists of two parts, namely a condensation bead outer wall and a condensation bead core, wherein the condensation bead core consists of a condensation bead membrane and a condensation bead inclusion compound. The outer wall of the condensed bead is formed by mixing crop straw powder, rhamnolipid and citric acid; the bead-forming film is a polyvinyl alcohol water-soluble film; the condensed bead inclusion compound is degradation bacterial liquid.
The specific composition of the activated degradation bead is as follows: based on the parts by weight, the weight ratio,
3-5 parts of crop straw powder; preferably, the straw powder is selected from corn straw, wheat straw and rice straw.
4-7 parts of rhamnolipid;
3-5 parts of citric acid;
5-8 parts of degraded bacterial liquid; preferably, the degrading bacterium liquid is bacillus subtilis, rhodococcus, aspergillus niger and mycobacterium flavum.
The invention also aims to provide a preparation method of the activated and degraded bead, which comprises the following specific steps:
s1, selecting and preparing a degrading bacterial liquid: selecting a degrading strain, placing the degrading strain in a liquid culture medium, culturing for 18h under a certain condition until the cell density of the degrading strain is not less than 1 x 108Per mL;
s2, filling and preparing the active degradation gel beads: conveying the degraded bacterium liquid to a bead-coagulating membrane aseptic packaging machine through an aseptic pipeline under aseptic condition, filling into a single aseptic polyvinyl alcohol water-soluble quantitative film, and performing hot-pressing injection and membrane sealing to obtain beads; a cooling device is adopted at the bottom of the membrane model to prevent the influence of hot pressing on the degraded bacteria liquid;
s3, preparing a coagulated bead outer wall material: grinding straw powder and citric acid into powder, and sieving with a 200-mesh sieve; uniformly mixing crop straw powder, rhamnolipid and citric acid in proportion, and uniformly stirring to obtain an outer wall material;
s4, uniformly coating a layer of glycerol on the surface layer of the coagulated beads obtained in the step S2, putting the coagulated beads into the outer wall material obtained in the step S3 to wrap an outer wall material, and then putting the coagulated beads into a sterile vacuum compression bag to be sealed and refrigerated at 4 ℃ for storage, thus obtaining the activated and degraded coagulated beads.
As an example of the present invention, in step S1, the strain is bacillus subtilis; and/or the culture medium is LB liquid culture medium; and/or the culture condition is 30 ℃ and 170 r/min.
In step S2, the amount of degraded bacterial liquid in each bead is 5 g; in step S4, the diameter of the beads is 1.5-2.0 cm.
As an embodiment of the present invention, in step S3, the crop straw powder, rhamnolipid and citric acid are mixed in a ratio of 3:4: 3.
The invention further aims to provide application of the activated degradation coagulated beads, which is particularly suitable for activated degradation of polycyclic aromatic hydrocarbon polluted soil.
The fourth purpose of the invention is to provide an activation degradation method of polycyclic aromatic hydrocarbon polluted soil, which comprises the following steps of adding the activation degradation beading into the polycyclic aromatic hydrocarbon polluted soil:
the activated and degraded coagulated beads are uniformly scattered into soil, then the soil is loosened, the outer wall of the coagulated beads and the coagulated bead membrane are dissolved by manually adjusting the water content of the soil to 70 percent or lowering the water, and nutrient solution can be added to improve the microbial living environment.
The addition amount of the microcapsule is determined according to the pollution degree of polycyclic aromatic hydrocarbon in soil. Calculating by taking the amount of degradation bacterium liquid in each coagulated bead as 5g, and when the first-class land screening value is less than the polycyclic aromatic hydrocarbon concentration and less than the second-class land screening value, applying 200-300 particles of degradation bacterium liquid per cubic meter of land; when the concentration of the polycyclic aromatic hydrocarbon is larger than the screening value of the second-class land, the application amount of the polycyclic aromatic hydrocarbon is 400-600 particles per cubic meter of land.
The technical scheme provided by the invention has the beneficial effects that at least:
the activation degradation coagulated bead is an activation-degradation integrated material which is simple to operate and environment-friendly, and can activate polycyclic aromatic hydrocarbon in soil, repair polluted soil and improve the microenvironment of the soil. The effective components can be sequentially released according to the requirement and the time sequence while the good degradation performance of the degrading strain in the soil is kept. The activated and degraded coagulated beads disclosed by the invention are environment-friendly, can be biologically degraded, cannot cause secondary pollution, cannot damage the soil structure, are simple and convenient to use and operate, can act only by scattering the coagulated beads into the soil and adjusting the water content of the soil, and are free from recovery. In particular, the method comprises the following steps of,
(1) the activating and degrading bead provided by the invention can realize the function of activating-degrading integration; the activated and degraded coagulated beads adopt a double structure of coagulated bead outer walls and coagulated bead cores, rhamnolipid and citric acid in the coagulated bead outer walls can activate pollutants, and crop straw powder can provide a good growth environment for microorganisms; the degrading bacteria in the syneresis nucleus can effectively degrade the polycyclic aromatic hydrocarbon in the soil;
(2) the activated and degraded coagulated beads provided by the invention can effectively reduce the content of polycyclic aromatic hydrocarbon in soil, prevent migration and transformation of the polycyclic aromatic hydrocarbon to a certain extent, and have the advantages of simple preparation, low cost, capability of realizing factory quantitative production, easy popularization and the like;
(3) the activated and degraded coagulated beads provided by the invention adopt degrading bacteria to remove polycyclic aromatic hydrocarbons in soil, and degradation products cannot cause secondary pollution;
(4) the preparation method of the activated and degraded coagulated beads provided by the invention combines activation and degradation on the basis of the prior art, optimizes the repair procedure, is convenient to apply, can function only by uniformly scattering the coagulated beads into soil and then adjusting the water content and the content of nutrient elements, can realize the efficient removal of polycyclic aromatic hydrocarbon in the soil polluted by a coking site by using the material provided by the invention, and the used materials are all environment-friendly materials, can be biodegraded and cannot generate secondary pollution.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in further detail below.
In the following examples, Bacillus subtilis (Bacillus subtilis) is selected as a degradation strain, purchased from Guangdong province microorganism strain preservation center, naphthalene, pyrene and the like can be used as main carbon sources and energy sources in the growth process, and degradation is realized through dechlorination and ring opening.
LB medium of Bacillus subtilis was purchased from Guangdong province culture Collection of microorganisms.
Example 1 preparation of activated degradable beads
S1, selecting and preparing a degrading bacterial liquid: selecting Bacillus subtilis with microbial inoculum GDM number of 1.181, placing Bacillus subtilis strain in LB liquid culture medium, culturing at 30 deg.C and 170r/min in constant temperature shaking table for 18h until cell density is 1 × 108When the strain is one/mL, the strain is ready for use;
s2, filling and preparing activated and degraded beads: conveying the degraded bacterium liquid to a polyvinyl alcohol water-soluble film sterile packaging machine through a sterile pipeline under the sterile condition, filling the sterile bacterium liquid into a single sterile polyvinyl alcohol water-soluble quantitative film, and performing hot-pressing injection and film sealing to obtain coagulated beads; a cooling device is adopted at the bottom of the membrane model to prevent the influence of hot pressing on the degraded bacteria liquid; the amount of the degrading bacteria liquid in each coagulated bead is 5 g;
s3, preparing a coagulated bead outer wall material: grinding straw powder and citric acid into powder, and sieving with a 200-mesh sieve; uniformly mixing crop straw powder, rhamnolipid and citric acid in proportion, and uniformly stirring to obtain an outer wall material; wherein the crop straw powder, the rhamnolipid and the citric acid are mixed in a ratio of 3:4: 3;
s4, uniformly coating a layer of glycerol on the surface layer of the coagulated beads obtained in the step S2, putting the coagulated beads into the outer wall material obtained in the step S3 to wrap an outer wall material, and then putting the coagulated beads into a sterile vacuum compression bag to be sealed and refrigerated at 4 ℃ for storage, so as to obtain the activated and degraded coagulated beads; the diameter of the activated degradation bead is about 1.8 cm.
Example 2 degradation of simulated polycyclic aromatic Hydrocarbon contaminated soil Using activated degradation beads
Adding methanol solution of naphthalene and pyrene 40 mg/kg-1Adding the mixture into a sterilized soil sample, uniformly mixing, and placing in a fume hood for 24 hours to naturally volatilize the solvent to obtain the simulated contaminated soil.
And (3) putting 40g of the simulated polycyclic aromatic hydrocarbon contaminated soil into a 250mL conical flask, adding sterile water, adjusting the water content of the soil to be 70%, adding 2 activated and degraded beads prepared in the example 1, fully stirring, and culturing at room temperature for 5 days.
And (3) after extracting the polycyclic aromatic hydrocarbon remained in the soil, carrying out quantitative detection and analysis on the result by adopting a liquid chromatograph, wherein the mobile phase A is ultrapure water, and the mobile phase B is acetonitrile. The gradient elution program was set up as:
0-2.0 min, mobile phase A, B volume ratio VA:VB＝3:7；
2.0～28.0min，VA:VB＝1:9；
28.0～28.5min，VA:VB＝3:7；
28.5～35.0min，VA:VB＝3:7。
The sample volume is 10 mu L, the column temperature is 35 ℃, and the flow rate is 1 mL/min.
Meanwhile, a blank control experiment is established, 40g of the simulated polycyclic aromatic hydrocarbon contaminated soil is put into a 250mL conical flask, sterile water is added, the water content of the soil is adjusted to 70%, and the soil is cultured for 5 days at room temperature after being fully stirred. And (3) after extracting the polycyclic aromatic hydrocarbon remained in the soil, carrying out quantitative detection and analysis on the result by adopting a liquid chromatograph under the same gradient elution procedure.
Comparative example 1 degradation of simulated polycyclic aromatic hydrocarbons contaminated soil directly using Bacillus subtilis liquid
And (3) putting 40g of the simulated polycyclic aromatic hydrocarbon-polluted soil described in the example 2 into a 250mL conical flask, adding sterile water, adjusting the water content of the soil to 70%, adding 10g of bacillus subtilis suspension, fully stirring, and culturing at room temperature for 5 days. After extraction of the polycyclic aromatic hydrocarbons remaining in the soil, the results were quantitatively analyzed by liquid chromatography under the same gradient elution procedure as in example 2.
Specific results are shown in table 1.
TABLE 1
| Rate of degradation | Naphthalene | Pyrene |
| Addition of activated degradation beads (example 2) | 39.4％ | 39.9％ |
| Bacillus subtilis liquid (comparative example 1) | 20.5％ | 19.9％ |
| Without addition of activated degradation beads (blank control) | 1.8％ | 1.6％ |
Example 3 degradation of polycyclic aromatic hydrocarbons contaminated soil in coking field Using activated degradation beads
Placing 40g of polycyclic aromatic hydrocarbon contaminated soil in a coking field into a 250mL conical flask for sterilization, adding sterile water, adjusting the water content of the soil to 70%, adding 2 activated and degraded beads prepared in the example 1, fully stirring, and culturing at room temperature for 5 days.
And (3) after extracting the polycyclic aromatic hydrocarbon remained in the soil, carrying out quantitative detection and analysis on the result by adopting a liquid chromatograph, wherein the mobile phase A is ultrapure water, and the mobile phase B is acetonitrile. The gradient elution program was set up as:
0-2.0 min, mobile phase A, B volume ratio VA:VB＝3:7；
2.0～28.0min，VA:VB＝1:9；
28.0～28.5min，VA:VB＝3:7；
28.5～35.0min，VA:VB＝3:7。
The sample volume is 10 mu L, the column temperature is 35 ℃, and the flow rate is 1 mL/min.
Meanwhile, a blank control experiment is established, 40g of the polycyclic aromatic hydrocarbon contaminated soil in the coking field is put into a 250mL conical flask for sterilization, sterile water is added, the water content of the soil is adjusted to 70%, and the soil is cultured for 5 days at room temperature after being fully stirred. Extracting the polycyclic aromatic hydrocarbon remained in the soil. And (3) carrying out quantitative detection and analysis on the result by adopting a liquid chromatograph under the same gradient elution procedure. Specific results are shown in table 1.
Comparative example 2 degradation of polycyclic aromatic hydrocarbons contaminated soil in coking field by directly using Bacillus subtilis solution
And (3) putting 40g of the polycyclic aromatic hydrocarbon polluted soil of the coking field in the example 3 into a 250mL conical flask for sterilization, adding sterile water, adjusting the water content of the soil to be 70%, adding 10g of bacillus subtilis bacterial suspension, fully stirring, and culturing at room temperature for 5 days. After extraction of the polycyclic aromatic hydrocarbons remaining in the soil, the results were quantitatively analyzed by liquid chromatography under the same gradient elution procedure as in example 3.
TABLE 2
| Rate of degradation | Naphthalene | Pyrene |
| Addition of activated degradation beads (example 3) | 36.3％ | 35.6％ |
| Bacillus subtilis liquid (comparative example 2) | 18.6％ | 18.9％ |
| Without addition of activated degradation beads (blank control) | 1.3％ | 1.4％ |
The experiments show that the final pollutant degradation rate is obviously improved by 18-20% compared with the degradation rate of directly adding the degradation bacterial liquid when the polycyclic aromatic hydrocarbon polluted soil is degraded by adopting the activated degradation coagulated beads.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. An activated degradation bead, comprising: the bead core consists of a bead membrane and a bead inclusion compound; the condensed bead inclusion compound is a degrading bacterial liquid.
2. The activated degradation beads of claim 1, wherein: the outer wall of the condensed bead is formed by mixing crop straw powder, rhamnolipid and citric acid; based on the parts by weight, the weight ratio,
preferably, the straw powder is selected from any one of corn straw, wheat straw and rice straw;
preferably, the degrading bacterial liquid is any one of bacillus subtilis, rhodococcus, aspergillus niger and mycobacterium flavum.
3. The activated degradation beads of claim 1, wherein: the beading film is selected from a polyvinyl alcohol water-soluble film.
4. A method of making the activated degradation beads of any one of claims 1 to 3, wherein: the method comprises the specific steps of carrying out,
s1, selecting and preparing a degrading bacterial liquid: selecting degrading strain, placing in liquid culture medium, and culturing until the cell density of the degrading strain is not less than 1 × 108Per mL;
s2, filling and preparing activated and degraded beads: conveying the degraded bacterium liquid to a bead-coagulating membrane aseptic packaging machine through an aseptic pipeline under aseptic condition, filling into a single aseptic polyvinyl alcohol water-soluble quantitative film, and performing hot-pressing injection and membrane sealing to obtain beads;
s3, preparing a coagulated bead outer wall material: grinding straw powder and citric acid into powder, and sieving with a 200-mesh sieve; uniformly mixing crop straw powder, rhamnolipid and citric acid in proportion, and uniformly stirring to obtain an outer wall material;
s4, uniformly coating a layer of glycerol on the surface layer of the coagulated beads obtained in the step S2, putting the coagulated beads into the outer wall material obtained in the step S3 to wrap an outer wall material, and then putting the coagulated beads into a sterile vacuum compression bag to be sealed, refrigerated and stored, so that the activated and degraded coagulated beads are obtained.
5. The method of preparing activated degradable beads according to claim 4, wherein: in step S1, the strain is bacillus subtilis; and/or the culture medium is LB liquid culture medium; and/or the culture condition is 30 ℃ and 170 r/min.
6. The method of preparing activated degradable beads according to claim 4, wherein: in step S3, the crop straw powder, rhamnolipid and citric acid are mixed in a ratio of 3:4: 3.
7. The activated degraded beads of any one of claims 1 to 3 or the activated degraded beads obtained by the method of any one of claims 4 to 7, for use in the activated degradation of polycyclic aromatic hydrocarbon contaminated soil.
8. A method for activating and degrading polycyclic aromatic hydrocarbon polluted soil is characterized by comprising the following steps: the method comprises the steps of adding the activated and degraded coagulated beads in any one of claims 1 to 3 or the activated and degraded coagulated beads prepared by the method in any one of claims 4 to 7 into polycyclic aromatic hydrocarbon contaminated soil, adjusting the water content of the soil to 70% to dissolve the outer wall of the coagulated beads, and improving the microbial living environment by adding a nutrient solution.
9. The method for activating and degrading polycyclic aromatic hydrocarbon-contaminated soil according to claim 8, wherein: the amount of the degrading bacteria liquid in each coagulated bead is 5g, and when the first-class land screening value is less than the polycyclic aromatic hydrocarbon concentration and less than the second-class land screening value, the application amount of the degrading bacteria liquid in each cubic meter of land is 200-300.
10. The method for activating and degrading polycyclic aromatic hydrocarbon-contaminated soil according to claim 8, wherein: and calculating by taking the amount of the degrading bacteria liquid in each coagulated bead as 5g, wherein when the concentration of the polycyclic aromatic hydrocarbon is greater than the screening value of the second-class land, the application amount of the degrading bacteria liquid is 400-600 particles per cubic meter of land.
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