CN116673319A - Degradation method of polystyrene in soil - Google Patents

Degradation method of polystyrene in soil Download PDF

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Publication number
CN116673319A
CN116673319A CN202310945240.8A CN202310945240A CN116673319A CN 116673319 A CN116673319 A CN 116673319A CN 202310945240 A CN202310945240 A CN 202310945240A CN 116673319 A CN116673319 A CN 116673319A
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degradation
soil
liquid
temperature
groups
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CN116673319B (en
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熊静
刘亚茹
李书鹏
郭丽莉
韩亚萌
李嘉晨
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BCEG Environmental Remediation Co Ltd
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BCEG Environmental Remediation Co Ltd
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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B09DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09CRECLAMATION OF CONTAMINATED SOIL
    • B09C1/00Reclamation of contaminated soil
    • B09C1/10Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02WCLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00Technologies for solid waste management
    • Y02W30/50Reuse, recycling or recovery technologies
    • Y02W30/62Plastics recycling; Rubber recycling

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  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Mycology (AREA)
  • Soil Sciences (AREA)
  • Environmental & Geological Engineering (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to the field of polystyrene degradation, in particular to a degradation method of polystyrene in soil, which comprises the steps of dividing a microbial inoculum containing heat-resistant pseudomonas aeruginosa bacteria liquid into at least two times, injecting the microbial inoculum into soil containing polystyrene, wherein the temperature of the soil is 25-30 ℃, and keeping the temperature for 10-15 hours; then raising the temperature of the soil to 40-45 ℃ and maintaining the temperature until the next time before injecting the microbial inoculum for 1-3 hours; according to the invention, the in-situ injection of the microbial inoculum and/or the degradation liquid is realized by adopting the high-efficiency low-carbon thermal stabilization system, and the system can ensure that the soil temperature is kept at 40-45 ℃ when the polystyrene in the soil is degraded, so that the degradation efficiency of the polystyrene in the soil is improved.

Description

Degradation method of polystyrene in soil
Technical Field
The invention relates to the field of polystyrene degradation, in particular to a degradation method of polystyrene in soil.
Background
Polystyrene is a polymer formed by polymerizing styrene monomers through free radicals, and is a thermoplastic plastic with wide application. The polystyrene main chain is macromolecular saturated hydrocarbon polymer, and the lateral group is a rigid benzene ring structure with larger volume, so that the polystyrene main chain has the advantages of transparency, high rigidity, excellent electrical property, acid and alkali resistance, easy molding, low price and the like, and is widely applied to the fields of packaging materials, electrons, building materials, automobile parts, electric appliances, toys and the like.
The global plastic production in 2019 was reported to be 3.68 hundred million tons, and china accounts for 31% of the total world production. 9% of the plastics produced worldwide are recovered, 12% are incinerated, and the remaining 79% are released into the environment. As a third major class of resin plastics, which is inferior to polyethylene and polyvinyl chloride, polystyrene is rapidly and largely accumulated in the environment due to its wide use, poor management and difficult degradability, and polystyrene released into the environment is photodegradation, embrittled and broken down under the effect of long-time physical, chemical and biological degradation, slowly decomposed into plastic particles of smaller size, and finally widely distributed in the natural environment. Due to the small size, the microplastic polystyrene can be directly or indirectly ingested by organisms at various nutrient levels in the ecosystem to produce toxic effects; for example, rossi et al (2014) demonstrated that nano-polystyrene readily penetrates into lipid membranes, resulting in changes in membrane structure, significantly reducing molecular diffusion, further affecting cell function; the garden Wen Ke (2020) considers that the micro-plastic polystyrene can be rapidly and largely adsorbed on the surface of the aquatic plant crude-pedunculate pteridophyte spores to inhibit the germination of the sporophytes and the development of gametophytes; liu Yang (2022) research shows that the microplastic polystyrene has strong genotoxicity and oxidation damage to soybean roots, can reduce the activity of the roots, and further influence the absorption of the roots to nutrients. As can be seen, the management of microplastic polystyrene has become an environmental issue that needs to be addressed.
The existing polystyrene degradation modes comprise physical degradation, chemical degradation and biological degradation, wherein the microbial degradation method has the advantages of high efficiency, small pollution, reproducibility and the like, and has a relatively high application prospect in the pollution treatment of polystyrene microplastic. Peixoto (2017) separates escherichia coli, citrobacter, alcaligenes and shortwave monad, and all show depolymerizing enzyme activity and have certain degradation capacity on polystyrene; pseudomonas and Bacillus strains isolated from soil by Mohan (2016) promote degradation of aliphatic carbon chains; pseudomonas aeruginosa separated in China patent CN114058558B can effectively degrade polystyrene plastic films and microplastic. The strain can degrade polystyrene, but has lower degradation efficiency, and the conventional strain is difficult to survive at a higher temperature, so that the degradation rate of the strain applied to soil remediation is not high.
Therefore, how to optimize and improve the polystyrene degradation process, and improve the degradation efficiency of polystyrene, so that the polystyrene can be applied in engineering, is a technical problem to be solved in the field.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the defect that the degradation efficiency of the biodegradation method in the prior art on polystyrene is low, so that the degradation method of polystyrene in soil is provided by optimizing the degradation process conditions, and the degradation efficiency of polystyrene in soil can be improved.
The invention provides a degradation method of polystyrene in soil, which comprises the steps of dividing a microbial inoculum containing heat-resistant pseudomonas aeruginosa bacterial liquid into at least two times, injecting the microbial inoculum into soil containing polystyrene, wherein the temperature of the soil is 25-30 ℃, and keeping the temperature for 10-15 hours; then raising the temperature of the soil to 40-45 ℃ and maintaining the temperature until the next time before injecting the microbial inoculum for 1-3 hours;
the preparation method of the heat-resistant pseudomonas aeruginosa bacterial liquid comprises the following steps: s1, taking pseudomonas aeruginosa with the preservation number of CGMCC No.23974, and culturing at 25-30 ℃ to logarithmic phase; the method comprises the steps of (1) increasing the culture temperature of a strain in a logarithmic growth phase to 40-45 ℃ according to the temperature difference of 1-2 ℃ in each domestication period to obtain a primary screening strain; s2, culturing the primary screening strain at 40-45 ℃, re-screening, and purifying the strain to obtain heat-resistant pseudomonas aeruginosa; s3, inoculating the heat-resistant pseudomonas aeruginosa into a culture medium, and fermenting for 48-56 hours at the temperature of 25-30 ℃.
Wherein the domestication period is the time of culturing the strain at a culture temperature.
The microbial inoculum also comprises degradation liquid; the degradation liquid comprises the following components: k (K) 2 HPO 4 0.4 g/L~0.6 g/L、KH 2 PO 4 3.5 g/L~4 g/L、(NH 4 ) 2 HPO 4 0.5 g/L~1 g/L、MgCl 2 ·6H 2 O 18 mg/L~22 mg/L、FeCl 3 0.1 mg/L~0.3 mg/L、CaCl 2 1 mg/L~3 mg/L、Na 2 SO 4 180 mg/L~220 mg/L、KNO 3 1.1 g/L-1.5 g/L, wherein the pH value of the degradation liquid is 7.0-7.2; and/or the number of the groups of groups,
the bacterial activity of the heat-resistant pseudomonas aeruginosa bacterial liquid is 1.0x10 8 CFU/mL~1.0×10 9 CFU/mL。
The culture medium adopted in any one of the steps S1-S3 comprises glucose 4 g/L-6 g/L, beef extract 8 g/L-12 g/L, peptone 8 g/L-12 g/L, naCl 4 g/L-6 g/L, and the pH value of the culture medium is 7.0-7.2; and/or the number of the groups of groups,
each domestication period in the step S1 is 12-24 hours; and/or the number of the groups of groups,
the culture time in the step S2 is 24-48 hours; and/or the number of the groups of groups,
the volume ratio of the heat-resistant pseudomonas aeruginosa bacterial liquid to the degradation liquid in the microbial inoculum is 1: 2-5; and/or the number of the groups of groups,
the degradation method further includes the step of injecting the degradation liquid separately into the soil.
The injection interval of the microbial inoculum at the early stage of degradation treatment is 7 d-9 d, and the injection interval of the microbial inoculum at the later stage of degradation treatment is 16 d-30 d; during degradation treatment, the degradation liquid is independently injected after the bacterial agent is injected each time, the time between the independent injection of the degradation liquid and the injection of the bacterial agent in the early stage of the degradation treatment is 3 d-5 d, the time between the independent injection of the degradation liquid and the injection of the bacterial agent in the later stage of the degradation treatment is 8 d-29 d, wherein the early stage of the degradation treatment is 1 d-24 d from the beginning of the degradation treatment, and the later stage of the degradation treatment is 25 th d and later from the beginning of the degradation treatment; and/or the number of the groups of groups,
the single injection amount of the microbial inoculum and the soil volume ratio are 50L/m 3 ~200 L/m 3 The method comprises the steps of carrying out a first treatment on the surface of the And/or the number of the groups of groups,
when the degradation liquid is singly injected, the single injection amount of the degradation liquid and the soil volume ratio are 30L/m 3 ~50 L/m 3
The degradation method of polystyrene in soil provided by the invention further comprises the steps of adopting a micro-plastic online monitoring system based on Raman spectrum to monitor the degradation condition of polystyrene in the soil in real time, and adopting a high-efficiency low-carbon thermal stabilization system to realize in-situ injection of the microbial inoculum and/or the degradation liquid; the high efficiency low carbon thermal stabilization system comprises: the heat source generating module, the heat exchanger, the controller and the temperature acquisition device, wherein,
the heat source generating module and the heat exchanger are mutually communicated through a pipeline to form a loop, and a power device is arranged on the pipeline and provides power for medium circulation in the heat source generating module and the heat exchanger;
the heat exchanger comprises a plurality of heating units which are arranged side by side, and the heating units are vertically arranged;
the temperature acquisition device is positioned between two adjacent heating units and is used for acquiring the temperature of soil between the heating units, and the temperature acquisition device and the power device are electrically connected with the controller;
the controller controls the power device to accelerate the medium flow speed in the heat source generating module and the heat exchanger when the temperature of the soil between the heating units is lower than a threshold value.
The heating unit comprises a plurality of heating groups which are arranged in parallel, and the water inlet of each heating group is communicated with the water outlet of the power device; the water outlet of the heating group is communicated with the water inlet of the power device; each heating group comprises a communicating pipe which is horizontally arranged and a capillary which is vertically arranged, wherein the communicating pipe is communicated with the capillary, and the inner diameter of the communicating pipe is larger than that of the capillary.
The capillary comprises an outer sleeve and an inner sleeve, and the inner sleeve is sleeved in the outer sleeve; the outer sleeve is provided with an outer liquid outlet, the position, corresponding to the outer liquid outlet, of the inner sleeve is provided with an inner liquid outlet, and the outer liquid outlet and the inner liquid outlet are identical in structure; the top cover of the outer sleeve is provided with a perforation, the inside of the inner sleeve is connected with a connecting rod, the connecting rod and the inner sleeve are coaxially arranged, the connecting rod is connected with the inner wall of the inner sleeve through a supporting rod, and the connecting rod penetrates through the perforation; one end of the connecting rod is positioned outside the outer sleeve, and a gear is arranged at one end of the connecting rod positioned outside the outer sleeve and is connected with a driving motor through a driving belt;
the medium in the heat source generating module and the heat exchanger is the microbial inoculum and/or the degradation liquid for repairing soil, the driving motor is controlled to drive the gear to rotate, so that the inner sleeve and the outer sleeve relatively rotate, the inner liquid outlet on the inner sleeve and the outer liquid outlet on the outer sleeve are overlapped, the microbial inoculum and/or the degradation liquid is filled into the soil through the inner liquid outlet and the outer liquid outlet, the driving motor is controlled to enable the inner liquid outlet and the outer liquid outlet to be staggered after filling, and the microbial inoculum and/or the degradation liquid cannot pass through the inner liquid outlet and the outer liquid outlet.
The upper end of the outer sleeve is provided with a first bearing, the lower end of an outer ring of the first bearing is fixed on the outer wall of the outer sleeve, and the connecting rod is fixed on an inner ring of the first bearing; the bottom in the outer sleeve is provided with a second bearing, an outer ring of the second bearing is fixed on the inner wall of the outer sleeve, and the lower end of the connecting rod is fixed on an inner ring of the second bearing; and/or the number of the groups of groups,
the outer sleeve is coated with non-woven fabrics, and the non-woven fabrics cover the outer liquid outlet; the capillaries are arranged in rows, and the gears of the capillaries in the same row are connected with the driving motor through the same driving belt.
The distance between the adjacent heating units is 0.5 m-2 m, and the distance between the adjacent capillaries is 0.1 m-0.5 m; and/or the heat source generating module is a solar heat collector, the inner diameter of the communicating pipe is 10-20 cm, the inner diameter of the capillary pipe is 1-2 cm, and the power device is a water pump.
Compared with the prior art, the technical scheme of the invention has the following advantages:
1. the degradation method of polystyrene in soil provided by the invention comprises the steps of dividing a microbial inoculum containing heat-resistant pseudomonas aeruginosa bacteria liquid into at least two times, injecting the microbial inoculum into soil containing polystyrene, wherein the temperature of the soil is 25-30 ℃, and keeping the temperature for 10-15 hours; then raising the temperature of the soil to 40-45 ℃ and maintaining the temperature until the next time before injecting the microbial inoculum for 1-3 hours; the heat-resistant pseudomonas aeruginosa bacterial liquid is obtained by fermenting domesticated strains. When the soil temperature reaches 40-45 ℃, a suitable environment is provided for degrading the polystyrene by microorganisms, the degradation rate of the polystyrene is remarkably improved, and the environmental soil polystyrene micro-plastic pollution is reduced. According to the invention, pseudomonas aeruginosa is slowly domesticated at a temperature difference of 1-2 ℃ in each domestication period, and the heat-resistant pseudomonas aeruginosa liquid with stable performance is obtained through screening and optimization, and the heat-resistant pseudomonas aeruginosa liquid can show higher activity in a temperature environment with a temperature of 40-45 ℃, so that the degradation efficiency of polystyrene is improved.
2. The degradation method of polystyrene in soil provided by the invention is characterized in that the time for in-situ injection of the microbial inoculum containing the heat-resistant pseudomonas aeruginosa bacterial liquid in the early and late degradation treatment and the time for independent injection of the degradation liquid in the early and late degradation treatment are respectively limited, namely, the method of injecting the microbial inoculum in the early degradation treatment by high frequency is adopted to ensure the number of microorganisms planted and survived in the soil, and the injection frequency of the microbial inoculum is reduced in the late degradation treatment, so that the aim of supplementing the microbial inoculum is fulfilled; meanwhile, after each time of injection of the microbial inoculum, the degradation liquid is injected for a period of time, so that necessary nutrition sources are provided for survival of the heat-resistant pseudomonas aeruginosa in the microbial inoculum, and insufficient nutrition of the microbial inoculum is avoided, and growth and propagation of the microbial inoculum in soil are influenced. Based on the method, the degradation effect of the plastic can be ensured, the dosage of the microbial inoculum can be reduced, and secondary pollution to the environment can be avoided.
3. The degradation method of polystyrene in soil provided by the invention comprises the steps of adopting a high-efficiency low-carbon thermal stabilization system to realize in-situ injection of the microbial inoculum and/or the degradation liquid, keeping the temperature of the soil within a certain range, particularly keeping the temperature of the soil at 25-30 ℃ during in-situ injection of the microbial inoculum, keeping the temperature for 10-15 h, raising the temperature of the soil to 40-45 ℃, keeping the temperature until 1-3 h before the next injection of the microbial inoculum, and keeping the temperature of the soil at 25-30 ℃ before 1-3 h before the next injection of the microbial inoculum; the high efficiency low carbon thermal stabilization system comprises: the device comprises a heat source generating module, a heat exchanger, a controller and a temperature acquisition device, wherein the heat source generating module and the heat exchanger are mutually communicated through a pipeline to form a loop, a power device is arranged on the pipeline, and the power device provides power for medium circulation in the heat source generating module and the heat exchanger; the heat exchanger comprises a plurality of heating units which are arranged side by side, and the heating units are vertically arranged; the temperature acquisition device is positioned between two adjacent heating units and is used for acquiring the temperature of soil between the heating units, and the temperature acquisition device and the power device are electrically connected with the controller; the controller controls the power device to accelerate the medium flow speed in the heat source generating module and the heat exchanger when the temperature of the soil between the heating units is lower than a threshold value. According to the invention, the soil temperature can be kept at 40-45 ℃ by adopting the high-efficiency low-carbon thermal stabilization system, so that the heat-resistant pseudomonas aeruginosa can be ensured to better play a role in degradation. The high-efficiency low-carbon heat stabilization system has two functions of in-situ heating and in-situ injection, the microbial inoculum is in-situ injected into the soil through the high-efficiency low-carbon heat stabilization system, and the temperature of the soil is kept through the thermal circulation of the medium, so that the degradation efficiency of the heat-resistant pseudomonas aeruginosa in the soil on the polystyrene is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of a high-efficiency low-carbon thermal stabilization system according to embodiment 1 of the present invention;
FIG. 2 is an enlarged view of section A of the structure of the high efficiency low carbon thermal stabilization system of example 1 of the present invention;
FIG. 3 is a schematic view of the structure of the capillary tube and the communicating tube in the high-efficiency low-carbon thermal stabilization system according to embodiment 1 of the present invention;
FIG. 4 is a schematic view of the structure of the outer sleeve in the high efficiency low carbon thermal stabilization system of example 1 of the present invention;
FIG. 5 is a schematic view showing the structure of an inner sleeve in the high-efficiency low-carbon thermal stabilization system according to embodiment 1 of the present invention;
FIG. 6 is a flow chart of a method for preparing a heat-resistant Pseudomonas aeruginosa liquid according to example 2 of the present invention;
wherein, the reference numerals are as follows:
the device comprises a 1-temperature acquisition device, a 2-heating unit, a 3-controller, a 4-power device, a 5-heat source generating module, a 6-heating group, a 7-heat exchanger, an 8-driving belt, a 9-communicating pipe, a 10-capillary tube, an 11-first bearing, a 12-perforation, a 13-top cover, a 14-external liquid outlet, a 15-gear, a 16-connecting rod, a 17-supporting rod, an 18-internal sleeve, a 19-internal liquid outlet and a 20-external sleeve.
Detailed Description
The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
The original pseudomonas aeruginosa (Pseudomonas aeruginosa) adopted in the examples and the comparative examples is purchased from China general microbiological culture collection center (CGMCC) No.23974.
The test is located in a certain city of the North China area and is suburbd and influenced by factors such as human activities, industrial and agricultural activities and the like, so that soil in the area is polluted by polystyrene.
Example 1
Referring to fig. 1, the present invention provides a high-efficiency low-carbon thermal stabilization system:
the high-efficiency low-carbon thermal stabilization system is used for in-situ injection of a microbial inoculum containing heat-resistant pseudomonas aeruginosa bacteria liquid, and comprises: the device comprises a heat source generating module 5, a heat exchanger 7, a controller 3 and a temperature acquisition device 1, wherein the heat source generating module 5 and the heat exchanger 7 are mutually communicated to form a loop through a pipeline, a power device 4 is arranged on the pipeline, and the power device 4 provides power for medium circulation in the heat source generating module 5 and the heat exchanger 7; the heat exchanger 7 comprises a plurality of heating units 2 which are arranged side by side, and the heating units 2 are arranged vertically; the temperature acquisition device 1 is positioned between two adjacent heating units 2, the temperature acquisition device 1 is used for acquiring the temperature of soil between the heating units 2, and the temperature acquisition device 1 and the power device 4 are electrically connected with the controller 3; when the temperature of the soil between the heating units 2 is lower than a threshold value, the controller 3 controls the power device 4 to accelerate the medium flow speed in the heat source generating module 5 and the heat exchanger 7;
specifically, as shown in fig. 1 and 2, the high-efficiency low-carbon thermal stabilization system of the present invention further includes the heating unit 2, where the heating unit 2 includes a plurality of heating groups 6 arranged in parallel, and a water inlet of each heating group 6 is communicated with a water outlet of the power device 4; the water outlet of the heating group 6 is communicated with the water inlet of the power device 4; each heating group 6 comprises a communicating pipe 9 which is horizontally arranged and a capillary tube 10 which is vertically arranged, the communicating pipe 9 is communicated with the capillary tube 10, and the inner diameter of the communicating pipe 9 is larger than that of the capillary tube 10.
Referring to fig. 3-5, the capillary tube 10 includes an outer sleeve 20 and an inner sleeve 18, wherein the inner sleeve 18 is sleeved in the outer sleeve 20; the outer sleeve 20 is provided with an outer liquid outlet 14, the inner sleeve 18 is provided with an inner liquid outlet 19 at a position corresponding to the outer liquid outlet 14, and the outer liquid outlet 14 and the inner liquid outlet 19 have the same structure; the top cover 13 of the outer sleeve 20 is provided with a through hole 12, the inside of the inner sleeve 18 is connected with a connecting rod 16, the connecting rod 16 and the inner sleeve 18 are coaxially arranged, the connecting rod 16 is connected with the inner wall of the inner sleeve 18 through a supporting rod 17, and the connecting rod 16 passes through the through hole 12; one end of the connecting rod 16 is positioned outside the outer sleeve 20, a gear 15 is arranged at one end of the connecting rod 16 positioned outside the outer sleeve 20, and the gear 15 is connected with a driving motor through a driving belt 8; the medium in the heat source generating module 5 and the heat exchanger 7 is the microbial inoculum and/or the degradation liquid for repairing the soil, the driving motor is controlled to drive the gear 15 to rotate, so that the inner sleeve 18 and the outer sleeve 20 relatively rotate, the inner liquid outlet 19 on the inner sleeve 18 and the outer liquid outlet 14 on the outer sleeve 20 are overlapped, the microbial inoculum and/or the degradation liquid is fed into the soil through the inner liquid outlet 19 and the outer liquid outlet 14, after the feeding is finished, the driving motor is controlled to stagger the inner liquid outlet 19 and the outer liquid outlet 14, and the microbial inoculum and/or the degradation liquid cannot pass through the inner liquid outlet 19 and the outer liquid outlet 14;
specifically, as shown in fig. 3-5, the upper end of the outer sleeve 20 is provided with a first bearing 11, the lower end of the outer ring of the first bearing 11 is fixed on the outer wall of the outer sleeve 20, and the connecting rod 16 is fixed on the inner ring of the first bearing 11; the bottom in the outer sleeve 20 is provided with a second bearing, an outer ring of the second bearing is fixed on the inner wall of the outer sleeve 20, and the lower end of the connecting rod 16 is fixed on an inner ring of the second bearing; the outer sleeve 20 is coated with non-woven fabrics, and the non-woven fabrics cover the outer liquid outlet 14; the capillaries 10 are arranged in rows, and the gears 15 of the capillaries 10 in the same row are connected with a driving motor through the same driving belt 8.
Specifically, the distance between the adjacent heating units 2 is 0.5 m to 2 m, and the distance between the adjacent capillaries 10 is 0.1 m to 0.5 m; and/or the heat source generating module 5 is a solar collector, the inside diameter of the communication pipe 9 is 10 to 20 cm, the inside diameter of the capillary tube 10 is 1 to 2 cm, and the power unit 4 is a water pump.
Example 2
The invention provides a degradation method of polystyrene in soil, which comprises the following specific steps:
the preparation method of the heat-resistant pseudomonas aeruginosa bacterial liquid comprises the following steps: referring to FIG. 6, the original strain Pseudomonas aeruginosa is cultivated at 28 ℃ to logarithmic phase, the temperature difference is increased by taking 18 h as domestication period and 1 ℃ as temperature difference, the cultivation temperature is gradually increased, namely, the strain Pseudomonas aeruginosa which can normally grow at 29 ℃ is cultivated at 18 h, the strain Pseudomonas aeruginosa which can normally grow at 29 ℃ is transferred to new culture medium, 18 h is cultivated at 30 ℃, the strain Pseudomonas aeruginosa which can normally grow at 30 ℃ is transferred to new culture medium, and so on until the strain which can normally grow at 45 ℃ is domesticated and selected, the high temperature resistant strain obtained by primary screening is inoculated to 36 h under 45 ℃ for re-screening and strain purification, and finally the strain Pseudomonas aeruginosa which can resist high temperature and heat and degrade polystyrene is obtained; fermenting the obtained heat-resistant Pseudomonas aeruginosa strain at 28deg.C for 52 h to obtain heat-resistant Pseudomonas aeruginosa liquid with bacterial activity of 5.0X10 8 CFU/mL; the culture medium formula is the same in each step of the preparation method of the heat-resistant pseudomonas aeruginosa bacteria liquid, and specifically adopts glucose 5 g/L, beef extract 10 g/L, peptone 10 g/L, naCl 5 g/L and pH 7.1;
according to K 2 HPO 4 0.5 g/L、KH 2 PO 4 3.815 g/L、(NH 4 ) 2 HPO 4 0.825 g/L、MgCl 2 ·6H 2 O 20 mg/L、FeCl 3 0.2 mg/L、CaCl 2 2 mg/L、Na 2 SO 4 200 mg/L、KNO 3 1.2625 g/L; preparing degradation liquid with the pH value of 7.1;
according to the volume ratio of the heat-resistant pseudomonas aeruginosa bacteria liquid to the degradation liquid of 1:3, preparing a microbial inoculum;
on the soil to be repaired, the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1 of the invention is buried in the soil to be repaired, so that the upper surface of the high-efficiency low-carbon thermal stabilization system is positioned on the surface layer of the ground, at the moment, the liquid outlet positions of the capillaries of each heating group are in the range of 10 meters on the surface and underground, and the temperature of the soil is kept at 45 ℃ when the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1 of the invention is used for degrading polystyrene microplastic.
The microbial inoculum containing the heat-resistant pseudomonas aeruginosa bacterial liquid is injected into the reactor at a single injection rate of 100L/m in the early stage of degradation treatment (1 d, 9 d, 17 d from the beginning of degradation treatment) and in the later stage of degradation treatment (33 d, 49 d, 79 d, 109 d from the beginning of degradation treatment) 3 The soil was injected into the soil to be remediated in situ by the high-efficiency low-carbon thermal stabilization system provided in example 1 of the present invention, and the injection amount was 40L/m at the early stage of degradation treatment (5 th d, 13 d th from the start of degradation treatment) and at the later stage of degradation treatment (25 th d, 41 d, 64 d, 94 d, 138 d th from the start of degradation treatment), respectively, according to a single injection amount 3 The degradation liquid is independently injected into the soil to be repaired through the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1, the temperature of the soil is kept to be 28 ℃ when the microbial inoculum is injected each time, and the heat-resistant pseudomonas aeruginosa can grow and propagate rapidly at the temperature, so that the number of the bacterial strains in the soil is increased, after the microbial inoculum is injected into 12 h, the temperature of the soil to be repaired is increased to 45 ℃, the polystyrene micro-plastics are degraded at the temperature, and the temperature of the soil to be repaired is reduced to 28 ℃ before the next microbial inoculum is injected into 2 h. The early stage of degradation treatment is 1 d-24 d from the beginning of degradation treatment, and the later stage of degradation treatment is 25 th d from the beginning of degradation treatment and beyond.
Example 3
The invention provides a degradation method of polystyrene in soil, which comprises the following specific steps:
the preparation method of the heat-resistant pseudomonas aeruginosa bacterial liquid comprises the following steps: culturing the original strain pseudomonas aeruginosa to logarithmic growth phase at 25 ℃, taking 24 h as a domestication period, taking 2 ℃ as a temperature difference value, increasing the culture temperature gradually, namely culturing at 27 ℃ for 24 h, transferring the pseudomonas aeruginosa strain which can normally grow at 27 ℃ to a new culture medium, culturing at 29 ℃ for 24 h, transferring the pseudomonas aeruginosa strain which can normally grow at 29 ℃ to the new culture medium, and so on until the strain which can normally grow at 40 ℃ is domesticated and screened out, culturing at 40 ℃ for 24 h after the high temperature resistant strain obtained by primary screening is inoculated, performing rescreening and strain purification, and finally obtaining the heat resistant pseudomonas aeruginosa strain which can resist high temperature and degrade polystyrene; fermenting the obtained heat-resistant Pseudomonas aeruginosa strain at 25deg.C for 56 h to obtain heat-resistant Pseudomonas aeruginosa liquid with bacterial activity of 1.0X10 9 CFU/mL; the culture medium formula is the same in each step of the preparation method of the heat-resistant pseudomonas aeruginosa bacteria liquid, and specifically, glucose 4 g/L, beef extract 8 g/L, peptone 12 g/L, naCl 6 g/L and pH value of 7.0 are adopted;
according to K 2 HPO 4 0.4 g/L、KH 2 PO 4 3.5 g/L、(NH 4 ) 2 HPO 4 1 g/L、MgCl 2 ·6H 2 O 18 mg/L、FeCl 3 0.1 mg/L、CaCl 2 3 mg/L、Na 2 SO 4 180 mg/L、KNO 3 1 g/L; preparing degradation liquid with the pH value of 7.0;
according to the volume ratio of the heat-resistant pseudomonas aeruginosa bacteria liquid to the degradation liquid of 1:5, preparing a microbial inoculum;
on the soil to be repaired, the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1 of the invention is buried in the soil to be repaired, so that the upper surface of the high-efficiency low-carbon thermal stabilization system is positioned on the surface layer of the ground, at the moment, the liquid outlet positions of the capillaries of each heating group are in the range of 10 meters on the surface and underground, and the temperature of the soil is kept at 40 ℃ when the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1 of the invention is used for degrading polystyrene microplastic.
The microbial inoculum containing the heat-resistant pseudomonas aeruginosa bacterial liquid is injected into the reactor at a single injection rate of 200L/m in the early stage of degradation treatment (1 d, 9 d, 17 d from the beginning of degradation treatment) and in the later stage of degradation treatment (33 d, 49 d, 79 d, 109 d from the beginning of degradation treatment) 3 The soil was injected into the soil to be remediated in situ by the high-efficiency low-carbon thermal stabilization system provided in example 1 of the present invention, and the injection amount was 30L/m at the early stage of degradation treatment (5 th d, 13 d th from the start of degradation treatment) and at the later stage of degradation treatment (25 th d, 41 d, 64 d, 94 d, 138 d th from the start of degradation treatment), respectively, in a single injection 3 The degradation liquid is independently injected into the soil to be repaired through the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1, the temperature of the soil is kept to be 25 ℃ when the microbial inoculum is injected each time, and the heat-resistant pseudomonas aeruginosa can grow and propagate rapidly at the temperature, so that the number of the bacterial strains in the soil is increased, after the microbial inoculum is injected into 10 h, the temperature of the soil to be repaired is increased to 40 ℃, the polystyrene micro-plastics are degraded at the temperature, and the temperature of the soil to be repaired is reduced to 25 ℃ before the next microbial inoculum is injected into 1 h. The early stage of degradation treatment is 1 d-24 d from the beginning of degradation treatment, and the later stage of degradation treatment is 25 th d from the beginning of degradation treatment and beyond.
Example 4
The invention provides a degradation method of polystyrene in soil, which comprises the following specific steps:
the preparation method of the heat-resistant pseudomonas aeruginosa bacterial liquid comprises the following steps: firstly, an original strain pseudomonas aeruginosa is cultivated to a logarithmic growth phase at 30 ℃, a domestication period is taken as 12 h, a temperature difference is taken as 1 ℃, a gradient is increased, the cultivation temperature is gradually increased, namely, the pseudomonas aeruginosa strain which can normally grow at 31 ℃ is cultivated to 12 h, the pseudomonas aeruginosa strain which can normally grow at 31 ℃ is transferred to a new culture medium, the pseudomonas aeruginosa strain which can normally grow at 32 ℃ is cultivated to 12 h, the pseudomonas aeruginosa strain which can normally grow at 32 ℃ is transferred to the new culture medium, and so on, until the strain which can normally grow at 43 ℃ is domesticated and screened out, the high temperature resistant strain obtained by primary screening is inoculated to 48 h which is cultivated at 43 ℃ for re-screening and strain purificationThen obtaining a heat-resistant pseudomonas aeruginosa strain which is resistant to high temperature and degrading polystyrene; fermenting the obtained heat-resistant Pseudomonas aeruginosa strain at 30deg.C for 48 h to obtain heat-resistant Pseudomonas aeruginosa liquid with bacterial activity of 1.0X10 8 CFU/mL; the culture medium formula is the same in each step of the preparation method of the heat-resistant pseudomonas aeruginosa bacteria liquid, and specifically adopts glucose 6 g/L, beef extract 12 g/L, peptone 8 g/L, naCl 4 g/L and pH 7.2;
according to K 2 HPO 4 0.6 g/L、KH 2 PO 4 4 g/L、(NH 4 ) 2 HPO 4 0.5 g/L、MgCl 2 ·6H 2 O 22 mg/L、FeCl 3 0.3 mg/L、CaCl 2 1 mg/L、Na 2 SO 4 220 mg/L、KNO 3 1.5 g/L; preparing degradation liquid with the pH value of 7.2;
according to the volume ratio of the heat-resistant pseudomonas aeruginosa bacteria liquid to the degradation liquid of 1:2, preparing a microbial inoculum;
on the soil to be repaired, the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1 of the invention is buried in the soil to be repaired, so that the upper surface of the high-efficiency low-carbon thermal stabilization system is positioned on the surface layer of the ground, at the moment, the liquid outlet positions of the capillaries of each heating group are in the range of 10 meters on the surface and underground, and the temperature of the soil is kept at 43 ℃ when the high-efficiency low-carbon thermal stabilization system provided by the embodiment 1 of the invention is used for degrading polystyrene microplastic.
The microbial inoculum containing the heat-resistant pseudomonas aeruginosa bacterial liquid is injected at a single injection rate of 50L/m in the early stage of degradation treatment (1 d, 9 d, 17 d from the beginning of degradation treatment) and in the later stage of degradation treatment (33 d, 49 d, 79 d, 109 d from the beginning of degradation treatment) 3 The soil was injected into the soil to be remediated in situ by the high-efficiency low-carbon thermal stabilization system provided in example 1 of the present invention, and the injection amount was 50L/m at the early stage of degradation treatment (5 th d, 13 d th from the start of degradation treatment) and at the later stage of degradation treatment (25 th d, 41 d, 64 d, 94 d, 138 d th from the start of degradation treatment), respectively, according to a single injection amount 3 In-situ injection of degradation liquid into soil to be repaired through the efficient low-carbon thermal stabilization system provided in the embodiment 1 of the inventionThe temperature of the soil is kept at 30 ℃ when the microbial inoculum is injected each time, and the heat-resistant pseudomonas aeruginosa can grow and reproduce rapidly at the temperature, so that the number of the strain in the soil is increased, after the microbial inoculum is injected into the soil, 15 h, the temperature of the soil to be repaired is increased to 43 ℃, the polystyrene micro-plastics are degraded at the temperature, and 3 h before the next microbial inoculum injection, the temperature of the soil to be repaired is reduced to 30 ℃. The early stage of degradation treatment is 1 d-24 d from the beginning of degradation treatment, and the later stage of degradation treatment is 25 th d from the beginning of degradation treatment and beyond.
Comparative example 1
The degradation method of polystyrene in soil provided in this comparative example is substantially the same as the procedure of example 2, the only difference being that the soil temperature is maintained at 50℃when the polystyrene microplastic is degraded.
Comparative example 2
The degradation method of polystyrene in soil provided in this comparative example is substantially the same as the procedure of example 2, the only difference being that the soil temperature is maintained at 30℃when the polystyrene microplastic is degraded.
Comparative example 3
The degradation method of polystyrene in soil provided by the comparative example is basically the same as the step of example 2, and the only difference is that the original strain is not subjected to heat-resistant domestication, namely the adopted bacterial liquid is obtained by fermenting 52 h of pseudomonas aeruginosa with the preservation number of CGMCC No.23974 at 28 ℃.
Experimental example 1
The degradation rates of polystyrene in the soil of examples 2 to 4 and comparative examples 1 to 3 were detected by using a microplastic online monitoring system based on raman spectrum, respectively, at the beginning of degradation, 5 th d, 10 th d, 20 th d, 40 th d, 80 th d, 140 th d and 180 th d, and the detection results are shown in table 1.
TABLE 1 degradation rate of polystyrene in soil
As can be seen from Table 1, the degradation of polystyrene in soil by example 2 to example 4 is excellent compared with comparative examples 1 to 4, and in particular, the degradation rate of polystyrene in soil to be repaired by example 2 reaches 61.7% at 180 d of the degradation treatment, and the degradation rate of polystyrene in soil to be repaired is improved by 60.7% compared with comparative example 1, which means that the degradation efficiency of the degradation method of the present invention is the highest when the soil temperature is maintained at 45 ℃. Also in the 180 th d of degradation treatment, when the soil temperature is increased to 50 ℃, the soil temperature exceeds the tolerance range of the strain after heat-resistant domestication, so that the degradation rate is reduced; the degradation rate of the example 2 is improved by 131.1% compared with that of the comparative example 2, which shows that the degradation effect of the domesticated strain at a proper temperature is far higher than that at normal temperature; the degradation rate of example 2 is increased by 134.6% compared with that of comparative example 3, which shows that the degradation effect of the domesticated strain on polystyrene in soil is much higher than that of the non-domesticated strain.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.

Claims (10)

1. The degradation method of polystyrene in soil is characterized by comprising the following steps:
dividing a microbial inoculum containing heat-resistant pseudomonas aeruginosa bacterial liquid into at least two times, injecting the microbial inoculum into soil containing polystyrene, wherein the temperature of the soil is 25-30 ℃, and keeping the temperature for 10-15 hours; then raising the temperature of the soil to 40-45 ℃ and maintaining the temperature until the next time before injecting the microbial inoculum for 1-3 hours;
the preparation method of the heat-resistant pseudomonas aeruginosa bacterial liquid comprises the following steps:
s1, taking pseudomonas aeruginosa with the preservation number of CGMCC No.23974, and culturing at 25-30 ℃ to logarithmic phase; the method comprises the steps of (1) increasing the culture temperature of a strain in a logarithmic growth phase to 40-45 ℃ according to the temperature difference of 1-2 ℃ in each domestication period to obtain a primary screening strain;
s2, culturing the primary screening strain at 40-45 ℃, re-screening, and purifying the strain to obtain heat-resistant pseudomonas aeruginosa;
s3, inoculating the heat-resistant pseudomonas aeruginosa into a culture medium, and fermenting for 48-56 hours at the temperature of 25-30 ℃.
2. The degradation method according to claim 1, wherein the microbial agent further comprises a degradation liquid; the degradation liquid comprises the following components: k (K) 2 HPO 4 0.4 g/L~0.6 g/L、KH 2 PO 4 3.5 g/L~4 g/L、(NH 4 ) 2 HPO 4 0.5 g/L~1 g/L、MgCl 2 ·6H 2 O 18 mg/L~22 mg/L、FeCl 3 0.1 mg/L~0.3 mg/L、CaCl 2 1 mg/L~3 mg/L、Na 2 SO 4 180 mg/L~220 mg/L、KNO 3 1.1 g/L-1.5 g/L, wherein the pH value of the degradation liquid is 7.0-7.2; and/or the number of the groups of groups,
the bacterial activity of the heat-resistant pseudomonas aeruginosa bacterial liquid is 1.0x10 8 CFU/mL~1.0×10 9 CFU/mL。
3. The degradation method according to claim 2, wherein the composition of the culture medium adopted in any one of the steps S1 to S3 comprises glucose 4 g/L to 6 g/L, beef extract 8 g/L to 12 g/L, peptone 8 g/L to 12 g/L, naCl 4 g/L to 6 g/L, and the pH value of the culture medium is 7.0 to 7.2; and/or the number of the groups of groups,
each domestication period in the step S1 is 12-24 hours; and/or the number of the groups of groups,
the culture time in the step S2 is 24-48 hours; and/or the number of the groups of groups,
the volume ratio of the heat-resistant pseudomonas aeruginosa bacterial liquid to the degradation liquid in the microbial inoculum is 1: 2-5; and/or the number of the groups of groups,
the degradation method further includes the step of injecting the degradation liquid separately into the soil.
4. The degradation method according to claim 3, wherein the injection interval of the microbial inoculum is 7 d-9 d in the early stage of the degradation treatment, and the injection interval of the microbial inoculum is 16 d-30 d in the later stage of the degradation treatment; during degradation treatment, the degradation liquid is independently injected after the bacterial agent is injected each time, the time between the independent injection of the degradation liquid and the injection of the bacterial agent in the early stage of the degradation treatment is 3 d-5 d, the time between the independent injection of the degradation liquid and the injection of the bacterial agent in the later stage of the degradation treatment is 8 d-29 d, wherein the early stage of the degradation treatment is 1 d-24 d from the beginning of the degradation treatment, and the later stage of the degradation treatment is 25 th d and later from the beginning of the degradation treatment; and/or the number of the groups of groups,
the single injection amount of the microbial inoculum and the soil volume ratio are 50L/m 3 ~200 L/m 3 The method comprises the steps of carrying out a first treatment on the surface of the And/or the number of the groups of groups,
when the degradation liquid is singly injected, the single injection amount of the degradation liquid and the soil volume ratio are 30L/m 3 ~50 L/m 3
5. The degradation method according to any one of claims 1 to 4, further comprising monitoring degradation of polystyrene in the soil in real time by using a micro-plastic online monitoring system based on raman spectroscopy.
6. The degradation method according to claim 4, wherein the in-situ injection of the microbial inoculum and/or the degradation liquid is achieved by using a high-efficiency low-carbon thermal stabilization system;
the high efficiency low carbon thermal stabilization system comprises: a heat source generating module (5), a heat exchanger (7), a controller (3) and a temperature acquisition device (1), wherein,
the heat source generation module (5) and the heat exchanger (7) are mutually communicated through a pipeline to form a loop, a power device (4) is arranged on the pipeline, and the power device (4) provides power for medium circulation in the heat source generation module (5) and the heat exchanger (7);
the heat exchanger (7) comprises a plurality of heating units (2) which are arranged side by side, and the heating units (2) are vertically arranged;
the temperature acquisition device (1) is positioned between two adjacent heating units (2), the temperature acquisition device (1) is used for acquiring the temperature of soil between the heating units (2), and the temperature acquisition device (1) and the power device (4) are electrically connected with the controller (3);
when the temperature of the soil between the heating units (2) is lower than a threshold value, the controller (3) controls the power device (4) to accelerate the medium flow speed in the heat source generating module (5) and the heat exchanger (7).
7. The degradation method according to claim 6, characterized in that the heating unit (2) comprises a plurality of heating groups (6) arranged in parallel, the water inlet of each heating group (6) being communicated with the water outlet of the power plant (4); the water outlet of the heating group (6) is communicated with the water inlet of the power device (4); each heating group (6) comprises a communicating pipe (9) which is horizontally arranged and a capillary tube (10) which is vertically arranged, the communicating pipe (9) is communicated with the capillary tube (10), and the inner diameter of the communicating pipe (9) is larger than the inner diameter of the capillary tube (10).
8. The degradation method according to claim 7, characterized in that the capillary tube (10) comprises an outer sleeve (20) and an inner sleeve (18), the inner sleeve (18) being nested within the outer sleeve (20); the outer sleeve (20) is provided with an outer liquid outlet (14), the inner sleeve (18) is provided with an inner liquid outlet (19) at a position corresponding to the outer liquid outlet (14), and the outer liquid outlet (14) and the inner liquid outlet (19) have the same structure; the top cover (13) of the outer sleeve (20) is provided with a perforation (12), the inside of the inner sleeve (18) is connected with a connecting rod (16), the connecting rod (16) and the inner sleeve (18) are coaxially arranged, the connecting rod (16) is connected with the inner wall of the inner sleeve (18) through a supporting rod (17), and the connecting rod (16) penetrates through the perforation (12); one end of the connecting rod (16) is positioned outside the outer sleeve (20), a gear (15) is arranged at one end of the connecting rod (16) positioned outside the outer sleeve (20), and the gear (15) is connected with a driving motor through a driving belt (8);
the medium in the heat source generating module (5) and the heat exchanger (7) is the microbial inoculum and/or the degradation liquid for repairing the soil, the driving motor is controlled to drive the gear (15) to rotate, so that the inner sleeve (18) and the outer sleeve (20) rotate relatively, the inner liquid outlet (19) on the inner sleeve (18) and the outer liquid outlet (14) on the outer sleeve (20) are overlapped, the microbial inoculum and/or the degradation liquid is fed into the soil through the inner liquid outlet (19) and the outer liquid outlet (14), and after the feeding is finished, the driving motor is controlled to enable the inner liquid outlet (19) and the outer liquid outlet (14) to be staggered, and the microbial inoculum and/or the degradation liquid cannot pass through the inner liquid outlet (19) and the outer liquid outlet (14).
9. The degradation method according to claim 8, characterized in that the upper end of the outer sleeve (20) is provided with a first bearing (11), the lower end of the outer ring of the first bearing (11) is fixed on the outer wall of the outer sleeve (20), and the connecting rod (16) is fixed on the inner ring of the first bearing (11); the bottom in the outer sleeve (20) is provided with a second bearing, an outer ring of the second bearing is fixed on the inner wall of the outer sleeve (20), and the lower end of the connecting rod (16) is fixed on an inner ring of the second bearing; and/or the number of the groups of groups,
the outer sleeve (20) is coated with non-woven fabrics, and the non-woven fabrics cover the outer liquid outlet (14); the capillaries (10) are arranged in rows, and the gears (15) of the capillaries (10) in the same row are connected with a driving motor through the same driving belt (8).
10. The degradation method according to any one of claims 7 to 9, characterized in that a distance between adjacent heating units (2) is 0.5 m to 2 m, and a distance between adjacent capillaries (10) is 0.1 m to 0.5 m; and/or the heat source generating module (5) is a solar collector, the inner diameter of the communicating pipe (9) is 10 to 20 cm, the inner diameter of the capillary tube (10) is 1 to 2 cm, and the power device (4) is a water pump.
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