CN116179529B - Method for degrading TBBPA by composting humus microbial agent and application - Google Patents
Method for degrading TBBPA by composting humus microbial agent and application Download PDFInfo
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- CN116179529B CN116179529B CN202310161243.2A CN202310161243A CN116179529B CN 116179529 B CN116179529 B CN 116179529B CN 202310161243 A CN202310161243 A CN 202310161243A CN 116179529 B CN116179529 B CN 116179529B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/80—Separation, elimination or disposal of harmful substances during the treatment
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
- C02F2101/345—Phenols
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/36—Organic compounds containing halogen
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2305/00—Use of specific compounds during water treatment
- C02F2305/06—Nutrients for stimulating the growth of microorganisms
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- 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
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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Abstract
The invention relates to the technical field of environmental protection, and discloses a composting humus microbial agent and application thereof, wherein the microbial agent is prepared by the following steps: s1: preparing wood chips and fresh cow dung; s2: preparing mixed humus by composting; s3: separating and mixing humus; s4: preparing Y17 bacterial liquid; s5: preparing a microbial degradation microbial agent; the application is degradation of organic pollutants: and (3) sowing the degrading microbial inoculum into soil or water containing the organic pollutant TBBPA, and adsorbing and degrading the organic pollutant TBBPA in the soil or water. The microbial agent disclosed by the invention can be simultaneously applied to degradation of TBBPA in soil or water, can support synchronous phytoecological restoration, can accelerate the ecological restoration, shorten the time, lengthen the continuous restoration action time, can meet the ecological management requirements of various organic environmental pollutants, and can greatly utilize agricultural and forestry wastes such as fruit tree branches and the like, and reduce the comprehensive management cost.
Description
Technical Field
The invention relates to the technical field of microorganism and environmental protection, in particular to a method for degrading TBBPA by utilizing a composting humus microbial agent and application thereof.
Background
In recent years, researches on biological treatment technology of organic pollution in the environment are rapidly developed, and various pollutant environment remediation technologies are proposed. At present, the environmental remediation of pollutants can be performed by physical methods, chemical methods and biological methods. Bioremediation has been favored by more and more environmental researchers over the last two decades, and has become a promising approach for polluted environmental remediation. But it also has its own drawbacks compared to conventional repair methods: (1) The repairing time is long, and the repairing method is generally not suitable for small-range serious pollution caused by sudden accidents; (2) The repairing effect is not as thorough as chemical repairing and physical repairing; (3) The method is greatly influenced by environmental factors, and the repair conditions are not easy to be controlled manually.
Therefore, bioremediation is a technology for purifying water and soil by using microbial degradation to purify toxic and harmful pollutants, and has strong advantages. Bioremediation involves biostimulation and bioscaling, which generally avoids dredging to control costs, and is considered a cost-effective and efficient technique. Bioremediation (Bioremediation) is a process of accelerating the removal of pollutants in the environment by utilizing the functions of absorption, metabolism, degradation, etc. of environmental pollutants by organisms. It is generally directed to pollution in the natural environment, either a controlled process or a spontaneous process. Generally including microbial repair and ecological phytoremediation.
However, existing bioremediation techniques involve a number of limitations, such as slow growth of contaminant-degrading microorganisms and low contaminant-degrading biomass, and due to O in the body of water 2 The slow rate of transport limits the growth rate of contaminant degrading microorganisms, and thus bioremediation requires enhanced removal of contaminants by microorganisms through biostimulation and biological amplification. Biostimulation refers to the stimulation of the potential to stimulate indigenous microorganisms to degrade pollutants or the co-metabolism to degrade target pollutants by the addition of suitable electron donors/acceptors or nutrients. However, these stimuli are based on the ability of indigenous microbial flora to degrade contaminants, and thus there is a lot of uncertainty in the biological stimulus. Whether the polluted water contains microorganisms capable of degrading pollutants or not is not determined or the time required for generating enough microorganisms for the polluted water is possibly long; in addition, the existing microorganism repairing technology does not support synchronous plant ecological repairing, so that the ecological repairing speed is low, the time is long, and the continuous repairing action time is short.
In order to solve the problem of microbial remediation of TBBPA in water, the inventor group provides a method for removing TBBPA in water, a microbial strain and a microbial agent in ZL202210029345.4, wherein the microbial strain is domesticated Burkholderia cepacia Burkholderia cepaciaA strain designated Y17 with deposit No. GDMCC No.62153; the microbial agent and the method for removing TBBPA in the water body are characterized in that a Y17 strain is planted on the surface and in a pore canal of biochar, TBBPA in the water body is used as a carbon source, air and water dissolved oxygen are used as oxygen sources, the biochar provides a microscopic growth environment for degrading TBBPA in the water body for the strain, the strain grows aerobically in the water body, and the degradation reaction continuously occurs to carry out biological enhanced degradation on TBBPA in the water body. However, the microbial strain and microbial agent provided by the invention have high cost, and can not support synchronous ecological restoration of plants, and the restoration speed needs to be further improved.
China is a large country of agriculture and forestry, and the resources of the agriculture and forestry waste generated based on the planting of fruit trees are rich. For example, apple tree planting is widely distributed nationally, and apple tree branches with average yield of 4297.5-7500 kg per hectare in apple orchards in China are difficult to treat according to statistics of related institutions. Aerobic composting is widely applied to resource recovery of agriculture and forestry waste and cow dung, and the production of humus from agriculture and forestry waste (fruit tree branches) through decomposition treatment is an advantageous measure for realizing resource utilization of agriculture and forestry waste. The agricultural and forestry waste can be used as a conditioner to improve the humification degree of the compost products, and lignocellulose can generate humus through microbial activity, so that cow dung compost is improved. However, agricultural and forestry waste having a difficult degradation lignocellulose structure is difficult to biodegrade during aerobic composting, and thus degradation speed becomes slow. However, apple wood is taken as a typical lignocellulose biomass, and although the lignocellulose structure can be destroyed through a certain pretreatment to accelerate the decomposition of agricultural and forestry wastes, the process and the products are still limited to obtain decomposed organic fertilizer.
The decomposed organic fertilizer prepared by composting in the prior art is a fertilizer which is harmless, is used more rapidly, safely and effectively, and is generally prepared by fermenting fresh livestock manure, gasket materials, stems and leaves of plants and straws at high temperature or normal temperature. The compost-decomposed organic fertilizer can effectively supplement nutrients required by crop growth, improve soil structure and promote plant yield increase. The decomposed organic fertilizer can treat organic waste, form ecological circulation and effectively reduce environmental pollution; however, the decomposed organic fertilizer formed by the existing compost cannot be further applied to ecological environment pollution control by a microbial degradation method.
In the prior art, wood chips and fresh livestock manure are subjected to composting to obtain decomposed organic fertilizer, so that on one hand, the livestock manure and nitrogenous fertilizer make up for the defect of too low nutrient of the wood chips, the decomposition of the wood chips is accelerated to form some humus, the volume of the wood chips is reduced, and on the other hand, the absorption effect of the wood chips can absorb easily-decomposed and soluble components in the organic fertilizer, so that the nutrient loss of the livestock manure is reduced. The humus obtained by the composting technology in the prior art aims at accelerating the decomposition of wood chips and improving fertility, and the obtained humus is unstable in structure, is difficult to prepare into a microbial inoculum for treating soil or water pollution, and cannot meet the requirements of in-situ ecological restoration of polluted soil or water.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a composting humus microbial agent, application of the composting humus microbial agent in degradation of TBBPA and ecological management, three substances are separated from humus prepared by composting after pretreatment of waste fruit tree branches, and Y17 microbial agent is further prepared by taking structurally stable humins as carriers, so that the microbial agent can be simultaneously applied to degradation of TBBPA in soil or water, and can support synchronous plant ecological restoration, so that the ecological restoration speed is increased, the time is shortened, the continuous restoration action time is prolonged, and the ecological management requirements of various organic environmental pollutants can be met; and a large amount of agricultural and forestry wastes such as fruit tree branches can be utilized in a recycling way, and the comprehensive treatment cost is reduced.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows:
the composting humus microbial agent is characterized by being prepared by the following steps:
s1: preparing wood chips and fresh cow dung: preparing waste fruit tree branches into wood chip particles, pretreating the wood chip particles, and then air-drying the pretreated wood chips until the water content is not more than 20%, so as to obtain pretreated wood chips for later use; collecting fresh cow dung in a farm, so that the water content of the cow dung is not higher than 70%, and obtaining the fresh cow dung for later use;
S2: preparing mixed humus by composting: uniformly mixing the standby pretreated wood chips and fresh cow dung according to a set weight ratio to obtain a pile; controlling the pile body, composting, and obtaining mixed humus with a plurality of components mixed with each other after the pile body is continued for a set time;
s3: separating and mixing humus: separating soluble fulvic acid, humic acid and insoluble humins from the mixed humus, and respectively collecting and storing the fulvic acid, the humic acid and the humins with rough surfaces, rich pores and stable structure;
s4: preparing Y17 bacterial liquid: adding a nutrition culture medium into the Y17 strain for rejuvenation and expansion culture, rotating at 35 ℃ and 160r/min, and performing expansion culture for 12 hours to obtain Y17 bacterial liquid;
s5: preparing a microbial degradation microbial agent: and (3) adding the Y17 bacterial liquid into the humins separated in the step (S3) according to a set proportion, and performing field planting and culture for a set time by taking the humins as carriers, so that the Y17 bacterial strain is planted on the rough surface and rich pore structures of the humins, thereby preparing the microbial degradation microbial inoculum.
The application of the composting humus microbial agent is characterized in that the microbial agent is used for degrading TBBPA, and the method comprises the following steps:
S6: degrading organic pollutants: according to the set proportion, the microbial degradation microbial inoculum is sown into soil or water containing organic pollutant TBBPA, and oxygen contained in a humin and a pore structure thereof provides growth conditions and microenvironment for Y17 strain to aerobically grow in the soil or water so as to adsorb and degrade the organic pollutant TBBPA in the soil or water;
s7: synergistic plant ecological restoration: the fulvic acid and the humin acid solution with required concentrations are synchronously added into the soil or the water body containing the organic pollutant TBBPA, so that the growth of the Y17 strain is promoted, the degradation speed of the TBBPA is accelerated, and the degraded components and the components which do not participate in the degradation can provide organic nutrient substances for plants growing in the soil or the water body, so that the plant growth and ecological restoration speed are accelerated.
The ecological restoration method of the TBBPA polluted soil adopts the steps of composting humus microbial agent and degrading TBBPA, the microbial degradation microbial agent is sown into the TBBPA polluted soil, and the microbial restoration and the plant restoration of the polluted soil are synchronously carried out in situ under the combined action of Y17 strain and humins.
The ecological restoration method of the TBBPA polluted water body adopts the steps of composting humus microbial inoculum and degrading TBBPA, the microbial degradation microbial inoculum is sown into the TBBPA polluted water body, and the microbial restoration and the phytoremediation of the polluted water body are synchronously carried out in situ under the combined action of Y17 strain, humins, fulvic acid and humic acid.
1. The composting humus microbial agent and the application thereof provided by the invention aim at simultaneously supporting the degradation of TBBPA in soil or water and simultaneously carrying out microbial remediation and phytoremediation, simplify the process steps, adopt a large amount of agricultural and forestry wastes, and can meet the requirements of large-scale preparation and application. According to the invention, three substances are separated from humus prepared by pretreatment and composting of waste fruit tree branches, and the humins with stable structures are further used as carriers to prepare the Y17 microbial inoculum, so that the microbial inoculum can be simultaneously applied to degradation of TBBPA in soil or water, and can support synchronous plant ecological restoration, so that the ecological restoration speed is increased, the time is shortened, the continuous restoration action time is prolonged, and the ecological management requirements of various organic environmental pollutants can be met.
2. The composting humus microbial agent and the application thereof provided by the invention can consume a large amount of agricultural and forestry wastes such as waste fruit trees, cow dung and the like and recycle the agricultural and forestry wastes, humus is prepared by pretreatment and composting, and is combined with Y17 strain, and the humus is used as a microbial carrier to be prepared into a microbial agent with low cost and a Y17 strain in a large scale, and other suitable microbial strains can be loaded, and can be used as a biodegradation agent and conditioner of soil or water, so that the dual repair of microbial repair and plant repair can be supported, the composting agent can be widely applied to the treatment of various pollutants including TBBPA in the soil and the water, the in-situ biological repair of polluted environment is accelerated, and the composting agent is particularly suitable for artificial green land and worker forestation purification repair engineering of in-situ microbial repair+synchronous planting ecological plant washing of polluted soil; the method is also suitable for the constructed wetland purification and repair engineering of in-situ microorganism repair and aquatic plant repair of the polluted water body, and has the effects of low cost, quick response, long duration and the like.
3. The invention provides a composting humus microbial agent and application thereof, and also respectively researches the performance and basic mechanism of pretreatment of apple tree branch particles by adopting three physicochemical pretreatment technologies (including acid, alkali and high-temperature treatment) so as to change microstructure, promote the processes of composting, sterilization, disinsection and the like in the composting process, and ensure that humus (humins in the composting process) obtained after composting has better surface roughness and higher porosity so as to be suitable for further preparation into the microbial agent; the hard fruit tree branches including apple tree branches are subjected to different pretreatment, so that lignocellulose can be better degraded more through the composting process, more biodegradable humus can be obtained, humification of the hard fruit tree branches and fresh cow dung and other materials in the composting process is accelerated, and the lignocellulose can be obviously degraded and humified, so that the humins with stable structures can be obtained.
4. The invention provides a composting humus microbial agent and application thereof, which adopts woody fiber wood chips (agricultural and forestry waste taking woody fibers as a main body, such as apple branches and the like) of waste fruit and wood branches with specific dimensions to carry out pretreatment, in particular to alkalization treatment, so as to reduce the crystallinity of the fruit and wood branches, then the woody fiber wood chips and raw materials such as fresh cow dung (taking herbaceous fibers as a main body) are mixed according to a set proportion, the free airspace of the wood chips and the comprehensive conditioner effect of alkali concentration are increased after pretreatment, under the condition of static natural ventilation composting, various favorable conditions are created in the cow dung composting process, the growth of aerobic and anaerobic microorganisms is promoted in stages, the degradation and humification of woody and herbaceous lignocellulose are synchronously accelerated, and finally the decomposed soluble organic fertilizer and the humins with stable structures are obtained. In the embodiment of the invention, wood branches of apple trees are used for preparing wood chip particles, cow dung is used for composting, and the degradation rate of lignocellulose is improved by carrying out different pretreatment (including acid, alkali and high temperature) on the apple trees, so that the structural strength and stability of insoluble humins in the prepared humus can be improved.
5. The microbial degradation microbial agent is prepared by mutual cooperation of a humus structure and microbial strains, is sown in soil or water containing organic pollutant TBBPA, provides growth conditions and microenvironment for aerobic growth in the soil or water for the Y17 strain by using humins so as to adsorb and degrade the organic pollutant TBBPA in the soil or water, and ensures that the degraded components provide organic nutrient substances for plants growing in the soil or water, supports double ecological restoration, has high degradation efficiency for pollutants, is environment-friendly and has low comprehensive cost, and can meet the requirements of large-scale popularization and application.
6. Practical application verification and test show that after composting treatment, the surfaces of the wood chip particles subjected to alkalization pretreatment are rougher, more pores are displayed, the surface area of the wood chip particles is increased, and the adhesion of microorganisms on the surfaces of the wood chip particles is facilitated; the crystallinity index (CrI value) of the waste is obviously reduced, lignin and hemicellulose are effectively removed to expose cellulose, then the cellulose is promoted to be converted into an amorphous structure, the functional group of apple tree is obviously changed, the Y17 strain can be better fixed, better aerobic growth conditions and micro-environments can be obtained in soil or water, and TBBPA in the soil or water can be accelerated to be degraded.
7. The composting humus microbial agent and the application thereof provided by the invention have the advantages of good degradation effect on various organic pollutants in soil and water, environmental friendliness, easiness in preparation, low cost, convenience in use and the like, and can be widely popularized and applied to treatment of organic pollutants TBBPA in soil or water. Through experiments and calculation, the total cost of treatment by adopting the technology of the invention is only about 20% of that of the chemical treatment method; the method has the advantages that the method is cooperated with plants for repairing, and the speed is increased by more than 30 percent compared with the existing single microorganism repairing method.
Drawings
FIG. 1 is a SEM comparative view of pretreated apple wood particles according to an embodiment of the invention;
FIG. 2 is a FTIR Fourier infrared spectrum of apple wood particles pretreated in accordance with an embodiment of the present invention;
FIG. 3 is a graph showing different pre-treatment apple wood grain size composting temperatures in accordance with an embodiment of the present invention;
FIG. 4 shows composting temperatures for different mixing ratios according to an embodiment of the invention;
FIG. 5 shows the temperature of pre-treatment compost for different pre-treatment of apple wood particles in accordance with an embodiment of the present invention;
FIG. 6 is a three-dimensional fluorescence graph of different pretreated apple wood particle mixed composting humus according to the embodiment of the invention;
FIG. 7 is a scanning electron microscope image of a microbial degradation agent (containing a small amount of an established strain) according to an embodiment of the present invention.
The technical scheme of the invention is described in detail below with reference to the accompanying drawings and examples.
Detailed Description
Example 1:
referring to fig. 1-7, the application of the composting humus microbial agent and the degradation TBBPA provided by the embodiment comprises the following steps:
a composting humus microbial agent is prepared by the following steps:
s1: preparing wood chips and fresh cow dung: preparing waste fruit tree branches into wood chip particles, pretreating the wood chip particles, and then air-drying the pretreated wood chips until the water content is not more than 20%, so as to obtain pretreated wood chips for later use; collecting fresh cow dung in a farm, so that the water content of the cow dung is not higher than 70%, and obtaining the fresh cow dung for later use;
s2: preparing mixed humus by composting: uniformly mixing the standby pretreated wood chips and fresh cow dung according to a set weight ratio to obtain a pile; controlling the pile body, composting, and obtaining mixed humus with a plurality of components mixed with each other after the pile body is continued for a set time;
s3: separating and mixing humus: separating soluble fulvic acid, humic acid and insoluble humins from the mixed humus, and respectively collecting and storing the fulvic acid, the humic acid and the humins with rough surfaces, rich pores and stable structure;
S4: preparing Y17 bacterial liquid: adding a nutrition culture medium into the Y17 strain for rejuvenation and expansion culture, rotating at 35 ℃ and 160r/min, and performing expansion culture for 12 hours to obtain Y17 bacterial liquid;
s5: preparing a microbial degradation microbial agent: and (3) adding the Y17 bacterial liquid into the humins separated in the step (S3) according to a set proportion, and performing field planting and culture for a set time by taking the humins as carriers, so that the Y17 bacterial strain is planted on the rough surface and rich pore structures of the humins, thereby preparing the microbial degradation microbial inoculum.
The application of the composting humus microbial agent comprises the following steps:
s6: degrading organic pollutants: according to the set proportion, the microbial degradation microbial inoculum is sown into soil or water containing organic pollutant TBBPA, and oxygen contained in a humin and a pore structure thereof provides growth conditions and microenvironment for Y17 strain to aerobically grow in the soil or water so as to adsorb and degrade the organic pollutant TBBPA in the soil or water;
s7: synergistic plant ecological restoration: the fulvic acid and the humin acid solution with required concentrations are synchronously added into the soil or the water body containing the organic pollutant TBBPA, so that the growth of the Y17 strain is promoted, the degradation speed of the TBBPA is accelerated, and the degraded components and the components which do not participate in the degradation can provide organic nutrient substances for plants growing in the polluted soil or the water body, so that the plant growth and ecological restoration speed are accelerated.
The ecological restoration method of the TBBPA polluted soil adopts the steps of composting humus microbial agent and degrading TBBPA, the microbial degradation microbial agent is sown into the TBBPA polluted soil, and the microbial restoration and the plant restoration of the polluted soil are synchronously carried out in situ under the combined action of Y17 strain and humins.
The ecological restoration method of the TBBPA polluted water body adopts the steps of composting humus microbial inoculum and degrading TBBPA, the microbial degradation microbial inoculum is sown into the TBBPA polluted water body, and the microbial restoration and the phytoremediation of the polluted water body are synchronously carried out in situ under the combined action of Y17 strain, humins, fulvic acid and humic acid.
The following describes the technical scheme of the present invention in detail by using more specific examples.
Example 2:
the composting humus microbial agent, the method for degrading TBBPA and the ecological treatment application provided by the embodiment of the invention are applied to in-situ ecological restoration of TBBPA polluted soil on the basis of the embodiment 1, and the Y17 strain microbial restoration and ecological greening plants (such as herbs or shrubs) are adopted to restore the TBBPA polluted soil together so as to accelerate the restoration speed, improve the restoration effect, obtain better and longer-acting ecological greening effect, and are suitable for ecological treatment projects such as artificial green land ecological treatment, worker forestation ecological treatment and the like of polluted soil (normal water content).
The method for preparing and degrading TBBPA by using the composting humus microbial agent comprises the following steps of:
s11: preparing wood chips: mechanically crushing the agricultural and forestry waste fruit and wood branches collected in batches, preparing into wood chip particles with the particle size of 0.5-2mm and 5-8mm, selecting alkalization to pretreat the wood chip particles, and then air-drying the pretreated wood chip until the water content is not more than 20%, so as to prepare pretreated wood chip for later use; in the embodiment, the agricultural and forestry waste fruit tree branches adopt apple tree branches generated by trimming or updating;
the alkalization pretreatment method comprises the following steps: mixing the apple sawdust particles with 0.5mol/L NaOH according to a mass ratio of 1:10, and soaking for 12 hours at 25 ℃; repeatedly washing the apple sawdust particles in the solution to neutrality, and washing off NaOH on the surfaces of the apple sawdust particles; and finally, air-drying the apple sawdust particles until the water content is about 20% (not more than 20%) for subsequent composting.
S12: fresh cow dung:
fresh cow dung collected in batches and collected in an intensive dairy farm is prepared into fresh cow dung for standby by reducing the moisture content of the fresh cow dung through a solid-liquid separator to ensure that the moisture content of the fresh cow dung is not higher than 70 percent.
The step S2 specifically comprises the following steps:
S21: preparing a pile: uniformly mixing the standby pretreated wood chips and fresh cow dung according to the following weight proportion: fresh cow dung = 1:1, obtaining a pile body;
s22: preparing mixed humus: and (3) carrying out composting condition control on the pile body, carrying out static composting, and carrying out pile turning frequency of 10 days/time for 20-40 days until the set composting time to obtain mixed humus with a plurality of components mutually mixed.
In other embodiments, other composting modes and methods in the conventional technology can be adopted, and mixed humus with multiple components mixed with each other can be obtained after the composting time is reached.
The step S3 specifically comprises the following steps:
s31: separating and mixing humus: separating soluble fulvic acid and humic acid from the filtrate by an alkali dissolution and acid precipitation method, separating insoluble humic acid from the filter residue, and then respectively collecting and storing the fulvic acid, the humic acid and the humic acid, wherein the humic acid has rough surface, rich pores and stable physical three-dimensional space structure;
the alkali dissolution and acid precipitation method comprises the following specific steps: 2g of the compost sample after cold drying was weighed into a 50mL centrifuge tube, and 35mL of an extract (0.1M sodium hydroxide: 0.1M sodium pyrophosphate=1:1) was added. Oscillating for 10min by a water bath constant temperature oscillator 240r/min, uniformly mixing, and heating at 60 ℃ for 2 hours. Centrifugation at 4000r/min for ten minutes followed by filtration. The filtrate is humus solution, and the filter residue is humin. Adding 20ml of humus solution into a 50ml centrifuge tube, adding 5ml of 1M hydrochloric acid to acidify the pH to below 2, centrifuging at 4000r/min for 10 minutes after a constant temperature water bath is carried out at 60 ℃ for 1h, filtering supernatant fluid, collecting the supernatant fluid, wherein the filtrate is a fulvic acid solution, and extracting filter residues by using an extracting solution, namely a humin acid solution.
S32: and respectively purifying the fulvic acid, the humic acid and the humin according to the conventional technology for standby.
The step S4 specifically comprises the following steps:
s41: preparing Y17 bacterial liquid, specifically adding a Y17 bacterial strain into a nutrient medium for rejuvenation and expansion culture: rotating at 35 ℃ and 160r/min, and performing expansion culture for 12 hours to obtain Y17 bacterial liquid; the nutrient medium is as follows: 3g of beef extract, 10g of peptone and 5g of NaCl are added into pure water, the volume is fixed to 1000mL, and the pH is regulated to 7.2.
The Y17 strain is an acclimatized Burkholderia cepacia Burkholderia cepacia strain, and is preserved in the Guangdong province microorganism strain collection center, wherein the preservation number is GDMCC No.62153, and the preservation date is 2021, 12 months and 21 days.
The step S5 specifically comprises the following steps:
s51: preparing a microbial degradation microbial agent: adding the Y17 bacterial liquid with the OD value of 0.45 prepared in the step S4 into the nutrient medium containing the humins of 1 g/L separated in the step S3 according to the addition amount of 5% by volume, performing field planting and culture for at least 12 hours at the normal temperature with the rotating speed of 160r/min, and using the humins as a carrier to enable the Y17 bacterial strain to be planted on the rough surface and rich pore structures of the humins, so as to prepare the Y17 microbial degradation bacterial agent for fixation of the humins, wherein an electron microscope image of the humins after the field planting is shown in the figure 7.
The step S6 specifically comprises the following steps:
s61: degrading organic pollutants: according to the prior art, the proportion of microbial degradation bacteria (for example, the mass ratio of the microbial degradation bacteria to the water-containing soil is 1:150-2500, and the microbial degradation bacteria are determined according to the concentration of pollutants) is set according to the concentration of TBBPA in the soil to be degraded, the microbial degradation bacteria in the proportion are physically sown into the soil containing organic pollutants TBBPA, the microbial degradation bacteria are uniformly distributed in a pollution layer of the soil, and oxygen contained in humins and pore structures thereof provides growth conditions and micro-environments for aerobic growth of Y17 strains in the soil or water so as to adsorb and degrade the organic pollutants TBBPA in the soil pollution layer. The organic pollutant can also be other organic pollutants with molecular formula and molecular weight similar to TBBPA.
As can be seen from a scanning electron microscope (figure 1), after the apple sawdust particles are pretreated, the surfaces of the apple sawdust particles are rougher, more pores are displayed, the surface area of the apple sawdust particles is increased, and the adhesion of microorganisms on the surfaces of the apple sawdust particles is facilitated. The crystallinity index (CrI value) is calculated by apple chip XDR analysis and is shown in table 1.
TABLE 1
As can be seen from table 1, pretreatment of the apple chip pellets significantly reduced the waste crystallinity index (CrI value), effectively removed lignin and hemicellulose exposing the cellulose and then promoted it to an amorphous structure, wherein the reduction in CrI was 24.39% and 26.27% respectively, as evidenced by the acidification and alkalization treatments.
As can be seen from functional group analysis (FIG. 2), the pre-treatment in the early stage can significantly change the functional groups of the apple wood particles, the acidification treatment and the high temperature treatment, 1250 cm -1 The absorption peaks rising at the positions are formed by stretching vibration of aliphatic C-OH and C=O, and aliphatic functional groups rise; all processing is at 1040 cm -1 Higher peaks are shown, which are caused by polysaccharide C-O stretching; alkalizing treatment at 1640 and 1640 cm -1 The peak at the position is obviously reduced, which indicates that the water-soluble benzene ring structure released by lignocellulose degradation is mineralized; alkalization 1730 and 1730 cm -1 The peak disappears, which is the absorption peak formed by stretching C=O in carboxylic acid, aldehyde and ketone, and reduces the functional groups such as carboxylic acid, aldehyde and ketone in apple tree.
In the embodiment, apple wood is respectively crushed into wood chip particles with the particle sizes of 0.5-2mm and 5-8 mm, and mixed composting is carried out in a box according to the ratio of cow dung to apple wood in terms of dry matter mass of 1:1. All the compost is subjected to static composting, and manual turning is performed once every 10 days in the composting process. The composting mixed by apple wood particles with the particle size of 5-8 mm is used, the temperature is not 50 ℃ in the whole composting process, the highest temperature is only 46.2 ℃, and the composting is not completed within 40 days; and the composting mixed by apple wood particles with the particle size of 0.5-2mm has 6 days in the high temperature period exceeding 50 ℃ in the whole composting process, and the highest temperature reaches 52.1 ℃. The particle size of the apple wood particles was chosen to be 0.5-2mm.
The composting temperatures of different apple wood particle sizes are shown in figure 3.
In the embodiment, apple sawdust particles with the particle size of 0.5-2mm are used, and mixed composting is carried out in a box according to the dry mass of apple wood and cow dung of 1:1 and 1:2 respectively. All the compost is subjected to static composting, and manual turning is performed once every 10 days in the composting process. Mixed composting with 1:1 has a high temperature period exceeding 50 ℃ for 6 days during the whole composting process, and the highest temperature reaches 52.1 ℃. Mixed composting with 1:2 has a high temperature period exceeding 50 ℃ for 8 days in the whole composting process, and the highest temperature reaches 56.9 ℃. So that cow dung and apple sawdust particles are selected to be mixed and composted according to dry mass of 1:2 respectively.
The composting temperatures of different mixing ratios are shown in figure 4.
The ecological restoration method for the TBBPA contaminated soil provided by the invention is a method for degrading TBBPA by adopting the composting humus microbial agent, wherein the microbial degradation microbial agent is sowed into the TBBPA contaminated soil, and the microbial restoration and the plant restoration of the soil are synchronously carried out in situ by the combined action of a Y17 strain and humins, so that the method can be widely applied to ecological management projects such as artificial green land ecological management, worker forestation ecological management and the like of the TBBPA contaminated soil (normal water content), such as in-situ restoration of old site soil of a chemical plant and the like.
The microbial degradation microbial inoculum prepared by the embodiment is mainly suitable for the pollutant treatment of dry land soil, the microbial degradation microbial inoculum prepared by taking humus as a carrier is sown into the soil, and is a microbial remediation agent on one hand, a soil fertility agent on the second hand, and simultaneously, a conditioner of physical structure, air permeability and the like of a soil layer, so that the pollutant treatment, the soil structure improvement, the soil fertility improvement and the like can be organically combined, on the basis of the restoration treatment of TBBPA polluted soil, the restoration speed can be increased, the restoration effect can be improved, the better and longer-acting ecological greening effect can be obtained, and the microbial degradation microbial inoculum is widely suitable for ecological treatment engineering such as artificial green land ecological treatment, artificial forestation ecological treatment and the like of the dry polluted soil (normal water content), the treatment cost is greatly reduced, and various comprehensive benefits are obtained.
Example 3:
the composting humus microbial agent, the method for degrading TBBPA and the application of ecological restoration are applied to in-situ ecological restoration of TBBPA polluted water based on the embodiments 1 and 2, and the Y17 strain microbial restoration and ecological greening plants (such as aquatic herbaceous plants) are adopted to restore the TBBPA polluted water together so as to accelerate the water restoration speed, improve the restoration effect, obtain better and longer-acting ecological greening effect, and are suitable for water ecological purification treatment projects of polluted water (such as reservoirs, purifying ponds, artificial wetlands, natural ponds, river surges and the like).
The technical scheme of this embodiment is basically the same as embodiments 1 and 2, except that:
the preparation of the composting humus microbial agent and the method for degrading TBBPA comprise the following steps of:
s11: preparing wood chips: preparing waste fruit tree branches (specifically selecting pear tree) into wood chip particles with the particle size of 0.5-2mm, selecting acidification to pretreat the wood chip particles, and then air-drying the pretreated wood chip until the water content is not more than 20%, so as to prepare pretreated wood chip for later use; in the embodiment, the agricultural and forestry waste fruit tree branches adopt pear tree branches generated by trimming or updating.
The acidification pretreatment method comprises the following steps: mixing wood chip particles of fruit trees with 0.5mol/L HCl according to a mass ratio of 1:10, and soaking for 12 hours at 25 ℃; repeatedly washing fruit wood chip particles in the solution to neutrality, and washing off HCl on the surfaces of the fruit wood chip particles; finally, the wood dust particles of the fruit trees are air-dried until the water content is about 20 percent and used for subsequent composting.
The step S2 specifically comprises the following steps:
s21: preparing a pile: uniformly mixing the standby pretreated wood chips and fresh cow dung according to the following weight proportion: fresh cow dung = 1:2, obtaining a pile body;
The step S6 specifically comprises the following steps:
s61: degrading organic pollutants: according to the concentration of TBBPA in the water body to be degraded and the throwing proportion of the microbial degradation microbial agent, the microbial degradation microbial agent is sown in the water body containing the organic pollutant TBBPA in proportion to ensure that the microbial degradation microbial agent is uniformly distributed in the water body, and the oxygen contained in the humins and the pore structures thereof provides the growth conditions and the microenvironment for the Y17 strain to aerobically grow in the water body so as to adsorb and degrade the organic pollutant TBBPA molecules in the water body and decompose the organic pollutant TBBPA molecules into small molecular organic matters which can be absorbed by aquatic plants.
S71: synergistic plant ecological restoration: the organic pollutant TBBPA-containing water body is synchronously or asynchronously added with the fulvic acid and the humidify acid solution with required concentration (the specific concentration and the addition amount can be calculated according to the requirement), so that the growth of the Y17 strain and aquatic plants is promoted, the degradation speed of the TBBPA is accelerated, the degraded components and the components which do not participate in degradation can provide organic nutrient substances for plants growing in the water body, the growth and ecological restoration speed of the plants are accelerated, the plants can be maintained for a long time, the water body is purified, and the adding times and the total adding amount of microbial degradation microbial inoculum are reduced.
The ecological restoration method for the TBBPA polluted water body provided by the embodiment adopts the composting humus microbial agent and the method for degrading TBBPA, the microbial degradation microbial agent is sown into the TBBPA polluted water body, and the microbial restoration and the phytoremediation of the polluted water body are synchronously carried out in situ through the combined action of Y17 strain, humins, fulvic acid and humic acid.
The method and the device for removing the TBBPA from the water body improve the effect of removing the TBBPA from the water body, simultaneously aim to reduce the total consumption and the sowing times of a material system, improve the adaptability of the strain and the microbial inoculum to the water body environment and the symbiosis of the strain and the microbial inoculum with aquatic plants for purifying the water body, greatly reduce the sewage treatment cost and facilitate large-area popularization. Through practical tests, the microbial degradation microbial inoculum provided by the invention can be applied once and degraded for 7-8 days to effectively remove TBBPA in water body, so as to reach the set standard; compared with the traditional technology, the overall water treatment scheme of the invention can save more than 80% of total cost and can improve more than 50% of degradation efficiency.
Example 4:
the composting humus microbial agent and the method for degrading TBBPA and the application thereof provided by the embodiment of the invention are applied to in-situ ecological restoration of a TBBPA polluted muddy water mixture on the basis of the embodiments 1-3, and the Y17 strain microbial restoration and ecological greening plants are adopted to restore the TBBPA polluted muddy water mixture together so as to accelerate the restoration speed of the muddy water mixture, improve the restoration effect, obtain better and longer-acting ecological greening effect and are suitable for ecological purification treatment engineering of polluted wetland and river bottom mud.
The embodiment focuses on the ecological treatment of the soil (wetland) with high water content for degrading TBBPA pollution or the bottom mud of the water body, namely, the double ecological treatment of the mud and water mixture. The mud-water mixture is one of the most difficult fields to treat, but by adopting the technical scheme of the invention, the synchronous treatment of mud and water can be conveniently realized.
The technical scheme of the embodiment is basically the same as that of the embodiments 1-3, and is characterized in that the method adopts litchi branches generated by trimming or updating, and comprises the following specific steps:
s11: preparing wood chips: preparing waste litchi branches into wood chip particles with the particle size of 0.5-2mm, pretreating the wood chip particles by selecting a high-temperature treatment mode, and then air-drying the pretreated wood chip particles until the water content is not more than 20%, so as to prepare pretreated wood chips for later use;
the high-temperature pretreatment method comprises the following steps: mixing litchi sawdust particles with water according to a ratio of 1:10, and carrying out high-temperature treatment for 2 hours at 110 ℃; repeatedly washing the litchi sawdust particles in the solution to be neutral, and washing off the surface substances of the litchi sawdust particles; and finally, air-drying the fruit wood chip particles until the water content is about 20% for subsequent composting.
And then carrying out static composting on the compost for 30-40 days to obtain humus.
The microbial agent prepared from the compost humus is used for degrading TBBPA, the microbial degradation microbial agent is sowed into a TBBPA polluted muddy water mixture, and the microbial restoration and the plant restoration of the muddy water mixture are synchronously carried out in situ under the combined action of the Y17 strain and the humins, so that the method can be widely applied to ecological management projects such as artificial green land ecological management, river sediment ecological management and the like of TBBPA pollution, such as in-situ restoration of old site soil, water body, wetland, sediment and the like of chemical plants.
Comparative example
For comparison of the verification effect, the following comparative examples of 5 different stack material ratios were designed.
Pure cow dung was individually composted as comparative example 1; and mixing and composting the non-pretreated apple wood chip particles (i.e. the apple wood chip particles which are not subjected to acidification, alkalization and high temperature pretreatment) in a box according to the dry matter mass ratio of cow dung to the apple wood chip particles of 2:1, and obtaining comparative examples 2-5.
Namely: comparative example 1 is pure cow dung compost, comparative example 2 is cow dung: apple chip particles = 2:1 compost; comparative example 3 is cow dung: acidizing apple wood chip particles = 2:1; comparative example 4 is cow dung: alkalization apple wood chip particles = 2:1; comparative example 5 is cow dung: high temperature treatment of apple chip particles = 2:1. all the composting is carried out according to the same conditions and process, and the composting is carried out manually once every 10 days.
The temperature of the compost for each of the same comparative examples (including different apple chip particle pretreatments) is shown in figure 5.
The lignocellulose in the fruit sawdust particles in the above embodiment of the present invention mainly comprises cellulose, hemicellulose and lignin. Taking the example of the apple wood chip particles, a large amount of hemicellulose can be removed in the pretreatment process, wherein the concentration of hemicellulose in the compost after the alkalization treatment of the apple wood chip particles is mixed is at least 330.39 mg/g in the initial stage. Compared with the mixed composting of the apple sawdust particles, the composting of the acidized apple sawdust particles and the composting of the apple sawdust particles treated at high temperature are respectively reduced by 18.0 percent, 7.7 percent and 22.3 percent. The alkalization treatment is more remarkable in hemicellulose removal capacity.
In the embodiment of the invention, a large amount of lignin can be removed from the apple sawdust particles in the pretreatment process, wherein the lignin concentration of the compost after the mixing of the alkalization-treated apple sawdust particles in the initial stage is at least 184.92 mg/g. Compared with the mixed composting of the apple sawdust particles, the composting of the acidized apple sawdust particles and the composting of the apple sawdust particles treated at high temperature are respectively 8.8 percent, 20.4 percent and 9.3 percent lower. The alkalization treatment is more remarkable in lignin removal ability.
In the composting process, the cellulose of the apple chip particles shows a decreasing trend. Because of the excellent removal of hemicellulose and lignin by the alkalization treatment, there is a higher concentration of cellulose in the mass of the mass mixed by the alkalized apple chip particles, but more cellulose is exposed, while the crystallinity of cellulose is reduced, while the pore structure is increased, creating favorable conditions for subsequent carriers as a microbial inoculum. After composting, the concentration of cellulose is the lowest, and only 220.31 g/kg, the removal rate reaches 53.24%, so that the structure of the separated humins is stable.
The alkalization treatment has remarkable removal effect on the lignin of the apple sawdust particles, so that the connection between cellulose and lignin is weakened, the crystallinity of the cellulose can be reduced, and the removal of hemicellulose exposes more cellulose, and meanwhile, the alkalization treatment increases the pore structure, is favorable for the attachment of cellulose and improves the accessibility of enzymes to the cellulose. The different pretreated apple chip particles were mixed to compost for lignocellulose content as shown in table 2.
TABLE 2
In the composting process of the pile body of the embodiment, the content of Humus (HA) is firstly reduced and then increased, except that the HS concentration of the pure cow dung compost is reduced by 3.68%, the rest of compost is increased by 14.20%, 27.4%, 22.09% and 15.41%, and the contents of the compost reach 158.92 mg/g, 152.51 mg/g, 157.86 mg/g, 145.12 mg/g and 153.93mg/g respectively, which indicates that the yield of humus can be increased finally by pretreatment.
The content of HA in the early stage is reduced mainly because the activity of microorganisms in the early stage of composting is higher, and part of humic acid is decomposed by microorganisms. After composting, the HA content of the pure cow dung is only increased by 1.05 percent, which is far lower than that of a pile body added with apple sawdust particles, which indicates that the addition of the apple sawdust particles can promote the HA production in the composting process, the alkalization treatment is more beneficial to the synthesis of humic acid, and the concentration of the humic acid after the alkalization treatment is up to 105.23 mg/g. The alkalization treatment HAs stronger capacity of promoting the generation of humus in the composting process, and more HA is formed.
All treatments continued to decrease in Fulvic Acid (FA) content. FA is a component with smaller molecular weight in humus, is easy to biodegrade and has higher activity, and can be used as a carbon source for microbial metabolism to produce humic acid with more stable property in the composting process. All compost FA was reduced by 19.61-36.28%, with an alkalization reduction of 22.69%, but since the initial FA concentration of the alkalized apple mixed heap was only 40.13mg/g at the minimum, the FA concentration was also the lowest after the end of composting, only 31.02 mg/g.
FA consists of organic compounds that are readily biodegradable and are readily converted to stable HA. FA can be used as a carbon source for microbial metabolism, producing more stable Humic Acid (HA) during composting. Thus, the ratio of Hu Fu (HA/FA) (i.e., degree of Polymerization (DP)) for all treatments continues to increase during composting. Compared with pure cow dung treatment, the pretreated agricultural and forestry waste has a higher DP value at the end of composting, wherein the highest polymerization Degree (DP) of 3.39 is obtained by alkalization treatment in all pretreatment, the polymerization degree of the alkalization treatment is increased from 1.97 to 3.39 by 71.63%, the humification degree is enhanced, and the pore structure of the humins is increased and stabilized.
Therefore, the alkalization pretreatment adopted in the embodiment of the invention can obviously enhance the humification process of the composting process, thereby promoting the generation of humus, accelerating the formation of HA, improving the polymerization Degree (DP), increasing and stabilizing the pore structure of the humins and ensuring that the prepared composting humus microbial agent HAs better physicochemical properties. The content of the mixed composting humus of the different pretreated apple wood chip particles is shown in table 3.
TABLE 3 Table 3
The three-dimensional fluorescence diagram of the humus after the compost is characterized by three-dimensional fluorescence is divided into I (tyrosine class), II (tryptophan class), III (fulvic acid), IV (soluble microorganism metabolite) and V (humic acid). As composting proceeds, the intensity of the I and II areas is reduced, which shows that the aromatic protein substances with fragile structure and high bioavailability are obviously degraded. V (humic acid) showed higher concentration after alkalization treatment than other treatments, and the same iii (fulvic acid) was also shown to be the lowest. This also shows that the pile body after mixing and alkalizing the apple sawdust particles generates more humic acid after 40 days of fermentation, improves the polymerization degree and has more stable humus structure. The specific three-dimensional fluorescence diagram of the mixed composting humus of the apple tree treated in the same way is shown in figure 6.
In summary, in the embodiment of the invention, the apple sawdust particles and 0.5mol/L NaOH are mixed according to the mass ratio of 1:10, and are soaked for 12 hours at 25 ℃ for alkalization treatment, and the mass of the treated apple sawdust particles is 2:1 is mixed as a pile body, and the composting effect is best when composting is carried out for 40 days. Of course, on the basis of the method, better composting effect can be obtained by changing composting process conditions.
The three-dimensional fluorescence picture of the humus material prepared by the pretreatment of the wood chip particles is shown in figure 6.
As can be seen from the various results of the above examples of the present invention, a proper mixing ratio can improve the decomposition capacity of the pile; the smaller particle size of the apple sawdust is beneficial to improving the surface area and accessibility of microorganisms; different pretreatment can make the surface of the apple sawdust particles coarser, and show more pores, so that the surface area of the apple sawdust particles is increased, and the adhesion of microorganisms on the surface of the apple sawdust particles is facilitated; and the crystallinity index (CrI value) of the waste can be obviously reduced by different pretreatment, lignin and hemicellulose are effectively removed to expose cellulose, and then the cellulose is promoted to be converted into an amorphous structure; functional group analysis shows that the pretreatment process can significantly change the functional groups of the apple wood chip particles.
The prepared humus material is separated and respectively prepared into the humins, the humic acid and the fulvic acid material, and the three materials are respectively applied to different ecological treatment steps so as to exert different functions to the greatest extent. Wherein the humins are used for preparing compost humus microbial inoculum, and the specific surface area of the humins can reach 5.756 m 2 The strain Y17 is cultured for 12 hours at the temperature of 35 ℃ and the pH value of 6, and then is mixed with humins as a carrier for immobilization, the immobilization culture temperature is 35 ℃, the immobilization culture pH value is 7, and the charcoal immobilization culture time is 12 hours, so that the suspension type microbial inoculum is prepared; the Y17 strain is planted on the surface of the humins and forms a liquid TBBPA microbial degradation microbial agent in the pore canal; humic acid and richThe linac acid is added in a proper proportion and at a proper timing as an accelerating component for promoting the growth of microorganisms and plants.
According to the test surface of the embodiment of the invention, the liquid microbial degradation microbial agent is put into a water body containing TBBPA according to the concentration of 0.1g/L, and the humins provide the growth conditions and the microenvironment of aerobic growth in the water body for the Y17 strain so as to degrade the TBBPA in the water body. Compared with free Y17 strain, the microbial inoculum has 42.2 percent higher degradation rate of TBBPA. The microbial inoculum can be continuously added with 50 mg/L in the degradation process, so that the degradation rate of TBBPA of the microbial inoculum can be continuously improved by 31.5% and 28.7% respectively.
After composting, each kilogram of non-separated germ material (or separated humins) is mixed with 1g of Y17 strain and then applied to soil, and the organic matter content of the soil is improved, so that the growth of the Y17 strain can be rapidly expanded, the degradation of TBBPA can be promoted, the growth of higher plants can be promoted, and the effects of ecological management, ecological beautification and the like can be jointly realized.
In other embodiments, the fruit tree branches, or woody waste fruit tree branches of other woody fruit trees with hard woody fibers, such as peach, pear, jujube, walnut, chestnut, plum, etc., may be specifically selected according to practical situations.
According to the various technical schemes provided by the embodiment of the invention, the composting humus microbial agent can degrade TBBPA in soil, water and muddy water mixtures through biological amplification, so that the degradation efficiency is high, the degradation is less influenced by the environment, and the composting humus microbial agent can meet the popularization and application requirements of large range and low cost due to the adoption of double bioremediation, stable repairing effect, environmental friendliness and low cost.
The invention breeds dominant strain Y17 grown in soil, and the strain is domesticated to be used as a microorganism for degrading TBBPA in soil, water and muddy water mixture; after the strain is domesticated, the environmental adaptability, TBBPA degradation capability and stress resistance of the strain are obviously improved. In order to enable the Y17 strain to grow in a water body, the strain is planted on humins to prepare a special liquid microbial degradation microbial agent, and the microbial agent is used for creating a microscopic environment in soil, water body and sediment, so that the strain can grow; through continuous aerobic growth and breeding of Y17, TBBPA is continuously degraded, and degradation products thereof and other components of humus provide nutrition for the growth of plants (higher plants), so that double ecological treatment is realized, the degradation effect is enhanced, the degradation time is shortened, and the method is suitable for various pollutants and various ecological treatment scenes.
The method improves the effect of removing the pollutant TBBPA in the water body, simultaneously aims at reducing the total consumption and the sowing times of a material system, improves the adaptability of the strain and the microbial inoculum to the water body environment, greatly reduces the sewage treatment cost and is convenient for large-area popularization. Through practical tests, the method can realize effective removal of TBBPA in the water body after one-time sowing and 7-8 days of degradation, and reaches the set standard; compared with the traditional technology, the overall water treatment scheme of the invention can save more than 70% of total cost and can improve more than 30% of degradation efficiency.
In other embodiments of the present invention, the technical effects described in the present invention may be achieved by performing specific selection of other different schemes within the ranges of the preparation and application steps, components, proportions and process parameters of the composting humus microbial agent described in the present invention, so the present invention is not listed one by one.
The above description is only of the preferred embodiment of the present invention, and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solution of the present invention or modifications to equivalent embodiments using the methods and technical contents disclosed above, without departing from the scope of the technical solution of the present invention. All equivalent changes of the components, proportions and processes according to the invention are covered in the protection scope of the invention.
Claims (4)
1. The composting humus microbial agent is characterized by being prepared by the following steps:
s1: preparing wood chips and fresh cow dung: preparing waste fruit tree branches into wood chip particles, pretreating the wood chip particles in an alkalization mode, and then air-drying the pretreated wood chips until the water content is not more than 20%, so as to obtain pretreated wood chips for later use; collecting fresh cow dung in a farm, so that the water content of the cow dung is not higher than 70%, and obtaining the fresh cow dung for later use;
the pretreatment step by adopting an alkalization mode comprises the following steps: mixing the apple sawdust particles with 0.5mol/L NaOH according to a mass ratio of 1:10, and soaking for 12 hours at 25 ℃; repeatedly washing the apple wood chip particles in the solution to neutrality, washing off NaOH on the surfaces of the apple wood chip particles to obviously reduce the crystallinity index of the apple wood chip particles, obviously changing apple wood functional groups, removing lignin and hemicellulose to expose cellulose, and promoting the cellulose to be converted into an amorphous structure; the method specifically comprises the following steps:
s11: preparing wood chips: preparing waste fruit tree branches into wood chip particles with the particle size of 0.5-2mm, pretreating the wood chip particles in an alkalizing mode, and then air-drying the pretreated wood chip until the water content is not more than 20%, so as to prepare pretreated wood chip for later use;
S12: fresh cow dung:
reducing the moisture content of fresh cow dung collected in a cow farm through a solid-liquid separator to ensure that the moisture content is not higher than 70%, and preparing the fresh cow dung for later use;
s2: preparing mixed humus by composting: uniformly mixing the standby pretreated wood chips and fresh cow dung according to a set weight ratio to obtain a pile; controlling the pile body, composting, and obtaining mixed humus with a plurality of components mixed with each other after the pile body is continued for a set time; the method specifically comprises the following steps:
s21: preparing a pile: uniformly mixing the standby pretreated wood chips and fresh cow dung according to the following weight proportion: fresh cow dung = 1: (1-2) obtaining a pile body;
s22: preparing mixed humus: the composting conditions of the pile body are controlled, static composting is carried out, the pile turning frequency is 10 days/time, and the composting time is set to 20-40 days, so that mixed humus with a plurality of components mixed with each other is obtained;
s3: separating and mixing humus: separating soluble fulvic acid, humic acid and insoluble humins from the mixed humus, and respectively collecting and storing the fulvic acid, the humic acid and the humins with rough surfaces, rich pores and stable structure; the method specifically comprises the following steps:
S31: separating and mixing humus: separating soluble fulvic acid and humic acid from the filtrate by an alkali dissolution and acid precipitation method, separating insoluble humic acid from the filter residue, and then respectively collecting and storing the fulvic acid, the humic acid and the humic acid, wherein the humic acid has rough surface, rich pores and stable physical three-dimensional space structure;
s32: purifying fulvic acid, humic acid and humin respectively for later use;
s4: preparing Y17 bacterial liquid: adding a nutrition culture medium into the Y17 strain for rejuvenation and expansion culture to obtain Y17 bacterial liquid; the method specifically comprises the following steps:
s41: preparing Y17 bacterial liquid, specifically adding a Y17 bacterial strain into a nutrient medium for rejuvenation and expansion culture: rotating at 35 ℃ and 160r/min, and performing expansion culture for 12 hours to obtain Y17 bacterial liquid; the nutrient medium is as follows: adding 3g of beef extract, 10g of peptone and 5g of NaCl into pure water, fixing the volume to 1000mL, and adjusting the pH to 7.2;
s5: preparing a microbial degradation microbial agent: adding Y17 bacterial liquid into the humins separated in the step S3 according to a set proportion, and performing field planting and culture for a set time by taking the humins as carriers, so that the Y17 bacterial strain is planted on the rough surface and rich pore structures of the humins to prepare the microbial degradation microbial agent, wherein the microbial degradation microbial agent specifically comprises the following components:
S51: preparing a microbial degradation microbial agent: and (3) adding the Y17 bacterial liquid with the OD value of 0.45 prepared in the step (S4) into the nutrient medium containing the humins of 1 g/L separated in the step (S3) according to the addition amount of 5% by volume, performing field planting and culture for at least 12 hours at the normal temperature with the rotating speed of 160r/min, and using the humins as carriers to perform field planting on the Y17 bacterial strain on the rough surface and rich pore structures of the humins to prepare the Y17 microbial degradation bacterial agent fixed by the humins.
2. The use of the composting humic substance microbial agent according to claim 1 for degrading TBBPA, which comprises the following steps:
s6: degrading organic pollutants: according to the set proportion, the microbial degradation microbial inoculum is sown into soil or water containing organic pollutant TBBPA, and oxygen contained in the humins and pore structures thereof provides growth conditions and microenvironment for Y17 strain to aerobically grow in the soil or water so as to adsorb and degrade the organic pollutant TBBPA in the soil or water.
3. The use of a composting humic substance microbial agent according to claim 2 wherein it further comprises the steps of:
s7: growth of synergistic plants: the fulvic acid and the humin acid solution with required concentrations are synchronously added into the soil or the water body containing the organic pollutant TBBPA, so that the growth of the Y17 strain is promoted, the degradation speed of the TBBPA is accelerated, and the degraded components and the components which do not participate in the degradation can provide organic nutrient substances for plants growing in the soil or the water body, so that the plant growth and ecological restoration speed are accelerated.
4. The application of the composting humic substance microbial inoculant according to claim 2, wherein the step S6 comprises the following steps:
s61: degrading organic pollutants: according to the concentration of TBBPA in soil or water to be degraded and the proportion of microbial degradation bacteria, the microbial degradation bacteria are spread in the soil or water containing organic pollutant TBBPA, so that the microbial degradation bacteria are uniformly distributed in the soil or water, and the oxygen contained in the humins and the pore structures thereof provides the Y17 strain with the growth conditions and the microenvironment for aerobic growth in the soil or water so as to adsorb and degrade the organic pollutant TBBPA in the soil or water.
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