CN117402795B - Composite microbial inoculum and application thereof in aerobic composting and plastic degradation - Google Patents
Composite microbial inoculum and application thereof in aerobic composting and plastic degradation Download PDFInfo
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- CN117402795B CN117402795B CN202311713607.XA CN202311713607A CN117402795B CN 117402795 B CN117402795 B CN 117402795B CN 202311713607 A CN202311713607 A CN 202311713607A CN 117402795 B CN117402795 B CN 117402795B
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- composting
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- aerobic composting
- microbial inoculum
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
- B09B3/60—Biochemical treatment, e.g. by using enzymes
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- 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
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- 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
- C05F17/70—Controlling the treatment in response to process parameters
-
- 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
-
- C—CHEMISTRY; METALLURGY
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE
- B09B2101/00—Type of solid waste
- B09B2101/75—Plastic waste
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE
- B09B2101/00—Type of solid waste
- B09B2101/75—Plastic waste
- B09B2101/78—Plastic waste containing foamed plastics, e.g. polystyrol
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
-
- 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
Abstract
The invention relates to the technical field of aerobic composting, in particular to a composite microbial inoculum and application thereof in aerobic composting and plastic degradation. The composite microbial inoculum provided by the invention has a certain decomposing capacity on cellulose; in addition, the single strain has great degradation effect on specific microplastic, and has strong pertinence, strong decomposing ability of bacillus subtilis to PET and strong decomposing ability of bacillus stearothermophilus to PS. Compared with the conventional microbial agent, the thermophilic microbial agent is added in the aerobic composting process, so that the high-temperature period of the compost can be prolonged, the phytotoxicity is reduced, the humification process is promoted, the microbial community structure is optimized, and the quality and the efficiency of the compost are improved. The interaction of the mixed microbial inoculum enables cellulose and protease to be rapidly decomposed, so that the decomposition rate is improved, and the degradation rate is accelerated through the change of the structure of Microplastic (MPs).
Description
Technical Field
The invention relates to the technical field of aerobic composting, in particular to a composite microbial inoculum and application thereof in aerobic composting and plastic degradation.
Background
The agricultural waste refers to organic matters discarded in the whole agricultural production process, and mainly is biomass residues generated in agricultural, forest, pasturing and fishery production processes, including crop straws, livestock and poultry manure, vegetable tails, dead livestock and poultry and the like, wherein the maximum production amount of the crop straws and the livestock and poultry manure is also most common.
At present, aerobic composting is a main means for treating agricultural wastes, but the traditional aerobic composting method has the problems of low composting quality, low humification degree and incapability of degrading Microplastic (MPs).
Disclosure of Invention
In order to solve the problems, the invention provides a composite microbial inoculum and application thereof in aerobic composting and plastic degradation. The composite microbial inoculum provided by the invention can not only improve the degradation rate of the micro-plastics, but also optimize the microbial community structure, effectively prolong the high-temperature period of the compost, reduce the phytotoxicity, promote the humification process and improve the quality and efficiency of the compost.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a composite microbial inoculant which comprises the following components in parts by volume: 10-15 parts of bacillus subtilis (Bacillus subtilis), 10-30 parts of olive soil bacteria (Olivibacter soli) and 10-45 parts of geobacillus stearothermophilus (Geobacillus stearothermophilus); the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253.
Preferably, the volume ratio of the bacillus subtilis, the olivetobacter soil and the geobacillus stearothermophilus is 1:2:1.
the invention provides application of the composite microbial inoculum in one or more of the following aspects, which comprises the following steps:
1) The efficiency of aerobic composting is improved; 2) Promoting degradation of carbohydrates in the compost raw material; 3) Promoting degradation of lignocellulose in the compost raw material; 4) Inhibiting and/or killing pathogenic bacteria in the compost raw material; 5) Degrading the plastic; the plastic comprises one or more of polyethylene, polystyrene, and polyethylene terephthalate.
The invention provides an aerobic composting method for degrading plastics, which comprises the following steps:
mixing the composite microbial inoculum and the composting raw material in the technical scheme and performing aerobic composting.
Preferably, the composting raw material and the composite microbial inoculum are pretreated before being mixed; the pretreatment comprises the following steps: and heating the compost raw material to 70-80 ℃, maintaining the temperature for 60-80 min, and cooling to 35-40 ℃.
Preferably, the water content of the compost raw material before heating is 60% -70%.
Preferably, the aerobic composting method comprises the following steps: after aerobic fermentation for 15-20 d, adding an initiator to perform secondary aerobic fermentation; the initiator comprises diisopropyl peroxydicarbonate.
Preferably, the method of the second aerobic fermentation comprises the following steps: and (3) adding an initiator, heating to 56-60 ℃, maintaining for 2-3 days, naturally cooling, and decomposing and fermenting for 8-10 days.
Preferably, the aerobic composting is preceded by mixing a mixture of the composite microbial inoculum and the composting raw material with biochar.
Preferably, the mass ratio of the composite microbial inoculum to the compost raw material is 1: 100-120.
The beneficial effects are that:
the invention provides a composite microbial inoculant which comprises the following components in parts by volume: 10-15 parts of bacillus subtilis, 10-30 parts of olive shaped soil bacteria and 10-45 parts of geobacillus stearothermophilus; the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253. The composite microbial inoculum provided by the invention comprises normal temperature and thermophilic bacteria, and is active in the main composting fermentation stages at the temperature rising and the high temperature respectively, so that the microbial decomposition and degradation capacity of the stage reaches the highest efficiency; all three strains have better decomposing ability on lignocellulose which is a main component difficult to decompose in compost raw materials; in addition, the single strain has great degradation effect on specific microplastic, and has strong pertinence, strong decomposing ability of bacillus subtilis to PET and strong decomposing ability of bacillus stearothermophilus to PS. Compared with the conventional microbial agent, the thermophilic microbial agent is added in the aerobic composting process, so that the high-temperature period of the compost can be prolonged, the phytotoxicity is reduced, the humification process is promoted, the microbial community structure is optimized, and the quality and the efficiency of the compost are improved. The interaction of the mixed microbial inoculum enables cellulose and protease to be rapidly decomposed, so that the decomposition rate is improved, and the degradation rate is accelerated through the change of MPs structure.
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 required to be used in the embodiments will be briefly described below.
FIG. 1 illustrates the moisture step-by-step characteristics of stacks under different treatments;
FIG. 2 is a graph of composition and relative abundance of bacterial populations falling at the gate level during composting;
FIG. 3 shows the effect of different treatments on Hu Mingsuan during composting;
FIG. 4 shows the effect of different treatments on fulvic acid during composting.
Detailed Description
The invention provides a composite microbial inoculant which comprises the following components in parts by volume: 10-15 parts of bacillus subtilis, 10-30 parts of olive shaped soil bacteria and 10-45 parts of geobacillus stearothermophilus; the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253.
In the present invention, the volume ratio of the bacillus subtilis, the olivetobacter soil and the geobacillus stearothermophilus is preferably 1:1:1 or 1:2:1 or 1:2:3, more preferably 1:2:1. the volume parts of the composite microbial inoculum are preferably mixed according to the volume ratio after three strains are directly purchased.
The composite microbial inoculum comprises 10-15 parts by volume of bacillus subtilis, preferably 10 parts by volume. The bacillus subtilis is one of bacillus, is purchased from China general microbiological culture collection center, has a strain number of CGMCC 1.3358, has an active temperature of 15-85 ℃, and also has viable spores in polar environments such as high temperature, acid and alkali, and the like, so that the bacillus is suitable for the environment to survive; once the environment becomes suitable for growth and the nutrition is sufficient, the spores automatically enter the reproductive phase, and the spores regrow into bacillus subtilis; can secrete high-activity amylolytic enzyme, protein lytic enzyme, lipolytic enzyme, cellulolytic enzyme and other types of lytic enzymes, has strong organic substance decomposing power, and can rapidly improve the temperature and the speed of composting; active substances such as subtilisin, polymyxin, nystatin, gramicidin and the like can be produced, and the inhibition effect on soil pathogenic bacteria is strong; the microbial enzymes produced by the bacillus subtilis are released from the polyolefin plastic, so that the growth and propagation of environmental indigenous bacteria on the plastic product body are induced, the accelerated aging of MPs is caused, and the degradation purpose is finally achieved.
Based on the volume parts of the bacillus subtilis, the composite microbial inoculum comprises 10-30 parts, preferably 10-20 parts, and more preferably 20 parts of olive shaped soil bacteria. The soil olivary fungus (strain 18B) is purchased from China center for type culture collection of agricultural microorganisms, and the strain number is ACCC 02948. The strain 18B can grow in the range of 12-48 ℃, the optimal growth temperature is 30-37 ℃, at this time, thermophilic microorganisms become dominant bacterial groups, residual or newly generated soluble organic matters in compost are continuously decomposed, and complex organic matters such as hemicellulose, cellulose and proteins are also strongly decomposed; strain 18B has genes capable of synthesizing cellulases, mainly endoglucanases and beta-glucanases, the endoglucanases mainly act on the amorphous region of cellulose, new ends are generated by random cutting, the products are oligosaccharides with indefinite length, and the beta-glucosidase is more prone to act on the non-reducing ends, and cellodextrin and cellobiose are hydrolyzed to generate glucose; the cellulose such as straw in the pile is rich, the cellulose is mainly derived from cellulose degradation strains, and the cellulose degradation capability of the strain is stronger.
Based on the volume parts of the bacillus subtilis, the composite microbial inoculum comprises 10-45 parts, preferably 10-30 parts, and more preferably 10 parts of geobacillus stearothermophilus. The Geobacillus stearothermophilus disclosed by the invention is purchased from China center for type culture collection of agricultural microorganisms, and the strain number is ACCC 10253. In the process of treating the microplastic, the thermophilic geobacillus stearothermophilus firstly promotes the C-H and C=C structures on the benzene ring of the microplastic to be depolymerized or broken under the action of microorganisms, changes the C-H and C=C structures into low molecular mass substances, and then increases the C-O bond, C=O bond, C-OH and other oxygen-containing structures on the surface of the microplastic through biological oxidation, so that the hydrophilicity of the surface of the MPs is improved, the microbial colonization is facilitated, and the biodegradability of the MPs is improved. The geobacillus stearothermophilus can grow by taking MPs as a carbon source, and forms a biological film with higher activity on the surface of the MPs, and free oxygen is provided for other microorganisms, so that the activity and diversity of the microorganisms are improved; polystyrene (PS) belongs to a substance difficult to degrade, and the Geobacillus stearothermophilus provided by the invention has a strong effect on the structure of PS, and the degradation rate of PS reaches 4.2%.
The invention provides application of the composite microbial inoculum in one or more of the following aspects, which comprises the following steps:
1) The efficiency of aerobic composting is improved; 2) Promoting degradation of carbohydrates in the compost raw material; 3) Promoting degradation of lignocellulose in the compost raw material; 4) Inhibiting and/or killing pathogenic bacteria in the compost raw material; 5) A degradable plastic comprising one or more of Polyethylene (PE), polystyrene, and polyethylene terephthalate (PET).
The invention provides an aerobic composting method for degrading plastics, which comprises the following steps:
mixing the composite microbial inoculum and the composting raw material in the technical scheme and performing aerobic composting.
The aerobic composting method is preferably carried out in composting equipment, and the composting equipment is preferably purchased from Galaxy mechanical and electrical appliances limited company in New county, model: HYJF; the composting equipment comprises a composting bin, a heating and heat preserving system, a stirring system and an aeration system; the outside of the composting bin is provided with an insulating layer and a heating layer which are used for heating and insulating materials in the bin; a stirrer is arranged in the bin, the stirring speed is 1-2 r/min, the stirring mode is that the stirring is operated for 2min at intervals of 8min, the forward and reverse rotation is carried out alternately, and the stirring is adjusted to be operated for 1min at intervals of 14min after the stirring is operated for 40 h; the working time of a fan of the aeration system is 50s in every 130s of the compost heating period, 50s in every 80s of the high temperature period, and the air in the top space of the bin can be completely discharged in 50s, so that 1 ventilation is completed.
The invention preferably crushes and mixes the livestock manure and the straw agricultural waste to obtain the mixed material. In the invention, the mass ratio of the livestock manure to the dry matter of the straw agricultural waste is preferably 1-2: 2-3; the livestock manure preferably comprises one or more of pig manure, cow manure, horse manure and chicken manure; the straw agricultural waste preferably comprises one or more of corn straw, wheat straw, rice straw, cotton straw, vegetable tails and fruit tree pruning; the carbon-nitrogen ratio of the mixed material is preferably 25-35: 1, more preferably 25:1.
after the mixed material is obtained, the mixed material is preferably subjected to pretreatment (thermophilic pretreatment) to obtain a compost raw material; the pretreatment preferably includes: and (3) adjusting the water content of the mixed material to 60% -70%, heating to 70-80 ℃, maintaining the temperature for 60-80 min, and cooling to 35-40 ℃. In the invention, the water content of the mixture is preferably 70%; the temperature of the pretreatment is preferably 80℃and the holding time is preferably 80 minutes.
The invention promotes the transformation of high binding energy form water in the pile body to low binding energy form water (the transformation of bound water to free water) through thermophilic pretreatment, is beneficial to the evaporation of water, enhances the air permeability of materials and avoids anaerobic fermentation; meanwhile, bacteria are rapidly propagated and metabolized vigorously, so that the time for entering compost can be shortened. Thermophilic pretreatment can destroy the MPs structure through rapid temperature rise, promote the cell lysis of the primary microorganism and release the intracellular and extracellular polymer substances (protein, polysaccharide and nucleic acid) from the cells, accelerate the degradation of the large polymer structure (polyethylene and polyethylene terephthalate) of the MPs into oligomers, dimers and monomers (ethylene, ethylene glycol and terephthalic acid), and improve the depolymerization effect of the MPs. The high-concentration free ammonia generated in the composting process can inhibit the growth of nitrifying bacteria, and the substrate is insufficient, so that the nitrogen preservation of the compost is facilitated; the reducing sugar and total sugar produced after pretreatment of the material are increased, the molecular weight and polymerization degree of humus are increased in the composting process, and the humification level of the composting product is improved. In addition, mineral elements beneficial to plants can be effectively preserved in solid products (excluding Na) under thermophilic pretreatment conditions, while NH 3 、SO 2 、H 2 Harmful substances such as S, organic amine compounds and the like can be volatilized and discharged as gas in the process, and the method plays an important role in safe fertilizer utilization of solid products after composting.
When the temperature of the pretreated material is reduced to below 40 ℃, preferably, the biochar and the composite microbial inoculum according to the technical scheme are added into the compost raw material, and the mixture is uniformly mixed to obtain the fermentation material. In the invention, the mass ratio of the composite microbial inoculum to the compost raw material is preferably 1:100 to 120, more preferably 1:120; the mass ratio of the biochar to the compost raw material is preferably 5-510:100, more preferably 8:100; the biochar preferably comprises one or more of straw biochar, rice husk biochar, bagasse biochar and pine needle biochar. The porous structure of the biochar provides a good habitat for the growth of microorganisms, and the strong water-holding capacity and rich nutrient substances of a solid-liquid interface are also beneficial to the proliferation of the microorganisms, so that the high-temperature period is advanced by 2-3 days, the composting process is remarkably accelerated, and the composting quality is improved; the biochar can promote the conversion of nitrogen and reduce the volatilization of ammonia, on one hand, the addition of the biochar obviously increases the content of nitrate nitrogen in the composting process, reduces the content of ammonium nitrogen, is beneficial to the conversion of ammonium nitrogen into nitrate nitrogen, has good adsorptivity to ammonia, and reduces the accumulated volatilization of ammonia of a heap; on the other hand, the carbon-nitrogen ratio of the compost material is inversely related to the ammonia emission, the lower the carbon-nitrogen ratio is, the more serious the ammonia volatilizes, and the biochar is added during composting, so that the carbon-nitrogen ratio is improved, and the ammonia emission is reduced. Due to the composting process NH 3 Volatilization is a main way of nitrogen loss of the compost, and accounts for 46.8% -77.4% of the total nitrogen loss, and the nitrogen fixation capacity can be further improved by adding biochar. In addition, the addition of biochar can promote the conversion of fulvic acid in the water-soluble organic matter component into humic acid, thereby improving the stability of humus.
After the fermentation material is obtained, the invention preferably carries out aerobic fermentation, namely first composting fermentation, on the fermentation material. In the invention, the time of the first composting fermentation is preferably 15-20 d, more preferably 17d. At the initial stage of primary composting fermentation and composting, no spore bacteria, mold and other normal temperature bacteria are dominant at this stage, mesophilic microorganisms utilize soluble organic matters in the compost to carry out vigorous reproduction, and decompose organic matters (such as simple sugar, starch, protein and the like) which are easy to decompose, and the temperature at this stage is below 45 ℃ and the fermentation time is within 12 hours.
The soil olivary bacteria (strain 18B) are novel cellulose degrading bacteria, grow at 12-48 ℃, have genes capable of synthesizing cellulase, mainly endoglucose and beta-glucanase, and can efficiently degrade cellulose and hemicellulose in a pile, shorten degradation time and improve degradation efficiency. Extracellular cellulase produced by the bacterium can also act on MPs to accelerate degradation.
The active temperature of the bacillus subtilis is 15-85 ℃, and the bacillus subtilis has high-activity amylolytic enzyme, protein lytic enzyme, lipolytic enzyme, cellulolytic enzyme and other types of lytic enzyme, has strong organic substance decomposing capability, and has good degrading capability on various plastics such as PS (bacillus degrading bacteria), PET (lipase degrading), polyurethane (PUR, lipase and protease degrading) and the like. Because the spores can survive in polar environments such as high temperature, acid and alkali, and the like, the environment adaptability is strong. Can decompose organic substances with maximum efficiency. In addition, microbial enzymes produced by bacillus are convenient to release from polyolefin plastics, so that growth and propagation of environmental indigenous bacteria on the plastic product body are induced, and accelerated aging of MPs is promoted.
The thermophilic geobacillus stearothermophilus can grow by taking MPs as a carbon source and provide free oxygen for other microorganisms, so that the key problem of low decomposition efficiency caused by insufficient oxygen and reduced activity of a large number of strains in a high-temperature stage is solved, and the activity and diversity of the microorganisms are improved. PS belongs to a refractory substance, and the Geobacillus stearothermophilus has a strong effect on the structure of PS, and the degradation rate of PS reaches 4.2 percent.
As the temperature of the pile rises, the high temperature stage of the compost is entered when the temperature is higher than 45 ℃. Thermophilic fungi and actinomycetes (protozoon and artificially added thermophilic geobacillus stearothermophilus) are propagated in a large quantity, and organic matters such as cellulose, hemicellulose and pectic substances are decomposed and converted, and the high temperature is maintained at 50-65 ℃. The process can be reasonably stirred to avoid overhigh composting temperature, and the composting temperature is about 5-7 d.
Bacillus stearothermophilus can grow by using MPs as a carbon source, form a biological film with higher activity on the surface of WPs, provide free oxygen for other microorganisms, and improve the activity and diversity of microorganisms.
In the process of treating the microplastic, oxidation, dehydrogenation and C=C structural fracture on benzene rings of the microplastic (PS, PE) are firstly promoted to be depolymerized or broken due to the action of microorganisms (primordium and artificially added bacteria), the benzene rings are changed into low molecular mass substances, and then the oxygen-containing structures such as C-O bonds, C=O bonds and C-OH on the surface of the microplastic are increased through biological oxidation, so that the hydrophilicity of the surface of the MPs is improved, the benzene rings are more beneficial to the colonization of the microorganisms, and the degradation of the microorganisms (primordium and degrading bacteria) is improved.
When most of the organic matters are degraded and the temperature starts to be reduced gradually to the room temperature (20-25 ℃), the invention preferably adds 0.1-0.3 wt.% of diisopropyl peroxydicarbonate initiator, accelerates the condensation of polyphenol oxide quinone and amino acid or peptide, is favorable for forming humus, and then carries out secondary composting. At this time mesophilic microorganisms (bacillus subtilis) re-grow into a dominant species.
The secondary composting method of the invention preferably comprises the following steps: starting a heating system, enabling the temperature of the pile body to rise to 56-60 ℃, maintaining for 2-3 d, closing the heating system to enable the temperature of the pile body to naturally and slowly drop to be lower than 45 ℃, and entering a decomposition stage; the time of the decomposition stage is preferably 8-10 d. In the process, the ventilation and the stirring strengthening modes of the bin body are matched, so that the decomposition of materials in the pile body and the degradation of MPs are further promoted. Most of the microorganisms with good heat property in the high temperature stage die or enter a dormant state in a large quantity, which is beneficial to accelerating the decomposition of the compost. After primary composting, part of humus is formed, and rich nutrients and energy can be provided for microorganisms in the pile body in secondary composting, so that the decomposition rate which does not enter a decomposition state or a half-decomposition state is promoted, and the fertility is improved. In the primary composting stage, the accumulation of organic acid can lead to the decrease of pH, the heap is acidic, and in the secondary composting process, the microorganism further degrades the residual micromolecular organic acid to generate CO 2 And further degrading the nitrogen-containing organic matters to generate ammonia substances, and finally stabilizing the pH value of the pile body at 8.5-9.0 after secondary composting.
The invention can promote the staged growth and propagation of microorganisms (such as common spore bacteria form spores to resist stress at high temperature and become stress-resistant dormancy bodies, and the spores germinate and recover activity after the environmental conditions are recovered) by adding thermophilic pretreatment and secondary heating to create the intermittent heating and cooling effects, and improve the accumulation of humus (such as high temperature is beneficial to the formation of humic acid and the decomposition and conversion of fulvic acid, and low temperature is beneficial to the decomposition of humic acid and the accumulation of fulvic acid), and has better decomposition effect than the continuous heating.
The invention has the promotion effects of high-temperature pretreatment and secondary temperature rise on the decomposition of material compost and the formation of humus. The high-temperature pretreatment is favorable for evaporating water, so that the material reaches the optimal water condition of composting, and the micro plastic is primarily treated, so that the micro plastic enters a decomposition stage in advance, and the efficiency is improved. The secondary temperature rise promotes the substances which are not decomposed to continue to carry out the decomposition stage, so that the PH of the pile body is stabilized. The two ways are added to create temperature conditions of three heating and three cooling, which is beneficial to the diversity of microorganisms, thereby accelerating the degradation of the microplastic.
For further explanation of the present invention, a composite microbial agent and its application in aerobic composting and degradation of plastics provided by the present invention will be described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
Example 1
A composite microbial inoculum consists of the following components in parts by volume: 10 parts of bacillus subtilis, 10 parts of olive shaped soil and 10 parts of geobacillus stearothermophilus; the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253.
Example 2
A composite microbial inoculum consists of the following components in parts by volume: 10 parts of bacillus subtilis, 20 parts of olive shaped soil and 10 parts of geobacillus stearothermophilus; the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253.
Example 3
A composite microbial inoculum consists of the following components in parts by volume: 10 parts of bacillus subtilis, 20 parts of olive shaped soil and 30 parts of geobacillus stearothermophilus; the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253.
Example 4
Taking cow dung and corn straw waste as composting raw materials according to a dry matter mass ratio of 1:3, fully mixing, adjusting the carbon nitrogen ratio of the mixture to 25:1 and the water content to 70%, and putting the mixture into a composting bin for pretreatment.
The heating system was started and 3 treatments were set, respectively, to raise the temperature of the contents of the bin to 80 ℃ for 0.5h and to maintain the temperature at this temperature for 80min (C1), to raise the temperature of the contents of the bin to 80 ℃ for 1h (C2) and to maintain the temperature at this temperature for 80min and without warming (CK). 3 replicates were set for each treatment. The pretreatment water contents were measured, and the water contents of C1 and C2 were 68.0% and 63.2%, respectively, and the water content of CK was 70%.
After pretreatment is finished, the water distribution characteristics of the pile body under different treatments are measured, and the measuring method comprises the following steps: the literature 'influence of ultra-high temperature pretreatment on water holding characteristics of livestock manure and subsequent composting process', and the results are shown in Table 1 and FIG. 1.
TABLE 1 moisture distribution characteristics (g/g) of stacks under different treatments
| Mean value of | Initial moisture content | Free water content | Capillary water content | Adsorbed water content | Internal bound water content |
| CK | 3.358 | 0.042 | 2.100 | 1.250 | 0.051 |
| C1 | 2.769 | 0.037 | 2.020 | 1.127 | 0.045 |
| C2 | 2.550 | 0.037 | 1.945 | 1.001 | 0.041 |
As can be seen from table 1 and fig. 1, the thermophilic pretreatment promotes the conversion of high-binding energy form water to low-binding energy form water (conversion from bound water to free water) in the stack, which is beneficial to water evaporation, enhances the air permeability of the material and avoids anaerobic fermentation; meanwhile, bacteria are rapidly propagated and metabolized vigorously, so that the time for entering compost can be shortened.
After pretreatment, when the temperature of the material is reduced to below 40 ℃, 8wt.% of straw biochar is added into the compost raw material. 5 treatment groups were set, specifically as follows:
taking the raw materials which are not pretreated and the mixed microbial inoculum which is not added as a Control (CK);
the raw materials after pretreatment for 1h and the C2 group without adding mixed bacteria;
the composite microbial agent (T1) described in the embodiment 1 is added into the raw material subjected to pretreatment for 1h, and the mass ratio of the composite microbial agent to the raw material is 1:120;
the composite microbial agent (T2) described in the example 2 is added into the raw material subjected to pretreatment for 1h, and the mass ratio of the composite microbial agent to the raw material is 1:120;
the composite microbial agent (T2) described in the embodiment 3 is added into the raw material subjected to pretreatment for 1h, and the mass ratio of the composite microbial agent to the raw material is 1:120;
each treatment was repeated three times. And composting is carried out once in a composting bin. The fermentation period was 17d, and the composting temperatures at various times were measured and the results are shown in Table 2.
TABLE 2 compost temperature Change results (. Degree.C.)
| Composting time (d) | CK | C2 | T1 | T2 | T3 |
| 0 | 30.0 | 30.0 | 30.0 | 30.0 | 30.0 |
| 1 | 33.2 | 33.1 | 34.2 | 34.8 | 34.3 |
| 2 | 38.3 | 38.2 | 41.5 | 43.7 | 42.3 |
| 3 | 46.7 | 46.3 | 50.8 | 53.2 | 52.2 |
| 4 | 51.8 | 51.2 | 54.7 | 58.9 | 55.4 |
| 5 | 53.6 | 52.9 | 55.2 | 60.1 | 58.4 |
| 6 | 54.5 | 54.0 | 56.4 | 61.8 | 58.1 |
| 7 | 56.8 | 55.5 | 58.9 | 62.5 | 59.7 |
| 8 | 57.7 | 57.1 | 59.2 | 63.5 | 59.9 |
| 9 | 58.0 | 57.8 | 59.5 | 63.8 | 60.1 |
| 10 | 57.4 | 56.7 | 58.1 | 63.2 | 59.7 |
| 11 | 54.6 | 53.2 | 55.7 | 62.1 | 56.9 |
| 12 | 49.8 | 49.5 | 53.2 | 59.6 | 54.6 |
| 13 | 45.7 | 45.6 | 51.7 | 55.7 | 52.4 |
| 14 | 42.6 | 42.1 | 50.1 | 52.8 | 50.5 |
| 15 | 40.8 | 39.0 | 45.8 | 47.8 | 46.7 |
| 16 | 38.3 | 37.8 | 40.8 | 43.2 | 42.1 |
| 17 | 37.8 | 36.8 | 38.5 | 41.1 | 39.2 |
The metabolism of microorganisms during composting can produce and consume energy, thereby affecting the composting temperature. As can be seen from table 2, the temperature rise process of T1, T2, T3 is significantly accelerated on day 3 of composting, the temperature is gradually higher than C2, CK, and T2 reaches a peak value of 63.8 ℃ on day 9; and the peak temperatures of T3 and T1 are 60.1 ℃ and 59.5 ℃; CK. The peak temperature of C2 was 58.0℃and 57.8 ℃. In the whole composting process, the temperature of T2 is always higher than that of T3, T1, C2 and CK, and the duration (12 d) of the high-temperature period (more than 50 ℃) of T3, T2 and T1 is 4d more than that of C2 and CK, so that the microbial inoculum can prolong the high-temperature period and is beneficial to the completion of the decomposition of organic matters.
After the end of one compost, the composition and relative abundance of the bacterial groups of the different stacks at the portal level were determined by the following method: the literature "metabolic diversity of aerobic composting microorganisms and bacterial community structure", the results are shown in Table 3 and FIG. 2.
TABLE 3 composition and relative abundance of bacterial groups falling on the gate level after one compost (%)
| Treatment of | Thick wall fungus door | Proteus gate | Bacteroides door | Others |
| CK | 6.83 | 38.67 | 41.78 | 12.72 |
| C2 | 7.22 | 39.89 | 39.65 | 13.24 |
| T1 | 8.43 | 39.95 | 37.67 | 13.95 |
| T2 | 10.52 | 40.38 | 34.55 | 14.55 |
| T3 | 8.93 | 40.26 | 36.21 | 14.60 |
As can be seen from Table 3 and FIG. 2, the relative abundance of the phylum Thick-walled bacteria is further reduced, and the relative abundance of the phylum thick-walled bacteria is 7.22%, 8.43%, 10.52% and 8.93% in the treatment of C2, T1, T2 and T3 respectively, and the phylum thick-walled bacteria can secrete a plurality of extracellular enzymes to degrade macromolecular substrates such as protein, pectin and cellulose; whereas the relative abundance of Proteus was maintained at a higher level, 39.89%, 39.95%, 40.38% and 40.26% in C2, T1, T2, T3 treatments, respectively. The Proteus has good nitrogen fixation property, and can fix more nitrogen in compost than other bacteria. At this stage, the relative abundance of bacteroides increased, accounting for 39.65%, 37.67%, 34.55% and 36.21% in C2, T1, T2, T3 treatments, respectively. From the results, the inoculation of the microbial inoculum can promote degradation of carbohydrates and lignocellulose in the high-temperature stage of composting, inhibit or kill pathogenic bacteria in the composting, and finally improve the efficiency and quality of the composting.
In the composting process, the organic matters are continuously mineralized and humified, and the content is continuously reduced. In the composting process, the organic matter content of each treatment group is in a decreasing trend. CK. The final organic matter degradation rates of C2, T1, T2 and T3 are respectively 12.05%, 12.09% and 13.67%, 15.08% and 14.08%, and the determination method of the organic matter degradation rates is described in the national environmental protection standard of the people's republic of China. The inoculation of the microbial inoculum can promote the biodegradation of organic matters in the composting process.
When most of the organics were degraded, the temperature began to drop and gradually lowered to room temperature, yielding a first stack. And adding diisopropyl peroxydicarbonate into the 5 treatment groups, and then performing secondary composting, wherein the mass ratio of the diisopropyl peroxydicarbonate to the first pile is 0.002:1000. starting a heating system to enable the temperature of the secondary compost pile body to rise to 60 ℃ for 1h, maintaining for 2d, closing the heating system to enable the temperature of the pile body to naturally and slowly drop below 45 ℃ and enter a decomposition stage, and in the process, promoting the decomposition of materials in the pile body and the degradation of MPs by matching with a ventilation and reinforced stirring mode of the bin body, wherein the time of the decomposition stage is 8d.
After pretreatment, after the end of primary composting and after the end of secondary composting, the effect of different treatments on Hu Mingsuan and fulvic acid was determined by: the literature "influence of ultra-high temperature pretreatment on carbon-nitrogen conversion and loss in pig manure composting process" shows the results in tables 4-5 and figures 3-4.
TABLE 4 influence of different treatments on Hu Mingsuan during composting
| Treatment of | After pretreatment | Primary composting | Secondary compost |
| CK | 42.80 | 62.37 | 67.82 |
| C2 | 36.67 | 70.85 | 75.63 |
| T1 | 38.98 | 78.18 | 84.39 |
| T2 | 37.83 | 88.64 | 96.78 |
| T3 | 35.21 | 85.31 | 90.54 |
Table 5 shows the effect of different treatments on fulvic acid during composting
| Treatment of | After pretreatment | Primary composting | Secondary compost |
| CK | 104.40 | 33.75 | 25.40 |
| C2 | 85.28 | 33.68 | 28.22 |
| T1 | 82.94 | 35.76 | 29.40 |
| T2 | 72.75 | 38.21 | 32.15 |
| T3 | 71.86 | 36.79 | 31.11 |
As can be seen from tables 4 to 5 and FIGS. 3 to 4, as composting proceeds, the Hu Fu ratio (Hu Mingsuan: fulvic acid) of the inoculated group and the blank group increases gradually, indicating that the organic matter is converted to humus and the humification degree increases gradually. Finally, the CK, C2, T1, T2 and T3 treatments Hu Fubi are respectively 2.67, 2.68, 2.87, 3.01 and 2.91, which indicate that the stage changes of high temperature and low temperature can strengthen the humification degree of the compost.
Finally, carrying out size and particle distribution statistics through laser infrared imaging, an optical microscope and microscopic infrared, so as to obtain degradation rate, wherein the determination method refers to the soil microplastic analysis technology and research progress based on a laser infrared imaging system, the determination is carried out on the microplastic residues of each treatment, and the degradation rate of the T2 treatment on PS, PE, PET reaches the highest, namely 95%, 88% and 85% respectively. In the invention, the optimal combination of pretreatment time of 1h, microbial inoculum ratio of 1:2:1, addition of biochar, initiator and secondary composting comprehensive treatment is obtained, thereby providing a reference for promoting the aerobic composting degradation of the micro-plastics of the agricultural wastes.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (10)
1. The composite microbial inoculum is characterized by comprising the following components in parts by volume: bacillus subtilis @Bacillus subtilis) 10-15 parts of soil olive shaped bacteriaOlivibacter soli) 10-30 parts of Geobacillus stearothermophilusGeobacillus stearothermophilus) 10-45 parts of a lubricant; the strain number of the bacillus subtilis is CGMCC 1.3358, the strain number of the soil olivary bacteria is ACCC 02948, and the strain number of the geobacillus stearothermophilus is ACCC 10253.
2. The composite microbial inoculant according to claim 1, wherein the volume ratio of bacillus subtilis, olivopodiomycetes soil and geobacillus stearothermophilus is 1:2:1.
3. use of a composite microbial agent according to claim 1 or 2, comprising:
1) The efficiency of aerobic composting is improved; 2) Promoting degradation of carbohydrates in the compost raw material; 3) Promoting degradation of lignocellulose in the compost raw material; 4) Inhibiting and/or killing pathogenic bacteria in the compost raw material; 5) Degrading the plastic; the plastic comprises one or more of polyethylene, polystyrene, and polyethylene terephthalate.
4. An aerobic composting method for degrading plastics, which is characterized by comprising the following steps:
mixing the composite microbial inoculum of claim 1 or 2 with a composting material and aerobic composting.
5. The aerobic composting method of claim 4, wherein the composting material is pretreated prior to mixing with the composite microbial inoculant; the pretreatment comprises the following steps: and heating the compost raw material to 70-80 ℃, maintaining the temperature for 60-80 min, and cooling to 35-40 ℃.
6. The aerobic composting method according to claim 5, wherein the water content of the composting raw material before heating is 60% to 70%.
7. The aerobic composting method according to claim 4, wherein the aerobic composting method comprises: after aerobic fermentation for 15-20 d, adding an initiator to perform secondary aerobic fermentation; the initiator comprises diisopropyl peroxydicarbonate.
8. The aerobic composting method according to claim 7, wherein the method of the second aerobic fermentation comprises: and (3) adding an initiator, heating to 56-60 ℃, maintaining for 2-3 days, naturally cooling, and decomposing and fermenting for 8-10 days.
9. The aerobic composting method of claim 4 or 7, further comprising mixing a mixture of a composite microbial inoculant and a composting material with biochar prior to aerobic composting.
10. The aerobic composting method according to claim 4, wherein the mass ratio of the composite microbial inoculum to the composting raw material is 1: 100-120.
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