阅读说明：本技术 一种微生物土壤修复剂及其制备方法 (microbial soil remediation agent and preparation method thereof ) 是由 张海涛 张磊 徐长青 于 2019-11-19 设计创作，主要内容包括：一种微生物土壤修复剂及其制备方法,属于土壤修复技术领域。该土壤修复剂是由复合微生物菌剂5-15份、吸附载体5-10份、腐植酸10-20份、硅藻土30-40份制成。本发明所制备的微生物土壤修复剂可显著提高受污染土壤中有机质含量,调节土壤菌群结构及土壤酸碱性,提高土壤透气性,改善土壤理化性质。可以有效降解土壤中有机污染物及重金属铅、铬、砷等的含量,提高土壤肥力,作物增产增收效果显著。(The invention relates to a microbial soil remediation agent and a preparation method thereof, belonging to the technical field of soil remediation, wherein the soil remediation agent is prepared from 5-15 parts of a compound microbial agent, 5-10 parts of an adsorption carrier, 10-20 parts of humic acid and 30-40 parts of diatomite.)

The microbial soil remediation agent is characterized by being prepared from the following raw materials, by weight, 5-15 parts of a compound microbial agent, 5-10 parts of an adsorption carrier, 10-20 parts of humic acid and 30-40 parts of diatomite.

2. The microbial soil remediation agent of claim 1, wherein said complex microbial inoculant is prepared by the following method:

(1) under the aseptic condition, pseudoxanthomonas pentabasic CGMCC No: 1.10978 inoculating to liquid culture medium at 3% of the liquid culture medium, and shake culturing at 22-25 deg.C for 3 days to obtain seed bacteria liquid;

(2) inoculating the seed bacterial liquid obtained in the step (1) into a liquid culture medium according to 7% of the mass of the liquid culture medium, carrying out shake culture at 28-30 ℃ in the dark for 7-10 days to obtain a zymogen liquid, and directly freezing and drying to obtain bacterial powder A;

(3) and mixing the bacterial powder A and the bacillus subtilis according to the weight ratio of 5:1 to obtain the compound microbial agent.

3. The microbial soil remediation agent of claim 2 wherein said liquid medium is made of: 10-15 parts of peptone, 3-6 parts of yeast powder, 1-3 parts of monopotassium phosphate, 1-3 parts of nano composite additive and 120 parts of purified water.

4. The microbial soil remediation agent of claim 3, wherein said nanocomposite additive is prepared by the following method: completely crushing the fish leftovers, mixing the fish leftovers with water in equal weight ratio, adding trypsin into the mixture according to the total weight of 1000U/g for enzymolysis, adjusting the pH value to 6.0-7.5, heating to 42-45 ℃ for enzymolysis for 20-24h, and performing nanofiltration by using a nanofiltration membrane after the enzymolysis is finished to obtain polypeptide liquid; adding 1M sodium selenite solution into the polypeptide solution according to the volume ratio of 2:1, adjusting the pH value to 5.0-5.5, heating to 60-65 ℃ for chelation reaction for 4-6 hours to obtain polypeptide chelate solution; soaking the carbon nano tube in 5% dilute nitric acid for 12 hours, soaking the treated carbon nano tube in the polypeptide chelating solution, performing ultrasonic treatment for 30-40 minutes, and centrifuging to obtain solid filter residue, namely the nano composite additive.

5. The microbial soil remediation agent of claim 4 wherein said sonication frequency is 90-100 KHz.

6. The microbial soil remediation agent of claim 1 wherein said adsorbent carrier is prepared by the following method:

(a) soaking 100g of kapok in 100ml of a mixed solution of an aluminum nitrate ethanol solution and a ferric nitrate ethanol solution for 30 minutes, then performing ultrasonic treatment for 20 minutes, taking out and drying, and calcining at 450 ℃ for 1 hour to obtain a mixture;

(b) and (3) soaking the mixture prepared in the step (a) into 50ml of 25% manganese nitrate solution for 1 hour, taking out, drying, and calcining at 650 ℃ for 1 hour to obtain the final adsorption carrier.

7. The microbial soil remediation agent of claim 6 wherein said aluminum nitrate ethanol solution is at a concentration of 0.8mol/L and said ferric nitrate ethanol solution is at a concentration of 0.3 mol/L.

A method of microbial soil remediation according to any one of claims 1 to 7 to , comprising the steps of:

(1) completely crushing the fish leftovers, mixing the fish leftovers with water in equal weight ratio, adding trypsin into the mixture according to the total weight of 1000U/g for enzymolysis, adjusting the pH value to 6.0-7.5, heating to 42-45 ℃ for enzymolysis for 20-24h, and performing nanofiltration by using a nanofiltration membrane after the enzymolysis is finished to obtain polypeptide liquid; adding 1M sodium selenite solution into the polypeptide solution according to the volume ratio of 2:1, adjusting the pH value to 5.0-5.5, heating to 60-65 ℃ for chelation reaction for 4-6 hours to obtain polypeptide chelate solution; soaking a carbon nano tube in 5% dilute nitric acid for 12 hours, soaking the treated carbon nano tube in the polypeptide chelating solution, performing ultrasonic treatment for 30-40 minutes, and centrifuging to obtain solid filter residue, namely the nano composite additive, for later use;

(2) preparing a liquid culture medium: mixing 10-15 parts of peptone, 3-6 parts of yeast powder, 1-3 parts of monopotassium phosphate, 1-3 parts of nano composite additive and 120 parts of purified water, uniformly stirring, and then sterilizing at the temperature of 121 ℃ for 15-30 minutes under the pressure of 0.1MPa to obtain a liquid culture medium;

(3) preparing a compound microbial agent: under the aseptic condition, pseudoxanthomonas pentabasic CGMCC No: 1.10978 inoculating the liquid culture medium prepared in the step (2) according to 3% of the mass of the liquid culture medium, and performing shaking culture at 22-25 ℃ for 3 days to obtain seed bacterial liquid; inoculating the obtained seed bacterial liquid into a liquid culture medium according to 7% of the mass of the liquid culture medium, carrying out shake culture at 28-30 ℃ in the dark for 7-10 days to obtain a zymogen liquid, and directly freezing and drying to obtain bacterial powder A; mixing the bacterial powder A and bacillus subtilis according to a weight ratio of 5:1 to obtain a compound microbial agent;

(4) preparation of the adsorption carrier: soaking 100g of kapok in 100ml of a mixed solution of an aluminum nitrate ethanol solution and a ferric nitrate ethanol solution for 30 minutes, then performing ultrasonic treatment for 20 minutes, taking out and drying, and calcining at 450 ℃ for 1 hour to obtain a mixture; soaking the prepared mixture into 50ml of 25% manganese nitrate solution for 1 hour, taking out, drying, and calcining at 650 ℃ for 1 hour to obtain a final adsorption carrier for later use;

(5) weighing the compound microbial agent obtained in the step (3), the adsorption carrier obtained in the step (4), humic acid and diatomite according to the parts by weight, placing the mixture in a pre-mixer for 45r/min, mixing for 15-20 minutes, and subpackaging to obtain a finished product.

9. The method of claim 8 wherein said step (4) is carried out at a concentration of 0.8mol/L in an ethanol solution of aluminum nitrate and 0.3mol/L in an ethanol solution of ferric nitrate.

10. The method of claim 8 wherein the ultrasonic treatment frequency in step (1) is 90-100 KHz.

Technical Field

The invention relates to an microbial soil remediation agent and a preparation method thereof, belonging to the technical field of soil remediation.

Background

With the continuous acceleration of the industrialization process, unreasonable mining and smelting discharge of mineral resources, sewage irrigation and sludge application to soil for a long time, atmospheric sedimentation caused by artificial activities, application of chemical fertilizers and pesticides and the like cause serious soil pollution, soil is an important material basis of human social production activities and is an indispensable and difficult-to-regenerate natural resource, an untreated polluted site is a chemical timing bomb, large-area outbreak can cause an inestimable influence on national sustainable development, and therefore high attention must be paid to prevention of soil pollution and remediation of polluted soil.

Soil remediation refers to the physical, chemical and biological processes used to transfer, absorb, degrade and transform pollutants in soil to reduce their concentration to acceptable levels, or to transform toxic and harmful pollutants into harmless materials. Fundamentally, the technical principle of contaminated soil remediation may include: (1) changing the existing form of the pollutants in the soil or the combination mode of the pollutants and the soil, and reducing the mobility and bioavailability of the pollutants in the environment; (2) the concentration of harmful substances in the soil is reduced. At present, theoretically possible repair techniques include several major categories, such as phytoremediation techniques, microbial repair techniques, chemical repair techniques, physical repair techniques, and comprehensive repair techniques. In which microbial remediation techniques are receiving increasing attention with their own advantages.

Disclosure of Invention

The invention aims to provide microbial soil remediation agents, which can efficiently degrade organic pollutants in soil, reduce the content of heavy metals in the soil, improve the soil fertility, promote the growth of plants and improve the yield and quality of crops.

The invention also provides a preparation method of the microbial soil remediation agent.

The invention adopts the following technical scheme:

microbial soil remediation agent, which is prepared from the following raw materials, by weight, 5-15 parts of compound microbial agent, 5-10 parts of adsorption carrier, 10-20 parts of humic acid, and 30-40 parts of diatomite.

Wherein the compound microbial agent is prepared by adopting the following method:

(1) under the aseptic condition, pseudoxanthomonas pentabasic CGMCC No: 1.10978 inoculating to liquid culture medium at 3% of the liquid culture medium, and shake culturing at 22-25 deg.C for 3 days to obtain seed bacteria liquid;

(2) inoculating the seed bacterial liquid obtained in the step (1) into a liquid culture medium according to 7% of the mass of the liquid culture medium, carrying out shake culture at 28-30 ℃ in the dark for 7-10 days to obtain a zymogen liquid, and directly freezing and drying to obtain bacterial powder A;

(3) and mixing the bacterial powder A and the bacillus subtilis according to the weight ratio of 5:1 to obtain the compound microbial agent.

The liquid culture medium is prepared from the following components: 10-15 parts of peptone, 3-6 parts of yeast powder, 1-3 parts of monopotassium phosphate, 1-3 parts of nano composite additive and 120 parts of purified water.

The nano composite additive is prepared by adopting the following method: completely crushing the fish leftovers, mixing the fish leftovers with water in equal weight ratio, adding trypsin into the mixture according to the total weight of 1000U/g for enzymolysis, adjusting the pH value to 6.0-7.5, heating to 42-45 ℃ for enzymolysis for 20-24h, and performing nanofiltration by using a nanofiltration membrane after the enzymolysis is finished to obtain polypeptide liquid; adding 1M sodium selenite solution into the polypeptide solution according to the volume ratio of 2:1, adjusting the pH value to 5.0-5.5, heating to 60-65 ℃ for chelation reaction for 4-6 hours to obtain polypeptide chelate solution; soaking the carbon nano tube in 5% dilute nitric acid for 12 hours, soaking the treated carbon nano tube in the polypeptide chelating solution, performing ultrasonic treatment for 30-40 minutes, and centrifuging to obtain solid filter residue, namely the nano composite additive.

The ultrasonic treatment frequency is 90-100 KHz.

The adsorption carrier is prepared by adopting the following method:

(a) soaking 100g of kapok in 100ml of a mixed solution of an aluminum nitrate ethanol solution and a ferric nitrate ethanol solution for 30 minutes, then performing ultrasonic treatment for 20 minutes, taking out and drying, and calcining at 450 ℃ for 1 hour to obtain a mixture;

(b) and (3) soaking the mixture prepared in the step (a) into 50ml of 25% manganese nitrate solution for 1 hour, taking out, drying, and calcining at 650 ℃ for 1 hour to obtain the final adsorption carrier.

Preferably, the concentration of the aluminum nitrate ethanol solution is 0.8mol/L, and the concentration of the ferric nitrate ethanol solution is 0.3 mol/L.

The preparation method of microbial soil remediation agent comprises the following steps:

(1) completely crushing the fish leftovers, mixing the fish leftovers with water in equal weight ratio, adding trypsin into the mixture according to the total weight of 1000U/g for enzymolysis, adjusting the pH value to 6.0-7.5, heating to 42-45 ℃ for enzymolysis for 20-24h, and performing nanofiltration by using a nanofiltration membrane after the enzymolysis is finished to obtain polypeptide liquid; adding 1M sodium selenite solution into the polypeptide solution according to the volume ratio of 2:1, adjusting the pH value to 5.0-5.5, heating to 60-65 ℃ for chelation reaction for 4-6 hours to obtain polypeptide chelate solution; soaking a carbon nano tube in 5% dilute nitric acid for 12 hours, soaking the treated carbon nano tube in the polypeptide chelating solution, performing ultrasonic treatment for 30-40 minutes, and centrifuging to obtain solid filter residue, namely the nano composite additive, for later use;

(2) preparing a liquid culture medium: mixing 10-15 parts of peptone, 3-6 parts of yeast powder, 1-3 parts of monopotassium phosphate, 1-3 parts of nano composite additive and 120 parts of purified water, uniformly stirring, and then sterilizing at the temperature of 121 ℃ for 15-30 minutes under the pressure of 0.1MPa to obtain a liquid culture medium;

(3) preparing a compound microbial agent: under the aseptic condition, pseudoxanthomonas pentabasic CGMCC No: 1.10978 inoculating the liquid culture medium prepared in the step (2) according to 3% of the mass of the liquid culture medium, and performing shaking culture at 22-25 ℃ for 3 days to obtain seed bacterial liquid; inoculating the obtained seed bacterial liquid into a liquid culture medium according to 7% of the mass of the liquid culture medium, carrying out shake culture at 28-30 ℃ in the dark for 7-10 days to obtain a zymogen liquid, and directly freezing and drying to obtain bacterial powder A; mixing the bacterial powder A and bacillus subtilis according to a weight ratio of 5:1 to obtain a compound microbial agent;

(4) preparation of the adsorption carrier: soaking 100g of kapok in 100ml of a mixed solution of an aluminum nitrate ethanol solution and a ferric nitrate ethanol solution for 30 minutes, then performing ultrasonic treatment for 20 minutes, taking out and drying, and calcining at 450 ℃ for 1 hour to obtain a mixture; soaking the prepared mixture into 50ml of 25% manganese nitrate solution for 1 hour, taking out, drying, and calcining at 650 ℃ for 1 hour to obtain a final adsorption carrier for later use;

(5) weighing the compound microbial agent obtained in the step (3), the adsorption carrier obtained in the step (4), humic acid and diatomite according to the parts by weight, placing the mixture in a pre-mixer for 45r/min, mixing for 15-20 minutes, and subpackaging to obtain a finished product.

Preferably, in the step (4), the concentration of the aluminum nitrate ethanol solution is 0.8mol/L, and the concentration of the ferric nitrate ethanol solution is 0.3 mol/L.

Preferably, the ultrasonic treatment frequency in the step (1) is 90-100 KHz.

The pseudoxanthomonas campestris used in the invention is purchased from the common microorganism center of China Committee for culture Collection of microorganisms, and the number of the culture is CGMCC No: 1.10978. the bacillus subtilis is purchased from Shandong Taibang agriculture development Co., Ltd, and the number of effective viable bacteria is more than or equal to 10 hundred million/g.

The composite microbial agent is prepared by culturing a specific culture medium under specific conditions for 2 times, wherein pseudoxanthomonas pentandra is cultured in a special liquid culture medium, and a nano composite additive added in the liquid culture medium can provide a large amount of organic nutrients in the culture process of the strains and simultaneously increase the selectivity of the strains, is favorable for purifying the strains, and can also effectively adsorb certain dead bacteria and bacteria metabolites, so that the strain breeding environment is improved, the activity of the cultured strains is high, and after the strains are compounded with bacillus subtilis, a large amount of beneficial active microorganisms act on soil, so that the soil fertility can be obviously improved, the soil structure is improved, and the content of heavy metals and organic pollutants in the soil is reduced.

The invention has the beneficial effects that: the microbial soil remediation agent prepared by the invention can obviously improve the content of organic matters in the polluted soil, adjust the structure of soil flora and the pH value of the soil, improve the air permeability of the soil and improve the physical and chemical properties of the soil. Can effectively degrade the contents of organic pollutants and heavy metals such as lead, chromium, arsenic and the like in the soil, improve the soil fertility and have obvious effects of increasing the yield and the income of crops.

Detailed Description

The present invention is further illustrated in step by reference to specific examples.