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CN-121495752-B - Microbial liquid composite microbial inoculant and biological mineral fertilizer with coal gangue activation and heavy metal treatment functions

CN121495752BCN 121495752 BCN121495752 BCN 121495752BCN-121495752-B

Abstract

The invention belongs to the technical field of biological mineral fertilizers, and particularly relates to a microbial liquid composite microbial inoculant and a biological mineral fertilizer with coal gangue activation and heavy metal treatment. The microbial liquid compound bacterial agent is prepared by inoculating and fermenting functional strains such as pseudomonas putida, azotobacter chroococcus furiosus, azoospira baxi, phosphorus-dissolving bacteria, pseudomonas fluorescens, biocontrol bacteria bacillus subtilis, potassium-dissolving bacteria bacillus mucilaginosus, silicon-dissolving bacteria bacillus megaterium and synergistic bacteria trichoderma asperellum by using electrochemical low-hertz water in a three-stage time sequence. The biological mineral fertilizer is prepared from the microbial liquid composite microbial agent and coal gangue powder serving as a main raw material. The scientific compatibility of different strains provided by the invention realizes the efficient improvement of heavy metal polluted lands and the yield increase of crops, changes the waste coal gangue into valuable, solves the problem of pollution caused by the massive accumulation of the coal gangue, and has important economic and social values.

Inventors

  • ZHU ANNI
  • WANG SONGRUI

Assignees

  • 北京京师祺瑞科技有限公司

Dates

Publication Date
20260505
Application Date
20260108

Claims (7)

  1. 1. The microbial liquid composite microbial agent with the functions of coal gangue activation and heavy metal treatment is characterized by comprising the following functional strains, namely pseudomonas putida, azotobacter chroococcus, azotobacter bazedoxus, phosphorus-decomposing bacteria, pseudomonas fluorescens, biocontrol bacteria bacillus subtilis, potassium-decomposing bacteria bacillus mucilaginosus, bacillus megaterium and trichoderma echinocandii serving as repair bacteria, wherein the functional strains are prepared by using electrochemical low-hertz water through three-stage sequential inoculation fermentation, and the inoculation fermentation preparation comprises the following steps of: s1, adding electrochemical low-hertz water into a sterilized main fermentation tank, adding pre-mixed repair bacteria, azotobacter and growth promoting bacteria seed liquid, starting stirring, and adjusting the fermentation condition to a first time sequence fermentation condition, wherein the pH=6.5-7.0, the dissolved oxygen content is 20-30%, the temperature is 30+/-0.5 ℃, the humidity of the top space of the fermentation tank is 90-92%, and the fermentation is carried out for 23-25 hours, wherein the molecular group of the electrochemical low-hertz water is less than or equal to 6 molecules; S2, after the S1 stage is finished, adjusting the fermentation condition to a second time sequence, adding pre-mixed phosphorus-dissolving bacteria and biocontrol strain seed liquid into a main fermentation tank, adding coal gangue powder into the main fermentation tank, continuously stirring, adjusting the fermentation condition to the second time sequence fermentation, wherein the pH value is 6.8+/-0.2, the dissolved oxygen amount is 30-40%, the temperature is 30+/-0.5 ℃, the headspace humidity of the fermentation tank is 91-93%, and the fermentation is carried out for 17-19 hours; S3, after fermentation in the S2 stage is finished, adding the pre-mixed potassium-decomposing bacteria, silicon-decomposing bacteria and synergistic bacteria seed liquid into a main fermentation tank, continuously stirring, and adjusting to a third time sequence fermentation condition, wherein the pH=7.0+/-0.2, the dissolved oxygen amount is 40-50%, the temperature is 30+/-0.5 ℃, the headspace humidity of the fermentation tank is 85-88%, and the fermentation is carried out for 35-37 hours; When the total viable count in the main fermentation tank is more than or equal to 2 multiplied by 10 10 CFU/mL, ending the fermentation; the volume ratio of the seed solution of the repairing bacteria, the azotobacter, the growth promoting bacteria, the phosphate solubilizing bacteria, the biocontrol bacteria, the potassium decomposing bacteria, the silicon decomposing bacteria and the synergistic bacteria is 2:3:1:1:1:1:1:1; Adding nutrition factor citric acid and sodium molybdate and stress-resistance induction liquid low concentration Cr 6+ when fermentation is carried out in the S1 stage to the middle stage of the logarithmic growth phase, adding nutrition factor glycerin and coal gangue powder after the S2 stage fermentation is started and the pH value of the system is stable, and adding nutrition factor potassium feldspar powder and humic acid when the S3 stage is started; In the S1, citric acid, sodium molybdate and low-concentration Cr 6+ are added in pulses, wherein the adding amount of each pulse is 1/3 of the total required amount, and the interval is 2 hours, and the total adding amount is 3 times, wherein the pulse adding is completed within 10 minutes by adopting slow flow injection, and the slow flow injection of the low-concentration Cr 6+ liquid is completed within 1/3 of the total required amount of 5 minutes after 1 hour after each injection of the citric acid and the sodium molybdate; S2, synchronously adding glycerol and coal gangue powder, continuously injecting the glycerol by adopting a peristaltic pump, and feeding the coal gangue powder in batches by using a sterile crushing feeding device; and S3, synchronously adding potassium feldspar powder and humic acid, wherein the potassium feldspar powder is fed in batches through a sterile feeding device, and the humic acid is continuously injected through a peristaltic pump.
  2. 2. The microbial liquid composite microbial inoculant with both coal gangue activation and heavy metal treatment according to claim 1 is characterized in that the end of the S1 stage is marked by the total bacterial count in a main fermentation tank being equal to or greater than 1 multiplied by 10 9 CFU/mL, the heavy metal adsorption rate being equal to or greater than 60%, the activity of azotase being equal to or greater than 20nmolC 2 H 4 /(mL.h) and the IAA content being equal to or greater than 10mg/L, the end of the S2 stage is marked by the total bacterial count being equal to or greater than 1.5X10 9 CFU/mL, the effective phosphorus concentration of the supernatant being equal to or greater than 50mg/mL and the bacillus subtilis spore rate being equal to or greater than 90%, the end of the S3 stage is marked by the total viable bacterial count being equal to or greater than 2 multiplied by 10 CFU/mL, the potassium feldspar decomposition rate being equal to or greater than 35%, the silicon dissolution rate being equal to or greater than 25%, and the conversion rate of coal gangue organic matter being equal to or greater than 30%, and the total viable bacterial count comprises the fungal spore count and bacterial count.
  3. 3. The microbial liquid composite microbial inoculant with both gangue activation and heavy metal treatment according to claim 1, wherein the preparation and mixing process of the seed liquid in S1-S3 comprises the following steps: Step1, pretreatment, namely respectively inoculating each strain to a special culture medium, culturing at 30 ℃ for 24-48 hours to a logarithmic phase, culturing the repairing bacteria by using LB+0.01% cadmium ion culture medium, culturing the azotobacter by using Abental Bei Modan culture medium, culturing the phosphate-solubilizing bacteria by using KB culture medium, culturing the biocontrol bacteria by using LB culture medium, culturing the potassium-solubilizing bacteria by using potassium-solubilizing culture medium, culturing the silicon-solubilizing bacteria by using beef extract peptone+2% silicon mineral powder culture medium, culturing the trichoderma spinosum by using PDA culture medium containing 0.5% sand grains in an induction mode, preparing the special culture medium by using electrochemical low-hertz water, and sterilizing at 121 ℃ for 20 minutes; Step 2, culturing in a first-level seed tank, namely preparing independent first-level seed tanks, respectively adding the strains cultured in the step 1 into different independent first-level fermentation tanks, and culturing for a certain time to obtain seed liquid of a single strain, wherein the culture conditions of the repairing bacteria and the azotobacter are 30 ℃ plus or minus 0.5 ℃, 88-90% of the top space humidity of the fermentation tank, 20-30% of dissolved oxygen and 24 hours, the culture conditions of the phosphate-solubilizing bacteria, potassium-solubilizing bacteria and silicon-solubilizing bacteria are 30 ℃ plus or minus 0.5%, 90-92% of the top space humidity of the fermentation tank, 30-40% of dissolved oxygen, 85-88% of the top space humidity of the fermentation tank, 25-35% of dissolved oxygen and 40-50% of dissolved oxygen; And 3, culturing in a secondary premix tank, namely mixing the seed solutions of the single strains obtained in the step 2 according to a certain proportion, and preserving the temperature for 30 minutes at 30 ℃ to obtain the seed solution of the mixed strains.
  4. 4. The biological mineral fertilizer with both coal gangue activation and heavy metal treatment is characterized by being prepared by using the microbial liquid composite microbial agent with both coal gangue activation and heavy metal treatment according to any one of claims 1-3.
  5. 5. The biological mineral fertilizer with both gangue activation and heavy metal remediation of claim 4, wherein the preparation method of the biological mineral fertilizer comprises the following steps: A1, adding the microbial liquid composite microbial inoculum, 80-mesh coal gangue powder, wheat bran, soybean meal and electrochemical low-hertz water into a mixing and stirring system, and after uniformly mixing and stirring, conveying the mixture to a solid fermentation system production line through an intelligent material distribution system to perform primary solid fermentation, wherein the solid fermentation system automatically adjusts the temperature in the fermentation process through the linkage of a temperature sensor and intermittent micro-blasting frequency, wherein the temperature in the initial stage is 42-45 ℃, the temperature in the peak stage is maintained for 22-24 hours, the temperature in the peak stage is 60-65 ℃, the temperature in the peak stage is maintained for 46-48 hours, the temperature in the end stage is 40-45 ℃, and the temperature in the end stage is maintained for 22-24 hours; And A2, rearranging the materials obtained after the first solid state fermentation in the step A1 to an aging fermentation production line through an intelligent material distribution system for carrying out second aging fermentation, namely finishing deep decomposition of the materials which are piled up naturally by utilizing the residual heat of the materials and residual bacterial groups, and finishing fermentation after lasting for 10-12 days.
  6. 6. The biological mineral fertilizer with both gangue activation and heavy metal treatment according to claim 5, wherein the mark of the end of the first solid state fermentation in A1 is that the temperature is maintained at the final stage temperature within 24 hours continuously, the humidity of the material is less than or equal to 40%, the color of the material is dark brown, no peculiar smell exists, the hand is loose, and no bonding phenomenon exists.
  7. 7. The biological mineral fertilizer with both coal gangue activation and heavy metal treatment according to claim 5, wherein the mark of the end of the second aging fermentation in A2 is that the material is naturally dehumidified to 15-29% of water content, the color is even and dark brown, the texture is loose, the soil fishy smell is free of other peculiar smell, the pH is 6.5-7.5, the beneficial viable count is more than or equal to 2 hundred million/g, the total amount of the solid components in A1 is 100%, the solid components account for 90-95% of 80-mesh coal gangue powder, 3-8% of wheat bran and 4-10% of soybean meal, the initial water content of the coal gangue powder is 5-9% of the initial water content of the wheat bran and the soybean meal is 8-12%, after the solid components are uniformly mixed, electrochemical low-hertz water is added for full stirring, and simultaneously the microbial liquid compound microbial agent is synchronously added in the stirring process, wherein the addition amount is 0.8-1.2% of the total amount of the solid substrate, and the total water content of the material is finally 50-60%.

Description

Microbial liquid composite microbial inoculant and biological mineral fertilizer with coal gangue activation and heavy metal treatment functions Technical Field The invention belongs to the technical field of biological mineral fertilizers, and particularly relates to a microbial liquid composite microbial inoculant and a biological mineral fertilizer with coal gangue activation and heavy metal treatment. Background The gangue is used as a solid waste inevitably generated in the coal exploitation and processing process, and the mine waste gangue is piled for a long time to cause series of environmental pollution, including occupation of land resources, pollution of soil by heavy metal seepage, acidification and desertification of the soil caused by groundwater and high sulfur components, and ecological balance of a region is destroyed, however, the gangue contains aluminosilicate mineral frameworks and slow-release mineral elements (Si, K, ca and the like), so that the piled gangue occupies a large amount of land resources, and natural processes such as weathering, leaching and the like and possible spontaneous combustion phenomena of the gangue can cause a series of environmental pollution problems such as atmospheric pollution, ecological damage and the like. In view of this, the resource utilization of the gangue is particularly urgent and important. In recent years, how to change the coal gangue into valuable is actively explored in the industry to realize the recycling value, and the activation of aluminosilicate mineral frameworks and slow-release mineral elements in the coal gangue by microorganisms is reported to effectively utilize the minerals in the coal gangue, however, the utilization efficiency of the aluminosilicate mineral frameworks and slow-release mineral elements in the conventional coal gangue is not high. The coal gangue contains various heavy metal elements including toxic components ‌ such As lead (Pb), chromium (Cr), mercury (Hg), arsenic (As), cadmium (Cd) and the like, under the flushing of rainwater, the heavy metal soluble components can infiltrate into soil and underground water to cause heavy metal pollution of the soil and the underground water, organic sulfur is contained in the coal gangue, long-term accumulation is carried out, high sulfur components induce soil acidification and desertification to destroy ecological balance of an area, in addition, dust generated by spontaneous combustion or accumulation of the coal gangue carries heavy metal, and enters the soil ‌ through dust fall or rainfall, so that the coal gangue accumulation area has high heavy metal pollution risk and occupies land resources, and if crops are planted on the land accumulated by the coal gangue, the heavy metal can enter a food chain through plant absorption to threaten human health. With the rapid development of modern agriculture, single-function microbial agents for treating heavy metal pollution or activating coal gangue have been reported, however, the single-function microbial agents for treating heavy metal pollution are generally only limited for specific heavy metals, the repairing capability of the single-function coal gangue activated strain is difficult to simultaneously meet multiple functions (such as nitrogen fixation, phosphorus dissolution, potassium dissolution and the like) required by the coal gangue activation, the activation efficiency is low, the coal gangue composition difference in different areas is large, the single microbial agents can not adapt to all physicochemical conditions (such as pH, heavy metal content and the like) to influence the stability and effect ‌ ‌, and in addition, the single microbial agents are difficult to colonize in complex soil environments and are easy to be subjected to competition inhibition of indigenous microorganisms, so that the number of active bacteria is rapidly reduced. Aiming at the defects of a single microbial inoculum, it is reported that a composite microbial inoculum is adopted to treat a coal gangue polluted area, the functional complementation is realized through the cooperation of multiple strains, the environmental adaptability and stability are enhanced, and the system restoration capability is improved. In addition, the growth habits (such as the demands on pH, humidity and nutrients) of different strains in the composite microbial inoculum are greatly different, and blind mixing can cause mutual inhibition among the strains, reduce the overall activity and influence the repairing effect. In addition, the industrial production of the composite microbial inoculum has serious challenges that if each strain is fermented independently, the problems of complex process flow, large equipment investment and high energy consumption exist, and if mixed fermentation is performed, the uneven growth rate is easily caused due to the difference of the requirements of different strains on the components of a culture medium and fermentation