A method for repairing mine rock slopes using microorganisms

Abstract

The present invention discloses a method for restoring rocky slopes in mines using microorganisms, belonging to the field of slope ecological restoration. The method utilizes an irrigation system to spray a cultured bacterial solution onto an ecological slope protection substrate on the slope, thereby reinforcing the ecological slope protection substrate and completing the ecological restoration of the rocky slope. The present invention utilizes microorganisms to prepare calcium carbonate, utilizing abundant microbial resources, achieving efficient reactions, and being environmentally friendly and pollution-free. Solid waste alkali residue is reused as a calcium source, achieving not only resource recycling but also significantly reducing environmental pressure, enabling a greater breakthrough in the application of microorganisms in the field of slope restoration.

Description

Technical Field

[0001] The present invention relates to the field of ecological restoration of rock slopes, and in particular to a method for restoring mine rock slopes by utilizing microorganisms. Background Art

[0002] For rock slopes in mines, due to their steep slopes and smooth slope surfaces, it is difficult for ecological restoration substrates to stabilize on the rock side for plant seeds to germinate and take root. In order to address the problems of loose ecological slope protection substrates and difficulty in attaching to the surface of rock slopes during rock slope restoration, the current main technology used is Microbially Induced Calcite Precipitation (MICP). MICP technology uses urease-producing microorganisms to hydrolyze urea, which combines with free calcium ions to form calcium carbonate precipitates. This calcium carbonate produced by MICP technology can fill the pores in the sudden pores, cementing soil particles together and giving the sudden pores a certain strength. At present, this technology has made great breakthroughs in engineering applications.

[0003] A Chinese invention patent document (CN111997039B) discloses a soil reinforcement method using W-shaped interlocking piles based on MICP technology. This method utilizes MICP technology to design W-shaped interlocking piles, which improves the soil reinforcement effect of MICP technology. However, the substrate for vegetation growth is difficult to adhere to the slope surface for a long time, which means that the problem of long-term stable reinforcement is not solved.

[0004] A Chinese invention document (CN116695740A) discloses a mine slope repair system and method. This method uses anchor rods to anchor a straw grid to the slope. The straw grid has plant growth grooves, effectively stabilizing the ecological substrate required for plant growth on the slope. However, the calcium source used is calcium chloride, which is relatively expensive and unsuitable for large-scale rock slope repair.

[0005] Therefore, in order to solve the above problems, the present invention is based on MICP technology and uses alkali slag as a calcium source to invent a method for repairing mine rock slopes using microorganisms. The invention of this method has important engineering practical significance and theoretical research value for the ecological restoration of rock slopes. Summary of the Invention

[0006] The technical problem to be solved by the present invention is the drawback of the existing technology. This invention provides a method for remediating rock slopes in mines using microorganisms. The method uses urease produced by microorganisms to hydrolyze urea to produce carbonate ions, which combine with free calcium ions in alkali residue to form calcium carbonate precipitates. This reinforces and stabilizes the ecological slope protection base material, while also utilizing the solid waste of alkali residue and reducing environmental pressure.

[0007] The technical solution adopted in the present invention is as follows:

[0008] A method for repairing a rocky slope in a mine using microorganisms, wherein the microorganism is Bacillus subtilis, is used to reinforce an ecological slope protection substrate to complete the repair of the rocky slope in the mine. Specifically, the method comprises the following steps:

[0009] Step 1, preparation of bacterial solution

[0010] In step 1.1, prepare the culture medium using the American Type Culture Collection's recommended formula, ATCC 1376 NH4-YE. Sterilize the culture medium in an autoclave for 20 to 30 minutes at a temperature of 101 to 123 degrees Celsius. Place the culture medium on a clean bench for ultraviolet sterilization and ventilation. Use the culture medium after it cools to room temperature.

[0011] In step 1.2, after the temperature of the culture solution has dropped to room temperature, the culture solution is placed in a container and Bacillus pastoris is added to the culture solution using a sterile pipette to obtain a bacterial solution, wherein the volume ratio of Bacillus pastoris to the culture solution is 1:100; the bacterial solution is placed in an incubator for cultivation, wherein the incubator temperature is set to 28°C to 32°C, the oscillation frequency is set to 180 times / minute to 220 times / minute, and the incubation time is set to 48 hours to 72 hours;

[0012] Step 1.3: Measure the absorbance of the bacterial solution obtained in step 1.2 at a wavelength of 600 nm, i.e., OD 600 Value, in OD 600 =0.8~1.2 when it is taken out for use;

[0013] Step 2: Preparation of ecological slope protection substrate

[0014] The ecological slope protection substrate is prepared by mixing planting soil, habitat substrate improver, organic fertilizer, compound fertilizer, seeds, calcium source and urea, and the mass proportions thereof are 80-120:4-6:2-4:1-2:0.1-0.5:40-60:1-2 in the above order;

[0015] Step 3: Repair of rock slopes

[0016] Add a certain amount of water to the ecological slope protection base material obtained in step 2 according to the set moisture content A, stir it thoroughly, and then spray it evenly on the rock slope, and ensure that the thickness of the ecological slope protection base material on the rock slope reaches 8cm-12cm;

[0017] 3-5 days after the ecological slope protection substrate is sprayed, the bacterial solution prepared in step 1 is poured on the slope using an irrigation system at a rate of 6L-12L per square meter to complete the reinforcement of the ecological slope protection substrate.

[0018] Preferably, the calcium source is alkali slag.

[0019] Preferably, the slope of the mine rock slope is 30° to 60°.

[0020] Preferably, the culture medium prepared with the American Type Culture Collection recommended formula ATCC 1376 NH4-YE is composed of: 20g yeast extract, 10g (NH4)2So4, 0.13mol / L Tris Buffer, deionized water is added to 1L, and the culture medium pH is adjusted to 9.0 with 1mol / LHCl.

[0021] Preferably, the moisture content A=20%-25%.

[0022] Compared with the existing technology, this technology has the following beneficial effects:

[0023] 1. The present invention utilizes MICP technology to cause calcium carbonate precipitation to be generated in the soil pores, which can better cement the soil particles together, making it have greater strength and better cementing the ecological slope protection base material to the slope.

[0024] 2. The present invention uses alkali residue as a calcium source for MICP technology, allowing solid waste to be reused, achieving resource utilization and reducing environmental pressure. Alkali residue refers to alkaline waste residue discharged during alkali production and alkali treatment in industrial production. This includes waste residue discharged during the ammonium alkali process and alkaline waste residue discharged from other industrial production processes. Alkali residue is rich in calcium chloride and other calcium salts, which can be used as a calcium source. Using alkali residue as a calcium source greatly reduces environmental pressure. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a flow chart of the present invention. DETAILED DESCRIPTION

[0026] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0027] Figure 1 It is a flow chart of the present invention, consisting of Figure 1 As can be seen, the present invention utilizes microorganisms to repair rock slopes in mines, achieving the restoration of rock slopes by reinforcing ecological slope protection substrates with microorganisms. The microorganism is Bacillus parviflorus. Specifically, the present invention utilizes urease-producing microorganisms to hydrolyze urea to produce carbonate ions. These ions combine with calcium ions produced by the hydrolysis of alkali residues, reacting in the pores of the soil to produce calcium carbonate precipitation, cementing the soil particles together and imparting a certain strength.

[0028] Example 1.

[0029] Step 1, preparation of bacterial solution

[0030] In step 1.1, prepare the culture medium using the American Type Culture Collection's recommended formula, ATCC 1376NH4-YE. Sterilize the culture medium in an autoclave for 20 to 30 minutes at a temperature of 101 to 123 degrees Celsius. Place the culture medium on a clean bench for ultraviolet sterilization and ventilation. Use the culture medium after it cools to room temperature.

[0031] In step 1.2, after the temperature of the culture solution drops to room temperature, the culture solution is placed in a container, and Bacillus pastoris is added to the culture solution using a sterile pipette to obtain a bacterial solution, wherein the volume ratio of Bacillus pastoris to the culture solution is 1:100; the bacterial solution is placed in an incubator for cultivation, and the incubator temperature is set to 28°C to 32°C, the oscillation frequency is set to 180 times / minute to 220 times / minute, and the incubation time is set to 48 hours to 72 hours.

[0032] Step 1.3: Measure the absorbance of the bacterial solution obtained in step 1.2 at a wavelength of 600 nm, i.e., OD 600 Value, in OD 600 =0.8~1.2 when it is taken out for use.

[0033] Step 2: Preparation of ecological slope protection substrate

[0034] The ecological slope protection substrate is prepared by mixing planting soil, habitat substrate improver, organic fertilizer, compound fertilizer, seeds, calcium source and urea, and the mass proportions thereof are 80-120:4-6:2-4:1-2:0.1-0.5:40-60:1-2 in the above order.

[0035] In this embodiment, the calcium source is alkali slag.

[0036] Step 3: Repair of rock slopes

[0037] Add a certain amount of water to the ecological slope protection base material obtained in step 2 according to the set moisture content A, stir it thoroughly, and then spray it evenly on the rock slope, and ensure that the thickness of the ecological slope protection base material on the rock slope reaches 8cm-12cm.

[0038] 3-5 days after the ecological slope protection substrate is sprayed, the bacterial solution prepared in step 1 is poured on the slope using an irrigation system at a rate of 6L-12L per square meter to complete the reinforcement of the ecological slope protection substrate.

[0039] In this embodiment, the moisture content A=20%-25%.

[0040] Example 2

[0041] Step 1, preparation of bacterial solution

[0042] In step 1.1, prepare the culture medium using the American Type Culture Collection's recommended formula, ATCC 1376NH4-YE. Sterilize the culture medium in an autoclave for 25 minutes at 123°C. Place the culture medium on a clean bench for UV sterilization and ventilation. Use the culture medium after it cools to room temperature.

[0043] In step 1.2, after the temperature of the culture solution drops to room temperature, the culture solution is placed in a container and Bacillus pastoris is added to the culture solution using a sterile pipette to obtain a bacterial solution, wherein the volume ratio of Bacillus pastoris to the culture solution is 1:100; the bacterial solution is placed in an incubator for cultivation, the incubator temperature is set to 30 degrees Celsius, the oscillation frequency is set to 200 times / minute, and the incubation time is set to 60 hours.

[0044] Step 1.3: Measure the absorbance of the bacterial solution obtained in step 1.2 at a wavelength of 600 nm, i.e., OD 600 Value, in OD 600 =1.0, take it out for use.

[0045] Step 2: Preparation of ecological slope protection substrate

[0046] Take 120kg of planting soil, 4kg of habitat substrate improver, 4kg of organic fertilizer, 2kg of compound fertilizer, 0.1kg of seeds, 50kg of alkali residue, and 2kg of urea and mix them thoroughly to obtain the ecological slope protection substrate.

[0047] Step 3: Repair of rock slopes

[0048] Add 45.5 kg of water to the ecological slope protection base material obtained in step 2, mix it thoroughly to ensure that its moisture content is 25%, and then spray it evenly on the rock slope, and ensure that the thickness of the ecological slope protection base material on the rock slope reaches 10 cm.

[0049] Three days after the ecological slope protection substrate was sprayed, the bacterial solution prepared in step 1 was sprayed at 7L / m 2 The amount is poured onto the slope using the irrigation system to complete the reinforcement of the ecological slope protection base material.

[0050] Example 3

[0051] Step 1 is the same as Example 2.

[0052] Step 2: Preparation of ecological slope protection substrate

[0053] Take 110kg of planting soil, 4kg of habitat substrate improver, 3kg of organic fertilizer, 2kg of compound fertilizer, 0.5kg of seeds, 60kg of alkali residue, and 2kg of urea, mix them thoroughly to obtain the ecological slope protection substrate.

[0054] Step 3: Repair of rock slopes

[0055] Add 45.25 kg of water to the ecological slope protection base material obtained in step 2, mix thoroughly to ensure that its moisture content is 25%, and then evenly spray it on the rock slope, ensuring that the thickness of the ecological slope protection base material on the rock slope reaches 12 cm.

[0056] Three days after the ecological slope protection substrate is sprayed, the bacterial solution cultured in step 1 is heated at 7L / m 2 The amount is sprayed on the ecological slope protection base material using the irrigation system to complete the reinforcement and use of the ecological slope protection base material.

[0057] In the three examples above, the culture medium prepared using the American Type Culture Collection's recommended formula, ATCC 1376 NH4-YE, consisted of 20 g yeast extract, 10 g (NH4)2SO4, and 0.13 mol / L Tris Buffer, with deionized water added to 1 L. The pH of the culture medium was adjusted to 9.0 with 1 mol / L HCl. The slope of the rocky mine slope ranged from 30° to 60°.

[0058] In this embodiment, a habitat substrate modifier produced by Hubei Runzhi Ecological Technology Co., Ltd. is selected. This modifier can adjust soil texture, enhance soil water retention and aeration, adjust soil acidity (pH value) and increase soil nutrient content, promote the growth and reproduction of soil microorganisms, and improve the growth environment of plant roots.

Claims

1. A method for repairing mine rock slopes using microorganisms, characterized in that: The restoration of the rock slope of the mine is completed by reinforcing the ecological slope protection base material with microorganisms. The microorganism is Bacillus subtilis. Specifically, the process includes the following steps: Step 1, preparation of bacterial solution In step 1.1, prepare the culture medium using the American Type Culture Collection's recommended formula, ATCC 1376 NH4-YE. Sterilize the culture medium in an autoclave for 20 to 30 minutes at a temperature of 101 to 123 degrees Celsius. Place the culture medium on a clean bench for ultraviolet sterilization and ventilation. Use the culture medium after it cools to room temperature. In step 1.2, after the temperature of the culture solution has dropped to room temperature, the culture solution is placed in a container and Bacillus pastoris is added to the culture solution using a sterile pipette to obtain a bacterial solution, wherein the volume ratio of Bacillus pastoris to the culture solution is 1:100; the bacterial solution is placed in an incubator for cultivation, wherein the incubator temperature is set to 28°C to 32°C, the oscillation frequency is set to 180 times / minute to 220 times / minute, and the incubation time is set to 48 hours to 72 hours; Step 1.3: Measure the absorbance of the bacterial solution obtained in step 1.2 at a wavelength of 600 nm, i.e., OD 600 Value, in OD 600 =0.8~1.2 when it is taken out for use; Step 2: Preparation of ecological slope protection substrate The ecological slope protection substrate is prepared by mixing planting soil, habitat substrate improver, organic fertilizer, compound fertilizer, seeds, calcium source and urea, and the mass proportions thereof are 80-120:4-6:2-4:1-2:0.1-0.5:40-60:1-2 in the above order; Step 3: Repair of rock slopes Add a certain amount of water to the ecological slope protection base material obtained in step 2 according to the set moisture content A, stir it thoroughly, and then spray it evenly on the rock slope, and ensure that the thickness of the ecological slope protection base material on the rock slope reaches 8cm-12cm; 3-5 days after the ecological slope protection substrate is sprayed, the bacterial solution prepared in step 1 is poured on the slope using an irrigation system at a rate of 6L-12L per square meter to complete the reinforcement of the ecological slope protection substrate.

2. The method for repairing a mine rock slope using microorganisms according to claim 1, characterized in that: The calcium source is alkali slag.

3. The method for repairing mine rock slopes using microorganisms according to claim 1, characterized in that: The slope of the mine rock slope is 30° to 60°.

4. The method for repairing a mine rock slope using microorganisms according to claim 1, characterized in that: The composition of the culture medium prepared with the American Type Culture Collection's recommended formula ATCC 1376 NH4-YE is: 20 g yeast extract, 10 g (NH4)2SO4, 0.13 mol / L Tris Buffer, deionized water is added to 1 L, and the culture medium pH is adjusted to 9.0 with 1 mol / L HCl.

5. The method for repairing mine rock slopes using microorganisms according to claim 1, characterized in that: The moisture content A=20%-25%.

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