Method for repairing rock slope of mine by using microorganisms

Abstract

The invention discloses a method for repairing a rock slope of a mine by utilizing microorganisms, and belongs to the field of ecological restoration of slopes. According to the method, cultured bacterial liquid is sprayed on an ecological slope protection base material on a side slope through an irrigation system, the ecological slope protection base material is reinforced, and the ecological restoration work of the rock side slope is completed. According to the method, the calcium carbonate is prepared by utilizing microorganisms, the utilized microorganisms are rich in resource, efficient in reaction, environment-friendly and pollution-free, and the solid waste alkaline residues are used as a calcium source, so that resource reutilization is realized, the environmental protection pressure is greatly reduced, and the microorganisms make a greater breakthrough in the field of slope restoration.

Description

Technical Field

[0001] The invention relates to the field of ecological restoration of rock slopes, and in particular to a method for restoring rock slopes in mines 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 is microbially induced calcium carbonate precipitation (MICP). MICP technology uses urease-producing microorganisms to hydrolyze urea and combine it with free calcium ions to generate calcium carbonate precipitation. This calcium carbonate produced by MICP technology can fill in the pores of the sudden, cementing soil particles together and giving the sudden a certain strength. At present, this technology has made a huge breakthrough in engineering applications.

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

[0004] Chinese invention document (CN116695740A) discloses a mine slope repair system and method, which uses anchor rods to anchor a straw grid on the slope. The straw grid has a plant growth groove, which can better stabilize the ecological substrate required for plant growth on the slope. However, its calcium source is also calcium chloride, which is relatively expensive and is not suitable for large-scale mine 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 defect of the prior art. The present invention provides a method for repairing rock slopes in mines by using microorganisms, using urease produced by microorganisms to hydrolyze urea to produce carbonate ions, which are combined with free calcium ions in alkali slag to generate calcium carbonate precipitation, thereby reinforcing the ecological slope protection base material and stabilizing the ecological slope protection base material. At the same time, the solid waste of alkali slag is utilized to reduce environmental pressure.

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

[0008] A method for repairing a rocky slope in a mine using microorganisms, wherein the repair of the rocky slope in a mine is completed by reinforcing an ecological slope protection substrate with microorganisms, wherein the microorganism is Bacillus subtilis, and specifically comprises the following steps:

[0009] Step 1, preparation of bacterial solution

[0010] Step 1.1, use the formula recommended by the American Type Culture Collection ATCC1376NH 4 -YE prepares the culture solution, and then sterilizes the culture solution in an autoclave, the sterilization time is set to 20 minutes to 30 minutes, the temperature of the autoclave is set to 101 degrees Celsius to 123 degrees Celsius, and then places it on a clean bench for ultraviolet sterilization and ventilation, and waits for the temperature of the culture solution to drop to room temperature before use;

[0011] Step 1.2, after the temperature of the culture solution drops to room temperature, take the culture solution into a container, add Bacillus pastoris into the culture solution with a sterile pipette to obtain a bacterial solution, wherein the volume ratio of Bacillus pastoris to the culture solution is 1:100; place the bacterial solution in an incubator for culturing, wherein the incubator temperature is set at 28 degrees Celsius to 32 degrees Celsius, the oscillation frequency is set at 180 times / minute to 220 times / minute, and the culturing time is set at 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 600nm, i.e. OD 600 Value, in OD 600 =0.8~1.2, take it out for use;

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

[0014] The ecological slope protection substrate is mixed with planting soil, habitat substrate improver, organic fertilizer, compound fertilizer, seeds, calcium source and urea, and the mass fractions 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 substrate obtained in step 2 according to the set moisture content A, stir it evenly, and then spray it evenly on the rock slope, and ensure that the thickness of the ecological slope protection substrate on the rock slope reaches 8cm-12cm;

[0017] 3-5 days after the spraying of the ecological slope protection substrate is completed, the bacterial solution prepared in step 1 is poured on the slope using an irrigation system at a dosage 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 American Type Culture Collection recommends formula ATCC 1376NH 4 -YE prepared culture solution composition: 20g yeast extract, 10g (NH 4 ) 2 So 4 , 0.13 mol / L Tris Buffer, add deionized water to 1 L, and adjust the culture medium pH to 9.0 with 1 mol / L HCl.

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

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

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

[0024] 2. The present invention uses alkali residue as the calcium source of MICP technology, so that solid waste can be reused, resource utilization is achieved, and environmental pressure is reduced. Alkali residue refers to the alkaline waste residue discharged during alkali production and alkali treatment in industrial production. It includes waste residue discharged during the ammonium alkali method alkali production process and alkaline waste residue discharged from other industrial production processes. Alkali residue contains rich calcium chloride and other calcium salts, which can be used as a calcium source. 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 is further described below in conjunction with the accompanying drawings and specific embodiments.

[0027] Figure 1 Is a flow chart of the present invention, consisting of Figure 1 It can be seen that the method of repairing rock slopes in mines by using microorganisms in the present invention completes the repair of rock slopes in mines by reinforcing the ecological slope protection substrate with microorganisms, and the microorganism is Bacillus subtilis. Specifically, the present invention utilizes urease-producing microorganisms to hydrolyze urea to produce carbonate ions, which are combined with calcium ions produced by hydrolysis of alkali residues to react in the pores of the soil to produce calcium carbonate precipitation, cementing the soil particles together to give it a certain strength.

[0028] Example 1.

[0029] Step 1, preparation of bacterial solution

[0030] Step 1.1, use the formula recommended by the American Type Culture Collection ATCC1376NH 4 -YE prepares the culture medium, and then places the culture medium in the autoclave for sterilization. The sterilization time is set to 20 minutes to 30 minutes, and the temperature of the autoclave is set to 101 degrees Celsius to 123 degrees Celsius. Then place it on the clean bench for ultraviolet sterilization and ventilation. Wait until the temperature of the culture medium drops to room temperature before use.

[0031] Step 1.2, after the temperature of the culture solution drops to room temperature, take the culture solution into a container, use a sterile pipette to add Bacillus pastoris to the culture solution to obtain a bacterial solution, the volume ratio of Bacillus pastoris to the culture solution is 1:100; put the bacterial solution into an incubator for culturing, the incubator temperature is set to 28 degrees Celsius to 32 degrees Celsius, the oscillation frequency is set to 180 times / minute to 220 times / minute, and the culturing 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 600nm, 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 mixed with 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 evenly, 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 spraying of the ecological slope protection substrate is completed, the bacterial solution prepared in step 1 is poured on the slope using an irrigation system at a dosage 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] Step 1.1, use the formula recommended by the American Type Culture Collection ATCC1376NH 4 -YE prepares the culture medium, and then places the culture medium in the autoclave for sterilization. The sterilization time is set to 25 minutes, and the temperature of the autoclave is set to 123 degrees Celsius. Then place it on the clean bench for ultraviolet sterilization and ventilation. Wait until the temperature of the culture medium drops to room temperature before use.

[0043] Step 1.2, after the temperature of the culture solution drops to room temperature, take the culture solution into a container, use a sterile pipette to add Bacillus pastoris to the culture solution to obtain a bacterial solution, the volume ratio of Bacillus pastoris to the culture solution is 1:100; put the bacterial solution into an incubator for culturing, the incubator temperature is set to 30 degrees Celsius, the oscillation frequency is set to 200 times / minute, and the culturing 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 600nm, i.e. OD 600 Value, in OD 600 =1.0, take it out for later 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 evenly 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 and evenly 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 and evenly 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 and evenly, and ensure that its moisture content is 25%. 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 12 cm.

[0056] Three days after the ecological slope protection substrate was sprayed, the bacterial solution cultured in step 1 was 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 above three embodiments, the American Type Culture Collection recommends the formula ATCC 1376NH 4 -YE prepared culture solution composition: 20g yeast extract, 10g (NH 4 ) 2 SO 4 , 0.13 mol / L Tris Buffer, add deionized water to 1 L, and adjust the pH of the culture solution to 9.0 with 1 mol / L HCl. The slope of the rock slope of the mine is 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 rock slopes in mines using microorganisms, characterized in that: The ecological slope protection substrate is reinforced by microorganisms to complete the repair of the rock slope of the mine. The microorganism is Bacillus subtilis. Specifically, the following steps are included: Step 1, preparation of bacterial solution Step 1.1, prepare the culture solution using the formula ATCC1376NH4-YE recommended by the American Type Culture Collection, then sterilize the culture solution in an autoclave, set the sterilization time to 20 minutes to 30 minutes, set the autoclave temperature to 101 degrees Celsius to 123 degrees Celsius, then place it on a clean bench for ultraviolet sterilization and ventilation, and use it when the temperature of the culture solution drops to room temperature; Step 1.2, after the temperature of the culture solution drops to room temperature, take the culture solution into a container, add Bacillus pastoris into the culture solution with a sterile pipette to obtain a bacterial solution, wherein the volume ratio of Bacillus pastoris to the culture solution is 1:100; place the bacterial solution in an incubator for culturing, wherein the incubator temperature is set at 28 degrees Celsius to 32 degrees Celsius, the oscillation frequency is set at 180 times / minute to 220 times / minute, and the culturing time is set at 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, take it out for use; Step 2: Preparation of ecological slope protection substrate The ecological slope protection substrate is mixed with planting soil, habitat substrate improver, organic fertilizer, compound fertilizer, seeds, calcium source and urea, and the mass fractions 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 substrate obtained in step 2 according to the set moisture content A, stir it evenly, and then spray it evenly on the rock slope, and ensure that the thickness of the ecological slope protection substrate on the rock slope reaches 8cm-12cm; 3-5 days after the spraying of the ecological slope protection substrate is completed, the bacterial solution prepared in step 1 is poured on the slope using an irrigation system at a dosage of 6L-12L per square meter to complete the reinforcement of the ecological slope protection substrate.

2. A method for repairing rock slopes in mines using microorganisms according to claim 1, characterized in that: The calcium source is alkali slag.

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

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

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

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