Mine ecological restoration liquid, preparation method and application thereof

Abstract

The application relates to a mine ecological restoration liquid and a preparation method and application thereof. The preparation method of the mine ecological restoration liquid comprises the following steps: collecting a soil sample and mine gushing water in a mine to be restored; inoculating the soil sample in a first algal culture medium, carrying out enrichment and separation and purification treatment, and obtaining a microalgal strain; inoculating the microalgal strain into a culture medium containing the mine gushing water for culture, and preparing a mine ecological restoration liquid. The above preparation method can screen a local microalgal strain by using the soil environment of the mine to be restored, and can expand the culture by using the local mine gushing water as a culture medium, so that the mine ecological restoration liquid with excellent mine ecological restoration efficiency can be prepared at low cost.

Description

Technical Field

[0001] This application relates to the field of mine ecological restoration technology, and in particular to mine ecological restoration fluids, their preparation methods and applications. Background Technology

[0002] Mining environments are typically characterized by extreme pH levels, high concentrations of heavy metals, and high salinity, making traditional bioremediation technologies often ineffective in practical applications. For example, the activity of microorganisms is highly susceptible to fluctuations in water quality, making it difficult to maintain a stable and continuous effect; plants, on the other hand, have long growth cycles and low survival rates in barren and arid mining environments, failing to meet the urgent needs of mine ecological restoration.

[0003] Currently, algal bioremediation has attracted widespread attention due to its relatively good environmental tolerance and growth rate. However, algae still have limited tolerance to environmental conditions such as high salinity and high heavy metal content. In the special environment of mines, their growth and metabolic activities are also inhibited, resulting in limited ecological improvement effects and high costs. Summary of the Invention

[0004] To address the above issues, this application provides a mine ecological restoration fluid with high resource utilization, good ecological restoration effect and high restoration efficiency, as well as its preparation method and application.

[0005] The first aspect of this application provides a method for preparing a mine ecological restoration fluid.

[0006] A method for preparing a mine ecological restoration fluid includes the following steps:

[0007] Collect soil samples and mine water samples from the mine to be restored;

[0008] The soil sample was inoculated in the first algae culture medium, and after enrichment and purification, microalgae strains were obtained.

[0009] The microalgae strain was inoculated into a culture medium containing the mine inflow water for cultivation to prepare a mine ecological restoration solution.

[0010] In some implementations, the cultivation includes the following steps:

[0011] Microalgae strains were inoculated into the first culture vessel, and the culture temperature was adjusted to 20℃~30℃. After primary culture, primary algae strains were obtained. The first culture vessel contained a second algal culture medium.

[0012] In some embodiments, the cultivation further includes the following steps:

[0013] The primary algal strain was inoculated into the second culture vessel, and the airflow was adjusted to 0.6 L / min~1.5 L / min, with a light intensity of 60 μmol / (m²). 2 ·s)~130μmol / (m 2 •s), the culture temperature is 20℃~30℃, and after two-stage culture, a secondary algal strain is obtained. The second culture vessel contains a third algal culture medium.

[0014] In some embodiments, the cultivation further includes the following steps:

[0015] The secondary algal strain was inoculated into a photobioreactor, and the mine water was introduced. The ambient airflow was adjusted to 2 L / min~4 L / min, and the light intensity was 150 μmol / (m²). 2 ·s)~300μmol / (m 2 •s), the culture temperature is 20℃~30℃, and after three-stage culture, the third-stage algal strain is obtained.

[0016] In some embodiments, the cultivation further includes the following steps:

[0017] The tertiary algal strain and the mine inflow water were mixed to prepare a culture medium, which was then introduced into the raceway pool. The natural light intensity was adjusted to 100 μmol / (m²). 2 ·s)~150μmol / (m 2 •s), the culture temperature is 25℃~30℃, and after four stages of culture, the mine ecological restoration solution is obtained.

[0018] In some embodiments, the first algal culture medium, the second algal culture medium, and the third algal culture medium each independently include BG11 culture medium; the BG11 culture medium may or may not contain NaNO3.

[0019] In some embodiments, the dry weight content of algae in the mine ecological restoration solution is 1.5 g / L to 2 g / L.

[0020] In a second aspect, this application provides a mine ecological restoration fluid.

[0021] A mine ecological restoration fluid is prepared using the above-mentioned method for preparing mine ecological restoration fluid.

[0022] In a third aspect, this application provides a method for preparing the above-mentioned mine ecological restoration fluid, or the application of the above-mentioned mine ecological restoration fluid in the restoration of mine ecosystems.

[0023] The above-described preparation method utilizes the soil environment of the mine to be restored to screen for localized microalgae strains, and then uses mine water from that region as a culture medium for expansion, enabling the low-cost preparation of a mine ecological restoration solution with excellent mine ecological restoration efficiency. Specifically, the microalgae strains enriched and purified from the local mine soil naturally possess excellent tolerance to the specific water quality and environmental stresses of the mine, effectively solving the problem of their adaptability in extreme environments. Using mine water as a culture medium not only utilizes the nitrogen, phosphorus, and other nutrients in the water to support microalgae growth without the need for additional large-scale culture media, but also allows for further screening of rapidly growing algae strains with high stress resistance. Through the synergistic combination of these methods, this application effectively enhances the adaptability of the mine ecological restoration solution to the special environment of the mine while reducing resource consumption, achieving a dual improvement in resource recycling and treatment efficiency during the mine ecological restoration process. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 This is a flowchart of the preparation method of the mine ecological restoration fluid in one embodiment of this application. Detailed Implementation

[0026] To make the above-mentioned objectives, features, and advantages of this application more apparent and understandable, a detailed description of specific embodiments of this application is provided below. Many specific details are set forth in the following description to provide a thorough understanding of this application. However, this application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of this application. Therefore, this application is not limited to the specific embodiments disclosed below.

[0027] In the description of this application, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified. In this application, "at least one" means one or more, such as one, two, or more than two. "Multiple" or "several" means at least two, such as two, three, etc.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0029] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0030] Unless otherwise specified, all steps in this application may be performed sequentially or randomly. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the method may also include step (c), indicating that step (c) may be added to the method in any order. For example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0031] In this application, "above" or "below" includes the stated number. For example, "below 1" includes 1.

[0032] Unless otherwise specified, the viscosity parameters in this application refer to the viscosity measured at 25°C.

[0033] Unless otherwise specified, the average molecular weight of polymers used in this application refers to weight-average molecular weight.

[0034] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0035] Mining environments are typically characterized by extreme pH levels, high concentrations of heavy metals, and high salinity, making traditional bioremediation technologies often ineffective in practical applications. Currently, in algae-based mine ecological restoration applications, the limited tolerance of algae to high salinity and heavy metal environments inhibits their growth and metabolic activities, resulting in limited improvement on the mine ecosystem and high application costs.

[0036] To address the aforementioned problems, the first aspect of this application provides a method for preparing a mine ecological restoration fluid with high resource utilization, good ecological restoration effect, and high restoration efficiency. For an example, please refer to [link to example]. Figure 1 , Figure 1 This is a flowchart of the preparation method of the mine ecological restoration fluid in one embodiment of this application.

[0037] In some embodiments, the preparation method of the mine ecological restoration fluid includes the following steps:

[0038] S1. Collect soil samples and mine water samples from the mine to be restored;

[0039] S2. Soil samples were inoculated into the first algae culture medium, and microalgae strains were obtained after enrichment and purification.

[0040] S3. Microalgae strains are inoculated into a culture medium containing mine water and cultured to prepare a mine ecological restoration solution.

[0041] The above-described preparation method utilizes the soil environment of the mine to be restored to screen for localized microalgae strains, and then uses mine water from that region as a culture medium for expansion, enabling the low-cost preparation of a mine ecological restoration solution with excellent mine ecological restoration efficiency. Specifically, the microalgae strains enriched and purified from the local mine soil naturally possess excellent tolerance to the specific water quality and environmental stresses of the mine, effectively improving their adaptability to extreme environments. Using mine water as a culture medium not only utilizes the nitrogen, phosphorus, and other nutrients in the water to support microalgae growth without the need for additional large-scale culture media, but also allows for further screening of rapidly growing algae strains with high stress resistance. Through the synergistic combination of these methods, this application effectively enhances the adaptability of the mine ecological restoration solution to the special environment of the mine while reducing resource consumption, achieving a dual improvement in resource recycling and treatment efficiency during the mine ecological restoration process.

[0042] In the above preparation method, microalgae are directly cultured using mine water, reducing the consumption of water resources and nutrient solutions. Furthermore, the growth of microalgae primarily relies on natural photosynthesis, eliminating the need for complex equipment and energy-intensive processing techniques, further reducing energy consumption and remediation costs.

[0043] In some implementations, the method for obtaining mine water inflow is as follows:

[0044] A water inflow pool is set up at the mine outlet. The water is filtered by a filtration device in the pool and then diverted to a water source and a collection pool to obtain mine water.

[0045] In some embodiments, the filtration device includes a multi-stage filter. Optionally, the pore size of the filter mesh is 0.1 mm to 5 mm. Optionally, the pore size of the filter mesh can be, but is not limited to, 0.1 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, or other values ​​within the range of 0.1 mm to 5 mm.

[0046] In some embodiments, the turbidity of the filtered mine water is less than 300 NTU. Optionally, the turbidity of the filtered mine water is 80 NTU to 300 NTU.

[0047] In some implementations, enrichment includes the following steps:

[0048] Prepare a soil suspension containing soil samples, inoculate the soil suspension into the first algal culture medium, and adjust the light intensity to 60 μmol / (m²). 2 ·s)~130μmol / (m 2 The algae were cultured at a temperature of 20℃~30℃ and then allowed to stand for a period of time to obtain an enriched algal solution.

[0049] Optionally, during static incubation, the light intensity can be, but is not limited to, 60 μmol / (m²). 2 ·s), 80μmol / (m 2 ·s), 100μmol / (m 2 ·s), 130μmol / (m 2 ·s) or 60μmol / (m 2 ·s)~130μmol / (m 2 Other values ​​within the range of ·s).

[0050] In some embodiments, the static incubation period is 7 to 15 days. Optionally, the static incubation period can be, but is not limited to, 7, 8, 9, 10, 11, 12, 13, 14, 15 days or other values ​​within the range of 7 to 15 days.

[0051] Optionally, during static incubation, the incubation temperature can be, but is not limited to, 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, or other values ​​within the range of 20℃ to 30℃.

[0052] In some embodiments, the separation and purification process includes: gradient dilution of the enriched algal solution, plate inoculation, separation culture, and pure culture to obtain microalgae strains.

[0053] In some implementations, the cultivation includes the following steps:

[0054] Microalgae strains were inoculated into the first culture vessel, and the culture temperature was adjusted to 20℃~30℃. After primary culture, primary algae strains were obtained. The first culture vessel contained a second algal culture medium.

[0055] In some embodiments, the second algal culture medium occupies 40% to 75% of the working volume of the first culture vessel.

[0056] In some implementations, the primary culture time is 3 to 10 days. Optionally, the primary culture time can be, but is not limited to, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, or other values ​​within the range of 3 to 10 days.

[0057] Optionally, during the primary culture, the culture temperature can be, but is not limited to, 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, or other values ​​within the range of 20℃ to 30℃.

[0058] In some implementations, the cultivation process further includes the following steps:

[0059] The primary algal strain was inoculated into the second culture vessel, and the airflow was adjusted to 0.6 L / min~1.5 L / min, with a light intensity of 60 μmol / (m²). 2 ·s)~130μmol / (m 2 •s), the culture temperature is 20℃~30℃, and after two-stage culture, a secondary algal strain is obtained. The second culture vessel contains a third algal culture medium.

[0060] Optionally, during secondary culture, the light intensity can be, but is not limited to, 60 μmol / (m²). 2 ·s), 80μmol / (m 2 ·s), 100μmol / (m 2 ·s), 130μmol / (m 2 ·s) or 60μmol / (m 2 ·s)~130μmol / (m 2 Other values ​​within the range of ·s).

[0061] Optionally, during secondary culture, the culture temperature can be, but is not limited to, 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, or other values ​​within the range of 20℃ to 30℃.

[0062] In some embodiments, the third algal culture medium occupies 70% to 100% of the working volume of the second culture vessel. It can be understood that when the third algal culture medium occupies 100% of the working volume of the second culture vessel, the volume of the third algal culture medium is equal to that of the second culture vessel.

[0063] In some implementations, the secondary culture time is 6 to 10 days. Optionally, the secondary culture time can be, but is not limited to, 6 days, 7 days, 8 days, 9 days, 10 days, or other values ​​within the range of 6 to 10 days.

[0064] In some implementations, the cultivation process further includes the following steps:

[0065] Secondary algal strains were inoculated into the photobioreactor, mine water was introduced, and the ambient airflow was adjusted to 2 L / min~4 L / min, with a light intensity of 150 μmol / (m²). 2 ·s)~300μmol / (m 2 •s), the culture temperature is 20℃~30℃, and after three-stage culture, the third-stage algal strain is obtained.

[0066] Optionally, during tertiary culture, the light intensity can be, but is not limited to, 150 μmol / (m²). 2 ·s), 200μmol / (m 2 ·s), 250μmol / (m 2 ·s), 300μmol / (m 2 ·s) or 150μmol / (m 2 ·s)~3000μmol / (m 2 Other values ​​within the range of ·s).

[0067] Optionally, during tertiary culture, the culture temperature can be, but is not limited to, 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, or other values ​​within the range of 20℃ to 30℃.

[0068] In some implementations, the tertiary culture time is 6 to 10 days. Optionally, the tertiary culture time can be, but is not limited to, 6 days, 7 days, 8 days, 9 days, 10 days, or other values ​​within the range of 6 to 10 days.

[0069] In some implementations, the cultivation process further includes the following steps:

[0070] The tertiary algal strains and mine inflow water were mixed to prepare a culture medium, which was then introduced into the raceway pool. The natural light intensity was adjusted to 100 μmol / (m²). 2 ·s)~150μmol / (m 2 •s), the culture temperature is 25℃~30℃, and after four stages of culture, the mine ecological restoration solution is obtained.

[0071] Optionally, during quaternary incubation, the intensity of natural light can be, but is not limited to, 100 μmol / (m²). 2 ·s), 110μmol / (m 2 ·s), 130μmol / (m 2 ·s), 150μmol / (m 2 ·s) or 100μmol / (m 2 ·s)~150μmol / (m 2 Other values ​​within the range of ·s).

[0072] Optionally, during the fourth-stage culture, the culture temperature can be, but is not limited to, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, or other values ​​within the range of 25°C to 30°C.

[0073] In some embodiments, a four-stage culture is performed with stirring. Optionally, the stirring rate is 15 rpm to 40 rpm.

[0074] In some embodiments, the dry weight content of tertiary algal strains in the culture medium is 0.3 g / L to 0.5 g / L. Optionally, the dry weight content of tertiary algal strains in the culture medium can be, but is not limited to, 0.3 g / L, 0.35 g / L, 0.4 g / L, 0.45 g / L, 0.5 g / L, or other values ​​within the range of 0.3 g / L to 0.5 g / L.

[0075] In some embodiments, the dry weight content of algae in the mine ecological restoration solution is 1.5 g / L to 2 g / L. Optionally, the dry weight of algae in the mine ecological restoration solution can be, but is not limited to, 1.5 g / L, 1.6 g / L, 1.7 g / L, 1.8 g / L, 1.9 g / L, 2 g / L, or other values ​​within the range of 1.5 g / L to 2 g / L.

[0076] The second aspect of this application provides a mine ecological restoration fluid prepared using the above-described preparation method.

[0077] In a third aspect, this application provides a mine ecological restoration solution prepared by the above-described preparation method or the application of the above-described mine ecological restoration solution in the restoration of mine ecosystems.

[0078] The present application will be further described in detail below with reference to specific embodiments.

[0079] Unless otherwise specified, the raw materials used in the following specific embodiments and comparative examples are all commercially available products; the instruments used are all commercially available products; and the processes used are all conventionally selected by those skilled in the art unless otherwise specified.

[0080] Example 1

[0081] This embodiment provides a mine ecological restoration fluid.

[0082] The preparation method of the mine ecological restoration fluid is as follows:

[0083] A large water collection pool is set up at the mine outlet. The water flows into the collection pool after passing through a pretreatment filtration device. The pretreatment filtration device uses a multi-layer filter screen with mesh sizes of 5mm, 1mm and 0.5mm to remove large suspended particles. The turbidity of the resulting mine water is approximately 200 NTU.

[0084] Prepare a NaNO3-free BG11 medium and use it as the first, second, and third algal culture media. The specific formula is as follows: add 0.052 g of dipotassium hydrogen phosphate trihydrate, 0.075 g of magnesium sulfate heptahydrate, 0.036 g of calcium chloride dihydrate, 0.02 g of sodium carbonate, 0.006 g of citric acid, 0.006 g of ferric ammonium citrate, 0.001 g of EDTA, and 1 mL / L of trace element solution A5 per liter of water. The components of trace element solution A5 are: 2.86 g / L of boric acid, 1.86 g / L of manganese chloride tetrahydrate, 0.22 g / L of zinc sulfate heptahydrate, 0.39 g / L of sodium molybdate, 0.08 g / L of copper sulfate pentahydrate, and 0.05 g / L of cobalt nitrate hexahydrate.

[0085] Add 5 ml of NaNO3-containing, autoclaved algal culture medium (121°C, 30 min) to an Erlenmeyer flask. Collect soil samples from the mine to be remediated, add 150 ml of culture medium and 8 g of soil sample to a 250 ml Erlenmeyer flask, and stir for 30 min to obtain a soil suspension. Control the light intensity at 100 μmol / (m²) using a cool white fluorescent lamp. 2 ·s), under periodic shaking, the test tube was placed at 25°C and cultured for 14 days to obtain an enriched algal solution.

[0086] Prepare a 3% (w / v) agar solution with distilled water and sterilize it; prepare a double concentration of BG11 medium and sterilize it; mix the agar solution and BG11 medium evenly at 50°C to prepare a fourth algae medium with an agar concentration of 1.5 wt% for later use.

[0087] After diluting the enriched algal solution, the plates were inoculated onto the fourth algal culture medium. After the algal fallout, the algal fallout was picked up with a sterile loop and cultured on fresh fourth algal culture medium for 5 days to obtain a pure algal suspension. The algal fallout from the pure algal suspension was picked up and inoculated 3 times to obtain the microalgal strain.

[0088] The microalgae strain was inoculated into a 250ml Erlenmeyer flask, and 150ml of secondary algal culture medium was added. The flask was shaken periodically to prevent the microalgae strain from settling to the bottom. After culturing for 7 days, the primary algal strain was obtained.

[0089] The primary algal strain was inoculated into a 5L Erlenmeyer flask, forming a 5L working volume of the third algal culture medium. An air compressor was used to adjust the airflow to 1L / min, and the light intensity was controlled at 100 μmol / (m²). 2 •s), cultured at 25℃ for 8 days to obtain secondary algal strains.

[0090] The secondary algal strain was inoculated into a 200L photobioreactor, and 180L of mine water was added. An air compressor was used to adjust the airflow to 3L / min, and the light intensity was 200μmol / (m²). 2 •s), cultured at 25℃ for 8 days to obtain tertiary algal strains.

[0091] Tertiary algal strains were inoculated into an open raceway tank (culture area 30m²) at a dry weight of 0.4 g / L. 2 The culture medium depth is 0.25m. The track pool is lined with a black plastic film at the bottom, and the perimeter is covered with plastic turf. The track pool faces north with a higher elevation and south with a lower elevation. A glass greenhouse is used to control the temperature at 28℃, utilizing natural light for illumination. A shade net is used to control the light intensity at 125μmol / (m²). 2 •s), control the stirrer speed at 30rpm, and culture until the dry weight content of algae reaches 1.5g / L to obtain the mine ecological restoration solution.

[0092] Comparative Example 1

[0093] This embodiment provides a mine ecological restoration fluid.

[0094] The difference compared to Example 1 is that standard BG11 culture medium was used instead of mine water.

[0095] Test case

[0096] Parallel soil remediation experiments were conducted to assess the soil remediation capabilities of the mine ecological restoration fluids from Example 1 and Comparative Example 1. The soil (0-20cm topsoil) from the mine to be remediated in Example 1 was used as the experimental subject, with an experimental area of ​​100m². 2 According to 2g / m 2 Mine ecological restoration liquid was applied as base fertilizer, and 0.25 L / m² of surface water was applied daily.2 Then apply 1.5g / m every 8 days. 2 The mine ecological restoration solution was tested at an ambient temperature of 20℃~25℃ and a natural light intensity of 150~300μmol / (m²). 2 The soil was irradiated for 16 hours daily. Nine sampling points were evenly selected, and the soil organic matter content (potassium dichromate titration method) and soil moisture content (oven drying method) were measured before the experiment and after 60 days of the experiment. The test results are shown in Table 1.

[0097] Table 1. Restoration capacity test of the mine ecological restoration fluid in the examples and comparative examples.

[0098]

[0099] As shown in Table 1, by using soil samples and mine water from the mine to be remediated as the culture base for the mine ecological remediation solution, the solution demonstrated superior soil remediation capabilities for the local mine. Even in the harsh environment of mine water, the microalgae in the mine ecological remediation solution of Example 1 still exhibited rapid growth and efficient pollutant absorption capabilities, with a fast reproduction rate. They were also able to absorb nutrients such as carbon dioxide, nitrogen, and phosphorus from the mine water through photosynthesis, significantly improving water quality by adsorbing and transforming pollutants such as heavy metals. Furthermore, they reduced greenhouse gas emissions through carbon sequestration, contributing to the long-term stability of the mine's ecological environment and the restoration of the ecosystem. In addition, the nutrients in the mine ecological remediation solution also possess soil-improving capabilities.

[0100] It is evident that, compared to traditional biological treatment methods, the use of the mine ecological restoration liquid of this application for mine ecological restoration can effectively improve the soil organic matter content and soil moisture content. On the basis of soil improvement (continuous application for 15 days, and the soil condition can be effectively improved after 6 months), more tolerant native vegetation can be planted, such as crested wheatgrass, red bean grass, hairy rabbit lip flower, solitary greens, needlegrass, camel thorn, and small-flowered oxalis.

[0101] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0102] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this invention patent should be determined by the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. A method for preparing a mine ecological restoration fluid, characterized in that, Includes the following steps: Collect soil samples and mine water samples from the mine to be restored; The soil sample was inoculated in the first algae culture medium, and after enrichment and purification, microalgae strains were obtained. The microalgae strain is inoculated into a culture medium containing the mine inflow water for cultivation to prepare the mine ecological restoration solution.

2. The method for preparing the mine ecological restoration fluid according to claim 1, characterized in that, The cultivation process includes the following steps: The microalgae strain was inoculated into a first culture vessel, and the culture temperature was adjusted to 20℃~30℃. After primary culture, a primary algae strain was obtained. The first culture vessel contained a second algal culture medium.

3. The method for preparing the mine ecological restoration fluid according to claim 2, characterized in that, The cultivation process also includes the following steps: The primary algal strain was inoculated into the second culture vessel, and the ambient airflow was adjusted to 0.6 L / min~1.5 L / min, with a light intensity of 60 μmol / (m²). 2 ·s)~130μmol / (m 2 •s), the culture temperature is 20℃~30℃, and after two-stage culture, a secondary algal strain is obtained. The second culture vessel contains a third algal culture medium.

4. The method for preparing the mine ecological restoration fluid according to claim 3, characterized in that, The cultivation process also includes the following steps: The secondary algal strain was inoculated into a photobioreactor, and the mine water was introduced. The ambient airflow was adjusted to 2 L / min~4 L / min, and the light intensity was 150 μmol / (m²). 2 ·s)~300μmol / (m 2 •s), the culture temperature is 20℃~30℃, and after three-stage culture, the third-stage algal strain is obtained.

5. The method for preparing the mine ecological restoration fluid according to claim 4, characterized in that, The cultivation process also includes the following steps: The tertiary algal strain and the mine inflow water were mixed to prepare a culture medium, which was then introduced into the raceway pool. The natural light intensity was adjusted to 100 μmol / (m²). 2 ·s)~150μmol / (m 2 •s), the culture temperature is 25℃~30℃, and after four stages of culture, the mine ecological restoration solution is obtained.

6. The method for preparing the mine ecological restoration fluid according to claim 5, characterized in that, The dry weight content of the tertiary algal strains in the culture medium is 0.3 g / L to 0.5 g / L.

7. The method for preparing the mine ecological restoration fluid according to claim 5, characterized in that, The first algal culture medium, the second algal culture medium, and the third algal culture medium each independently include BG11 culture medium; the BG11 culture medium may or may not contain NaNO3.

8. The method for preparing the mine ecological restoration fluid according to any one of claims 1 to 7, characterized in that, The dry weight content of algae in the mine ecological restoration solution is 1.5g / L~2g / L.

9. A mine ecological restoration liquid, characterized in that, It is prepared using the method described in any one of claims 1 to 8 for preparing the mine ecological restoration fluid.

10. The application of a mine ecological restoration solution prepared by the method of any one of claims 1 to 8, or the mine ecological restoration solution of claim 9, in the restoration of a mine ecosystem.

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