Strain solidesulfovibrio magneticus sm-4 and applications thereof

Abstract

The present application relates to the technical field of microbial remediation, and provides a strain Solidesulfovibrio magneticus Sm-4 and its application. The strain Sm-4 has a reduction effect on SO4 2‑ and heavy metals, and has a solidification effect on heavy metals, thereby providing a bacterial resource for removing SO4 2‑ and heavy metals in AMD water bodies and heavy metal contaminated soil. The magnetic soil desulfurization vibrio Sm-4 provided by the present application can tolerate high-concentration SO4 2‑ and remove heavy metals, SO4 2‑ has a high reduction rate, and has a good treatment effect on SO4 2‑ and heavy metals in soil and water bodies, and has a wide application prospect in in-situ remediation technologies such as acid mine drainage, contaminated soil around waste mine areas, and heavy metal contaminated farmland soil.

Description

Technical Field

[0001] This invention relates to the field of microbial remediation technology, and more specifically, to strains. Solidesulfovibrio magneticus Sm-4 and its applications. Background Technology

[0002] The indiscriminate discharge of industrial wastewater, agricultural irrigation water, and domestic sewage is a major cause of water and surrounding soil pollution. Among the many pollution problems, the accumulation and discharge of acidic mine wastewater (AMD) is one of the main issues in mining areas, accounting for approximately 10% of my country's total industrial wastewater discharge. Since most natural metal and non-metal mineral deposits are composed of sulfide minerals, these sulfide minerals continuously react with oxygen and water in the environment during mining or stockpiling, leading to the formation of sulfates and the leaching of heavy metals, resulting in acidic mine wastewater. The heavy metal pollution caused by AMD leads to extremely serious water quality deterioration and surrounding soil pollution, characterized by high toxicity, difficulty in removal, and easy bioaccumulation. Once the wastewater overflows, the impact on the surrounding ecological environment is immeasurable.

[0003] To address water and soil pollution caused by acidic mine wastewater, researchers have continuously explored and proposed various remediation technologies to reduce its damage to the ecological environment. These include physical remediation technologies such as adsorption, ion exchange, and membrane treatment; chemical remediation technologies such as neutralization precipitation, constructed wetlands combined with lime precipitation, and continuous alkali production systems; and microbial in-situ remediation, an emerging technology for remediating AMD (Ammonium Acid) pollution. Physical remediation technologies are costly and have poor sustainability. Chemical remediation technologies require large amounts of neutralizing and precipitating agents, but the solidified heavy metals are prone to back-dissolution, causing secondary pollution. Microbial in-situ remediation, on the other hand, is simple to operate, has low investment costs, and provides long-lasting heavy metal solidification, showing promise for sustainable remediation. However, current challenges include a lack of resistance to acid and high concentrations of SO4. 2- Microbial strains to ensure SO4 2- It has a high removal rate and a long-lasting solidification of heavy metals. Summary of the Invention

[0004] The purpose of this invention is to provide strains Solidesulfovibrio magneticus Sm-4 and its applications.

[0005] To achieve the objectives of this invention, in a first aspect, this invention provides strain Sm-4, which was enriched and isolated from acidic mine drainage sediment samples from a copper mine in Yunnan Province, and is classified and named as follows: Solidesulfovibrio magneticusThe strain is now deposited at the China General Microbiological Culture Collection Center, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100101, China, accession number CGMCC No. 41191, deposit date April 17, 2024.

[0006] In a second aspect, the present invention provides a microbial preparation comprising one or more of the following: bacterial cells, bacterial powder, bacterial suspension, or bacterial extract selected from the strain Sm-4.

[0007] Thirdly, the present invention provides a method for preparing the microbial preparation, the method comprising the step of culturing the strain Sm-4.

[0008] Furthermore, the strain Sm-4 is cultured at 25-45°C and pH 4.5-7.0, preferably at 30-35°C and pH 6.0-7.0, and more preferably at 30°C and pH 6.0.

[0009] Furthermore, the strain Sm-4 was cultured using a modified Baars medium.

[0010] Preferably, the modified Baars medium contains 2 g / L MgSO4·7H2O, 5 g / L sodium citrate dihydrate, 1 g / L CaSO4·2H2O, 1 g / L NH4Cl, 0.5 g / L K2HPO4, and 3.5 g / L sodium lactate, prepared with deionized water, pH 6; 0.1 mL of filtered and sterilized 5% Fe(NH4)2•(SO4)2 is added before inoculation.

[0011] Preferably, the culture conditions are: culture at 30℃ for 7 days.

[0012] Fourthly, the present invention provides the application of the strain Sm-4 or the microbial preparation in any of the following:

[0013] (1) Used for sulfate reduction;

[0014] (2) Used for the remediation of drainage in acidic mines;

[0015] (3) Used for the remediation of soil contaminated with heavy metals;

[0016] (4) Used to prepare heavy metal curing agents.

[0017] Preferably, the application objectives of (2) and (3) are to increase the pH of acidic mine drainage and heavy metal contaminated soil in the mining area, and to solidify the heavy metals in the mine water and soil.

[0018] More preferably, the acidic mine drainage and heavy metal contaminated soil in the mining area are wastewater and contaminated soil generated from sulfur-containing minerals such as copper mines, iron mines, lead-zinc mines, antimony mines, and coal mines, and the heavy metals are one or more of copper, cadmium, chromium, and arsenic.

[0019] Fifthly, the present invention provides a sulfate reduction method, which utilizes the strain Sm-4 or the microbial preparation to reduce sulfate in the environment.

[0020] Preferably, the pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 5.1-7.2; more preferably 6.0-7.0; further preferably 6.0-6.5, and even more preferably 6.0.

[0021] Preferably, the C / S ratio of the environment in which the strain Sm-4 or the microbial preparation is used is 1:1-5:1; more preferably 2:1-5:1; further preferably 3:1-5:1, and even more preferably 3:1.

[0022] Furthermore, the sulfate concentration that the strain Sm-4 or the microbial preparation can tolerate is less than 50 g / L; preferably less than 25 g / L; more preferably less than 10 g / L.

[0023] Furthermore, the strain Sm-4 or the microbial preparation utilizes different carbon sources in the environment, including alcohols, small molecule organic acids and sugars, preferably alcohols and small molecule organic acids, more preferably ethanol (YC) and sodium lactate (RSN).

[0024] Furthermore, the strain Sm-4 or the microbial preparation utilizes different sulfur sources in the environment, including magnesium sulfate, sodium sulfate, sodium thiosulfate and calcium sulfate, preferably magnesium sulfate and sodium thiosulfate, more preferably sodium thiosulfate.

[0025] In a sixth aspect, the present invention provides a method for remediating acidic mine wastewater (AMD), which utilizes the strain Sm-4 or the microbial preparation to remediate the acidic mine wastewater.

[0026] Preferably, the amount of wastewater added is 0-100%, more preferably 25-50%, and most preferably 50%.

[0027] By employing the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0028] The novel sulfate-reducing bacterium Sm-4, which is tolerant to high concentrations of sulfate, provided by this invention, can efficiently reduce SO4 under weakly acidic conditions. 2- It further solidifies heavy metal ions, making it suitable for in-situ ecological remediation of acidic mine wastewater and its contaminated soil. This offers advantages in more economical and sustainable treatment of AMD and contaminated soil, and provides a solution for removing SO4 from AMD-contaminated water and heavy metal-contaminated soil.2- It provides microbial resources for the removal of heavy metals. Attached Figure Description

[0029] Figure 1 Electron micrographs of the colony and cell morphology of Vibrio Sm-4, a magnetic soil desulfurizing bacterium of the present invention.

[0030] Figure 2 This is a phylogenetic tree of the magnetic soil desulfurization Vibrio Sm-4 and similar type strains of the present invention.

[0031] Figure 3 The growth and desulfurization curves of magnetic soil desulfurizing Vibrio Sm-4 provided in Example 1 of this invention.

[0032] Figure 4 The desulfurization capacity of the magnetic soil desulfurizing Vibrio Sm-4 provided in Example 2 of this invention under different initial pH conditions.

[0033] Figure 5 The adaptability of the magnetic soil desulfurizing Vibrio Sm-4 provided in Example 3 of this invention to the initial C / S ratio.

[0034] Figure 6 The magnetic soil desulfurizing Vibrio Sm-4 provided in Example 4 of this invention is effective against different SO4 levels. 2- The reducing power at the initial concentration.

[0035] Figure 7 SO42- of the magnetic soil desulfurizing Vibrio Sm-4 provided in Example 5 of this invention under different initial carbon sources 2- Restorative ability.

[0036] Figure 8 The magnetic soil desulfurizing Vibrio Sm-4 provided in Example 6 of this invention under different initial sulfur sources SO4 2- Restorative ability.

[0037] Figure 9 This refers to the wastewater remediation capability of the magnetic soil desulfurization Vibrio Sm-4 provided in Example 7 of the present invention. Detailed Implementation

[0038] This invention provides a magnetic soil desulfurizing Vibrio strain Sm-4 and its applications. The strain Sm-4 belongs to the genus *Magnetic Soil Desulfurizing Vibrio*. Solidesulfovibrio sp.), through 16S rRNA sequence alignment in the Ezbiocloud database, this bacterium (sequencing length 1434 bp, sequence integrity 97.5%, 16S rRNA gene sequence of strain Sm-4 as shown in SEQ ID NO:1) is similar to *Vibrio magnetic soil desulfurization* (sp.). Solidesulfovibrio magneticus RS-1 TThe 16S rRNA sequence of the bacterium showed a maximum similarity of 99.6%. Combined with physiological and biochemical characteristics and phylogenetic analysis, this bacterium was identified as... Solidesulfovibrio magneticus Sm-4.

[0039] The present invention adopts the following technical solution:

[0040] This invention provides a strain of bacteria that is tolerant to acidic or weakly acidic conditions and efficiently reduces sulfate in acidic mine wastewater. Solidesulfovibrio magneticus ) and its application in wastewater remediation.

[0041] This invention provides a sulfate-reducing bacterium that is tolerant to high concentrations of sulfate. Solidesulfovibrio magneticus Sm-4, strain Sm-4, was enriched and isolated from acidic mine drainage sediment samples from a copper mine in Yunnan Province. This strain is effective against SO42-. 2- It has a reducing effect and a solidifying effect on heavy metals, making it suitable for removing SO4 from AMD-contaminated water bodies and heavy metal-polluted soil. 2- It provides microbial resources for the removal of heavy metals.

[0042] Strain Sm-4, cultured in modified Baars solid medium at 30℃ for 7 days, exhibited the following phenotypic characteristics: black, round colonies with neat edges; cells arranged singly or in pairs in an oval shape; apical flagella; size (0.8-1) μm × (1.8-5) μm; growth temperature 25-45℃; pH 5.0-7.0; and electron microscopy revealed oval cells arranged singly or in pairs. Figure 1 ).

[0043] The type strain with the highest 16S rRNA gene sequence similarity to strain Sm-4 is: ( Solidesulfovibrio magneticus RS-1), with a 99.6% full-length similarity to the 16S rRNA sequence. Soil-derived *Desulfovibrio* species (RS-1) were selected as similar. Solidesulfovibrio Phylogenetic tree constructed from the 16S rRNA gene sequence of the model strain is as follows: Figure 2 As shown, sequence similarity and evolutionary relationship prove that it is a member of the genus *Desulfuric Vibrio* in soil.

[0044] 1L of modified Baars medium contains: 2g / L MgSO4, 5g / L sodium citrate dihydrate, 1g / L CaSO4·2H2O, 1g / L NH4Cl, 0.5g / L K2HPO4 and 3.5g / L sodium lactate. It is prepared with deionized water and the pH is adjusted to 6. Before inoculation, 0.1mL of filtered and sterilized 5% Fe(NH4)2•(SO4)2 is added.

[0045] 1L of modified Baars solid medium contains: MgSO4 2g / L, sodium citrate dihydrate 5g / L, CaSO4·2H2O 1g / L, NH4Cl 1g / L, K2HPO4 0.5g / L and sodium lactate 3.5g / L, prepared with deionized water, 20g / L agar powder, pH adjusted to 6, and 0.1mL of filtered and sterilized 5% Fe(NH4)2•(SO4)2 added before pouring the plates.

[0046] The sulfate-reducing bacteria Sm-4 of the present invention in SO4 2- It can still grow in modified Baars medium with a concentration of 50 g / L, and the reduction rate reaches more than 70% after 7 days, which can greatly reduce SO4. 2- It can reduce the concentration of heavy metals in soil or water, increase the pH value of soil or water, and grows rapidly with low cost. It can be used on a large scale and quickly, which is beneficial to environmental protection.

[0047] Strain Sm-4 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 41191. This invention investigates the reduction effect of this bacterium on sulfate and the conditions under which it can provide strain resources for the reduction of sulfate and fixation of heavy metals in soil and water.

[0048] The present invention also provides a microbial preparation comprising one or more of the following: cells, powder, suspension or liquid extract of the magnetic soil desulfurizing Vibrio Sm-4.

[0049] The present invention also provides a method for preparing the aforementioned microbial preparation, the method comprising the step of culturing the magnetic soil desulfurization Vibrio Sm-4.

[0050] The preparation method of the microbial preparation includes culturing the desulfurized Vibrio Sm-4 at 25-45℃ and pH 4.5-7.0, preferably at 30-35℃ and pH 6.0-7.0, and more preferably at 30℃ and pH 6.0.

[0051] The magnetic soil desulfurization Vibrio Sm-4 was preferably cultured using a modified Baars medium. 1 L of modified Baars medium contained 2 g / L MgSO4·7H2O, 5 g / L sodium citrate dihydrate, 1 g / L CaSO4·2H2O, 1 g / L NH4Cl, 0.5 g / L K2HPO4, and 3.5 g / L sodium lactate, prepared with deionized water, pH 6. Before inoculation, 0.1 mL of filtered and sterilized 5% Fe(NH4)2•(SO4)2 was added. The medium was incubated at 30°C for 7 days.

[0052] This invention also provides the use of the magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation thereof in any one or more of the following:

[0053] (1) Remediation of acidic mine drainage or heavy metal contaminated soil;

[0054] (2) Heavy metal curing agent.

[0055] The preferred objective is to increase the pH of acidic mine drainage and heavy metal-contaminated soil in mining areas, and to solidify heavy metals in mine water and soil.

[0056] More preferably, the acidic mine drainage and heavy metal contaminated soil in the mining area are wastewater and contaminated soil generated from sulfur-containing minerals such as copper mines, iron mines, lead-zinc mines, antimony mines, and coal mines, and the heavy metals are one or more of copper, cadmium, chromium, and arsenic.

[0057] This invention also provides a sulfate reduction method:

[0058] The pH value of the environment in which the magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation is used is 5.1-7.2; preferably 6.0-7.0; more preferably 6.0-6.5, and even more preferably 6.0.

[0059] The C / S ratio of the magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation used in the environment is 1:1-5:1; preferably 2:1-5:1; more preferably 3:1-5:1, and even more preferably 3:1.

[0060] The magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation is tolerant to sulfate concentrations of 50 g / L or less; preferably 25 g / L or less; and more preferably 10 g / L or less.

[0061] The magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation can utilize different carbon sources, including alcohols, small molecule organic acids and sugars, preferably alcohols and small molecule organic acids, and more preferably ethanol (YC) and sodium lactate (RSN).

[0062] The magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation can utilize different sulfur sources, including magnesium sulfate, sodium sulfate, sodium thiosulfate and calcium sulfate, preferably magnesium sulfate and sodium thiosulfate, and more preferably sodium thiosulfate.

[0063] The present invention also provides a method for remediating acidic mine wastewater (AMD), using the magnetic soil desulfurizing Vibrio Sm-4 or the microbial preparation described above: preferably, the wastewater volume is 0-100%, more preferably, 25-50% of the wastewater is added, and most preferably, 50% of the wastewater is added.

[0064] The magnetic soil desulfurizing Vibrio Sm-4 provided by this invention can tolerate high concentrations of SO4. 2- And remove heavy metals, SO4 2- High reduction rate, effective for SO4 in soil and water. 2- It has a good treatment effect on both heavy metals.

[0065] By testing the sulfate-reducing capacity of strain Sm-4 under different conditions and its application in efficient desulfurization of acidic mine wastewater, the results showed that strain Sm-4 could effectively reduce sulfate in 50 g / L SO42-. 2- Growth and reduction of SO4 under certain conditions 2- Especially when the OD of the strain 600 When the value is greater than 1, the removal rate of sulfate can reach more than 80%, and the desulfurization rate is fast and effective. When a single sulfur source is used as the initial sulfur source, the single sulfur source that can be used is also gradually strengthening its ability to reduce sulfate. Therefore, it has broad application prospects in in-situ remediation technologies for acidic mine wastewater, polluted soil around abandoned mining areas, and farmland soil contaminated with heavy metals.

[0066] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0067] Example 1: Sulfate reducing ability of strain Sm-4

[0068] like Figure 3 As shown, the strain of the present invention Solidesulfovibrio magneticus Sm-4 was grown for 7 days in modified Baars medium at pH 6. Figure 3 Figure a shows that strain Sm-4 has a rapid growth ability, and the strain reaches its maximum OD on day 6. 600 It is 1.4; Figure 3 Results showed that strain Sm-4 had the ability to increase pH value. On the second day of growth, the pH value increased to 7.7, and the pH value gradually increased over time. Figure 3 The results showed that strain Sm-4 had a strong reducing ability, with an oxidation-reduction potential of about -400mV on the second day, which stabilized in this range over time. Figure 3 The results showed that strain Sm-4 achieved a sulfate reduction capacity of 53% on day 2 and stabilized at over 80% after day 4. This indicates that the strain of this invention possesses highly efficient SO42-reduction capacity. 2- Restorative ability.

[0069] Example 2: Sulfate reducing capacity of strain Sm-4 at different initial pH values

[0070] like Figure 4As shown, the strain of the present invention Solidesulfovibrio magneticus Sm-4 was grown for 14 days in modified Baars cultures at different pH values. Figure 4 The results showed that strain Sm-4 could grow at pH 5.5 and above, and after 14 days of culture, strain Sm-4 grew most vigorously in medium with pH 5.5. Figure 4 Results b showed that when pH ≥ 5.5, strain Sm-4 reached its maximum pH increase at 48-72 h, with strain Sm-4 exhibiting the strongest pH increase ability at pH = 6.5, reaching pH 8.0-8.3. Figure 4 The results showed that the rapid growth period of strain Sm-4 was 0-48 hours, during which its sulfate reduction capacity rapidly increased. When pH ≥ 5.5, the sulfate reduction rate reached 70-90% on the 5th day. This indicates that the strain of this invention can efficiently reduce sulfate at pH 5.5 and above.

[0071] Example 3: Sulfate reducing capacity of strain Sm-4 at different initial C / S ratios

[0072] like Figure 5 As shown, the strain of the present invention Solidesulfovibrio magneticus Sm-4 can grow normally at C / S ratios ranging from 1:1 to 5:1. Figure 5 The results showed that when the C / S ratio was 1:1, the OD of strain Sm-4 on day 4 was... 600 The value was higher than that of other groups, but significantly lower than that of other groups after day 4. On day 8, the C / S ratio was 3:1 for OD. 600 The value is higher than that of other treatment groups; Figure 5 Results showed that when the C / S ratio was 1:1, the sulfate reduction capacity of strain Sm-4 was significantly lower than that of other groups. When the C / S ratio was 1:1 and 2:1, the sulfate reduction capacity of strain Sm-4 reached its maximum on day 6 and then began to decline. However, when the C / S ratio was 3:1-5:1, the sulfate reduction rate of strain Sm-4 remained stable at over 95% after day 6. Figure 5 The results showed that strain Sm-4 had the lowest chemical oxygen demand (COD) at C / S ratios of 1:1 and 3:1. Considering both sulfate reducing capacity and strain growth, a C / S ratio of 3:1 was selected as the optimal C / S ratio for strain Sm-4.

[0073] Example 4: Strain Sm-4 under different initial SO4 2- sulfate reducing power at that time

[0074] The results are as follows Figure 6 As shown, the present invention Solidesulfovibrio magneticus Sm-4 in the initial SO4 2- It can reduce sulfate at concentrations ≤50g / L. Figure 6 The results showed that strain Sm-4, under different initial SO4...2- It can rapidly enter the logarithmic growth phase in media of all concentrations, SO4 2- When the concentration is ≤10g / L, the OD on day 7 600 It reaches 0.9-1.2, then enters its decline phase, OD 600 It decreased to 0.5-0.7; Figure 6 Results b show that the initial SO4 2- At concentrations ≤25 g / L, the pH-raising ability of strain Sm-4 showed a steady upward trend. On day 7, the pH increased to between 7.25 and 8.5, and on day 14, the pH value stabilized above 7.5. Figure 6 The results show that the initial SO4 2- The lower the concentration, the stronger the sulfate-reducing ability of strain Sm-4, and the lower the initial SO42- content. 2- When the concentration is ≤10g / L, its sulfate reduction rate is >40%; when the concentration is <10g / L, its sulfate reduction rate is >40%. 2 At ≤50 g / L, the sulfate reduction rate is 5%-28%. This indicates that the strain of this invention can reduce initial SO4 levels. 2- Growth reduction at concentrations ≤50g / L.

[0075] Example 5: Sulfate reducing capacity of strain Sm-4 under different carbon sources

[0076] The results are as follows Figure 7 As shown, the strain of the present invention Solidesulfovibrio magneticus Sm-4 can efficiently reduce sulfates under different carbon sources. Figure 7 The results showed that strain Sm-4 grew under 12 different carbon sources, with sodium formate (JSN) and sodium lactate (RSN) being the most suitable carbon sources, and maltose (MYT) and n-butanol (ZDC) not being used as carbon sources for growth. Figure 7 Figure b shows the results of the strain When Sm-4 uses sodium lactate as a carbon source, the pH increases most significantly, followed by ethanol (YC). Figure 7 The results showed that on day 5, strain Sm-4 achieved a sulfate reduction rate of >40% using 12 carbon sources, and on day 7, the sulfate reduction capacity using ethanol as the carbon source was optimal; therefore, the strain of this invention... Solidesulfovibrio magneticus The preferred carbon source for Sm-4 is ethanol, followed by small molecule organic acids.

[0077] Example 6: Sulfate reducing capacity of strain Sm-4 under different sulfur sources

[0078] The results are as follows Figure 8 As shown, the strain of the present invention Solidesulfovibrio magneticus Sm-4 can efficiently reduce sulfates under different sulfur sources. Figure 8 The results showed that the strain Solidesulfovibrio magneticusSm-4 can grow using magnesium sulfate, sodium sulfate, sodium thiosulfate, and calcium sulfate as sulfur sources. When grown with sodium sulfate as a substrate, the OD value after 14 days of culture is [missing value]. 600 The value can reach 0.9, but Sm-4 cannot grow with sodium sulfite and sodium dithionite as substrates; Figure 7 Results b showed that the strain Solidesulfovibrio magneticus When Sm-4 uses magnesium sulfate, sodium sulfate, sodium thiosulfate, and calcium sulfate as S sources, it can raise the pH of the solution from 6 to 7.5, but when sodium sulfite and sodium dithionite are used as S sources, the pH does not change significantly. Figure 7 The results showed that strain c Solidesulfovibrio magneticus Sm-4 can reduce SO4 in solution when magnesium sulfate, sodium sulfate, sodium thiosulfate, and calcium sulfate are used as sulfur sources. 2- Its reducing power is S4: sodium thiosulfate > S1: magnesium sulfate > S2: sodium sulfate > S6: calcium sulfate, but it does not reduce SO4 when sodium sulfite and sodium dithionite are used as the S source. 2- .

[0079] Example 7: Strain Sm-4's effect on SO4 in acidic mining wastewater 2- Highly efficient removal

[0080] The results are as follows Figure 9 As shown: When the pH of the acidic mine wastewater is 6, the strain of this invention... Solidesulfovibrio magneticus Sm-4 showed OD after 14 days of cultivation with 25% mineral water addition. 600 The pH value stabilized at 0.8, and the pH could be increased from 6 to around 7.5. The sulfate reduction rate reached 60%-70% after 14 days of cultivation with 50% mineral water added. 600 The pH value can reach 0.6, the pH can be increased from 6 to 7.8, and the sulfate reduction rate can reach 40%; with 75% mineral water addition, the OD value after 14 days of cultivation is [missing information]. 600 The pH value can reach 0.55, the pH can increase from 6 to 7.2, and the sulfate reduction rate is only 30%; with 100% mineral water addition, the OD value after 14 days of cultivation is... 600 When the pH value reaches 0.4, the pH can be increased from 6 to 7.1, but the sulfate reduction rate is only 32%. When the pH of the acidic mine wastewater is 4, the strain did not grow after 14 days of cultivation at 75% and 100% mine water addition, the pH remained acidic, and the sulfate reduction rate was only 10%-20%, with no obvious reduction effect.

[0081] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. Strains ( Solidesulfovibrio magneticus Sm-4, with accession number CGMCC No. 41191.

2. A microbial preparation, characterized in that, It contains one or more of the bacterial cells, bacterial powder, or bacterial suspension selected from the strain Sm-4 of claim 1.

3. The method for preparing the microbial preparation according to claim 2, characterized in that, The preparation method includes the step of culturing the strain Sm-4.

4. The preparation method according to claim 3, characterized in that, The strain Sm-4 was cultured at 25-45℃ and pH 4.5-7.

0.

5. The preparation method according to claim 4, characterized in that, The strain Sm-4 was cultured at 30-35℃ and pH 6.0-7.

0.

6. The preparation method according to claim 5, characterized in that, The strain Sm-4 was cultured at 30°C and pH 6.

0.

7. The preparation method according to any one of claims 3-6, characterized in that, The strain Sm-4 was cultured using a modified Baars medium. The modified Baars medium contained 2 g / L MgSO4·7H2O, 5 g / L sodium citrate dihydrate, 1 g / L CaSO4·2H2O, 1 g / L NH4Cl, 0.5 g / L K2HPO4, and 3.5 g / L sodium lactate, prepared with deionized water at pH 6; 0.1 mL of filtered and sterilized 5% Fe(NH4)2•(SO4)2 was added before inoculation. The culture conditions were: cultured at 30℃ for 7 days.

8. The strain Sm-4 of claim 1 or the microbial preparation of claim 2 may be used in any of the following applications: (1) Used for sulfate reduction; (2) Used for the remediation of drainage from acidic mines; among which, The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 5.1-7.

2.

9. The application according to claim 8, characterized in that, (2) The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.0-7.

0.

10. The application according to claim 9, characterized in that, (2) The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.0-6.

5.

11. The application according to claim 9, characterized in that, (2) The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.

0.

12. The application according to any one of claims 8-11, characterized in that, (2) The application goal is to increase the pH of acidic mine drainage and solidify heavy metals in the mine water.

13. The application according to claim 12, characterized in that, The acidic mine wastewater is wastewater generated from copper mines, iron mines, lead-zinc mines, antimony mines, and sulfur-containing minerals in coal mines, and the heavy metal is one or more of copper, cadmium, chromium, and arsenic.

14. A sulfate reduction method, characterized in that, The strain Sm-4 of claim 1 or the microbial preparation of claim 2 is used to reduce sulfate in the environment; Wherein, the pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 5.1-7.2; and / or, The C / S ratio of the strain Sm-4 or the environment in which the microbial preparation is used is 1:1-5:

1.

15. The method according to claim 14, characterized in that, The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.0-6.5; and / or, The C / S ratio of the strain Sm-4 or the environment in which the microbial preparation is used is 2:1-5:

1.

16. The method according to claim 15, characterized in that, The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.0; and / or, The C / S ratio of the strain Sm-4 or the environment in which the microbial preparation is used is 3:1-5:

1.

17. The method according to claim 16, characterized in that, The C / S ratio of the strain Sm-4 or the environment in which the microbial preparation is used is 3:

1.

18. The method according to any one of claims 14-17, characterized in that, The strain Sm-4 or the microbial preparation is tolerant to a sulfate concentration of 50 g / L.

19. The method according to claim 18, characterized in that, The strain Sm-4 or the microbial preparation is tolerant to sulfate concentrations below 25 g / L.

20. The method according to claim 19, characterized in that, The strain Sm-4 or the microbial preparation is tolerant to sulfate concentrations below 10 g / L.

21. The method according to any one of claims 14-17, characterized in that, The strain Sm-4 or the microbial preparation utilizes different carbon sources in the environment, including alcohols, small molecule organic acids, and sugars; The strain Sm-4 or the microbial preparation utilizes different sulfur sources in the environment, including magnesium sulfate, sodium sulfate, sodium thiosulfate, and calcium sulfate.

22. The method according to claim 21, characterized in that, The strain Sm-4 or the microbial preparation utilizes different carbon sources in the environment, namely alcohols and small molecule organic acids; The strain Sm-4 or the microbial preparation utilizes different sulfur sources in the environment, namely magnesium sulfate and sodium thiosulfate.

23. The method according to claim 22, characterized in that, The strain Sm-4 or the microbial preparation utilizes different carbon sources in the environment, namely ethanol and sodium lactate; The strain Sm-4 or the microbial preparation utilizes different sulfur sources in the environment, wherein the sulfur source is sodium thiosulfate.

24. A method for remediating acidic mine wastewater, comprising remediating acidic mine wastewater using the strain Sm-4 of claim 1 or the microbial preparation of claim 2; The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.0-6.

5.

25. The method according to claim 24, characterized in that, The pH value of the environment in which the strain Sm-4 or the microbial preparation is used is 6.

0.

26. The method according to claim 24 or 25, characterized in that, The amount of wastewater added is 25-50%.

27. The method according to claim 26, characterized in that, The wastewater addition rate is 50%.

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