WO2020217188A1 - Procédé pour le contrôle in situ de la contamination d'eaux souterraines par infiltration d'eaux de bassin de retenue des résidus - Google Patents
Procédé pour le contrôle in situ de la contamination d'eaux souterraines par infiltration d'eaux de bassin de retenue des résidus Download PDFInfo
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- WO2020217188A1 WO2020217188A1 PCT/IB2020/053811 IB2020053811W WO2020217188A1 WO 2020217188 A1 WO2020217188 A1 WO 2020217188A1 IB 2020053811 W IB2020053811 W IB 2020053811W WO 2020217188 A1 WO2020217188 A1 WO 2020217188A1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/30—Aerobic and anaerobic processes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K8/00—Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
- C09K8/42—Compositions for cementing, e.g. for cementing casings into boreholes; Compositions for plugging, e.g. for killing wells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- E—FIXED CONSTRUCTIONS
- E02—HYDRAULIC ENGINEERING; FOUNDATIONS; SOIL SHIFTING
- E02D—FOUNDATIONS; EXCAVATIONS; EMBANKMENTS; UNDERGROUND OR UNDERWATER STRUCTURES
- E02D3/00—Improving or preserving soil or rock, e.g. preserving permafrost soil
- E02D3/12—Consolidating by placing solidifying or pore-filling substances in the soil
Definitions
- the present invention refers to a process for the "in situ" control of groundwater contamination by infiltration of mining tailings dam waters.
- tailings correspond to a fine suspension of solids in liquid. These are fundamentally constituted by the same material present in-situ in the deposit, in addition to reagents and chemical elements used in the mineral obtaining processes.
- the tailings contain high concentrations of chemicals and elements that alter the environment, so they must be transported and stored in "dams or tailings deposits", where the contaminants slowly settle to the bottom and the water is recovered or evaporated. Due to the decrease in the ore grades of the deposits, it will be necessary to extract more and more tonnages of material to maintain their production levels, which will mean a proportional increase in the amount of waste that must be disposed of in the form of tailings.
- Tailings dams Long-term failure mechanisms in tailings dams include cumulative damage (eg, erosion or earthquakes), geological hazards (eg, landslides), liquefaction, and changing weather patterns (Chambers, DM, & Higman, B. (2011) . Long term risks of tailings dam failure. Center for Science in Public Participation, Bozeman, Montana) Although it has been described that the majority of tailings dam failures are due to combined causes (39%) (Rico , M., et al.
- tailings dams It is important to take into consideration that it has been estimated that the risk of failure in tailings dams is increasing, where it is directly related to the number of tailings accumulation dams greater than 5,000,000 cubic meters of capacity, necessary for allow the economic extraction of low grade minerals. Additionally, at least 11 catastrophic failures are expected worldwide between 2010 and 2019, with a total cost of approximately USD $ 6 billion (Mining (2015). Catastrophic mine waste spills increasing in frequency, severity and cost world-wide. Http: //www.mining.com/web/catastrophic- mine-waste-spills-increasing-in-frequency-severity-and-cost-world-wide /).
- tailings dams consider in their design and operation different monitoring and control measures of possible leaks or seeps to prevent them from contaminating groundwater.
- monitoring measures the use of monitoring wells downstream of drainage collecting pools, a system for measuring flows, groundwater levels and monitoring of surface waters can be considered.
- seepage collection wells installed downslope from the reservoir (Duda, R. (2014). The Influence of Drainage Wells Barrier on Reducing the Amount of Major Contaminants Migrating from a Very Large Mine Tailings Disposal Site. Archives of Environmental Protection, 40 (4), 87-99).
- This well is equipped with hydraulic pumps that send the seepage back to the tailings dam. These units can be used in conjunction with shear walls or trenches to minimize seepage downhill.
- the quality of the effluent is monitored to determine the possibility of leaks, since any leakage that occurs in the underground layers can cause complications in the operation of the mining site with the consequent associated economic loss. It is important to mention that depending on the quality of the effluent, the operation of the hydraulic pump can continue indefinitely (EPA, 1994). Consequently, the current treatment with the hydraulic pump does not completely solve the problem.
- the water that filters through the wall is recovered through drains and the channeling of the flow.
- the water from the wall can be returned to the dam, or it can be collected in a common pond with the water recovered from the lagoon for its recirculation to the beneficiation plant (SERNAGEOMIN (2003) Guide of good environmental practices for small mining; Levenick JL et al. (2009). Hydrogeological assessment of seepage through the Antamina tailings dam - Antamina copper / zinc mine, Peru, South America, International Mine Water Conference 19th - 23rd October 2009, Pretoria, South Africa).
- the water that infiltrates at the subsoil level is recirculated from a series of catchment wells located downstream and is monitored by independent drilling.
- a zone of low hydraulic conductivity is generated in the contaminated underground aquifer.
- the process occurs even in the absence of oxygen •
- the process is designed to operate at low temperature (for example at 19 ° C)
- the microbial consortium is adapted to groundwater contaminated with seepage from a tailings dam.
- the process allows additionally to reduce the sulfate concentration.
- the main object of the present invention is a method to reduce the hydraulic conductivity and generate the precipitation of insoluble minerals in an underground aquifer, which comprises at least the steps of:
- a) provide a culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated
- the underground aquifer is affected by surface water seepage.
- a particular embodiment refers to an underground aquifer that is affected by water seepage from a mining tailings dam.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated is produced with a method that comprises at least the steps: a) inoculate a culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated in a fixed-bed anaerobic bioreactor with a support material,
- the bioreactor support material is selected from the group of sand, silica, glass and ceramics.
- the injection of the underground aquifer with an enriched culture of microorganisms and the injection of the underground aquifer with an electron donor and nutrients suitable for the culture of microorganisms enriched in reducing bacteria, and one or more metal cations that favor the Insoluble mineral precipitation is done by one or more injection wells.
- the electron donor suitable for microorganism culture enriched in reducing bacteria is selected from the group of formate, formic acid, acetate and acetic acid.
- said nutrients suitable for the culture of microorganisms enriched in reducing bacteria comprise at least ammonium and phosphate.
- the nutrients suitable for the culture of microorganisms enriched in reducing bacteria further comprise a Complex nutrient, rich in vitamins, selected from the group of yeast extract and strained corn water.
- the insoluble mineral that precipitates in the solid material of the underground aquifer is calcium carbonate.
- the insoluble mineral that precipitates in the solid material of the underground aquifer is iron sulfide.
- the method for reducing the hydraulic conductivity and generating the precipitation of insoluble minerals in an underground aquifer also allows the removal of sulfate from the underground waters of said aquifer.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated is constituted, at least by bacteria of the Phylum Proteobacteria, Firmicutes and Bacteroidetes.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated is constituted, at least by bacteria from the Phylum Proteobacteria or Firmicutes.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated is constituted, at least by bacteria of the Gammaproteobacteria, Clostridia, Deltaproteobacteria and Bacteroidia Classes.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated is constituted, at least by bacteria of the Clostridia or Deltaproteobacteria Class.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated contains, at least, bacteria of the Desulfovibrionales Order.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated contains, at least, bacteria of the Order Desulfobacterales.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated contains at least bacteria of the Desulfuromonadal Order.
- the culture of microorganisms enriched in sulfate reducing bacteria and adapted to the groundwater of the aquifer to be treated contains, at least, bacteria of the Order Clostridiales.
- the culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated contains at least one genus of sulfate-reducing bacteria selected from the group consisting of Desulfomicrobium, Desulfovibrio, Desulfonema , Desulfuromonas, Desufutomaculum and Desulfosporosinus.
- the culture of microorganisms, enriched in sulfate-reducing bacteria adapted to the groundwater of the aquifer to be treated is obtained from samples taken from live microbialites from lagoons or saline lakes. Samples of these live microbialites are found, for example, in live thrombolites from Lake Sarmiento, or in live stromatolites from Website Amarga, both in Torres del Paine National Park, Chile, in live microbialites from Website from Madison Interna and La Website.
- the culture of microorganisms, enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated is obtained with a method that comprises at least the steps:
- the living microbialite is a living stromatolite. In another embodiment of the invention, the live microbialite is a live thrombolyte.
- Microbialites in this invention, the concept of microbialites is understood as the "organo-sedimentary deposits that have increased as a result of a benthic microbial community trapped and attached to detrital sediments and / or forming the site of mineral precipitation such as calcium carbonate
- the structures of microbialites can range from well-structured laminated stromatolites to coagulated thrombolites. Microbialites have been found in different environments, for example, hot springs, dolomitic ponds, hypersaline and / or alkaline lakes, rivers and lakes, environments open marine and hypersaline marine environments.
- Stromatolites they are microbialites that are characterized by being laminated organo-sedimentary structures (typically CaCÜ3) that grow attached to the substrate and emerge vertically from it, producing structures of great morphological, volumetric and biogeographic variety. Thrombolites: these are microbialites related to stromatolites, but they lack lamination and are characterized by macroscopic coagulated tissue.
- Living microbialites, stromatolites or thrombolites they are organo-sedimentary structures that are currently found in lake or marine systems and that present an active microbial community, whose metabolism participates in the mineralization process of these structures.
- Microbial consortium in this invention, the concept of microbial consortium is understood as a group of different microorganisms that act together. In a microbial consortium, microorganisms with different metabolic capacities can be found. In the particular case of the sulfate-reducing microbial consortium, it is composed, for example, of fermentative, acetogenic and sulfate-reducing anaerobic microorganisms.
- Bio-clogging is defined as the reduction in hydraulic conductivity and porosity of a saturated porous medium due to microbial growth.
- Biologically Induced Mineralization is the process in which the metabolic activity of microorganisms (for example, bacteria) produces favorable chemical conditions for the formation of minerals (for example, the precipitation of calcium carbonates induced by increased alkalinity).
- FIGURE 1 is a diagrammatic representation of FIG. 1 :
- This figure shows a schematic diagram of the sand-filled bioreactor.
- the bioreactors (1) were made of glass.
- an outer glass jacket (2) was used, fed with water from a thermoregulated bath.
- the bioreactor was filled with sterilized sand (3).
- a pair of glass piezometers located one in the lower part (4) and another in the upper part (5) of the bioreactor. The feeding was done by peristaltic pumps from the lower part (6) to the upper part (7) of the bioreactor.
- FIGURE 2 is a diagrammatic representation of FIGURE 1
- FIGURE 3 shows the parameters in bioreactor effluent inoculated with microorganisms from a live thrombolite from Lake Sarmiento.
- A pH values
- B sulfate concentrations (filled squares) and calcium concentrations (open squares).
- FIGURE 3 shows the parameters in bioreactor effluent inoculated with microorganisms from a live thrombolite from Lake Sarmiento.
- A pH values
- B sulfate concentrations (filled squares) and calcium concentrations (open squares).
- This figure shows the parameters in the effluent of a bioreactor inoculated with microorganisms from a live stromatolite from Website from Website Amarga.
- A pH values and oxidation-reduction potential
- B sulfate concentrations (filled squares) and hydrogen sulfide concentrations (open squares)
- C calcium concentration.
- FIGURE 4
- This figure shows schematically the design of the steel columns of 1.40 meters high and 25 centimeters in diameter, with lateral outlets at 8, 50, 90 and 130 centimeters from the base for the installation of pressure measuring instruments, used to measure the reduction of hydraulic conductivity in the sand bed by the enriched microbial consortia.
- FIGURE 5
- FIGURE 6 This figure shows the parameters in the effluent from column A inoculated with the microbial consortium enriched from the sample of the microbialite from Lake Sarmiento.
- A Microbial ATP in the effluent (open squares) and oxidation-reduction potential (solid squares),
- B sulfate concentrations (open diamonds) and hydrogen sulfide in the effluent (solid diamonds).
- the line indicates the sulfate concentration in the added culture medium.
- FIGURE 6 This figure shows the parameters in the effluent from column B inoculated with the microbial consortium enriched from the stromatolites sample from Website.
- A Microbial ATP in the effluent (open squares) and oxidation-reduction potential (solid squares),
- B sulfate concentrations (open diamonds) and hydrogen sulfide in the effluent (solid diamonds).
- the line indicates the sulfate concentration in the added culture medium.
- FIGURE 7 is a diagrammatic representation of FIGURE 7
- This figure shows the parameters in the effluent from column C with biocide.
- A Microbial ATP in the effluent (open squares) and oxidation-reduction potential (solid squares)
- the line indicates the sulfate concentration in the added culture medium.
- FIGURE 8
- This figure shows the pH measurements in bioreactors B1, B2 and B3.
- Bioreactors B1 and B3 were fed with culture medium A, but bioreactor B1 was switched to culture medium B on day 170.
- Bioreactor B2 was permanently fed with culture medium B.
- FIGURE 9 is a diagrammatic representation of FIGURE 9
- Bioreactors B1, B2 and B3 were fed with culture medium A, but bioreactor B1 was switched to culture medium B on day 170. Bioreactor B2 was permanently fed with culture medium B.
- FIGURE 10 is a diagrammatic representation of FIGURE 10
- This figure shows the sulfur (hydrogen sulfide) measurements in bioreactors B1, B2, and B3.
- Bioreactors B1 and B3 were fed with culture medium A, but bioreactor B1 was switched to culture medium B on day 170.
- Bioreactor B2 was permanently fed with culture medium B.
- FIGURE 11 This figure shows the remaining sulfate measurements in bioreactors B1, B2, and B3. Bioreactors B1 and B3 were fed with culture medium A, but bioreactor B1 was switched to culture medium B on day 170. Bioreactor B2 was permanently fed with culture medium B.
- FIGURE 12 This figure shows the remaining sulfate measurements in bioreactors B1, B2, and B3. Bioreactors B1 and B3 were fed with culture medium A, but bioreactor B1 was switched to culture medium B on day 170. Bioreactor B2 was permanently fed with culture medium B.
- FIGURE 12 This figure shows the remaining sulfate measurements in bioreactors B1, B2, and B3. Bioreactors B1 and B3 were fed with culture medium A, but bioreactor B1 was switched to culture medium B on day 170. Bioreactor B2 was permanently fed with culture medium B.
- FIGURE 12 This figure shows the remaining sulfate measurements in bioreactors B1, B2, and B3. Bioreactors B1
- This figure shows an example of the aquifer model design built on the basis of acrylic chambers. Shown is an acrylic chamber with a removable lid packed with quartz sand. In the figure you can distinguish the ports for inoculation and stimulation of microorganisms, and for taking samples. At their ends, a pipe was installed internally for the flow water inlet and another for the effluent outlet.
- FIGURE 13 is a diagrammatic representation of FIGURE 13
- FIGURE 15
- This figure shows the relative retention times of the aquifer models during the experimentation time.
- the retention times relative to the initial retention time of the aquifer models are shown.
- FIGURE 16 is a diagrammatic representation of FIGURE 16
- FIGURE 17 shows the change in tracer flow around a zone of lower permeability in the porous medium of the aquifer models.
- An example of the tracer passage in one of the aquifer models is shown on day 55 of operation. Tracer deviation is observed when meeting the blackened area of the porous medium (o): inoculation point.
- FIGURE 17 shows the change in tracer flow around a zone of lower permeability in the porous medium of the aquifer models.
- An example of the tracer passage in one of the aquifer models is shown on day 55 of operation. Tracer deviation is observed when meeting the blackened area of the porous medium (o): inoculation point.
- FIGURE 17 shows the change in tracer flow around a zone of lower permeability in the porous medium of the aquifer models.
- This figure schematically shows a pH map in the sand of the inoculated aquifer models at the end of the experiment (60 days).
- a zone of pH 8 is observed that is distributed from the inoculation zone and reaches the end where the effluent exits, as well as specific zones of pH 8.5 in one of the duplicates.
- a pH 7.5 was observed in all areas of the model.
- the circle represents the point of inoculation and stimulation with culture medium.
- FIGURE 18 is a diagrammatic representation of FIGURE 18
- This figure schematically shows the microbial ATP maps in the sand from the aquifer models at the end of the experiment.
- the black arrows indicate the direction of the hydraulic flow inside the aquifer models.
- the circle represents the point of inoculation and stimulation with culture medium.
- FIGURE 19 is a diagrammatic representation of FIGURE 19
- FIGURE 20 This figure is a photograph illustrating the biomineralization of quartz sand at the inoculation point of the aquifer models. A portion of biomineralized quartz sand that was located at the inoculation point of the inoculated models is shown. In areas more distant from the inoculation point, the formation of sand agglomerates similar to the one in the image was not observed.
- FIGURE 20
- This figure shows the microbial sulfate reduction (SCU 2- and H2S) and physicochemical parameters (pH and oxide-reduction potential) in the aquifer models stimulated with the different formate concentrations.
- SCU 2- and H2S microbial sulfate reduction
- physicochemical parameters pH and oxide-reduction potential
- FIGURE 21 is a diagrammatic representation of FIGURE 21.
- This figure shows the remaining formate concentration in the aquifer models stimulated with the different concentrations of the electron donor.
- FIGURE 22
- This figure shows the retention times relative to the initial time in the aquifer models stimulated with the different formate concentrations.
- FIGURE 23 is a diagrammatic representation of FIGURE 23
- FIGURE 24 This figure schematically shows the pH maps in the sand of the aquifer models stimulated with the different formate concentrations at the end of the experiment (60 days).
- the pH values in sand tend to increase as the concentration of formate in the growing medium is increased.
- specific areas of pH 8.5 were observed.
- the black arrows indicate the direction of the hydraulic flow inside the aquifer models.
- the circle (o) represents the point of inoculation and stimulation with culture medium.
- This figure schematically shows the microbial ATP maps in the sand from the aquifer models stimulated with the different formate concentrations at the end of the experiment.
- the ATP value (in URL / g of sand) is shown in the three zones of the sand of the aquifer models stimulated with the different formate concentrations.
- the black arrows indicate the direction of the hydraulic flow inside the aquifer models.
- FIGURE 25
- This figure shows the biologically induced mineralization of the quartz sand from the aquifer models inoculated with the different formate concentrations at the end of the experiment.
- Aquifer models uncovered at the end of the experiment are shown.
- the red line delimits the mineralized zone (of greater hardness compared to other zones) of the porous medium of the aquifer models. All the models (duplicates) used are shown (central column).
- the bioreactor on the right shows the percentage of the area occupied by the mineralized zone with respect to the total surface of the upper face of the models (Calculated using the ImageJ program).
- FIGURE 26 is a diagrammatic representation of FIGURE 26.
- FIGURE 27 shows the surface area of biomineralized sand in the aquifer models as a function of the formate concentration in the culture medium. A direct dependence of
- This figure shows the images obtained by scanning electron microscopy of the quartz sand from the aquifer models stimulated with the lowest and highest concentrations of formate (3 and 18 g / L) at the end of the experiment.
- A Crystalline CaCÜ3 mineral and bacterial clumps in a quartz sand sample from one of the models stimulated with 3 g / L formate.
- B CaCÜ3 crystalline mineral and bacterial clusters in a quartz sand sample from one of the models stimulated with 18 g / L of formate.
- ac quartz sand carb: carbonate. Arrows: bacterial clusters.
- FIGURE 29 is a diagrammatic representation of FIGURE 29.
- This figure illustrates the flow diagram of a particular application of the process to reduce hydraulic permeability and generate precipitation of insoluble minerals in situ from a sulfate-contaminated underground aquifer.
- Lake Sarmiento de Gamboa 51 ° 03O0 "S, 72 ° 45'0 W), Torres del Paine National Park, XII Region, Chile.
- Sarmiento Lake has a maximum depth of 312 m and an approximate area between 83 and 84 km 2.
- Lake Sarmiento de Gamboa is an alkaline lake with an average salinity of 1.9 mg / L, a pH between 8.3 and 8.7, and an average surface temperature of 6.2 ° C in winter and 12.2 ° C in summer.
- Lake Sarmiento has live microbialites, which have a non-laminated structure made up of carbonate clots which classifies them as thrombolites.
- the sampling process consisted of the manual extraction of live thrombolites located underwater on the shore of the Northeast sector of the lake, which were placed at 6 ° C in sterile plastic jars. Because it is a State Protected Wild Area, permission was obtained from the National Forestry Corporation (CONAF Authorization N ° 012/2014) to carry out the sampling.
- the bioreactors (1) were made of glass and had 173 mL capacity, 18 cm in length and 3.5 cm in internal diameter.
- an outer glass jacket (2) was used, fed with water from a thermoregulated bath at 18 ° C.
- the bioreactor was filled with sand (3).
- a filter was installed at the bottom of the bioreactor to prevent sand from running into the feed hoses. This filter consisted of a PVC tube covered with a plastic mesh, through the lower opening.
- the bioreactor was filled with sterile distilled water, taking care to remove all trapped air.
- the sand was sterilized and dried in an oven at 180 ° C for 2 hours.
- a pair of piezometers of glass located one at the bottom (4) and one at the top (5) of the bioreactor. The difference in the water height of the piezometers allowed determining the hydraulic conductivity.
- the feeding was done by peristaltic pumps (Cole-Palmer, model 7557-14, 1-100 rpm, USA) from the bottom (6) to the top (7) of the bioreactor, with an upward flow of 0.2 mL. / min.
- the external jacket of the bioreactor was connected to the thermoregulated bath with silicone hoses, while the other connections were made as follows: with silicone hoses, the inlets, outlets and installation of the piezometers; with Tygon hoses (impermeable to gaseous exchange) the feeding, recirculation and effluent outlet routes. All the hoses of the system, and their connectors, were previously sterilized in an autoclave. To generate the feed flow, a peristaltic pump (Cole-Palmer, model 7557-14, 1-100 rpm, USA) was used. To prevent changes in the internal temperature of the system, along with preventing the passage of light to the interior (simulating the conditions of the underground layer), the bioreactors were wrapped with a thermal insulator.
- the medium was adjusted to pH 7.9.
- the culture medium was prepared in water extracted groundwater hydraulic barrier of a tailings dam, whose main components are HCO3 ⁇ 217.7 mg / L, Ca 2+ 302 mg / L, Mg 2+ 76.6 mg / L and SCL 2- 1192.3 mg / L.
- Groundwater extracted from a contaminated aquifer was used to prepare the culture medium to enrich a microbial consortium adapted to the environmental conditions of the site to be treated.
- barium sulfate that is formed by adding the soluble salt of barium chloride (BaCh) in excess was used (American Public Health Association, American Water Works Association and Water Environment Federation. 1998a. "4500-S04-2 E”. In: "Standard methods for the examination of water and wastewater”. Ed. 20).
- the barium sulfate formed was determined by turbidimetry using a spectrophotometer at 450 nm (Dynamica spectrophotometer, model HALO RB-10, Great Britain).
- the concentration of calcium in solution was performed by absorption spectrometry, using the following protocol: 1 mL of effluent was centrifuged for 10 minutes at 11,600 g (Microcentaur MSB010.Cx2.5 centrifuge; Sanyo, Great Britain), 500 pL of supernatant was diluted in distilled water, to bring the determination within the measurable range. From the mentioned solution, 1 mL of solution was taken and added to a test tube with 8 mL of distilled water and 1 mL of a 4,000 mg / L Sr 2+ solution with 0.8N HCIO4. Finally the solution is measured using a Perkin Elmer atomic absorption spectrophotometer, model 3110, USA (American Public Health Association, 1992a; American Public Health Association, 1992b).
- the reduction in calcium concentration is explained by the mineralization of calcium carbonate inside the bioreactor (biologically induced mineralization).
- the effluent from this column which contains the sulfate-reducing microbial consortium from thrombolites from Lake Sarmiento, Torres del Paine, is used as an inoculum for a process that reduces hydraulic permeability and generates the precipitation of insoluble minerals in a contaminated aquifer. .
- this effluent is used to inoculate a new bioreactor of equal or greater size.
- Live stromatolites were sampled from Georgia Amarga (50 ° 58'27 "S, 72 ° 44'55” W), Torres del Paine, XII Region, Chile.
- Website Amarga is a shallow endorheic mesosaline lagoon (maximum depth 4 m) with an approximate area of 1. 9 km 2 .
- the average pH of the lake is 9.1, while the average salinity and temperature is 26.1 mg / L and 11.7 ° C, respectively.
- La Website Amarga presents live laminar microbialites or stromatolites, with bulbous, domed and elongated shapes (Solari et al.
- bioreactors filled with sand were used, according to what was previously described in Example 1.
- the inoculation of the bioreactors with a sample of stromatolite from the Website Amarga and the subsequent colonization of the sand was also carried out. as already described in Example 1.
- the culture medium was then fed to the bioreactor, without recirculation, with a flow rate of 30-90 mL / week.
- the determination of the sulfate and calcium concentrations in solution were carried out according to Example 1.
- Figure 3A shows the result of the enrichment of the microbial consortium.
- Figure 3B shows a significant reduction in the sulfate concentration in the bioreactor effluent, compared to the concentration added with the culture medium. In this case, it was reached sulfate concentrations less than 1500 mg / L.
- Figure 3B shows that from day 100 the hydrogen sulfide concentration increases significantly. This shows the activity of the sulfate reducing microorganisms in the enriched microbial pool.
- Figure 3C shows a progressive decrease in the concentration of calcium in the effluent compared to what is added in the culture medium.
- the effluent from this bioreactor which contains the sulfate-reducing microbial consortium enriched from stromatolites from the Website Amarga, is used as inoculum for a process that allows reducing hydraulic permeability and generating the precipitation of insoluble minerals in a contaminated aquifer.
- this effluent is used to inoculate a new bioreactor of equal or greater size.
- the column cover was placed and the hose connections made using Tygon hoses. Then a hose connected to a thermoregulated bath was wound along the columns and the structure was surrounded with a thermal insulator, in order to keep the interior of the columns at 18 ° C.
- the following culture medium was used: K2SO4 4.6 g / L; MgSO4 * 7H 2 0 0.06 g / L; NaHCOs 0.035 g / L; CaCl 2 * 2H 2 0 3.4; NhUCI 1 g / L; Na 2 (Si0 3 ) 0.009 g / L; KH 2 R0 4 0.05 g / L; 0.279 g / L sodium citrate; sodium acetate 4.5 g / L; yeast extract 1 g / L; sodium thioglycolate 0.099 g / L.
- the culture medium was prepared with groundwater extracted from the hydraulic barrier of a tailings dam, whose main components are HCO3 ⁇ 217.7 mg / L, Ca 2+ 302 mg / L, Mg 2+ 76.6 mg / L and 1192.3 mg SCL 2- / L.
- thioglycollate was also added, a reducing compound that gives the culture medium a suitable oxide reduction potential for the development of sulfate reducing bacteria.
- Example 2 The determination of sulfate and calcium concentrations was carried out according to Example 1 and the measurement of hydrogen sulfide (or sulfides) in solution, according to Example 2.
- the ATP concentration in the effluents was determined with the commercial ATP measurement system LixKit ® , Biohidrica, Chile, which is based on the reaction of ATP-dependent oxidation of luciferin catalyzed by the enzyme luciferase.
- the amount of light produced by the reaction was determined using a Kikkoman luminometer, model Lumitester PD-20, Japan. For this, 5 mL of effluent from the models were taken, from which the microorganisms were concentrated by filtration on nitrocellulose membranes (0.22 pm).
- the filtered microorganisms were collected with the torula provided in the system, which is inserted into the reaction rod containing the cell lysis liquid and the other lyophilized components for ATP-dependent oxidation of luciferin.
- the luminescence value obtained (in Relative Light Units, URL), was divided by 5 to obtain the value of URL / mL.
- Figure 6A shows high values of intracellular ATP in the effluent of the column at the beginning of the experiment, which decrease over time.
- the sulfate measurements from the effluent, shown in Figure 6B indicate that from the beginning of the experiment there was a decrease in the sulfate values in the effluent from column B compared to the sulfate concentration fed with the medium. cultivation.
- Figure 6B shows an active production of hydrogen sulfide from day 60, increasing its concentration in the effluent, to reach values of 450 mg / L on day 140.
- Hydraulic conductivity was determined at the end of the experiment in the columns, performing pressure measurements with pressure transmitters (flush diaphragm pressure transmitters model C9000156, Veto, Chile). Hydraulic conductivity was calculated using Darcy's equation, presented below:
- Hsat is the saturation hydraulic conductivity
- Q is the flow rate (feed rate of the column)
- L is the distance between the measuring stations
- A is the diameter area of the column
- DH is the difference between the heights of the measured water columns.
- bioreactors filled with sand of 250 ml of internal volume were mounted according to Example 1 and were named: B1, B2 and B3.
- Each of the bioreactors contains quartz sand as internal support material (particle size 297-841 pm) and are maintained under anaerobic conditions at a temperature of 18 ° C and periodically fed with culture media A and B described in the Table 3.
- the media differ because medium A contains formate, while medium B has acetate as major electron donor. Furthermore, medium A lacks citrate and has a lower concentration of yeast extract.
- These culture media were prepared in water from the tailings dam, which had a pH of 7.9 and a sulfate concentration of 1500 mg / L.
- the three bioreactors were inoculated with the effluent from the bioreactor of Example 1, which contains the sulfate-reducing microbial consortium enriched from Lake Sarmiento thrombolites.
- bioreactor B1 Two bioreactors were fed with culture medium A (B1 and B3), however, bioreactor B1 was switched to culture medium B on day 170. On the other hand, bioreactor B2 was permanently fed with culture medium B To each of the bioreactors, periodic measurements of physical-chemical parameters (pH, sulfate, sulfur,) and biological parameters (bacterial count, ATP) were carried out, in order to determine and establish possible conditions that occur in the assembled systems.
- physical-chemical parameters pH, sulfate, sulfur,
- biological parameters bacterial count, ATP
- Example 2 The determination of the concentrations of sulfate, calcium, according to Example 1, ATP according to Example 3 and hydrogen sulfide (or sulfides) in solution were carried out according to Example 2.
- the pH shows a clear increase (> 8.2) in the bioreactor B2, which contains formate as a carbon source (culture medium A).
- the B1 bioreactor when supplied with formate (from day 170 of operation), the pH increases considerably from 7.2 and exceeding 8.5.
- Bioreactor B3 maintains a constant pH, varying slightly between 7.1 and 7.6 ( Figure 8).
- the bacterial count ( Figure 9), maintains a variable rhythm until day 125, however, then reaches a steady state until day 170, maintaining a count of the 3 bioreactors between 1, 8 and 3.0 x 10 8 bacteria / mL. The same is observed in intracellular ATP measurements. A significant increase in the bacterial count (5.0 x 10 8 bacteria / mL) is observed in bioreactor B1 when switching from electron donor to formate on day 200 (culture medium A).
- an aquifer model or (sandbox) was designed in which the water flows horizontally through a sand bed.
- East Aquifer model has an inlet (infiltration well) for the injection of nutrients and microbial consortium and sampling ports.
- transparent acrylic chambers Panexiglas
- Figure 12 transparent acrylic chambers with internal dimensions of 32 x 12 x 1 cm were used; length, width and height respectively ( Figure 12). These were provided with a removable lid, which had holes where ports were installed for the inoculation and stimulation of microorganisms, and a sampling port located downstream with respect to the previous ones.
- an internal pipe was installed at one end, which spanned the entire width of the model, built with Tygon hose (MasterFlex) perforated every 1 cm, It was used for the water inlet that simulates the underground aquifer hydraulic flow.
- Tygon hose was installed for the effluent outlet of the models.
- the internal volume of the model (384 mL) was packed with 545 g of quartz sand with a particle size between 200-800 pm, which was previously washed with 10% HCl (v / v), rinsed with distilled water.
- a brilliant blue (AB) solution (as a tracer) at a concentration of 100 mg / L was passed through the water inlet of the models flow, and samples were taken from the sampling port every 30 minutes since its entry into the models was observed. This procedure was performed until the The concentration of the tracer in the sample port reached half the concentration in the initial solution (i.e., 50 mg / L), and that time was recorded as the time required for the tracer to travel through that model section (or time retention).
- the AB concentration was determined in a spectrophotometer at 630 nm (Dynamica spectrophotometer, model HALO RB-10, Great Britain).
- the effluent volume was collected during 5 min in a test tube previously tared. The value obtained was divided by 5 to obtain the effluent volume in mL per minute.
- the aquifer models were inoculated with the sulfate-reducing microbial consortium using the effluent from the bioreactor B1 of Example 4.
- the microbial composition of this consortium is detailed below in Example 6 (see Tables 4, 5, 6, 7 and 8).
- each inoculum pulse contained T10 9 microorganisms as calculated by microbial count.
- culture medium was supplied continuously at a rate of 0.2 mL / min.
- the composition of the culture medium per liter was: 0.035 g of NaHCC> 3, 0.03 g of MgSO7H 2 0, 2.4 g of K2SO4, 0.007 g FeSC> 4-7H 2 0.1 g of NH 4 CI, with 4.95 g of sodium formate, 0.01 g of KH 2 PC> 4, 1.72 g of CaCl 2 -2H 2 0, 0.1 g of yeast extract and 0.099 g of thioglycolic acid.
- the culture medium was prepared with groundwater extracted from the hydraulic barrier of a tailings dam, whose main components are HCO3 ⁇ 217.7 mg / L, Ca 2+ 302 g / L, 76.6 mg Mg 2+ / L and S04 2 to 1192.3 mg / L
- the effluent from the models was used to make the determinations of pH, potential, formate, microbial count and ATP.
- Example 1 ATP according to Example 3 and hydrogen sulphide (or sulphides) in solution were carried out according to Example 2. Microbial counting was carried out using the methodology described in Example 4.
- a protocol was used to determine the amount of ATP of the microorganisms adhering to the quartz sand of the aquifer models. For this, 200 mg of a sand sample was taken, which was washed with distilled water to remove the bacteria present in the pore volume, after which 1 mL of the cell lysis solution contained in the system was added. determination of ATP. Subsequently, the lysis supernatant was reacted with the other lyophilized components (luciferin-luciferase) to determine the amount of ATP by means of a Kikkoman luminometer, model Lumitester PD-20, Japan. The luminescence value obtained was expressed as URL / g of sand.
- Determination of pH in pore water Once the aquifer models were opened, the pH in the sand pore water was immediately determined in different areas of the models, wetting pH indicator strips. The colors of the strips were compared to the standard provided on the product package. The pH readings were used to make a pH map corresponding to the upper plane of the porous medium of the models.
- X-ray diffraction The sand samples were ground in a mortar to obtain a sample with adequate granulometry to perform X-ray diffraction (XRD). They were then analyzed using a D8 Advance X-ray diffraction machine, Bruker, Germany. The diffractogram obtained was compared with those corresponding to standard mineral samples with CaCC> 3.
- the analyzes of the experiments with the tracer on day 55 show that the blackened feathers represent an area of lower permeability compared to other areas of the porous medium, since it is observed that the dye deviates when it encounters this area, causing most of the hydraulic flow occurs through the non-blackened areas (Figure 16).
- the blackened areas of the plume are due to the formation of iron sulphide precipitates from the hydrogen sulphide generated in the inoculated models and the presence of traces of iron in the infiltrated culture medium.
- the aquifer models were disassembled to determine the pH, the microbial intracellular ATP, the relative abundance of the microorganisms and the carbonate minerals formed in the sand.
- the pH value in the sand pore water was determined by means of pH indicator rods. This allowed making a map of this parameter (Figure 17). Three zones were distinguished in the sand of the models: a) zone 1: first third located in the vicinity of the flow water inlet zone, b) zone 2: second third located in the central zone, and c) zone 3: third third located towards the effluent outlet zone. In all the inoculated aquifer models, a pH 8 zone was observed in zones 2 and 3. Point zones of pH 8.5 were also observed in one of the duplicates, which are located within the pH 8 zones. A lower pH value (close to 7.5) was observed in zone 1. High local pH values are of great importance to induce the precipitation of carbonates. Instead, in the No alkalinization was observed in the control model, since a pH of 7.5 was detected in all areas.
- the control model (not inoculated), showed considerably lower values in all zones compared to the inoculated ones ( Figure 18).
- This example shows that it is possible to establish an active sulfate reducing microbial consortium in the porous medium of a continuous hydraulic flow model aquifer, which consists of hydraulic barrier water from a sulfate-containing tailings dam.
- the microbial reduction of sulfate which is the product of the inoculation and stimulation of the microbial consortium, was able to biomineralize a portion of the quartz sand (porous medium), due to the accumulation of CaCÜ3 minerals, which according to the experiments carried out with the tracer corresponds to an area of less permeability to hydraulic flow.
- the massive sequencing of the 16S rRNA gene was used.
- the microorganisms of the inoculum were concentrated by centrifugation from a sample of the effluent of the bioreactor B1 of Example 4, used for the production of the inoculum.
- a sand sample was taken from the central zone (zone 2) of one of the inoculated duplicates. From these samples, the total genomic DNA of the microorganisms adhered to the sand was extracted for subsequent massive sequencing of the 16S rRNA gene by Illumin technology.
- Tables 4, 5, 6, 7 and 8 show the relative abundance of the 16S rRNA gene sequences in the DNA extracts of the microorganisms of the inoculum used, at the level of Phylum, Order, Class, Family and genus. respectively.
- the sequencing results showed that the main Phyla present in the inoculum microbial community were classified as Proteobacteria, Firmicutes and Bacteroidetes (Table 4).
- the inoculum microbial community was composed mainly of Gammaproteobacteria, Clostridia, Deltaproteobacteria, Betaproteobacteria, Alphaproteobacteria and Bacteroidia (Table 5).
- the predominant Orders corresponded to Oceanospirillales, Clostridiales, Enterobacteriales, Desulfovibrionales, Burkholderiales, Pseudomonadales, Bacteroidales y Caulobacterales (Table 6).
- the main microorganisms present were: Oceanospirillaceae, Peptococcaceae, Enterobacteriaceae, Desulfomicrobiaceae, Pseudomonadaceae, Comamonadaceae, Porphyromonadaceae, Clostridiaceae, Caulobacteraceae and [Tissierellaceae] (Table 7).
- the sequencing results showed that the main genera of the microbial community of the inoculum were classified into: Family Oceanospirillaceae, Desulfosporosinus, Desulfomicrobium, Pseudomonas, Citrobacter, Family
- the results of the sequencing showed that the main Phyla present in the microbial community adhered to the quartz sand were classified as Proteobacteria, Firmicutes and Bacteroidetes (Table 4).
- the microbial community of the sand was composed mainly of Gammaproteobacteria, Clostridia, Deltaproteobacteria and Bacteroidia (Table 5).
- the predominant Orders corresponded to Clostridiales, Desulfovibrionales, Pseudomonadales and Bacteroidales (Table 6).
- the main microorganisms present were: Desulfomicrobiaceae,
- the sequencing results showed that the main genera of the sand microbial community were classified into: Desulfomicrobium, Pseudomonas, Family Porphyromonadaceae, Family Clostridiaceae, Sedimentibacter, Tissierella_Soehngenia and Order Bacteroidales (Table 8).
- the microorganisms that showed the highest relative abundance were Desulfomicrobium (58.37%), Family Porphyromonadaceae (19.04%) and Pseudomonas (4.53%).
- Desulfomicrobium stands out among them for its ability to reduce sulfate biologically.
- Desulfomicrobium increased considerably with respect to the percentage composition of the inoculum community (Table 8). A significant increase in sequences corresponding to a genus of the Porphyromonadaceae family was also found. These differences can be attributed to the particular microenvironmental conditions of the sand.
- the analyzes of this example show that the enriched microbial consortium used as inoculum in Example 5 exhibits high bacterial diversity, among which some classes can be found that include sulfate-reducing bacteria, such as the Deltaproteobacteria. Among them, the Desulfosporosinus and Desulfomicrobium genera stand out for their ability to reduce sulfate biologically. When studying the microbial community adhered to the quartz sand at the end of the experiment in the aquifer model, it was found that this was qualitatively similar to that of the inoculum used, with some microorganisms that significantly increased their proportion. This is the case of the genus Desulfomicrobium.
- the aquifer models were disassembled to determine the pH, the microbial intracellular ATP, the relative abundance of the microorganisms and the carbonate minerals formed in the sand.
- the pH value in the sand pore water was determined by means of pH indicator rods. This allowed making a map of this parameter (Figure 23). Three zones were distinguished in the sand of the models: a) zone 1: first third located in the vicinity of the flow water inlet zone, b) zone 2: second third located in the central zone, and c) zone 3: third third located towards the effluent outlet zone.
- the pH values in sand tended to increase as the concentration of formate in the culture medium increased (Figure 23).
- ATP values in sand were higher in zones 2 and 3 of the models, while zone 1 (upstream to the inoculation point) was lower with respect to the other zones ( Figure 24).
- zones 2 and 3 a dependent increase of formate concentration in the culture medium of this parameter was observed, within the range between 3 and 12 g / L.
- a maximum value of ATP / g of sand is reached, since in the models with the highest amount of formate (18 g / L), these decrease to values similar to those of the test carried out with 6 g / L of formate.
- the increase in the concentration of formate in the culture medium produced an enrichment of the sequences corresponding to the sulfate reducing genus Desulfomicrobium, which predominated in all samples (Table 10), reaching 35, 48, 52 and 57% in the sand from the aquifer models stimulated with 3, 6, 12 and 18 g / L of formate, respectively.
- the increase in the electron donor also produced the notorious enrichment of the sequences corresponding to the genus Pseudomonas.
- the opposite effect on other components of the microbial community was also observed, since the sequences of the predominantly aerobic family were reduced.
- the process consists of a system to reduce hydraulic permeability and generate the precipitation of insoluble minerals in an aquifer with groundwater contaminated with sulfate and / or with sulfate and metals.
- the process consists of at least the steps of: a) providing a culture of microorganisms enriched in sulfate-reducing bacteria and adapted to the groundwater of the aquifer to be treated, b) injecting the aquifer with an enriched culture of microorganisms, c) injecting the aquifer with an electron donor and nutrients suitable for the culture of microorganisms enriched in sulfate-reducing bacteria and one or more metal cations that favor the precipitation of insoluble minerals, and d) allow the microorganisms to multiply and colonize the solid material of the aquifer, reducing the hydraulic permeability of the aquifer and generating the precipitation of insoluble minerals in the solid material of the aquifer.
- the process begins with the extraction of groundwater contaminated with sulfate and / or with sulfate and metals (1), from an underground aquifer, whose flow direction is indicated by arrows (2).
- One well or more wells are used to extract groundwater extraction (3), which are installed from the surface level of the ground (4) to extend below the water table (5).
- the well (s) In the water table sector, the well (s) have grooves, perforations or other permeable sections (6), which allow the extraction of groundwater.
- the extracted groundwater enters through the conduit (7) to the pond (8).
- conduit (9) an electron donor and nutrients suitable for the cultivation of microorganisms enriched in sulfate-reducing bacteria and one or more metal cations that favor the precipitation of insoluble minerals are added.
- the added compounds are stirred in the pond to obtain a solution
- the solution is entered into the fixed-bed anaerobic bioreactor (11), which has been previously inoculated with an enriched sulfate reducing microbial consortium, of according to the previous Examples.
- the well (s) In the water table sector, the well (s) have grooves, perforations or other permeable sections (14), which allow the injection of microorganisms and nutritive solutions into the groundwater.
- the solution from the pond (8) is also added directly, through the conduit (15) to one well or more injection wells (13), said solution is injected to groundwater through slots, perforations, or other permeable sections (14).
- Microorganisms proliferate in the pores and on the surface of the particulate matter in the underground aquifer, impeding the flow of water. Furthermore, the metabolic activity of microorganisms, especially the biological reduction of sulfate, produces the partial removal of sulfate and the alkalization of the solution. As a result of the above, it is possible to generate a zone of reduction of hydraulic permeability and of precipitation of insoluble minerals in situ (16) in an area downstream of the injection point in an underground aquifer contaminated with sulfates. Insoluble minerals are primarily metallic carbonates and sulfides.
- Carbonates such as calcite, aragonite, or vaterite, are formed in the environment alkaline, from bicarbonate produced by the anaerobic microbial degradation of organic matter and calcium added and / or present in groundwater.
- metallic sulphides particularly FeS, precipitate by the reaction between the biologically formed H2S and iron oxides, such as maghemite, magnetite, hematite or goethite commonly present in soils and in particulate matter of underground aquifers.
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Abstract
L'invention concerne un procédé in situ pour réduire la conductivité hydraulique et générer la précipitation de minerais insolubles dans un aquifère souterrain qui est affecté par des infiltrations d'eau de surface ou par des infiltrations d'eau d'un bassin de retenue des résidus miniers, qui comprend au moins les étapes consistant à : (a) fournir une culture de micro-organismes enrichi en bactéries réductrices de sulfate et adapté aux eaux souterraines de l'aquifère à traiter, (b) injecter, dans l'aquifère souterrain, une culture de micro-organismes enrichie, (c) injecter, dans l'aquifère souterrain, un donneur d'électrons et d'éléments nutritifs pour la culture de micro-organismes enrichi en bactéries réductrices et un ou plusieurs cations métalliques qui favorisent la précipitation de minerais insolubles, et (d) permettre que les micro-organismes se multiplient et colonisent le matériau du solide de l'aquifère souterrain, la perméabilité hydraulique de l'aquifère souterrain étant ainsi réduite et la précipitation de minerais insolubles dans le matériau solide de l'aquifère souterrain étant ainsi générée.
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EP4023353A1 (fr) * | 2020-12-31 | 2022-07-06 | Groundwater Technology B.V. | Procédé de précipitation minérale microbienne dans un support poreux doté d'une conductivité hydraulique |
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US20020166813A1 (en) * | 2001-05-14 | 2002-11-14 | Bartlett Robert W. | In situ anaerobic bioremediation of earth and sold waste contaminants using organic/water emulsions |
US20090324344A1 (en) * | 2006-05-09 | 2009-12-31 | Van Der Zon Wilhelmus Hendrikus | Biosealing |
US20170232490A1 (en) * | 2007-03-16 | 2017-08-17 | Jrw Bioremediation, Llc | Bioremediation enhancing agents and methods of use |
AU2018236858A1 (en) * | 2018-05-04 | 2019-11-21 | Korea Advanced Institute Of Science And Technology | Reinforcement method for water leaking of water facilities using soil microbes biostimulation and microparticles injection |
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US20020166813A1 (en) * | 2001-05-14 | 2002-11-14 | Bartlett Robert W. | In situ anaerobic bioremediation of earth and sold waste contaminants using organic/water emulsions |
US20090324344A1 (en) * | 2006-05-09 | 2009-12-31 | Van Der Zon Wilhelmus Hendrikus | Biosealing |
US20170232490A1 (en) * | 2007-03-16 | 2017-08-17 | Jrw Bioremediation, Llc | Bioremediation enhancing agents and methods of use |
AU2018236858A1 (en) * | 2018-05-04 | 2019-11-21 | Korea Advanced Institute Of Science And Technology | Reinforcement method for water leaking of water facilities using soil microbes biostimulation and microparticles injection |
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COTORAS TADIC, D., PROYECTO FONDEF DE INVESTIGACION Y DESARROLLO: BIOMINERALIZACION APLICADA EN MINERIA , CODIGO DEL PROYECTO CA 13110019, 4 April 2018 (2018-04-04), XP055752865, Retrieved from the Internet <URL:http://repositorio.conicyt.cl/bitstream/handle/10533/209771/CA13110019.pdf?sequence=1&isAllowed=y>> [retrieved on 20200616] * |
SINGH, R. ET AL.: "Removal of sulphate, COD and Cr(VI) in simulated and real wastewater by sulphate reducing bacteria enrichment in small bioreactor and FTIR study", BIORESOURCE TECHNOLOGY, vol. 102, 2011, pages 677 - 682, XP027581361, DOI: 1 0.1 016/j.biortech.201 0.08.041 * |
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EP4023353A1 (fr) * | 2020-12-31 | 2022-07-06 | Groundwater Technology B.V. | Procédé de précipitation minérale microbienne dans un support poreux doté d'une conductivité hydraulique |
NL2027257B1 (en) * | 2020-12-31 | 2022-07-21 | Groundwater Tech B V | Method of microbial mineral precipitation in a porous medium having hydraulic conductivity |
CN113617827A (zh) * | 2021-06-30 | 2021-11-09 | 中冶成都勘察研究总院有限公司 | 一种矿山修复土层结构改良方法 |
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