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By Caren Caldwell, Leslie Thompson

1.1 Biological Process Description Pintail Systems, Inc. (PSI) has developed and in partnership with their clients in the mining industry has applied biological remediation processes for detoxification of spent ore and process solutions from heap leach operations. Contaminants that are treated to mine closure levels or drinking water standards include cyanides (weak acid dissociable and total cyanides), thiocyanate, nitrates and heavy metals. Biological processes are both site-specific and ore-specific processes. PSI specializes in isolating indigenous microorgan­isms from spent ore and process solutions and augments them in the laboratory for enhanced bio-detoxification in the field. Bioaugmentation in the lab is a key to successful heap bio-detox in the field. In PSI's bioaugmentation processes native microorganisms are isolated from the spent ore environment and are tested for contaminant remediation capacity. A majority of the indigenous microbes will be tolerant of the spent ore environment but will not contribute to contaminant metabolism. Indigenous enrichment or biostimulation of these non-working microbes by adding trace nutrients can competitively inhibit the working microbes. For this reason PSI focuses on the bioaugmentation process where non-working microbes are discarded and the working population is enhanced through stress, mutation and exogenous enrichment before it is re-introduced to the heap and process solutions.

Jan 1, 1997

By Yu. Gurevich, M. Teremova, I. Prokopev, A. Razvjaznaya, N. Algebraistova

"The objective of this work is the scientific justification and development of efficient and environmentally-friendly method of bulk concentrate preparation for selection with application of microbiological practices. This research suggests to apply Pseudomonas Japonica bacterial culture to the bulk concentrate before the selection circuit in order to clear the surface of sulphides from the xanthate skin. It is also suggested to apply bacterial consortium Ochrobactrum ?nthropi and Pseudomonas aeruginosa JCM 5962 for desorption of diesel residual concentrations. High-efficient and environmentally friendly method of copper-molybdene and lead-zinc bulk concentrate selection was developed based on theoretical and experimental research work with application of microbiological practices. In comparison with conventional methods (steaming, sodium sulphide washing) the above suggested bio-technological method of bulk concentrate preparation before selection cycle will allow to reduce material and power expenditures, and it is also environmentally friendly. INTRODUCTION High-efficient selection of sulphide bulk concentrate is gained exclusively after collecting agent is removed from mineral surface. Therefore, the problem of bulk concentrate preparing before the selection cycle is acute, since the further technological parameters of the subsequent separation depends on the effectiveness of pulp preparation. Existing traditional ways of bulk concentrate preparation. such as washing or steaming, are characterized by high energy and material costs in industrial conditions [1,2]. The microbiological treatment before separation of the bulk concentrate into different kind of concentrates appear one of perspective processes for treatment of bulk concentrates [3,4]. At the present time, the world practice of processing mineral raw materials demonstrates the wide application of technologies using microorganisms: bacterial leaching and oxidation, purification of industrial wastewater. This is due to the fact that biotechnological methods are environmentally safe, low-cost, and suitable for refining complex ores and raw materials of industrial origin."

Jan 1, 2018

By C. W. Hancher, B. D. Patton, W. W. Pitt

"There are a number of nitrate-containing wastewater sources, as concentrated as 30 wt % NO3 and as large as 2000 m3/day, in the nuclear fuel cycle. The biological reduction of nitrate in waste water to gaseous nitrogen, accompanied by the oxidation of a nutrient carbon source to gaseous carbon dioxide, is an ecologically sound and cost-effective method of nitrate waste disposal. These nitrate-containing wastewater sources can be successfully biologically denitrified to meet discharge standards in the range of 10 to 20 g N(NO3-)/m3 by the use of a fluidized bed bioreactor. The denitrification bacteria are a mixed culture derived from garden soil; the major strain is Pseudomonas. In the fluidized-bed bioreactor, the bacteria are allowed to attach to 0.25- to 0.50-mm-diam. coal fluidization particles, which are then fluidized by the upward flow of influent wastewater. Maintaining the bacteria-to-coal weight ratio at approximately 1:10 results in a bioreactor bacteria loading of greater than 20,000 g/m3,This paper describes the results of a bio denitrification R&D program based on the use of fluidized bioreactors capable of operating at nitrate levels of up to 7000 g/m3 and achieving denitrification rates as high as 80 g N(N03-) per day per litre of empty bioreactor volume. IntroductionThe discharge of wastewater containing nitrate at concentrations greater than stipulated by the Environmental Protection Agency (EPA) creates an environmental hazard and constitutes a civil and/ or criminal legal liability.(I.2) At present, we have three options: (1) recycle and reuse of nitrate streams the total economic and technical balance should be Closely examined; (2) use of an alternative acid, which may pose an entirely different set of problems ; or (3) disposal of nonreusable nitrate sources by an environmentally acceptable procedure."

Jan 1, 1980

By Tina Maniatis

Applied Biosciences has developed a biological technology for removal of arsenic, nitrate, selenium, and other metals from mining and industrial waste waters.The ABMet® technology was implemented at a closed gold mine site in Canada for removing arsenic from tailings pond water. The system included six bioreactors that began treating water in the spring of 2004. Design criteria incorporated a maximum flow of 567 L/min (150 gallons per minute) and water temperatures ranging from 10°C to 15°C. Influent arsenic concentrations range from 0.5 mg/L to 1.5 mg/L. The ABMet® technology consistently removes arsenic to below detection limits (0.02 mg/L). Data from the full scale system will be presented, as well as regulatory requirements and site specific challenges. Keywords: Arsenic, Sulfate Reduction, Metal Sulfide Precipitate, Biological Water Treatment

Jan 1, 2005

By J. D. Isbister

Coal is the most abundant and most economical source of energy in the United States, however, combustion of coal results in release of pollutants to the environment. The release of sulfur dioxide and oxides of nitrogen during combustion of coal contributes to the deterioration of the environment. Energy independence in the United States must include the increased use of coal reserves. Increased reliance on coal for energy will dictate the necessity for making coal a cleaner fuel. New technologies employing a variety of techniques from harsh chemical treatment to mild physical separations to clean coal are rapidly becoming available. Success for any coal cleaning process is in economics which translate into efficient removal of sulfur with high recovery of combustibles as well as low capital and operating costs for the process. Conventional coal cleaning processes remove water soluble sulfates and a portion of the pyritic sulfur and ash from coals. Removal of additional sulfur from coals requires more advanced coal cleaning techniques. Coal contains many organic sulfur forms which are the most difficult of the sulfur types to remove from coal. The heteroaromatic sulfur species, such as the comlex thiophenes, are the most commonly found organic sulfur forms. These sulfur forms are resistant to chemical attack and are considered to be refractory compounds. Atlantic Research Corporation has developed an "unique" microorganism, CB1 (Coal Bug 1), capable of oxidizing thiophenic sulfur in coal to form water soluble sulfates.

Jan 1, 1986

By L B. Sukla, V N. Misra

Microbial desulfurisation of coal has significant advantages over physicochemical desulfurisation processes since the former process selectively removes the inorganic sulfur compounds and heavy metals without significant carbon loss. The purpose of this study was to improve the desulfurisation yield of a coal sample from northeastern coalfields, Assam, India using microbial technique. The coal sample contained 2.13 per cent of total sulfur and 2.59 per cent of ash. Stirred tank reactors of 40 L capacity were operated in batch mode. Initially, the reactor was fed with 40 L of slurry containing ten per cent (w/v) coal, ten per cent (v/v) inoculum of mixed culture of mesophilic acidophiles, native to coal itself and the requisite amounts of constituents for 9K medium without added ferrous sulfate. The percolate of the reactor was then used to seed the medium for the next experiment. The purpose was to obtain a better adapted and more active biomass which would give a better desulfurisation yield for coal. Further studies were conducted in the bioreactor, fed with 40 L of slurry containing ten per cent (w/v) pulp density of coal, ten per cent inoculum (derived from previous studies) and the medium, supplemented with ferrous sulfate. Results showed an increased total desulfurisation yield of 43 per cent in the latter treatment in comparison to the previous one, where the desulfurisation yield was only 23 per cent. The findings also demonstrated the convenience of using inocula derived from coal, where 50 - 60 per cent of pyritic sulfur was removed, while chemical desulfurisation is of little significance with the majority of coal types. The ash content was also reduced to 1.74 per cent as the low pH of the process caused dissolution of a number of common minerals present in the coal.

Jan 1, 2004

By Robert D. Rogers

The demand for phosphate will continue into the future, because phosphate is currently irreplaceable as a plant nutrient and in many chemical applications. Phosphate is not recycled, so the supply must be replenished through a process of mining and purification. Numerous microbial isolates. obtained from the environment were screened find those which demonstrated the greatest promise of solubilizing PO4 from the ore of interest. Screening was accomplished in a step wise manner an agar plate bioassay, shake flask studies, and semi continuous growth conditions. It was found that with the proper conditions of nutrient supply and solution pH; the bacteria and. fungus used in the semicontinuous process were able to solubilize in excess of 80% of an Idaho rock phosphate ore (RP) within nine days. A significant amount of solubilization occurred within the first six days. There was a 'reasonable correlation between the occurrence of soluble Ca and PO.4 during the processing of the RP thus providing evidence that the RP was being disintegrated by the organisms. In all cases, HPLC analysis of the solutions in which PO4 had been. extracted from RP showed that organic acids, which have been implicated as the mechanism of solubilization, were present.

Jan 1, 1991

By Daniel C. McLean

Extraction data for Cu, Zn and Fe were obtained from 4.5" and 10" columns using massive sulfide ore and natural acid mine water. The purpose was to determine if recycling of solution through the column would produce leach liquors of 4 g/l Cu and Zn at extraction rates of 1 g/l per pass at typical flow rates of 0.002 to 0.006 gpm/sq.ft. Test results exceeded expectations. Cu and Zn concentrations of 4 g/l were obtrained within 4 to 6 cycles. Concentrations greater than 6 g/l Cu and 8 g/l Zn were reached within 10 cycles. Concentrations of greater than 40 g/l of Fe and free sulfuric acid were obtained with no decrease in biological activity being observed. No reagents were used.

Jan 1, 1995

By K. A. Natarajan, S. Usha Padukone

"An industrial waste liquor having high sulfate concentrations was subjected to biological treatment using the sulfate-reducing bacteria (SRB) Desulfovibrio desulfuricans. Toxicity levels of different sulfate, cobalt and nickel concentrations toward growth of the SRB with respect to biological sulfate reduction kinetics was initially established. Optimum sulfate concentration to promote SRB growth amounted to 0.8 - 1 g/L. The strain of D. desulfuricans used in this study initially tolerated up to 4 -5 g/L of sulfate or 50 mg/L of cobalt and nickel, while its tolerance could be further enhanced through adaptation by serial subculturing in the presence of increasing concentrations of sulfate, cobalt and nickel. From the waste liquor, more than 70% of sulfate and 95% of cobalt and nickel could be precipitated as sulfides, using a preadapted strain of D. desulfuricans. Probable mechanisms involving biological sulfide precipitation and metal adsorption onto precipitates and bacterial cells are discussed.IntroductionIndustrial effluents from hydrometallurgical and mineral processing operations often contain high sulfate and metal concentrations posing significant disposal problems, which require urgent solution to avoid serious environmental contamination. In such waste liquors, sulfate and heavy metals such as cobalt, nickel, zinc and iron originate from the chemical or biological oxidation of exposed sulfide minerals. Reduction of sulfate and precipitation of toxic metal ions from industrial wastes is essential from the viewpoint of environmentally acceptable waste disposal and recovery of associated strategic metals.Many of these heavy metals are essential for metabolic activity of bacterial cells at low levels. But they exert toxic effects at concentrations encountered in polluted water. Removal of toxic metals through biological reduction of sulfates from industrial waste waters have been practiced for several decades. Metal remediation through common physicochemical techniques such as oxidation-reduction, chemical precipitation, filtration, electrochemical treatment, evaporation, ion exchange and reverse osmosis processes is expensive and not suitable in the case of voluminous effluents containing complexing organic matter and low metal concentrations. Further, strong and contaminating reagents are used for desorption, resulting in toxic sludge and secondary environmental pollution. Biotechnological approaches can be more efficient and cost effective for detoxification of such effluents. Biotreatment/ bioremoval offers several advantages over conventional chemical and physical treatment technologies, especially for diluted and mixed contaminants. Biological detoxification, which involves the use of microbes to detoxify and degrade environmental contaminants, has received increasing attention in recent years (Iwamoto and Nasu, 2001). In addition, valuable metals can be recovered from metal sulfide sludge (Boonstra et al., 1999)."

Jan 1, 2015

By G A. Ekama

This paper describes a novel system for the biological sulfate reduction (BSR) of acid mine drainage (AMD) using primary sewage sludge (PSS) as carbon source in an upflow anaerobic sludge bed (UASB) reactor configuration. A UASB reactor was operated at a temperature of 35¦C and it received PSS (1875 mg/L COD) augmented with sulfate (1500 mg/L SO42-). The experimental results indicate that a high treatment efficiency was achieved at more than 90 per cent sulfate reduction at a liquid hydraulic retention time (HRT) of 13.5 h. In this study, the effects of various operational parameters were also investigated. The effect of a biomass recycle stream from the top to the bottom of the sludge bed was found to initiate rapid BSR from the bottom of the bed. Profile tests showed that effective and immediate sulfate reduction was achieved as soon as the influent entered the reactor. From these results, it can be concluded that the UASB configuration using PSS as energy source would be a viable method for the BSR of AMD.

Jan 1, 2009

By A. L. DeVegt, H. G. Bayer, C. J. Buisman

During the last ten years, Paques Inc. has been engaged in the development and installation of treatment systems to remove sulfur compounds from water and gaseous streams using biotechnological processes. Metal and sulfate can be removed from mine waters in two biological steps. In the first step, sulfate-reducing bacteria converts sulfate to hydrogen sulfide (H2S). The HS reacts with the dissolved metals to form insoluble metal sulfide precipitates. In the second step, sulfide-oxidizing bacteria convert excess H2S to elemental sulfur. The Budelco Zinc Refinery in The Netherlands installed a groundwater treatment system to remove metals and sulfate using the biological steps described above. The system has been operating since May 1992 and has been treating 5,000 m3/d (180 thousand cu ft/day). A pilot plant began operation in November 1995 at Kennecott's Bingham Canyon Utah copper mine to develop processes for metal and sulfate removal from groundwater and for the selective copper recovery from leach water. The principle of the biotechnological methods, the full-scale experience and an overview of the test program at the mine are presented

Jan 1, 1999

By A. L. de Vegt

For the past ten years Paques has been engaged in the development and installation of treatment systems based on biotechnological processes to remove sulfur compounds from water, air and gaseous streams. Metal and sulfate can be removed from mine waters using two biological steps: 1. Sulfate reducing bacteria convert sulfate to hydrogen sulfide (H2S). The H2S reacts with the dissolved metals to form insoluble metal sulfide precipitates. 2. Sulfide oxidizing bacteria convert excess H2S to elemental sulfur. Paques has designed and installed a groundwater treatment system for the Budelco zinc refinery in the Netherlands to remove metals and sulfate as described above. The system has been operating since May 1992 treating a flow of 5000 m /d. A pilot plant started operation in November 1995 at Kennecott's Bingham Canyon Utah copper mine to develop the processes for metal and sulfate removal from groundwater and selective copper recovery from leach water. The principle of the biotechnological methods, full scale experience, and an overview of the test program at the copper mine are presented.

Jan 1, 1997

By B. Waterman, R. Collins, V. R. K. Paruchuri, P. Gonzalez, H. Dijkman

"INTRODUCTION Mining and processing of sulfide ore bodies has the potential to solubilize sulfate. Gypsum in the host rock, as well as oxidation of sulfides during milling and processing can solubilize sulfate. Acid rock drainage (ARD) also has the potential to generate waters high in sulfate. Even after treatment of the ARD with lime, the treated water can have sulfate present in gypsum saturation levels. Most mine-sites reuse sulfate impacted water but as a mine nears closure, reuse options may be limited and water treatment becomes an option. Furthermore, sulfate discharge requirements, particularly for anti-degredation narrative standards, can be very strict. Freeport has investigated many new technologies to economically remove sulfate from these types of water. A Dutch company, Paques B.V. developed one of these technologies and it utilized biological reactors to convert sulfate into elemental sulfur in a 2-step process. In 1992 the first industrial reference of the SULFATEQTM technology was commissioned at the Nystar Budel zinc refinery in the Netherlands. PILOT PLANT TECHNOLOGY AND DESCRIPTION As shown in Figure 1, the plant contains several process steps, which can mainly be divided into the following sections: 1. Anaerobic biological system in which sulfate is reduced to hydrogen sulfide by sulfate reducing bacteria. 2. Aerobic biological system in which hydrogen sulfide is oxidized to elemental sulfur by sulfide oxidizing bacteria. 3. Solid/liquid separation in which sulfur, calcium carbonate and biomass solids are separated and dewatered. Sulfate impacted ground water is pumped to the feed water tank through a plate and frame heat exchanger to maintain the solution temperature around 95°F. A custom mix of micronutrients is dosed to the feed tank. This nutrient mixture provides essential elements for the growth of anaerobic and aerobic biomass. Urea is added to the feed tank and provides the required nitrogen source to the biomasses. The pretreated feed water is pumped into the anaerobic reactor. The reactor works on the gas lift principle. Sulfate reducing bacteria utilize hydrogen gas as electron donors and reduce sulfate to hydrogen sulfide. This reaction increases the alkalinity of the solution. Carbon dioxide gas is added in order to control the pH within working range of the reactor. Due to the low solubility of the hydrogen gas, the anaerobic gas mixture is recycled through the system. The gas mixture recycle also provides the gas lift for mixing within the anaerobic reactor."

Jan 1, 2016

By C. J. N. Buisman

THIOPAQ technology developed and marketed by PAQUES Bio Systems in Balk, Netherlands, has been successfully used at commercial scale at the Budelco zinc refinery in the Netherlands for the treatment of contaminated groundwater since 1992. In essence, THIOPAQ technology consists of two biological process steps in series: sulfate reduction to hydrogen sulfide (anaerobic) and sulfide oxidation to elemental sulfur (aerobic). The biogenic sulfide produced can be employed for the chemical precipitation of metals in solution either inside the anaerobic reactor or in a separate vessel. Since the solubilities of most metal sulfides are much lower than of their respective hydroxides, considerably lower effluent metal concentrations can be achieved with THIOPAQ systems than in neutralization processes which immobilize metals predominantly by hydrolytic precipitation. In recent years, PAQUES has made significant advances in the design and operation of aerobic and anaerobic bioreactors for metal-sulfur systems. Moreover, the company has increased its technology portfolio to allow the development of more process oriented applications of THIOPAQ technology. A new PAQUES designed plant at Budelco treating sulfatecontaining proceswaters will be started up in 1999. The sulfate reduction is accomplished in a 500 m3 reactor in which hydrogen is used as electron donor. The endproduct, ZnS, is used for Zn and H2S04 production.

Jan 1, 1999

By Lyn Jones, Mike Bratty, Rick Lawrence, David Kratochvil

"The treatment of water is required at many mining operations to remove metals and prevent the discharge of contaminated water to the environment. In other cases, water treatment is used to provide better quality water for recycle to the processing plant, or to process bleed streams in hydrometallurgical operations in order to prevent deterioration in metallurgical performance or process equipment. BioteQ Environmental Technologies Inc. has developed and commercialized the BioSulphide® Process which can provide an alternative for these treatment needs and which can also be used for profitable metal recovery, particularly in cases where conventional metal winning techniques are not economical. The BioSulphide® Process is a high rate anaerobic biotechnology for on-site production of H2S from sulphur species, although for smaller flows and metal loadings, the use of chemical sulphide reagent might be more cost effective. Both new projects and existing commercial operations, used for removal and/or recovery of copper, nickel, and zinc, and which range in sulphide generating capacity from 50 kg/day to 3.7 tonnes/day, are discussed. Particular focus is placed on the results of operations at the Raglan Mine in Northern Quebec where dissolved nickel is removed from contaminated water at a high efficiency through the precipitation of nickel sulphide. Plant effluent is discharged directly to the receiving environment and the high-grade nickel sulphide product is combined with the nickel flotation concentrate produced at the mine. The plant at Raglan will replace the existing conventional water treatment plant and will eliminate the need for on-site storage of treatment plant sludge.INTRODUCTIONThe BioSulphide® Process, which produces low cost H2S for use in selective metal precipitation, offers a low capital cost alternative which can operate efficiently and cost-effectively within a wide range of flows and solution grades. Since the technology can be used to recover metals from solutions with low flows and metal grades, the technology is also applicable to environmental applications for water treatment, with the sale of recovered metals providing an offset to treatment costs. For environmental control, the technology has a distinct advantage in being able to meet strict effluent discharge criteria due to the very low solubility of metal sulphides. To date, BioteQ Environmental Technologies has designed and constructed three plants for metal sulphide precipitation, with another three projects in the advanced development stage."

Jan 1, 2006

R F Johnston 1 ABSTRACT The possibilities for bacterial leaching of refractory gold ores found in the Central Victorian region of Australia are examined. The behaviour of bacteria, especially 77iobacillus ferroaridans is outlined and the effects of these when they are exposed to pyrite, arsenopyrite and stibnite are examined. The factors which are particular to Central Victoria are outlined and the possibility of adapting a bacterial culture to suit the conditions is discussed. It is concluded that a bacterial leaching system is a feasible proposition and that it could enhance leaching rates and yields.

Jan 1, 1994

By J. L. Whitlock

An attached growth aerobic biological treatment process has been developed at Homestake Mining Co.'s Lead operation which not only oxidizes free and complexed cyanides, including the stable iron complexed cyanides, but also thiocyanate, and the oxidation byproduct ammonia. Through the employment of a mutant strain of bacteria which has been gradually and specifically acclimated to the waste, these potential pollutants are mineralized to relatively harmless sulfates, carbonates, and nitrates. The resultant effluent, through toxicological testing, has been shown compatible with the receiving stream, which serves as a cold water marginal trout fishery.

Jan 1, 1985

By Terry I. Mudder

Metal complexed cyanides in wastewaters form as a result of interactions of free cyanide with metals present in the wastewater and exhibit varying degrees of stability, toxicity, and treatability. Thiocyanate, a pollutant commonly found in cyanide bearing wastewaters, is formed through the interaction of free cyanide with sulfur containing species (i. e. pyrrhotite) both present in the wastewater. In certain industrial processes, such as the beneficiation of gold and silver, cyanide is an essential reagent. Since free cyanide, complexed cyanides, and thiocyanate are potentially toxic to humans and aquatic organisms, these compounds must be removed from wastewaters prior to their discharge into surface or ground waters serving as potential potable water sources, marine or fresh water habitats

Jan 1, 1984

By M. Balakrishnan, K. A. Natarajan, J. M. Modak, J. S. Gururaj Naik

The ability of various fungal species isolated from gold mines to uptake gold and base metals such as zinc, copper, iron and lead from gold-process cyanide effluents was investigated. Under identical treatment conditions, different strains showed wide variations in their ability to take up gold and other base metals. Aspergillus terreus exhibited as high as a 60% gold uptake. Almost complete removal of zinc was possible with various fungi and with the yeast Saccharomyces cerevisiae. Studies at various metal concentrations showed a higher gold uptake from dilute solutions, and studies with Aspergillus niger showed that gold uptake by living cells increased gradually as the contact time increased. Metal uptake capacities between living and nonliving cells were also compared. A waste biomass (ABM- I), obtained from a fermentation industry, wasalso identified as having a high gold uptake capacity. At 5% (W/v) solids, ABM-I could efficiently remove up to 83% of the gold contained in an effluent solution within 24 hrs.

Jan 1, 1995

By Brad Wahlquist

An ex-situ biological treatment system was implemented at a gold mine site in South Dakota. The system removes selenium and nitrate from a mixture of groundwater and surface water runoff at ambient water temperatures. The Applied Biosciences? system replaced a denitrification system that did not perform consistently, did not treat for selenium and required heating of the influent water. Installation was completed in phases starting with the selenium circuit being brought into operation in February, 2002 and the nitrate circuit in March, 2002. System design can accommodate average flows of 380 L/min (100 gallons/minute) and a maximum flow of 1,136 L/min (300 gallons/min). Four cells were designed to remove nitrate from approximately 30 mg/L to below 10 mg/L and two cells were designed to remove selenium from 0.01 mg/L and higher, to below 0.005 mg/L. No pretreatment is required to operate the system at ambient temperatures, which can fall as low as -12C in the winter, and treat water at temperatures ranging between 8C to 16C. Nutrient requirements are met using a sugar based nutrient mixture that includes a balanced C:N:P:S ratio, trace elements and vitamins. Since start up, the system has consistently been in compliance for nitrate and selenium, treating water to below detection limits for both contaminants. This paper discusses laboratory biotreatability testing of a system designed to remove selenium, arsenic, and nitrate and full-scale implementation and optimization of a system to remove nitrate and selenium.

Jan 1, 2003