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Microbial acquisition of iron from ferric iron bearing minerals

Description: This is the final report of a three-year, Laboratory Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). Iron is a universal requirement for all life forms. Although the fourth most abundant element in the geosphere, iron is virtually insoluble at physiological pH in oxidizing environments, existing mainly as very insoluble oxides and hydroxides. Currently it is not understood how iron is solubilized and made available for biological use. This research project addressed this topic by conducting a series of experiments that utilized techniques from both soil microbiology and mineral surface geochemistry. Microbiological analysis consisted of the examination of metabolic and physiological responses to mineral iron supplements. At the same time mineral surfaces were examined for structural changes brought about by microbially mediated dissolution. The results of these experiments demonstrated that (1) bacterial siderophores were able to promote the dissolution of iron oxides, (2) that strict aerobic microorganisms may use anaerobic processes to promote iron oxide dissolution, and (3) that it is possible to image the surface of iron oxides undergoing microbial dissolution.
Date: December 31, 1998
Creator: Hersman, L.E. & Sposito, G.
Partner: UNT Libraries Government Documents Department

Microbial recovery of metals from spent coal liquefaction catalysts. Quarterly report, April--June, 1992

Description: During April and May the authors began the rather laborious task of crushing each of the solvent-extracted catalyst samples for the determination of the effect of crushing on leaching rate and percent metal recovered. The crushing of these catalyst samples employed a ball mill and it took approximately 1 week to crush each sample. All catalyst samples, after crushing, were sieved to a particle size of >100 mesh. Samples of 10, and 40 mesh were also collected, but only the >100 mesh was subjected to bacterial leaching. These samples were tested using denitrifying bacteria for their ability to release Ni and Mo from these samples. The data are shown in the attached figures as well as comparisons for rates of Ni and Mo released in relation to catalyst solvent pretreatment. These data were obtained with citrate as a carbon and energy source for the microorganisms. It is quite clear that crushing the catalyst allows for a more rapid release of these metals from the catalyst samples, regardless of the organic solvent used to pretreat the catalyst. However, the effects of the different solvents are still evident, although not as striking, after the catalysts are crushed (see previous reports). There also still appears to be a greater ease in removing Mo over Ni, a case particularly prevalent if ethyl acetate, methanol or xylene is used as the pretreatment solvent. In any case, it appears that a variety of solvents will be useful and that an optimized system could yield >90% release and recovery of both Ni and Mo in as little as 10 days of treatment. Experimental data are enclosed.
Date: December 31, 1992
Creator: Sperl, P.L. & Sperl, G.T.
Partner: UNT Libraries Government Documents Department

Microbial recovery of metals from spent coal liquefaction catalysts. Final report

Description: This project was initiated on October 1, 1989, for the purpose of recovering metals from spent coal liquefaction catalysts. Two catalyst types were the subject of the contract. The first was a Ni-No catalyst support on alumina (Shell 324), the catalyst used in a pilot scale coal liquefaction facility at Wilsonville, Alabama. The second material was an unsupported ammonium molybdate catalyst used in a pilot process by the Department of Energy at the Pittsburgh Energy Technology Center. This material was obtained in late February 1990 but has not been pursued since the Mo content of this particular sample was too low for the current studies and the studies at the Pittsburgh Energy Technology Center have been discontinued. The object of the contract was to treat these spent catalysts with microorganisms, especially Thiobacillus ferrooxidans , but also other Thiobacillus spp. and possibly Sulfolobus and other potential microorganisms, to leach and remove the metals (Ni and Mo) from the spent catalysts into a form which could be readily recovered by conventional techniques.
Date: July 1, 1995
Creator: Sperl, P.L. & Sperl, G.T.
Partner: UNT Libraries Government Documents Department

Microbial recovery of metals from spent coal liquefaction catalysts. Final and quarterly report, July 1994--September 1994

Description: Research is reported on the recovery of molybdenum and nickel from spent coal liquefaction catalysts. Mo release from spent coal liquefaction catalysts has been shown to be dependent upon many parameters, but release is dominated by microbial growth. The microbial Mo release is a rapid process requiring less than one week for 90% of the releaseable Mo to be solubilized from whole washed (THF) catalyst. It could be expected that the rates would be even greater with crushed catalyst. Efforts were centered on optimizing the parameters that stimulate microbial growth and action and further efforts centered on catalyst pre-treatment prior to microbial bio-leaching. Recent experiments suggest that hydrogen peroxide promises to be an effective pre-treatment wash. Hydrogen peroxide was also found to be an effective and economical agent for metals solubilization per se and could promote solubilization without subjecting the catalyst to microbial growth.
Date: August 1, 1995
Creator: Sandbeck, K.A. & Cleveland, D.
Partner: UNT Libraries Government Documents Department

Microbial Ecology Assessment of Mixed Copper Oxide/Sulfide Dump Leach Operation

Description: Microbial consortia composed of complex mixtures of autotrophic and heterotrophic bacteria are responsible for the dissolution of metals from sulfide minerals. Thus, an efficient copper bioleaching operation depends on the microbial ecology of the system. A microbial ecology study of a mixed oxide/sulfide copper leaching operation was conducted using an "overlay" plating technique to differentiate and identify various bacterial consortium members of the genera Thiobacillus, �Leptospirillum�, �Ferromicrobium�, and Acidiphilium. Two temperatures (30C and 45C) were used to select for mesophilic and moderately thermophilic bacteria. Cell numbers varied from 0-106 cells/g dry ore, depending on the sample location and depth. After acid curing for oxide leaching, no viable bacteria were recovered, although inoculation of cells from raffinate re-established a microbial population after three months. Due to the low pH of the operation, very few non-iron-oxidizing acidophilic heterotrophs were recovered. Moderate thermophiles were isolated from the ore samples. Pregnant liquor solutions (PLS) and raffinate both contained a diversity of bacteria. In addition, an intermittently applied waste stream that contained high levels of arsenic and fluoride was tested for toxicity. Twenty vol% waste stream in PLS killed 100% of the cells in 48 hours, indicating substantial toxicity and/or growth inhibition. The data indicate that bacteria populations can recover after acid curing, and that application of the waste stream to the dump should be avoided. Monitoring the microbial ecology of the leaching operation provided significant information that improved copper recovery.
Date: June 1, 1999
Creator: Bruhn, D. F.; Thompson, D. N. & Noah, K.S .
Partner: UNT Libraries Government Documents Department

Effect of thiamine hydrochloride on the redox reactions of iron at pyrite surface. [Fourth quarterly techical progress report, September 1990--November 1990]

Description: The present investigation is a part of our studies on the electro chemical aspects of pyrite bioleaching involving Thiobacillus ferrooxidans. Previously (1,2) we have examined the effect of T. ferrooxidans and their metabolic products on the redox reactions of Fe{sup 2+}/Fe{sup 3+} couple at the pyrite surface. Results obtained suggest that beyond 1. 5 days during their growth in a batch fermenter, the bacteria and their metabolic products completely cover the pyrite surface and shut down all electron transfer across the electrode-solution interface. In addition, it has been observed that the bacteria serve as the nucleation site for jarosite formation, which is found detrimental to bioleaching. In the present work we have focussed on the effect of the presence of vitamins on the redox chemistry of iron. Our examination of the effect of the presence of thiamine hydrochloride in the redox behavior of Fe{sup 2+}/Fe{sup 3+} at the pyrite surface has revealed that thiamine hydrochloride does not undergo chemical interaction with ferrous or ferric iron. However, it may adsorb onto the pyrite surface causing polarization of the pyrite electrode.
Date: December 31, 1990
Creator: Pesic, B. & Oliver, D. J.
Partner: UNT Libraries Government Documents Department

Effect of Heavy metals on the iron oxidizing ability of Thiobacillus ferrooxidans: Part 1, Effect of silver. Technical progress report, July 1992--September 1992

Description: The effect of silver ions on the iron oxidizing ability of Thiobacillus ferrooxidans was studied using electrochemical and other physics-chemical techniques. Electrochemical investigation was conducted using a method based on redox potential change. Experiments were performed by adding an aliquot of separately prepared concentrate of the bacteria into the solution of ferrous ion and monitoring the redox potential for at least one hour. Pyrite was used as the indicator electrode. Parameters examined were pH, microbial cell density, ferrous, ferric and silver ion concentration, temperature and preconditioning period of the bacteria with silver ions, etc. Results obtained demonstrate that the rate of ferrous ion oxidation is dependent on pH (optimum pH range is 1.5--2.0) and the substrate (i.e. Fe(II)) to microbial cell concentration ratio. The mechanism of the bacteria mediated oxidation of ferrous iron is remarkably sensitive to temperature changes. At the vicinity of the optimum temperature (i.e. 25{degree}C), the reaction is likely to be controlled by the diffusion of Fe (II) ions through the cell wall of the bacteria, whereas below the range 18--25{degree}C, reaction kinetics may be the rate controlling factor. In the presence of 10 mg/L silver, the reaction may be kinetically controlled over the temperature range 5.5--25{degree}C. Inhibition of microbial FE(II) oxidation in the presence of silver may take place via a mixed mechanism in which silver may bind with both the enzyme and the enzyme-substrate complex.
Date: December 1, 1992
Creator: De, G. C. & Pesic, B.
Partner: UNT Libraries Government Documents Department

Electrochemistry of Thiobacillus ferrooxidans reactions with pyrite. Final report

Description: A cyclic voltammetry technique was used to study the interactions of pyrite during bioleaching with the bacterium Thiobacillus ferrooxidans. Potential effects of heavy metals (silver and mercury) and varying the pH on the iron oxidizing ability of the bacterium are reported. Redox potential techniques were used to study effect of ferrous sulfate concentration and pH on bacterial growth.
Date: January 20, 1993
Creator: Pesic, B.; Oliver, D. J.; Kim, Inbeum & De, G. C.
Partner: UNT Libraries Government Documents Department

Microbial recovery of metals from spent coal liquefaction catalysts. Quarterly report, July--September 1993

Description: The project objectives outlined in the previous reports involved defining conditions and cultures best suited to achieve the most effective metal release from spent coal liquefaction catalysts by microbial processes. The work initiated in the first quarter of 1993 was continued and expanded using solvent extraction systems defined by the multiple solvent tests used for washing the catalysts. To reduce the number of solvent systems the data were examined and two solvents selected for continued testing. The two solvent systems which were chosen and have been employed are the isopropanol and a tetrahydrofuran (THF) extractive solvents. In the present simplified extractive system, the catalysts are washed with the solvent with simple agitation to prevent catalyst breakage. Studies on the effect of catalyst surface area will be continued in conjunction with the progressive development of the optimized microbial metal releasing system. Thermophilic cultures (Bacillus stearothermophilus and Metallosphaera sedula) are being grown at 60 C for initial metal releasing studies. Since THF has proven to be a superior solvent system as judged by metal release, research has concentrated on using THF as the solvent system allowing work to proceed rapidly. However, additional work will still be directed toward defining any solvent system superior to THF in terms of aiding microbiological release of metals and economic feasibility. These studies include measurements of metal release at various catalyst-media ratios for both the tetrahydrofuran and isopropanol washed catalysts. The importance of the catalyst-media ratios is assuming a greater importance since it is desired to reduce the media volume to the lowest level possible while still retaining good metals release.
Date: December 31, 1993
Creator: Sandbeck, K. A. & Joffe, P. M.
Partner: UNT Libraries Government Documents Department

Electrochemistry of a semiconductor chalcopyrite concentrate leaching by Thiobacillus ferrooxidans

Description: Using carbon-paste-CuFeS{sub 2} electrodes and a cyclic voltammetric technique, it was found that a large number of intermediate electrochemical oxidation reactions were associated with the dissolution of chalcopyrite in presence and absence of bacteria. The effects of concentrations of copper, ferrous and ferric ions, as well as of agitation on the peaks of cyclic voltammograms were measured. It was established that chalcopyrite oxidation was solid-state controlled as suggested by the data of chronopotentiometric and chronoamperometric measurements. The activation energy of solid state diffusion of chalcopyrite leaching was determined by the Sand's method to be {triangle}E{sub a} = 20.5 kJ. The leaching mechanism is discussed in terms of solid-state properties (energy bonding) of the n-type semiconductor chalcopyrite and energy density states of redox systems of acidic bacterial leach media. A generalized model for the mechanism of chalcopyrite leaching in presence and absence of bacteria is presented. 23 refs., 10 figs.
Date: January 1, 1991
Creator: Torma, A.E.
Partner: UNT Libraries Government Documents Department

Effect of Heavy metals on the iron oxidizing ability of Thiobacillus ferrooxidans: Part 1, Effect of silver

Description: The effect of silver ions on the iron oxidizing ability of Thiobacillus ferrooxidans was studied using electrochemical and other physics-chemical techniques. Electrochemical investigation was conducted using a method based on redox potential change. Experiments were performed by adding an aliquot of separately prepared concentrate of the bacteria into the solution of ferrous ion and monitoring the redox potential for at least one hour. Pyrite was used as the indicator electrode. Parameters examined were pH, microbial cell density, ferrous, ferric and silver ion concentration, temperature and preconditioning period of the bacteria with silver ions, etc. Results obtained demonstrate that the rate of ferrous ion oxidation is dependent on pH (optimum pH range is 1.5--2.0) and the substrate (i.e. Fe(II)) to microbial cell concentration ratio. The mechanism of the bacteria mediated oxidation of ferrous iron is remarkably sensitive to temperature changes. At the vicinity of the optimum temperature (i.e. 25[degree]C), the reaction is likely to be controlled by the diffusion of Fe (II) ions through the cell wall of the bacteria, whereas below the range 18--25[degree]C, reaction kinetics may be the rate controlling factor. In the presence of 10 mg/L silver, the reaction may be kinetically controlled over the temperature range 5.5--25[degree]C. Inhibition of microbial FE(II) oxidation in the presence of silver may take place via a mixed mechanism in which silver may bind with both the enzyme and the enzyme-substrate complex.
Date: January 1, 1992
Creator: De, G.C. & Pesic, B.
Partner: UNT Libraries Government Documents Department

Molecular biological enhancement of coal biodesulfurization. Final technical report

Description: Progress is reported in understanding Thiobacillus molecular biology, specifically in the area of vector development. At the initiation of this program, the basic elements needed for performing genetic engineering in T. ferrooxidans were either not yet developed. Improved techniques are described which will make it easier to construct and analyze the genetic structure and metabolism of recombinant T. ferrooxidans. The metabolism of the model organic sulfur compound dibenzothiophene (DBT) by certain heterotrophic bacteria was confirmed and characterized. Techniques were developed to analyze the metabolites of DBT, so that individual 4S pathway metabolites could be distinguished. These techniques are expected to be valuable when engineering organic sulfur metabolism in Thiobacillus. Strain isolation techniques were used to develop pure cultures of T. ferrooxidans seven of which were assessed as potential recombinant hosts. The mixotrophic strain T. coprinus was also characterized for potential use as an electroporation host. A family of related Thiobacillus plasmids was discovered in the seven strains of P. ferrooxidans mentioned above. One of these plasmids, pTFI91, was cloned into a pUC-based plasmid vector, allowing it to propagate in E. coli. A key portion of the cloned plasmid was sequenced. This segment, which is conserved in all of the related plasmids characterized, contains the vegetative origin of DNA replication, and fortuitously, a novel insertion sequence, designated IS3091. The sequence of the DNA origin revealed that these Thiobacillus plasmids represent a unique class of replicons not previously described. The potentially useful insertion sequence IS3091 was identified as a new member of a previously undefined family of insertion sequences which include the E. coli element IS30.
Date: October 8, 1992
Creator: Litchfield, J. H.; Zupancic, T. J.; Kittle, J. D. Jr.; Baker, B.; Palmer, D. T.; Traunero, C. G. et al.
Partner: UNT Libraries Government Documents Department

Electrochemistry of Thiobacillus ferrooxidans reactions with pyrite

Description: A cyclic voltammetry technique was used to study the interactions of pyrite during bioleaching with the bacterium Thiobacillus ferrooxidans. Potential effects of heavy metals (silver and mercury) and varying the pH on the iron oxidizing ability of the bacterium are reported. Redox potential techniques were used to study effect of ferrous sulfate concentration and pH on bacterial growth.
Date: January 20, 1993
Creator: Pesic, B.; Oliver, D.J.; Kim, Inbeum & De, G.C.
Partner: UNT Libraries Government Documents Department

Effect of thiamine hydrochloride on the redox reactions of iron at pyrite surface

Description: The present investigation is a part of our studies on the electro chemical aspects of pyrite bioleaching involving Thiobacillus ferrooxidans. Previously (1,2) we have examined the effect of T. ferrooxidans and their metabolic products on the redox reactions of Fe[sup 2+]/Fe[sup 3+] couple at the pyrite surface. Results obtained suggest that beyond 1. 5 days during their growth in a batch fermenter, the bacteria and their metabolic products completely cover the pyrite surface and shut down all electron transfer across the electrode-solution interface. In addition, it has been observed that the bacteria serve as the nucleation site for jarosite formation, which is found detrimental to bioleaching. In the present work we have focussed on the effect of the presence of vitamins on the redox chemistry of iron. Our examination of the effect of the presence of thiamine hydrochloride in the redox behavior of Fe[sup 2+]/Fe[sup 3+] at the pyrite surface has revealed that thiamine hydrochloride does not undergo chemical interaction with ferrous or ferric iron. However, it may adsorb onto the pyrite surface causing polarization of the pyrite electrode.
Date: January 1, 1990
Creator: Pesic, B. & Oliver, D.J.
Partner: UNT Libraries Government Documents Department

Molecular biological enhancement of coal biodesulfurization

Description: Progress is reported in understanding Thiobacillus molecular biology, specifically in the area of vector development. At the initiation of this program, the basic elements needed for performing genetic engineering in T. ferrooxidans were either not yet developed. Improved techniques are described which will make it easier to construct and analyze the genetic structure and metabolism of recombinant T. ferrooxidans. The metabolism of the model organic sulfur compound dibenzothiophene (DBT) by certain heterotrophic bacteria was confirmed and characterized. Techniques were developed to analyze the metabolites of DBT, so that individual 4S pathway metabolites could be distinguished. These techniques are expected to be valuable when engineering organic sulfur metabolism in Thiobacillus. Strain isolation techniques were used to develop pure cultures of T. ferrooxidans seven of which were assessed as potential recombinant hosts. The mixotrophic strain T. coprinus was also characterized for potential use as an electroporation host. A family of related Thiobacillus plasmids was discovered in the seven strains of P. ferrooxidans mentioned above. One of these plasmids, pTFI91, was cloned into a pUC-based plasmid vector, allowing it to propagate in E. coli. A key portion of the cloned plasmid was sequenced. This segment, which is conserved in all of the related plasmids characterized, contains the vegetative origin of DNA replication, and fortuitously, a novel insertion sequence, designated IS3091. The sequence of the DNA origin revealed that these Thiobacillus plasmids represent a unique class of replicons not previously described. The potentially useful insertion sequence IS3091 was identified as a new member of a previously undefined family of insertion sequences which include the E. coli element IS30.
Date: October 8, 1992
Creator: Litchfield, J.H.; Zupancic, T.J.; Kittle, J.D. Jr.; Baker, B.; Palmer, D.T.; Traunero, C.G. et al.
Partner: UNT Libraries Government Documents Department

West Valley low-level radioactive waste site revisited: Microbiological analysis of leachates

Description: The abundance and types of microorganisms in leachate samples from the West Valley low-level radioactive waste disposal site were enumerated. This study was undertaken in support of the study conducted by Ecology and Environment, Inc., to assess the extent of radioactive gas emissions from the site. Total aerobic and anaerobic bacteria were enumerated as colony forming units (CFU) by dilution agar plate technique, and denitrifiers, sulfate-reducers and methanogens by the most probable number technique (MPN). Of the three trenches 3, 9, and 11 sampled, trench 11 contained the most number of organisms in the leachate. Concentrations of carbon-14 and tritium were highest in trench 11 leachate. Populations of aerobes and anaerobes in trench 9 leachate were one order of magnitude less than in trench 11 leachate while the methanogens were three orders of magnitude greater than in trench 11 leachate. The methane content from trench 9 was high due to the presence of a large number of methanogens; the gas in this trench also contained the most radioactivity. Trench 3 leachate contained the least number of microorganisms. Comparison of microbial populations in leachates sampled from trenches 3 and 9 during October 1978 and 1989 showed differences in the total number of microbial types. Variations in populations of the different types of organisms in the leachate reflect the changing nutrient conditions in the trenches. 14 refs., 3 figs., 4 tabs.
Date: October 1, 1990
Creator: Gillow, J.B. & Francis, A.J.
Partner: UNT Libraries Government Documents Department