The effects of Fe2+ on sulfur-oxidizing bacteria (SOB) driven autotrophic denitrification.

With the short-term exposure to Fe2+, the mechanism of autotrophic denitrification and sulfide oxidation and the correlation between microbial community changes and environmental factors have been explored in the ADSOB process. RSM was used to optimize conditions for the maximum nitrate reduction and sulfide oxidation. About 88% of nitrate could be autotrophically denitrified to nitrogen by utilizing sulfide as the electron donor with the molar ratio C/N of 1.14 and S/N of 0.99 at pH 7.1. Lower Fe2+ additions can reduce TDS inhibition with dissolved sulfide to form FeS precipitates, while high amount of Fe2+ limited the mass transfer of NO3- and intermediate products such as S0 may be generated. High-throughput sequencing and RDA analysis revealed the correlation between ferrous iron, environmental factors and microorganisms. Sulfurospirillum, Rhodanobacter, Thauera and Thiobacillus were all slightly promoted at NFL level and inhibited at NFH level. And the narrow angles of the arrows indicated that Thauera, Sulfurospirillum and Thiobacillus were positively correlated with SO42- concentrations, while large angles indicated these bacteria were inversely related with TDS and NO3- arrows, which further confirmed that these bacteria played a dominant role in the ADSOB process, and can reduce NO3- by the oxidation of TDS. The correlation further indicated that lower Fe2+ additions have a promoting effect, while high concentrations have an inhibiting effect.

Effects of sulfur application on cadmium bioaccumulation in tobacco and its possible mechanisms of rhizospheric microorganisms.

A potting experiment was conducted to investigate the effect of sulfur application on the bioaccumulation by tobacco and its mechanisms of rhizospheric microorganisms. Cadmium content in tobacco was analyzed using atomic absorption spectrometer, while bacterial community and related gene in soil were analyzed via high-throughput sequencing and quantitative PCR techniques, respectively. The obtained results indicated that tobacco had the ability to accumulate cadmium under no sulfur application conditions, with cadmium contents of 35.4, 23.6, and 26.3 mg kg-1 in leaves, stems, and roots, respectively. Under high-sulfur treatment, these values increased to 66.4, 46.1, and 42.6 mg kg-1, respectively, probably due to the increase of the available cadmium content (from 1.1 to 3.3 mg kg-1) in the soil through a decrease of the soil pH value, which was contributed by the sulfur oxidation reaction. dsrA and soxB genes might play an important role in sulfur oxidation, and Thiobacillus sp. was the dominant bacterial genus during the sulfur oxidation process. In addition, sulfur application exerted little effect on the diversity and structure of the soil bacterial community. The combined results indicate that sulfur application is an effective and safe method for Cd phytoextraction by tobacco.

Investigating the effect of design parameters on the response time of a highly sensitive microbial hydrogen sulfide biosensor based on oxygen consumption.

A novel hydrogen sulfide microbial biosensor was developed based on investigating the influence of four design parameters: cell concentration, immobilization bed type, hydrogen sulfide concentration, and geometrical shape of the biosensor. Thiobacillus thioparus was used as the recognition element and it was immobilized on sodium alginate as well as agarose bed. The results were optimized by the application of statistical optimization software based on response time of the system. Oxygen reduction was considered as the detection sign. Sodium alginate solution with a concentration of 2.3% (w/v) and optical density of 10 at 605 nm was found as the optimum conditions for immobilization with response time of 72s . Optimum response time of immobilized T. thioparus on agarose was also found equal to 120 s at agarose concentration of 1.2% (w/v) and optical density of 10.83. Performance of the biosensor in different temperatures, pH and agitation speeds was also analyzed. The designed biosensor could detect concentrations of hydrogen sulfide as low as 0.5 ppm. T. thioparus could retain 99% of the original activity in both systems, after ten days passing the fabrication. A fractal analysis was also done theoretically to investigate the diffusion of oxygen in immobilized cells which showed a satisfactory value of oxygen take up by the immobilized cells.

Isolation, identification and arsenic-resistance of Acidithiobacillus ferrooxidans HX3 producing schwertmannite.

Schwertmannite, a ubiquitous mineral present in iron oxyhydroxides formed in iron- and sulfate-rich acid media, favors incorporation of some toxic anions in its structure. We reported an iron-oxidizing bacterial strain HX3 from a municipal sludge that facilitates the formation of pure schwertmannite in cultures. Ferrous iron oxidation by the isolated strain HX3 was optimum at an initial pH of 2.0-3.3 and temperature of 28-35°C. Pure schwertmannite was found through bacterial oxidation of ferrous iron at an initial pH2.8 and temperature 28°C. Following 16S rDNA gene sequence analysis the bacterial strain HX3 was identified as Acidithiobacillus ferrooxidans. The arsenic-resistance A. ferrooxidans HX3 showed the potential of environmental application in arsenic removal from the As(III)- and iron-rich acid sulfate waters directly by As(III) adsorption or the formation of schwertmannite in the environment.

[Research on anti-corrosion of Thiobacillus for the geopolymer solidification MSWI fly ash].

In order to discuss the anti-Thiobacillus corrosion performance of geopolymer solidification MSWI fly ash, the research simulated the Thiobacillus corrosion process by experiment, investigated the change of mass, compressive strength, leaching concentration. The results showed that geopolymer had a good anti-corrosion ability: weight loss within 1%, the compressive strength still reached 21.88 MPa after 28 days, the corrosion resistance coefficient was above 0.9. The maximum leaching concentration of Cr, Cu, Zn, Cd, Hg, Pb were 107.7 microg x L(-1), 22.71 microg x L(-1), 39.18 microg x L(-1), 0.56 microg x L(-1), 34.84 microg x L(-1) and 3.03 microg x L(-1), respectively. And the leaching concentration of geopolymer reduced with the immersion time, showed a good anti-Thiobacillus corrosion performance. Through the X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscope spectra of geopolymer, we investigated the microstructure and mechanism of geopolymer anti-corrosion.

Effects of sulphur and Thiobacillus thioparus on cow manure aerobic composting.

A simulated aerobic composting experiment was used to explore the effects of sulphur and Thiobacillus thioparus during six manure composting treatments. The addition of sulphur led to a decrease of the pH level within the range 6-6.3, which was lower than the control treatment (CK). The concentration of ammonium nitrogen in T1 (0.25% sulphur), T2 (0.5% sulphur), T3 (0.25% sulphur + T. thioparus) and T4 (0.5% sulphur + T. thioparus) were much higher than the ammonium N in CK. The results indicated that addition of sulphur could increase the concentration of ammonium N and reduce loss of nitrogen. However, excess sulphur had a negative effect on temperature and GI. Addition of T. thioparus could increase concentration of available S, alleviate these negative influences and reduce compost biological toxicity.

Biological oxidation of hydrogen sulfide in mineral media using a biofilm airlift suspension reactor.

Hydrogen sulfide (H(2)S) removal in mineral media using Thiobacillus thioparus TK-1 in a biofilm airlift suspension reactor (BAS) was investigated to evaluate the relationship between biofilm formation and changes in inlet loading rates. Aqueous sodium sulfide was fed as the substrate into the continuous BAS-reactor. The reactor was operated at a constant temperature of 30 degrees C and a pH of 7, the optimal temperature and pH for biomass growth. The startup of the reactor was performed with basalt carrier material. Optimal treatment performance was obtained at a loading rate of 4.8 mol S(2-) m(-3) h(-1) at a conversion efficiency as high as 100%. The main product of H(2)S oxidation in the BAS-reactor was sulfate because of high oxygen concentrations in the airlift reactor. The maximum sulfide oxidation rate was 6.7 mol S(2-) m(-3) h(-1) at a hydraulic residence time of 3.3 h in the mineral medium. The data showed that the BAS-reactor with this microorganism can be used for sulfide removal from industrial effluent.

Development of a genetic system for the chemolithoautotrophic bacterium Thiobacillus denitrificans.

Thiobacillus denitrificans is a widespread, chemolithoautotrophic bacterium with an unusual and environmentally relevant metabolic repertoire, which includes its ability to couple denitrification to sulfur compound oxidation; to catalyze anaerobic, nitrate-dependent oxidation of Fe(II) and U(IV); and to oxidize mineral electron donors. Recent analysis of its genome sequence also revealed the presence of genes encoding two [NiFe]hydrogenases, whose role in metabolism is unclear, as the sequenced strain does not appear to be able to grow on hydrogen as a sole electron donor under denitrifying conditions. In this study, we report the development of a genetic system for T. denitrificans, with which insertion mutations can be introduced by homologous recombination and complemented in trans. The antibiotic sensitivity of T. denitrificans was characterized, and a procedure for transformation with foreign DNA by electroporation was established. Insertion mutations were generated by in vitro transposition, the mutated genes were amplified by the PCR, and the amplicons were introduced into T. denitrificans by electroporation. The IncP plasmid pRR10 was found to be a useful vector for complementation. The effectiveness of the genetic system was demonstrated with the hynL gene, which encodes the large subunit of a [NiFe]hydrogenase. Interruption of hynL in a hynL::kan mutant resulted in a 75% decrease in specific hydrogenase activity relative to the wild type, whereas complementation of the hynL mutation resulted in activity that was 50% greater than that of the wild type. The availability of a genetic system in T. denitrificans will facilitate our understanding of the genetics and biochemistry underlying its unusual metabolism.

The genome sequence of the obligately chemolithoautotrophic, facultatively anaerobic bacterium Thiobacillus denitrificans.

The complete genome sequence of Thiobacillus denitrificans ATCC 25259 is the first to become available for an obligately chemolithoautotrophic, sulfur-compound-oxidizing, beta-proteobacterium. Analysis of the 2,909,809-bp genome will facilitate our molecular and biochemical understanding of the unusual metabolic repertoire of this bacterium, including its ability to couple denitrification to sulfur-compound oxidation, to catalyze anaerobic, nitrate-dependent oxidation of Fe(II) and U(IV), and to oxidize mineral electron donors. Notable genomic features include (i) genes encoding c-type cytochromes totaling 1 to 2 percent of the genome, which is a proportion greater than for almost all bacterial and archaeal species sequenced to date, (ii) genes encoding two [NiFe]hydrogenases, which is particularly significant because no information on hydrogenases has previously been reported for T. denitrificans and hydrogen oxidation appears to be critical for anaerobic U(IV) oxidation by this species, (iii) a diverse complement of more than 50 genes associated with sulfur-compound oxidation (including sox genes, dsr genes, and genes associated with the AMP-dependent oxidation of sulfite to sulfate), some of which occur in multiple (up to eight) copies, (iv) a relatively large number of genes associated with inorganic ion transport and heavy metal resistance, and (v) a paucity of genes encoding organic-compound transporters, commensurate with obligate chemolithoautotrophy. Ultimately, the genome sequence of T. denitrificans will enable elucidation of the mechanisms of aerobic and anaerobic sulfur-compound oxidation by beta-proteobacteria and will help reveal the molecular basis of this organism's role in major biogeochemical cycles (i.e., those involving sulfur, nitrogen, and carbon) and groundwater restoration.

Nitrate promotes biological oxidation of sulfide in wastewaters: experiment at plant-scale.

Biogenic production of sulfide in wastewater treatment plants involves odors, toxicity and corrosion problems. The production of sulfide is a consequence of bacterial activity, mainly sulfate-reducing bacteria (SRB). To prevent this production, the efficiency of nitrate addition to wastewater was tested at plant-scale by dosing concentrated calcium nitrate (Nutriox) in the works inlet. Nutriox dosing resulted in a sharp decrease of sulfide, both in the air and in the bulk water, reaching maximum decreases of 98.7% and 94.7%, respectively. Quantitative molecular microbiology techniques indicated that the involved mechanism is the development of the nitrate-reducing, sulfide-oxidizing bacterium Thiomicrospira denitrificans instead of the direct inhibition of the SRB community. Denitrification rate in primary sedimentation tanks was enhanced by nitrate, being this almost completely consumed. No significant increase of inorganic nitrogen was found in the discharged effluent, thus reducing potential environmental hazards to receiving waters. This study demonstrates the effectiveness of nitrate addition in controlling sulfide generation at plant-scale, provides the mechanism and supports the environmental adequacy of this strategy.

Development of an automated water toxicity biosensor using Thiobacillus ferrooxidans for monitoring cyanides in natural water for a water filtering plant.

An on-line biosensor consisting of immobilized Thiobacillus ferrooxidans and an oxygen electrode was developed for automated monitoring of acute toxicity in water samples. T. ferrooxidans is an obligatory acidophilic, autotrophic bacterium and derives its energy by the oxidation of ferrous ion, elemental sulfur, and reduced sulfur compounds including metal sulfides. The assay is based on the monitoring of a current increase by addition of toxicoids, which is caused by the inhibition of bacterial respiration and decrease in oxygen consumption. Optimum cell number on the membrane was 5.0 x 10(8) cells. The steady-state current was obtained when concentration of FeSO4 was above 3.6 mM at pH 3. The sensor response of T. ferrooxidans immobilized membrane for 5.0 microM KCN was within an error of 10% for 30 membranes. A linear relationship was obtained at KCN concentration in the range of 0.5-3.0 microM in a flow-type monitoring system. Minimum detectable concentrations of KCN, Na2S, and NaN3 were 0.5, 1.2, and 0.07 microM, respectively. The monitoring system contained two biosensors and these sensors were cleaned with sulfuric acid (pH 1.5) twice a day. This treatment could remove fouling on microbial immobilized membrane by natural water and ferrous precipitation in the flow cell. This flow-type monitoring sensor was operated continuously for 5 months. Also, T. ferrooxidans immobilized membrane can be stored for one month at 4 degrees C when preserved with wet absorbent cotton under argon gas.

Volatilization of mercury by an iron oxidation enzyme system in a highly mercury-resistant Acidithiobacillus ferrooxidans strain MON-1.

A highly mercury-resistant strain Acidithiobacillus ferrooxidans MON-1, was isolated from a culture of a moderately mercury-resistant strain, A. ferrooxidans SUG 2-2 (previously described as Thiobacillus ferrooxidans SUG 2-2), by successive cultivation and isolation of the latter strain in a Fe2+ medium with increased amounts of Hg2+ from 6 microM to 20 microM. The original stain SUG 2-2 grew in a Fe2+ medium containing 6 microM Hg2+ with a lag time of 22 days, but could not grow in a Fe2+ medium containing 10 microM Hg2+. In contrast, strain MON-1 could grow in a Fe2+ medium containing 20 microM Hg2+ with a lag time of 2 days and the ability of strain MON-1 to grow rapidly in a Fe2+ medium containing 20 microM Hg2+ was maintained stably after the strain was cultured many times in a Fe2+ medium without Hg2+. A similar level of NADPH-dependent mercury reductase activity was observed in cell extracts from strains SUG 2-2 and MON-1. By contrast, the amounts of mercury volatilized for 3 h from the reaction mixture containing 7 microM Hg2+ using a Fe(2+)-dependent mercury volatilization enzyme system were 5.6 nmol for SUG 2-2 and 67.5 nmol for MON-1, respectively, indicating that a marked increase of Fe(2+)-dependent mercury volatilization activity conferred on strain MON-1 the ability to grow rapidly in a Fe2+ medium containing 20 microM Hg2+. Iron oxidizing activities, 2,3,5,6-tetramethyl-p-phenylenediamine (TMPD) oxidizing activities and cytochrome c oxidase activities of strains SUG 2-2 and MON-1 were 26.3 and 41.9 microl O2 uptake/mg/min, 15.6 and 25.0 microl O2 uptake/mg/min, and 2.1 and 6.1 mU/mg, respectively. These results indicate that among components of the iron oxidation enzyme system, especially cytochrome c oxidase activity, increased by the acquisition of further mercury resistance in strain MON-1. Mercury volatilized by the Fe(2+)-dependent mercury volatilization enzyme system of strain MON-1 was strongly inhibited by 1.0 mM sodium cyanide, but was not by 50 nM rotenone, 5 microM 2-n-heptyl-4-hydroxy-quinoline-N-oxide (HQNO), 0.5 microM antimycin A, or 0.5 microM myxothiazol, indicating that cytochrome c oxidase plays a crucial role in mercury volatilization of strain MON-1 in the presence of Fe2+.

Optimal conditions for bio-oxidation of ferrous ions to ferric ions using Thiobacillus ferrooxidans.

A chemo-biochemical process using Thiobacillus ferrooxidans for desulphurization of gaseous fuels and emissions containing hydrogen sulphide (H2S) has been developed. In the first stage, H2S present in fuel gas and emissions is selectively oxidized to elemental sulphur using ferric sulphate. The ferrous sulphate produced in the first stage of the process is oxidized to ferric sulphate using Thiobacillus ferrooxidans for recycle and reuse in the process. The effects of process variables, temperature, pH, total dissolved solids (TDS), elemental sulphur, ferric and magnesium ions on bio-oxidation of ferrous ions to ferric ions were investigated using flask culture experiments. The bio-oxidation of ferrous ions to ferric ions could be achieved efficiently in the temperature range of 20(+/-1)-44(+/-1) degrees C. A pH range of 1.8(+/-0.02)-2.2(+/-0.02) was optimum for the growth of culture and effective bio-oxidation of ferrous ions to ferric ions. The effect of TDS on bio-oxidation of ferrous ions indicated that a preacclimatized culture in a growth medium containing high dissolved solid was required to achieve effective bio-oxidation of ferrous ions. Elemental sulphur ranging from 1000 to 100,000 mg/l did not have any effect on efficiency of ferrous ion oxidation. The efficiency of bio-oxidation of ferrous ions to ferric ions was not affected in the presence of ferric ions up to a concentration of 500 mg/l while 3 mg/l of magnesium ion was optimal for achieving effective bio-oxidation.

ATP generation during reduced inorganic sulfur compound oxidation by Acidithiobacillus caldus is exclusively due to electron transport phosphorylation.

The synthesis of adenosine 5-triphosphate (ATP) (increase in phosphorylation potential) during the oxidation of reduced inorganic sulfur compounds was studied in the moderately thermophilic acidophileAcidithiobacillus caldus (strain KU) (formerly Thiohacillus caldus). The phosphorylation potential increased during the oxidation of all reduced inorganic sulfur compounds tested compared with resting cells. The generation of ATP in whole cells was inhibited by the F0F1 ATPase inhibitor oligomycin, electron transport chain inhibitors, valinomycin and potassium ions. There was no increase in the phosphorylation potential, nor synthesis of ATP. in the absence of electron transport. An apparent lack of substrate-level phosphorylation was indicated by the lack of adenosine 5-phosphosulfate reductase in tetrathionate-grown At. caldus. Studies were also performed on the synthesis of ATP by membrane vesicles of At. caldus when presented with an artificial proton gradient. Complete inhibition of ATP synthesis in these vesicles occurred when they were loaded with N,N-dicyclohexylcarbodiimide (DCCD), but not when they were loaded with oligomycin, vanadate or electron transport chain inhibitors. The data presented here suggest that during the oxidation of reduced inorganic sulfur compounds by At. caldus, all ATP is synthesized by oxidative phosphorylation via a membrane-bound F0F1 ATPase driven by a proton gradient.

Effect of organics on sulfur-utilizing autotrophic denitrification under mixotrophic conditions.

Sulfur-utilizing denitrification can be performed by denitrifying sulfur bacteria under autotrophic and heterotrophic conditions. To investigate the effect of organics (methanol and landfill leachate) on sulfur-utilizing denitrification, six laboratory-scale sulfur packed columns were operated under autotrophic, mixotrophic and heterotrophic conditions for approximately 1 year. The performance of the columns was monitored by measuring the pH, nitrate, nitrite, sulfate, sulfide, alkalinity dissolved organic carbon (DOC), and turbidity. These tests indicated that the mixotrophic column had a higher nitrate removal capacity than the autotrophic column. It was also found that under mixotrophic conditions, some portion of nitrate was removed heterotrophically and the remainder was denitrified by sulfur-utilizing autotrophic bacteria without inhibition by organics. In addition, sulfate production and alkalinity consumption were reduced under mixotrophic conditions. These results suggest that mixotrophic conditions that allow simultaneous reactions of autotrophic and heterotrophic denitrification provide significant advantages in terms of nitrate and DOC removal, sulfate production decrease, and alkalinity consumption over obligate autotrophic or heterotrophic denitrification.

Enhanced UV sensitivity of Thiobacillus ferrooxidans resulting from caffeine and acriflavine treatment of irradiated cells.

This study was aimed at identifying the roles of caffeine and acriflavine, two repair inhibitors, on UV sensitivity of iron-oxidizing Thiobacillus ferrooxidans ATCC 13728. The UV-dose response survival curve was inflected in nature, suggesting the population heterogeneity of the isolate. Caffeine and acriflavine potentiated the UV-induced killing of the organism. With the increase in concentrations of these compounds, the extent of survival decreased. Similarly, the inhibitory effects of caffeine and acriflavine increased with the increase in dose of UV-irradiation. The cells irradiated with 10 s (equivalent to 5.6 x 10(-5) J/m2/s) of UV-exposure tended to become resistant to the inhibitory effects of caffeine and acriflavine, as evidenced by the time course study of recovery. The cells appear to stage a dramatic recovery from UV damage in the presence of caffeine (3.0 mg/ml) and acriflavine (20 microg/ml) over a period of 25-30 h and 35-40 h respectively, when grown in the presence of energy sources.

Ferrous iron-dependent volatilization of mercury by the plasma membrane of Thiobacillus ferrooxidans.

Of 100 strains of iron-oxidizing bacteria isolated, Thiobacillus ferrooxidans SUG 2-2 was the most resistant to mercury toxicity and could grow in an Fe(2+) medium (pH 2.5) supplemented with 6 microM Hg(2+). In contrast, T. ferrooxidans AP19-3, a mercury-sensitive T. ferrooxidans strain, could not grow with 0.7 microM Hg(2+). When incubated for 3 h in a salt solution (pH 2.5) with 0.7 microM Hg(2+), resting cells of resistant and sensitive strains volatilized approximately 20 and 1.7%, respectively, of the total mercury added. The amount of mercury volatilized by resistant cells, but not by sensitive cells, increased to 62% when Fe(2+) was added. The optimum pH and temperature for mercury volatilization activity were 2.3 and 30 degrees C, respectively. Sodium cyanide, sodium molybdate, sodium tungstate, and silver nitrate strongly inhibited the Fe(2+)-dependent mercury volatilization activity of T. ferrooxidans. When incubated in a salt solution (pH 3.8) with 0.7 microM Hg(2+) and 1 mM Fe(2+), plasma membranes prepared from resistant cells volatilized 48% of the total mercury added after 5 days of incubation. However, the membrane did not have mercury reductase activity with NADPH as an electron donor. Fe(2+)-dependent mercury volatilization activity was not observed with plasma membranes pretreated with 2 mM sodium cyanide. Rusticyanin from resistant cells activated iron oxidation activity of the plasma membrane and activated the Fe(2+)-dependent mercury volatilization activity of the plasma membrane.

Thiobacillus ferrooxidans response to copper and other heavy metals: growth, protein synthesis and protein phosphorylation.

Respirometric experiments demonstrated that the oxygen uptake by Thiobacillus ferrooxidans strain LR was not inhibited in the presence of 200 mM copper. Copper-treated and untreated cells from this T. ferrooxidans strain were used in growth experiments in the presence of cadmium, copper, nickel and zinc. Growth in the presence of copper was improved by the copper-treated cells. However, no growth was observed for these cells, within 190 h of culture, when cadmium, nickel and zinc were added to the media. Changes in the total protein synthesis pattern were detected by two-dimensional polyacrylamide gel electrophoresis for T. ferrooxidans LR cells grown in the presence of different heavy metals. Specific proteins were induced by copper (16, 28 and 42 kDa) and cadmium (66 kDa), whereas proteins that had their synthesis repressed were observed for all the heavy metals tested. Protein induction was also observed in the cytosolic and membrane fractions from T. ferrooxidans LR cells grown in the presence of copper. The level of protein phosphorylation was increased in the presence of this metal.

Strain variability and the effects of organic compounds on the growth of the chemolithotrophic bacterium Thiobacillus ferrooxidans.

The effects of naturally-occurring organic compounds on ferrous iron oxidation by the bacterium Thiobacillus ferrooxidans were examined with a view to using these compounds to treat or prevent acid mine/rock drainage. The compounds glucose, cellobiose, galacturonic acid, and citric acid were added to the growth medium of five different strains of the bacterium and growth studies were done to determine whether or not strain differences existed with respect to organic compound sensitivity. The effects of these compounds were compared to the effects of sodium dodecyl sulfate (SDS) an anionic detergent. Each of the compounds tested had an inhibitory effect on the strains of the bacterium and sensitivity to these compounds was strain dependent. All strains appeared to be equally susceptible to SDS. Inhibitory concentrations ranged from 70 mM to >280 mM for glucose, 7.5 mM to 150 mM for cellobiose, 20 mM to 230 mM for galacturonic acid, and 50 mM to 130 mM for citric acid. SDS effectively inhibited iron oxidation for all strains at a concentration of 0.3 mM, the lowest concentration tested. Some naturally-occurring organic compounds, therefore, might be candidates for the growth control of T. ferrooxidans.

Selective inhibition of the oxidation of ferrous iron or sulfur in Thiobacillus ferrooxidans.

The oxidation of either ferrous iron or sulfur by Thiobacillus ferrooxidans was selectively inhibited or controlled by various anions, inhibitors, and osmotic pressure. Iron oxidation was more sensitive than sulfur oxidation to inhibition by chloride, phosphate, and nitrate at low concentrations (below 0.1 M) and also to inhibition by azide and cyanide. Sulfur oxidation was more sensitive than iron oxidation to the inhibitory effect of high osmotic pressure. These differences were evident not only between iron oxidation by iron-grown cells and sulfur oxidation by sulfur-grown cells but also between the iron and sulfur oxidation activities of the same iron-grown cells. Growth experiments with ferrous iron or sulfur as an oxidizable substrate confirmed the higher sensitivity of iron oxidation to inhibition by phosphate, chloride, azide, and cyanide. Sulfur oxidation was actually stimulated by 50 mM phosphate or chloride. Leaching of Fe and Zn from pyrite (FeS(2)) and sphalerite (ZnS) by T. ferrooxidans was differentially affected by phosphate and chloride, which inhibited the solubilization of Fe without significantly affecting the solubilization of Zn.