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Rajesh K Sani

Publications and source records attributed to Rajesh K Sani.

6 recordsLinked to original sources

Isolation and characterization of Cr(VI) reducing Cellulomonas spp. from subsurface soils: implications for long-term chromate reduction.

Microbial enrichments from Cr(VI) contaminated and uncontaminated US Department of Energy Hanford Site sediments produced Cr(VI) reducing consortia when grown in the presence of Cr(VI) with acetate, D-xylose or glycerol as a carbon and energy source. Eight of the nine isolates from the consortia were Gram positive and four of these were identified by 16S rRNA sequence homology and membrane fatty acid composition as belonging to the genus Cellulomonas. Two strains, ES6 and WS01, were further examined for their ability to reduce Cr(VI) under growth and non-growth conditions. During fermentative growth on D-xylose, ES6 and WS01 decreased aqueous Cr(VI) concentrations from 0.04 mM Cr(VI) to below the detection limit (0.002 mM Cr(VI)) in less than three days and retained their ability to reduce Cr(VI) even after four months of incubation. Washed ES6 and WS01 cells also reduced Cr(VI) under non-growth conditions for over four months, both with and without the presence of an exogenous electron donor. K-edge XANES spectroscopy confirmed the reduction of Cr(VI) to Cr(III). The ability to reduce Cr(VI) after growth had stopped and in the absence of an external electron donor, suggests that stimulation of these types of organisms may lead to effective long-term, in situ passive reactive barriers for Cr(VI) removal. Our results indicate that Cr(VI) reduction by indigenous Cellulomonas spp. may be a potential method of in situ bioremediation of Cr(VI) contaminated sediment and groundwater.

Cellulomonas↗

Toxic effects of uranium on Desulfovibrio desulfuricans G20.

The toxic effects of U(VI) were studied using Desulfovibrio desulfuricans G20 in a medium containing bicarbonate or 1,4-piperazinediethane sulfonic acid disodium salt monohydrate (PIPES) buffer (each at 30 mM and pH 7). Uranium(VI) toxicity was dependent on the medium buffer and was observed in terms of longer lag times and, in some cases, no measurable growth. The minimum inhibiting concentration was 140 microM U(VI) in PIPES-buffered medium. This is 36-fold lower than that reported previously for D. desulfuricans. For all cases in which D. desulfuricans G20 grew in the presence of U(VI), the final cell protein yield was equivalent to that of the U(VI)-free control. In 24 h, D. desulfuricans G20 (total cell protein, 40 mg/L) removed 50 FiM U(VI) from solution in PIPES buffer, as compared to 96 microM U(VI) in bicarbonate buffer under anaerobic, nongrowth conditions. Even though the solubility of U(VI) was significantly lower in PIPES buffer than in bicarbonate buffer, U(VI) was much more toxic in PIPES buffer than in bicarbonate buffer. Analysis of thin sections of D. desulfuricans G20 treated with 90 microM U(VI) in medium containing PIPES buffer revealed that only a very small fraction of cells had reduced U precipitates in the periplasmic spaces. In the presence of bicarbonate buffer, however, reduced U was observed not only in the periplasm but also in the cytoplasm. Selected-area electron diffraction patterns and crystallographic analysis of transmission-electron microscopic lattice fringe images confirmed the structure of precipitated U in the cell periplasm and cytoplasm as being that of uraninite. These results suggest that U(VI) toxicity and the detoxification mechanisms of D. desulfuricans G20 depend greatly on the chemical forms of U(VI) that are present.

Alkanesulfonic Acids↗

Reoxidation of reduced uranium with iron(III) (hydr)oxides under sulfate-reducing conditions.

In cultures of Desulfovibrio desulfuricans 620 the effects of iron(III) (hydr)oxides (hematite, goethite, and ferrihydrite) on microbial reduction and reoxidation of uranium (U) were evaluated under lactate-limited sulfate-reducing conditions. With lactate present, G20 reduced U(VI) in both 1,4-piperazinediethanesulfonate (PIPES) and bicarbonate buffer. Once lactate was depleted, however, microbially reduced U served as an electron donor to reduce Fe(III) present in iron(III) (hydr)oxides. With the same initial amount of Fe(III) (10 mmol/L) for each iron(III) (hydr)oxide, reoxidation of U(IV) was greater with hematite than with goethite orferrihydrite. As the initial mass loading of hematite increased from 0 to 20 mmol of Fe(III)/L, the rate and extent of U(IV) reoxidation increased. Subsequent addition of hematite [15 mmol of Fe(III)/L] to stationary-phase cultures containing microbially reduced U(IV) also resulted in rapid reoxidation to U(VI). Analysis by U L3-edge X-ray absorption near-edge spectroscopy (XANES) of microbially reduced U particles yielded spectra similar to that of natural uraninite. Observations by high-resolution transmission electron microscopy, selected area electron diffraction, and energy-dispersive X-ray spectroscopic analysis confirmed that precipitated U associated with cells was uraninite with particle diameters of 3-5 nm. By the same techniques, iron sulfide precipitates were found to have a variable Fe and S stoichiometry and were not associated with cells.

Alkanesulfonic Acids↗

Uranium immobilization by sulfate-reducing biofilms.

Hexavalent uranium [U(VI)] was immobilized using biofilms of the sulfate-reducing bacterium (SRB) Desulfovibrio desulfuricans G20. The biofilms were grown in flat-plate continuous-flow reactors using lactate as the electron donor and sulfate as the electron acceptor. U(VI)was continuously fed into the reactor for 32 weeks at a concentration of 126 microM. During this time, the soluble U(VI) was removed (between 88 and 96% of feed) from solution and immobilized in the biofilms. The dynamics of U immobilization in the sulfate-reducing biofilms were quantified by estimating: (1) microbial activity in the SRB biofilm, defined as the hydrogen sulfide (H2S) production rate and estimated from the H2S concentration profiles measured using microelectrodes across the biofilms; (2) concentration of dissolved U in the solution; and (3) the mass of U precipitated in the biofilm. Results suggest that U was immobilized in the biofilms as a result of two processes: (1) enzymatically and (2) chemically, by reacting with microbially generated H2S. Visual inspection showed that the dissolved sulfide species reacted with U(VI) to produce a black precipitate. Synchrotron-based U L3-edge X-ray absorption near edge structure (XANES) spectroscopy analysis of U precipitated abiotically by sodium sulfide indicated that U(VI) had been reduced to U(IV). Selected-area electron diffraction pattern and crystallographic analysis of transmission electron microscope lattice-fringe images confirmed the structure of precipitated U as being that of uraninite.

Biofilms↗

Toxicity of lead in aqueous medium to desulfovibrio desulfuricans G20.

The toxicity of Pb(II) to sulfate-reducing bacteria (SRB) was studied using Desulfovibrio desulfuricans G20 in a medium specifically designed to assess metal toxicity. The effects of Pb(II) toxicity were observed in terms of longer lag times, lower specific growth rates, and in some cases no measurable growth. With an increase in medium pH from 6 to 8, Pb(II) toxicity decreased. At all pH values, in the presence of Pb(II) concentrations ranging from 3 to 15 microM, specific growth rates decreased and lag times increased. The minimum inhibiting concentration (MIC) of Pb(II) causing a complete inhibition in growth at pH 6 was 10 microM, as compared to 15 microM at pH 7.2 and 8. These MIC values are 40 times lower than previously reported for SRB. Results also show that with increases in initial cell protein concentration (inoculum size), soluble Pb(II) removal rates increased and the degree to which Pb(II) caused increased lag times was reduced. In the presence of Pb(II), in all cases in which D. desulfuricans grew (even after a 312-h lag time), the final cell protein concentration was equivalent to that of the Pb-free control. Live/dead staining, based on membrane integrity, indicated that while Pb(II) inhibited growth, Pb(II) did not cause a loss of D. desulfuricans membrane integrity.

Anaerobiosis↗

Toxic effects of chromium(VI) on anaerobic and aerobic growth of Shewanella oneidensis MR-1.

Cr(VI) was added to early- and mid-log-phase Shewanella oneidensis (S. oneidensis) MR-1 cultures to study the physiological state-dependent toxicity of Cr(VI). Cr(VI) reduction and culture growth were measured during and after Cr(VI) reduction. Inhibition of growth was observed when Cr(VI) was added to cultures of MR-1 growing aerobically or anaerobically with fumarate as the terminal electron acceptor. Under anaerobic conditions, there was immediate cessation of growth upon addition of Cr(VI) in early- and mid-log-phase cultures. However, once Cr(VI) was reduced below detection limits (0.002 mM), the cultures resumed growth with normal cell yield values observed. In contrast to anaerobic MR-1 cultures, addition of Cr(VI) to aerobically growing cultures resulted in a gradual decrease of the growth rate. In addition, under aerobic conditions, lower cell yields were also observed with Cr(VI)-treated cultures when compared to cultures that were not exposed to Cr(VI). Differences in response to Cr(VI) between aerobically and anaerobically growing cultures indicate that Cr(VI) toxicity in MR-1 is dependent on the physiological growth condition of the culture. Cr(VI) reduction has been previously studied in Shewanella spp., and it has been proposed that Shewanella spp. may be used in Cr(VI) bioremediation systems. Studies of Shewanella spp. provide valuable information on the microbial physiology of dissimilatory metal reducing bacteria; however, our study indicates that S. oneidensis MR-1 is highly susceptible to growth inhibition by Cr(VI) toxicity, even at low concentrations [0.015 mM Cr(VI)].

Adaptation, Physiological↗