Biomedical subjects

Brent M Peyton

Publications and source records attributed to Brent M Peyton.

16 recordsLinked to original sources

PubMed2006

Solubilization, solution equilibria, and biodegradation of PAH's under thermophilic conditions.

Biodegradation rates of PAHs are typically low at mesophilic conditions and it is believed that the kinetics of degradation is controlled by PAH solubility and mass transfer rates. Solubility tests were performed on phenanthrene, fluorene and fluoranthene at 20 degrees C, 40 degrees C and 60 degrees C and, as expected, a significant increase in the equilibrium solubility concentration and of the rate of dissolution of these polycyclic aromatic hydrocarbons (PAHs) was observed with increasing temperature. A first-order model was used to describe the PAH dissolution kinetics and the thermodynamic property changes associated with the dissolution process (enthalpy, entropy and Gibb's free energy of solution) were evaluated. Further, other relevant thermodynamic properties for these PAHs, including the activity coefficients at infinite dilution, Henry's law constants and octanol-water partition coefficients, were calculated in the temperature range 20-60 degrees C. In parallel with the dissolution studies, three thermophilic Geobacilli were isolated from compost that grew on phenanthrene at 60 degrees C and degraded the PAH more rapidly than other reported mesophiles. Our results show that while solubilization rates of PAHs are significantly enhanced at elevated temperatures, the biodegradation of PAHs under thermophilic conditions is likely mass transfer limited due to enhanced degradation rates.

Bacillaceae↗

PubMed2006

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↗

PubMed2006

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↗

PubMed2005

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↗

PubMed2005

Nitrincola lacisaponensis gen. nov., sp. nov., a novel alkaliphilic bacterium isolated from an alkaline, saline lake.

A novel alkaliphilic bacterium, strain 4CAT, was isolated from decomposing wood taken from the shore of Soap Lake, a saline, alkaline lake in Grant County, WA, USA. Cells of the isolate were Gram-negative, asporogenous, short, motile rods that utilized only a limited range of organic acids as sole carbon and energy sources. In addition to oxygen, the strain possessed the ability to reduce in the presence of acetate. Strain 4CAT was oxidase- and catalase-positive; it degraded Tween 60, but not DNA, urea, gelatin or starch. It grew at pH values from 7.5 to 11.0, with optimum growth occurring at pH 9.0, and growth was observed in NaCl concentrations of 0.2-1.3 M, with optimum growth at 0.8 M NaCl. The optimum temperature for growth was 37 degrees C. Strain 4CAT was resistant to erythromycin, bacitracin, novobiocin, polymyxin B, neomycin, gentamicin, streptomycin, carbenicillin, rifampicin and tetracycline, and was susceptible to nalidixic acid, chloramphenicol, ampicillin and penicillin. The isolate's 16S rRNA gene sequence indicated that it belonged to the gamma-Proteobacteria, showing 90-94 % similarity to its closest relatives. Maximum-likelihood phylogenetic inferences placed strain 4CAT within a novel lineage related to the marine bacterial genera Neptunomonas and Marinobacterium. The DNA G+C content of the isolate was 47.4 mol%. On the basis of genotypic and phenotypic characterization, it was concluded that strain 4CAT should be placed in a separate taxon as a novel genus and species, with the proposed name Nitrincola lacisaponensis gen. nov., sp. nov. The type strain is 4CAT (=ATCC BAA-920T=DSM 16316T).

Base Composition↗

PubMed2004

Uranium complexes formed at hematite surfaces colonized by sulfate-reducing bacteria.

Modeling uranium (U) transport in subsurface environments requires a thorough knowledge of mechanisms likely to restrict its mobility, such as surface complexation, precipitation, and colloid formation. In closed systems, sulfate-reducing bacteria (SRB) such as Desulfovibrio spp. demonstrably affect U immobilization by enzymatic reduction of U(VI) species (primarily the uranyl ion, UO2(2+), and its complexes) to U(IV). However, our understanding of such interactions under chronic U(VI) exposure in dynamic systems is limited. As a first step to understanding such interactions, we performed bioreactor experiments under continuous flow to study the effect of a biofilm of the sulfate-reducing bacterium Desulfovibrio desulfuricans attached to specular hematite (alpha-Fe2O3) surfaces on surface-associated U(VI) complexation, transformation, and mobility. Employing real-time microscopic observation and X-ray photoelectron spectroscopy (XPS), we show that the characteristics of the U(VI) complex(es) formed at the hematite surface are influenced by the composition of the bulk aqueous phase flowing across the surface and bythe presence of surface-associated SRB. The XPS data further suggest higher levels of U associated with hematite surfaces colonized by SRB than with bacteria-free surfaces. Microscopic observations indicate that at least a portion of the U(VI) that accumulates in the presence of the SRB is exterior to the cells, possibly associated with the extracellular biofilm matrix. The U4f7/2 core-region spectrum and U5f2 valence-band spectrum provide preliminary evidence that the SRB-colonized hematite surface accumulates both U(VI) and U(IV) phases, whereas only the U(VI) phase(s) accumulates on uncolonized hematite surfaces. The results suggest that mineral surfaces exposed to a continuously replenished supply of U(VI)-containing aqueous phase will accumulate U phases that may be more representative of those that exist in U-contaminated aquifers than those which accumulate in closed experimental systems. These phases should be considered in models attempting to predict U transport through subsurface environments.

Biological Availability↗

PubMed2004

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↗

PubMed2004

Composition and diversity of microbial communities recovered from surrogate minerals incubated in an acidic uranium-contaminated aquifer.

Our understanding of subsurface microbiology is hindered by the inaccessibility of this environment, particularly when the hydrogeologic medium is contaminated with toxic substances. In this study, surrogate geological media contained in a porous receptacle were incubated in a well within the saturated zone of a pristine region of an aquifer to capture populations from the extant communities. After an 8-week incubation, the media were recovered, and the microbial community that developed on each medium was compared to the community recovered from groundwater and native sediments from the same region of the aquifer, using 16S DNA coding for rRNA (rDNA)-based terminal restriction fragment length polymorphism (T-RFLP). The groundwater and sediment communities were highly distinct from one another, and the communities that developed on the various media were more similar to groundwater communities than to sediment communities. 16S rDNA clone libraries of communities that developed on particles of a specular hematite medium incubated in the same well as the media used for T-RFLP analysis were compared with those obtained from an acidic, uranium-contaminated region of the same aquifer. The hematite-associated community formed in the pristine area was highly diverse at the species level, with 25 distinct phylotypes identified, the majority of which (73%) were affiliated with the beta-Proteobacteria. Similarly, the hematite-associated community formed in the contaminated area was populated in large part by beta-Proteobacteria (62%); however, only 13 distinct phylotypes were apparent. The three numerically dominant clones from the hematite-associated community from the contaminated site were affiliated with metal- and radionuclide-tolerant or acidophilic taxa, consistent with the environmental conditions. Only two populations were common to both sites.

Base Sequence↗

PubMed2003

Phenol and catechol biodegradation by the haloalkaliphile Halomonas campisalis: influence of pH and salinity.

Removal of aromatic compounds from alkaline and/or saline industrial wastewater is an environmental concern for industry. In addition, aromatics may be accumulating in soda lakes, unique natural systems, where the fate and toxicity of these contaminants is unknown. To determine the feasibility of aromatic compound biodegradation in saline and alkaline conditions, the effect of pH and salinity on the biodegradation of phenol as a model aromatic waste compound by the haloalkaliphilic bacterium Halomonas campisalis was examined. Phenol was degraded as a source of carbon and energy at pH 8-11 and 0-150 g/L NaCl. Metabolic intermediates catechol, cis,cis-muconate, and (+)-muconolactone were identified, thus indicating that phenol was degraded via the beta-ketoadipate metabolic pathway. Although phenol and catechol were completely degraded in all cases, small amounts of cis,cis-muconate accumulated proportionally to increases in pH. There was no noticeable influence of salinity on cis,cis-muconate accumulation except at 0 g/L NaCl where it was completely degraded. These results indicate that it may be feasible to use haloalkaliphiles forthe treatment of aromatics present in saline and/or alkaline systems. This is the first report of phenol and catechol biodegradation under combined saline and alkaline conditions.

Biodegradation, Environmental↗

PubMed2003

Modeling chromate reduction in Shewanella oneidensis MR-1: development of a novel dual-enzyme kinetic model.

Chromate (Cr(VI)) reduction tests were performed with nitrate- and fumarate-grown stationary phase cultures of Shewanella oneidensis MR-1 (henceforth referred to as MR-1) and disappearance of Cr(VI) was monitored over time. A rapid initial decrease in Cr(VI) concentration was observed, which was followed by a slower, steady decrease. These observations appear to be consistent with our previous results indicating that Cr(VI) reduction in MR-1 involves at least two mechanisms (Viamajala et al., 2002b). Modeling of metal reduction kinetics is often based on single-enzyme Michaelis-Menten equations. However, these models are often developed using initial rates and do not always match actual reduction profiles. Based on the hypothesis that multiple Cr(VI) reduction mechanisms exist in MR-1, a model was developed to describe the kinetics of Cr(VI) reduction by two parallel mechanisms: (1) a rapid Cr(VI) reduction mechanism that was deactivated (or depleted) quickly, and (2) a slower mechanism that had a constant activity and was sustainable for a longer duration. Kinetic parameters were estimated by fitting experimental data, and model fits were found to correspond very closely to quantitative observations of Cr(VI) reduction by MR-1.

Cell Culture Techniques↗

PubMed2003

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↗

PubMed2002

Chromate/nitrite interactions in Shewanella oneidensis MR-1: evidence for multiple hexavalent chromium [Cr(VI)] reduction mechanisms dependent on physiological growth conditions.

Inhibition of hexavalent chromium [Cr(VI)] reduction due to nitrate and nitrite was observed during tests with Shewanella oneidensis MR-1 (previously named Shewanella putrefaciens MR-1 and henceforth referred to as MR-1). Initial Cr(VI) reduction rates were measured at various nitrite concentrations, and a mixed inhibition kinetic model was used to determine the kinetic parameters-maximum Cr(VI) reduction rate and inhibition constant [V(max,Cr(VI)) and K(i,Cr(VI))]. Values of V(max,Cr(VI)) and K(i,Cr(VI)) obtained with MR-1 cultures grown under denitrifying conditions were observed to be significantly different from the values obtained when the cultures were grown with fumarate as the terminal electron acceptor. It was also observed that a single V(max,Cr(VI)) and K(i,Cr(VI)) did not adequately describe the inhibition kinetics of either nitrate-grown or fumarate-grown cultures. The inhibition patterns indicate that Cr(VI) reduction in MR-1 is likely not limited to a single pathway, but occurs via different mechanisms some of which are dependent on growth conditions. Inhibition of nitrite reduction due to the presence of Cr(VI) was also studied, and the kinetic parameters V(max,NO2) and K(i,NO2) were determined. It was observed that these coefficients also differed significantly between MR-1 grown under denitrifying conditions and fumarate reducing conditions. The inhibition studies suggest the involvement of nitrite reductase in Cr(VI) reduction. Because nitrite reduction is part of the anaerobic respiration process, inhibition due to Cr(VI) might be a result of interaction with the components of the anaerobic respiration pathway such as nitrite reductase. Also, differences in the degree of inhibition of nitrite reduction activity by chromate at different growth conditions suggest that the toxicity mechanism of Cr(VI) might also be dependent on the conditions of growth. Cr(VI) reduction has been shown to occur via different pathways, but to our knowledge, multiple pathways within a single organism leading to Cr(VI) reduction has not been reported previously.

Chromium↗

PubMed2002

Kinetics of phenol biodegradation in high salt solutions.

Biological treatment of high-salinity industrial wastewaters using halophilic bacteria can be used to remove organic compounds without first decreasing the salt concentration. While halophilic degradation of phenol and other organics has been investigated, there exists a general absence of kinetic data in current literature to allow evaluation of this treatment alternative. Liquid, soil and sediment samples were collected from three distinct saline environments in the western United States. Samples were enriched in media containing 10% (w/v) NaCl at pH 7.0, with phenol as a substrate. Mixed culture batch experiments were conducted at 30 degrees C with initial phenol concentrations of 50 mg/L. Evaluation of phenol degradation and corresponding cell growth data with Monod and Andrews models indicated that the kinetics were zero-order with respect to phenol. Zero-order specific growth rates ranged from 0.22 to 0.32 h(-1), while observed cell yields were 0.18-0.28 mg cell protein/mg phenol for the five cultures. For one of the cultures, phenol degradation rates were also quantified at concentrations of up to 320 mg/L. Under these conditions, specific growth rates ranged from 0.09 to 0.22 h(-1), decreasing with increasing initial phenol concentrations. Cell yields at these higher initial phenol concentrations ranged from 0.20 to 0.29 mg cell protein/mg phenol.

Biodegradation, Environmental↗

PubMed2002

A review of spectroscopic methods for characterizing microbial transformations of minerals.

Over the past decade, advances in surface-sensitive spectroscopic techniques have provided the opportunity to identify many new microbiologically mediated biogeochemical processes. Although a number of surface spectroscopic techniques require samples to be dehydrated, which precludes real-time measurement of biotransformations and generate solid phase artifacts, some now offer the opportunity to either isolate a hydrated sample within an ultrahigh vacuum during analysis or utilize sources of radiation that efficiently penetrate hydrated specimens. Other nondestructive surface spectroscopic techniques permit determination of the influence of microbiological processes on the kinetics and thermodynamics of geochemical reactions. The ability to perform surface chemical analyses at micrometer and nanometer scales has led to the realization that bacterial cell surfaces are active sites of mineral nucleation and propagation, resulting in the formation of both stable and transient small-scale surface chemical heterogeneities. Some surface spectroscopic instrumentation is now being modified for use in the field to permit researchers to evaluate mineral biotransformations under in situ conditions. Surface spectroscopic techniques are thus offering a variety of opportunities to yield new information on the way in which microorganisms have influenced geochemical processes on Earth over the last 4 billion years.

Bacteria↗

PubMedDate not supplied

Chromate reduction in Shewanella oneidensis MR-1 is an inducible process associated with anaerobic growth.

Cr(VI) reduction was observed during tests with Shewanella oneidensis MR-1 (previously named S. putrefaciens MR-1) while being grown with nitrate or fumarate as electron acceptor and lactate as electron donor. From the onset of anoxic growth on fumarate, we measured a gradual and progressive increase in the specific Cr(VI) reduction rate with incubation time until a maximum was reached at late exponential/early stationary phase. Under denitrifying conditions, the specific Cr(VI) reduction rate was inhibited by nitrite, which is produced during nitrate reduction. However, once nitrite was consumed, the specific reduction rate increased until a maximum was reached, again during the late exponential/early stationary phase. Thus, under both fumarate- and nitrate-reducing conditions, an increase in the specific Cr(VI) reduction rate was observed as the microorganisms transition from oxic to anoxic growth conditions, presumably as a result of induction of enzyme systems capable of reducing Cr(VI). Although Cr(VI) reduction has been studied in MR-1 and in other facultative bacteria under both oxic and anoxic conditions, a transition in specific reduction rates based on physiological conditions during growth is a novel finding. Such physiological responses provide information required for optimizing the operation of in situ systems for remediating groundwater contaminated with heavy metals and radionuclides, especially those that are characterized by temporal variations in oxygen content. Moreover, such information may point the way to a better understanding of the cellular processes used by soil bacteria to accomplish Cr(VI) reduction.

Anaerobiosis↗

PubMedDate not supplied

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↗