Molybdenum Availability Is Key to Nitrate Removal in Contaminated Groundwater Environments

The concentrations of molybdenum (Mo) and 25 other metals were measured in groundwater samples from 80 wells on the Oak Ridge Reservation (ORR) (Oak Ridge, TN), many of which are contaminated with nitrate, as well as uranium and various other metals. The concentrations of nitrate and uranium were in the ranges of 0.1 μM to 230 mM and <0.2 nM to 580 μM, respectively. Almost all metals examined had significantly greater median concentrations in a subset of wells that were highly contaminated with uranium (≥126 nM). They included cadmium, manganese, and cobalt, which were 1,300- to 2,700-fold higher. A notable exception, however, was Mo, which had a lower median concentration in the uranium-contaminated wells. This is significant, because Mo is essential in the dissimilatory nitrate reduction branch of the global nitrogen cycle. It is required at the catalytic site of nitrate reductase, the enzyme that reduces nitrate to nitrite. Moreover, more than 85% of the groundwater samples contained less than 10 nM Mo, whereas concentrations of 10 to 100 nM Mo were required for efficient growth by nitrate reduction for two Pseudomonas strains isolated from ORR wells and by a model denitrifier, Pseudomonas stutzeri RCH2. Higher concentrations of Mo tended to inhibit the growth of these strains due to the accumulation of toxic concentrations of nitrite, and this effect was exacerbated at high nitrate concentrations. The relevance of these results to a Mo-based nitrate removal strategy and the potential community-driving role that Mo plays in contaminated environments are discussed.     

Draft genome sequence for Microbacterium laevaniformans strain OR221, a bacterium tolerant to metals, nitrate, and low pH

Microbacterium laevaniformans strain OR221 was isolated from subsurface sediments obtained from the Field Research Center (FRC) in Oak Ridge, TN. It was characterized as a bacterium tolerant to heavy metals, such as uranium, nickel, cobalt, and cadmium, as well as nitrate and low pH. We present its draft genome sequence.     

Applying Mental Models to Qualitative Risk Assessment at the Tar Creek Superfund Site

Re-suspension of heavy metal residuals generated by former mining activities conducted at the Tar Creek Superfund site may result in exposures of nearby residents. Currently, remediation and mitigation activities include removal of yard soil containing high concentrations of heavy metals, removal of selected chat piles, and providing assistance for families with young children. Although these are important activities to reduce potential risks to Tar Creek residents, a qualitative assessment of the site that uses mental models identifies other potential contaminants of concern, transport, and exposure pathways that may require further investigation. Mental models structure judgments about hazards and risk by visually representing a contaminated site through the use of influence diagrams. A mental model was applied to the Mayer Ranch area of the Tar Creek Superfund site in Ottawa County, Oklahoma, to support a qualitative assessment of the health risks from heavy metals exposure. Documents were reviewed to develop influence diagrams illustrating the processes, interactions, and potential exposures that could subject a hypothetical individual recreating at Mayer Ranch to various health risks. The diagrams were modified to reflect expert judgments. The mental model includes four influence diagrams: source term, fate, and transport processes that may influence human health risks due to exposure to heavy metals; natural wetland processes that could remove heavy metals; net alkaline constructed wetland processes that could remove heavy metals; and potential future impacts from the removal of sediment and land development that could increase risk. Policies, regulations, and technologies that could most reduce the probability of exposures can be determined and evaluated by focusing on the environmental transport pathways and contaminants that pose the greatest potential risks and determining the difference between the risks. Also evident are the knowledge gaps with respect to the processes within wetlands.     

Screening Evaluation of the Ecological Risks to Terrestrial Wildlife Associated with a Coal Ash Disposal Site

Between 1955 and 1989, coal ash was deposited within an impounded watershed on the U.S. Department of Energy's Oak Ridge Reservation in Tennessee, creating the 3.6?ha Filled Coal Ash Pond (FCAP). The site has subsequently become vegetated wildlife habitat. To evaluate risks that metals in ash may pose to wildlife; ash, surface water, small mammal, and vegetation samples were collected and metal residues were determined. Metal concentrations, As and Se in particular, were elevated in ash, surface water, plant foliage, and small mammals relative to reference materials. Estimates of metal exposures were calculated for short-tailed shrews, white-footed mice, white-tailed deer, red fox, and red-tailed hawks. While shrews and mice were assumed to reside exclusively at and receive 100% exposure from the site, exposure experienced by deer, fox, and hawks was assumed to be proportional to the size of the site relative to their home range. Because deer had been observed to consume ash, presumably for its high sodium content, exposure experienced by deer consuming ash to meet sodium requirements was also estimated. Exposure estimates were compared to body-size adjusted toxicity data for each metal. These comparisons suggest that metals at the site may be detrimental to reproduction and survivorship of mice, shrews, and deer consuming ash for sodium; fox and hawks do not appear to be at risk     

Physicochemical and Mineralogical Characterization of Uranium-Contaminated Soils

Physicochemical and mineralogical properties of the contaminants should be taken into account to decide a remediation strategy for a given radionuclide because development and optimization of soil remedial technologies are based on physicochemical and mineralogical separation techniques. The objectives of this study are to (1) demonstrate how a priori physicochemical and mineralogical characterization of soil contaminants can direct the development of remediation strategies and their performance evaluation for soil treatments and (2) understand the nature of uranium contamination and its association with the soil matrix by chemical extractions. This study examined two U-contaminated sites (K311 and K1300) at the DOE K-25 site, presently located at East Tennessee Technology Park, Oak Ridge, Tennessee. Uranium concentrations of the soils ranged from 1499 to 216,413 Bq kg^?1 at both sites. Scanning electron microscopy with backscattered electron spectroscopy and X-ray diffraction analysis showed that the dominant U phases are U oxides (schoepite), U-Ca-silicate (uranophane) and U silicate (coffinite) from the K311 site soils, whereas U-Ca-oxide and U-Ca-phosphate dominate in the K1300 site soils. Sodium carbonate/bicarbonate leaching was effective on the K1300 site soils, whereas citric acid leaching is effective on the K311 site soils. Sequential leaching showed that the majority of the uranium in the contaminated soils was contained in carbonate minerals (45%) and iron oxides (40%). Conventional leaching showed that citric acid treatment was most effective on the K311 site soils, whereas the sodium carbonate/ bicarbonate treatment was most effective on the K1300 site soils.     

Iron- and aluminium-induced depletion of molybdenum in acidic environments impedes the nitrogen cycle

Anthropogenic nitrate contamination is a serious problem in many natural environments. Nitrate removal by microbial action is dependent on the metal molybdenum (Mo), which is required by nitrate reductase for denitrification and dissimilatory nitrate reduction to ammonium. The soluble form of Mo, molybdate (MoO₄²⁻), is incorporated into and adsorbed by iron (Fe) and aluminium (Al) (oxy) hydroxide minerals. Herein we used Oak Ridge Reservation (ORR) as a model nitrate-contaminated acidic environment to investigate whether the formation of Fe- and Al-precipitates could impede microbial nitrate removal by depleting Mo. We demonstrate that Fe and Al mineral formation that occurs as the pH of acidic synthetic groundwater is increased, decreases soluble Mo to low picomolar concentrations, a process proposed to mimic environmental diffusion of acidic contaminated groundwater. Analysis of ORR sediments revealed recalcitrant Mo in the contaminated core that co-occurred with Fe and Al, consistent with Mo scavenging by Fe/Al precipitates. Nitrate removal by ORR isolate Pseudomonas fluorescens N2A2 is virtually abolished by Fe/Al precipitate-induced Mo depletion. The depletion of naturally occurring Mo in nitrate- and Fe/Al-contaminated acidic environments like ORR or acid mine drainage sites has the potential to impede microbial-based nitrate reduction thereby extending the duration of nitrate in the environment.     

Toxicity evaluation of complex metal mixtures using reduced metal concentrations: application to iron oxidation by Acidithiobacillus ferrooxidans

In this study, we investigated the inhibition effects of single and mixed heavy metal ions (Zn2+, Ni2+, Cu2+, and Cd2+) on iron oxidation by Acidithiobacillus ferrooxidans. Effects of metals on the iron oxidation activity of A. ferrooxidans are categorized into four types of patterns according to its oxidation behavior. The results indicated that the inhibition effects of the metals on the iron oxidation activity were noncompetitive inhibitions. We proposed a reduced inhibition model, along with the reduced inhibition constant (alphai), which was derived from the inhibition constant (KI) of individual metals and represented the tolerance of a given inhibitor relative to that of a reference inhibitor. This model was used to evaluate the toxicity effect (inhibition effect) of metals on the iron oxidation activity of A. ferrooxidans. The model revealed that the iron oxidation behavior of the metals, regardless of metal systems (single, binary, ternary, or quaternary), is closely matched to that of any reference inhibitor at the same reduced inhibition concentration, [I]reduced, which defines the ratio of the inhibitor concentration to the reduced inhibition constant. The model demonstrated that single metal systems and mixed metal systems with the same reduced inhibitor concentrations have similar toxic effects on microbial activity.     

Optimization and/or acclimatization of activated sludge process under heavy metals stress

The present study aimed to overcome the toxicity of the heavy metals load, discharged with the industrial effluents into Alexandria sewerage network, on the activated sludge treatment system through effective acclimation for organic matter and heavy metals removal. Optimization and/or acclimatization of the activated sludge process in the presence of Cu, Cd, Co and Cr contaminating mixed domestic-industrial wastewater was investigated. Acclimatization process was performed through abrupt and stepwise addition of tested metals using sequencing batch reactors treatment approach and evaluated as microbial oxygen uptake rate (OUR), dehydrogenase activity (DHA), organic matter (COD) and heavy metals removal. Abrupt addition of metals adversely affected sludge bioactivity leading to decline in the removal efficiency of the targeted contaminants and loss of floc structure. Metals IC₅₀ confirmed that copper possessed the highest toxicity towards the OUR, DHA activity and COD removal with orders Cu > Cd > Cr > Co; Cu > Cd > Co = Cr and Cu > Cd > Cr > Co, respectively. The highest metal removal was recorded for Cd followed by Co, Cu and finally Cr, most of which was retained in the dissolved influent. However, controlled stepwise application of the tested metals exhibited high sensitivity of DHA and OUR activities only at the highest metal concentrations although enhanced at the lowest concentrations while COD removal was not significantly affected. In conclusion, this approach resulted in adaptation of the system where sludge microbes acquired and developed natural resistance to such metals leading to remarkable enhancement of both organic matter and heavy metals removal.     

Microarray analysis of a microbe–mineral interaction

The weathering of volcanic minerals makes a significant contribution to the global silicate weathering budget, influencing carbon dioxide drawdown and long‐term climate control. Basalt rocks may account for over 30% of the global carbon dioxide drawdown in silicate weathering. Micro‐organisms are known to play a role in rock weathering yet the genomics and genetics of biological rock weathering are unknown. We apply DNA microarray technology to determine putative genes involved in weathering using the heavy metal‐resistant organism, Cupriavidus metallidurans CH34; in particular we investigate the sequestering of iron. The results show that the bacterium does not depend on siderophores. Instead, the up‐regulation of porins and transporters which are employed concomitantly with genes associated with biofilm formation suggests that novel passive and active iron uptake systems are involved. We hypothesize that these mechanisms induce rock weathering by changes in chemical equilibrium at the microbe–mineral interface, reducing the saturation state of iron. We also demonstrate that low concentrations of metals in the basalt induce heavy metal‐resistant genes. Some of the earliest environments on the Earth were volcanic. Therefore, these results not only elucidate the mechanisms by which micro‐organisms might have sequestered nutrients on the early Earth but also provide an explanation for the evolution of multiple heavy metal resistance genes long before the creation of contaminated industrial biotopes by human activity.     

Interaction of metals with peroxidase--mediated luminol-enhanced, chemiluminescence (PLmCL).

The peroxidase-mediated luminol-enhanced chemiluminescence (PLmCL) method has been used to study the in vitro effect of contaminants such as heavy metals on the reactive oxygen species production by immunocytes. We were interested to know whether metals could directly affect peroxidase-mediated luminescence, taking horseradish peroxidase (HRP) as a model enzyme, since this could contribute to the inhibition of immunocyte LmCL. Copper inhibited PLmCL in a dose-dependent manner, while cadmium, iron, silver and lead only partly decreased the signal in the concentration range tested. In contrast, zinc enhanced the signal at high concentrations. Eventually, chromium, mercury and aluminium did not affect PLmCL. It is suggested that these effects reflect the ability of the metals to interact with the active site of the peroxidase. These results demonstrate that such interactions have to be considered when interpreting the effects of metals on immunocytes using the LmCL method.     

Horizontal gene transfer of PIB-type ATPases among bacteria isolated from radionuclide- and metal-contaminated subsurface soils

Aerobic heterotrophs were isolated from subsurface soil samples obtained from the U.S. Department of Energy's (DOE) Field Research Center (FRC) located at Oak Ridge, Tenn. The FRC represents a unique, extreme environment consisting of highly acidic soils with co-occurring heavy metals, radionuclides, and high nitrate concentrations. Four hundred isolates obtained from contaminated soil were assayed for heavy metal resistance, and a smaller subset was assayed for tolerance to uranium. The vast majority of the isolates were gram-positive bacteria and belonged to the high-G+C- and low-G+C-content genera Arthrobacter and Bacillus, respectively. Genomic DNA from a randomly chosen subset of 50 Pb-resistant (Pb(r)) isolates was amplified with PCR primers specific for P(IB)-type ATPases (i.e., pbrA/cadA/zntA). A total of 10 pbrA/cadA/zntA loci exhibited evidence of acquisition by horizontal gene transfer. A remarkable dissemination of the horizontally acquired P(IB)-type ATPases was supported by unusual DNA base compositions and phylogenetic incongruence. Numerous Pb(r) P(IB)-type ATPase-positive FRC isolates belonging to the genus Arthrobacter tolerated toxic concentrations of soluble U(VI) (UO(2)(2+)) at pH 4. These unrelated, yet synergistic, physiological traits observed in Arthrobacter isolates residing in the contaminated FRC subsurface may contribute to the survival of the organisms in such an extreme environment. This study is, to the best of our knowledge, the first study to report broad horizontal transfer of P(IB)-type ATPases in contaminated subsurface soils and is among the first studies to report uranium tolerance of aerobic heterotrophs obtained from the acidic subsurface at the DOE FRC.     

Heavy metal pollution negatively correlates with anuran species richness and distribution in south‐eastern Australia

Heavy metal pollution has likely played an important role in global biodiversity decline, but there remains a paucity of information concerning the effects of metals on amphibian diversity. This study assessed anuran species richness and distribution in relation to sediment metal content and water chemistry in wetlands located along the Merri Creek corridor in Victoria, south‐eastern Australia. Anurans were present in 60% (21/35) of study sites, with a total of six species detected: the eastern common froglet (Crinia signifera), the eastern sign‐bearing froglet (Crinia parinsignifera), the southern brown tree frog (Litoria ewingii), the growling grass frog (Litoria raniformis), the eastern banjo frog (Limnodynastes dumerilii) and the spotted marsh frog (Limnodynastes tasmaniensis). Mean species richness was 1.77 ± 0.32 per site, and species richness ranged from zero to six species per site. Across sites, species richness correlated negatively with sediment concentrations of six heavy metals: copper, nickel, lead, zinc, cadmium and mercury. Species richness also correlated negatively with wetland water electrical conductivity (a proxy for salinity) and concentrations of orthophosphate. Distributions of the three most commonly observed frog species (C. signifera, L. tasmaniensis and L. ewingii) were significantly negatively associated with the total level of metal contamination at individual sites. The study is the first to provide evidence for an association between metal contamination and anuran species richness and distribution in the southern hemisphere, adding to a small but growing body of evidence that heavy metal pollution has contributed to global amphibian decline.     

Heavy metals dynamics in seaweeds and seagrasses in Bahía Magdalena,B.C.S., México

The process of metal bioaccumulation in marine food chains is poorly understood because very little data is available on metal concentration at different trophic levels and their temporal or spatial variation. Because of that, we were interested to (1) determine the concentration range of heavy metals in seaweed and seagrasses species in Magdalena Bay; (2) describe the spatial and temporal variation of heavy metal concentrations in the seaweeds and seagrasses. Seasonal collections were done at Estero Banderitas in November 2004, February, and April 2005 wherein we divided the estuary into three major areas (upper, middle, and lower), and within each area, two sites were selected. Our results showed that iron, copper, and magnesium were the most significant metals found in seagrasses, red, and green algae. We found significant more variation in temporal heavy metal concentrations in relation to the maximum abundance in the samples and spatial variation in relation to the studied taxa suggesting that hervibores have a differential intake of the metals. Also, our results suggest that heavy metals might be incorporated regularly in the diet of many herbivorous animals with severe consequences to their health. Management strategies for these species should consider monitoring the levels of metals.     

Multiple Metal Resistance and Co-resistance in Acer pseudoplatanus L. (Sycamore) Callus Cultures

In vitro growth of Acer pseudoplatanus L. (sycamore) callustissue derived from shoot-tip explants and screened on metal-enrichedmedia was studied in an attempt to identify resistance traitswhich may explain the survival of trees at metal-contaminatedsites. Copper and Cd-resistance traits were identified in celllines originating from trees at a site with a relatively recenthistory of severe contamination by these metals, and Cd- andZn-resistance were identified in cell lines originating frommature trees at a mining spoil site with a much longer historyof exposure to elevated concentrations of these metals. In Zn-resistantcell lines, co-resistance to Ni was also found, even thoughthis metal was not elevated at the study site. This is the firstreport of multiple resistance and co-resistance to metals occurringat the cellular level in trees. The mechanisms of the measuredresistance traits remain unclear, although there was evidenceof reduced Cu and Ni uptake by resistant cell lines. It is concludedthat facultative adaptations allowing acclimation to metal stressmay be particularly significant for survival of mature trees;induction of metal resistance probably occurs in vivo in treesat metal-contaminated sites.Copyright 1995, 1999 Academic Press Acer pseudoplatanus, sycamore, callus, heavy metals, metal resistance, trees     

Stream Macroinvertebrate Community Affected by Point‐Source Metal Pollution

The impacts of mining activities on aquatic biota have been documented in many stream ecosystems. In mining streams, point‐source heavy metal pollution often appears in the stream. We hypothesize that this pollution is toxic to macroinvertebrates owing to high concentrations of metals and therefore affects macroinvertebrate community structure. We investigated macroinvertebrate community structure in mountain streams, including heavy metal‐polluted sites and neutral‐pH streams, to determine the relationship between community structure and environmental factors such as low pH and heavy metal concentrations. Based on multidimensional scaling ordination, the macroinvertebrate community at heavy metal pollution sites was remarkably different from that at the other sites. Inductively coupled plasma mass spectrometry revealed high concentrations of aluminum and iron in surface water at the polluted sites. Macroinvertebrate community structure at the metal pollution sites was significantly different from that at other sites in the same stream and in neutral‐pH streams. Thus, point‐source metal pollution may reduce the density and diversity of in situ macroinvertebrates. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Metallothionein induction in human peripheral blood lymphocytes by heavy metals

Human peripheral blood lymphocytes have the capacity to produce metallothioneins (MTs) as a protective response to cadmium exposure. To define the range of metal species inducing lymphocyte MTs, cellular proteins synthesized after exposure to each of 11 heavy metals were analyzed by gel electrophoresis. Toxic metals such as cadmium, mercury and silver were found to induce thioneins (apoproteins of MTs) at relatively low concentrations (maximum at approximately 10 microM), whereas less toxic metals such as zinc, copper and nickel were inductive at relatively high concentrations (maximum at approximately 200 microM). Tin, lead, iron, cobalt, and manganese did not induce thioneins. The heavy metal specificity of MT induction in the lymphocyte resembles that in the liver, and the regulatory mechanism of MT production seems to be similar in both of these tissues. In the cells exposed to highly toxic metals such as cadmium and mercury, expression of cytotoxicity (represented by decline of cysteine uptake) was remarkable at the metal concentrations higher than those saturating thionein induction, supporting the protective role of MTs against heavy metals.     

Trace elements in the Upper Fly River, Papua New Guinea

SUMMARY. Measurements were made of calcium, magnesium and eight trace elements (Cu, Zn, Fe, Mn, Ni, Pb, Cd, Co) in waters, sediment and biota at eight sites in the Upper Fly River, Papua New Guinea. Determinations from streams draining an area rich in copper ore showed relatively low values for soluble copper and this was attributed to the small amount of ore exposed to weathering, heavy rainfall and high pH of water associated with the limestone bedrock. More iron was transported in a soluble form down a headwater tributary (Alice River) than in the Fly River but higher iron concentrations were associated with particulate matter in the Fly than in the Alice. The geochemistry of the catchment and the distance from the headwaters were important influences on the amount of particulate iron and its distribution between soluble and particulate forms.
Levels of metals in the biota generally reflected the background concentrations of metals encountered at each site. Concentrations of Cu, Cd, and Zn were lower in starved invertebrates compared with unstarved individuals but no consistent changes were observed in Fe or Mn content.     

Potential for In Situ Bioremediation of a Low-pH,High-Nitrate Uranium-Contaminated Groundwater

The potential for stimulating microbial U(VI) reduction as an in situ bioremediation strategy for uranium-contaminated groundwater was evaluated in uranium-contaminated sediment from the FRC, Oak Ridge, TN. Sediment was at low pH (pH 4) and contained high (55 mM) concentrations of nitrate. The addition of organic electron donors resulted in a slow removal of ca. 20% of the nitrate over 120 days with a concurrent increase in pH. Uranium precipitated during nitrate reduction. This precipitation of U(VI) was not due to its reduction to U(IV) because over 90% of the uranium in the sediments remained as U(VI). Studies in which the pH of the sediments was artificially raised suggested that an increase in pH alone could not account for the precipitation of the U(VI) during nitrate reduction. Metal-reducing bacteria were recovered from the sediments in enrichment cultures, but molecular analysis of the sediment demonstrated that the addition of electron donors did not stimulate the growth of these metal reducers. Thus, although U(VI) was precipitated from the groundwater with the simple addition of electron donors, most of the uranium in the sediments was in the form of U(VI), and thus was not effectively immobilized.     

Evaluation of a new peat-based sorbent for metals capture

A new peat-based sorbent was evaluated for the capture of heavy metals from waste streams. The media is a pelletted blend of organic humic material targeted for the capture of soluble metals from industrial waste streams and stormwater. The metals chosen for the media evaluation were Cd, Cu, Ni, and Zn due to their occurrence and abundance in waste streams and runoff. Sorption tests included an evaluation of the rate and extent of metals capture by the media, single versus multicomponent metals uptake, pH, anion influence, leaching effects and the effect of media moisture content on uptake rate and capacity. Isotherms of the sorption results showed that the presence of multiple metals increased the total sorption capacity of the media compared to the single component metal capacity; a result of site selectivity within the media. However the capacity for an individual metal in a multicomponent metal matrix was reduced compared to its single component capacity, due to competition for sites. Evidence of ion exchange behavior was observed but did not account for all metals capture. The media also provided a buffering action to counter the pH drop typically associated with metals capture.