Range Of Cu Concentrations Research Articles

Microflow Injection System for Efficient Cu(II) Detection across a Broad Range

In this study, a modular, multi-step, photometric microflow injection analysis (micro-FIA) system for the automatic determination of Cu(II) in a bioreactor was developed. The system incorporates diverse 3D-printed modules, including a platform formed by a mixer module to mix Cu(II) with hydroxylamine, which reduces Cu(II) to Cu(I) linked to a diluter module via a Tesla valve, a chelation mixer module, a disperser module, and a detector module provided by an LED light source at λ = 455 nm and a light dependence resistor (LDR) as a light intensity detector. The system measures the color intensity resulting from the chelation between Cu(I) and neocuproine. The micro-FIA system demonstrated good capability for automatic and continuous Cu(II) determination, in a wide range of Cu concentrations, from 34 to 2000 mg L−1. The device exhibits a good repeatability (coefficient of variation below 2% across the measured concentration range), good reproducibility, and has an accuracy of around 100% between 600 and 1900 mg L−1. Real samples were analyzed using both the micro-FIA system and an atomic absorption spectroscopy method, revealing no statistically significant differences. Additionally, a Tesla valve located before the detector substituted a 3-way solenoid valve, eliminating the need for moving parts.

Genetical and Biochemical Basis of Methane Monooxygenases of Methylosinus trichosporium OB3b in Response to Copper

Over the past decade, copper (Cu) has been recognized as a crucial metal in the differential expression of soluble (sMMO) and particulate (pMMO) forms of methane monooxygenase (MMO) through a mechanism referred to as the “Cu switch”. In this study, we used Methylosinus trichosporium OB3b as a model bacterium to investigate the range of Cu concentrations that trigger the expression of sMMO to pMMO and its effect on growth and methane oxidation. The Cu switch was found to be regulated within Cu concentrations from 3 to 5 µM, with a strict increase in the methane consumption rates from 3.09 to 3.85 µM occurring on the 6th day. Our findings indicate that there was a decrease in the fold changes in the expression of methanobactin (Mbn) synthesis gene (mbnA) with a higher Cu concentration, whereas the Ton-B siderophore receptor gene (mbnT) showed upregulation at all Cu concentrations. Furthermore, the upregulation of the di-heme enzyme at concentrations above 5 µM Cu may play a crucial role in the copper switch by increasing oxygen consumption; however, the role has yet not been elucidated. We developed a quantitative assay based on the naphthalene–Molisch principle to distinguish between the sMMO- and pMMO-expressing cells, which coincided with the regulation profile of the sMMO and pMMO genes. At 0 and 3 µM Cu, the naphthol concentration was higher (8.1 and 4.2 µM, respectively) and gradually decreased to 0 µM naphthol when pMMO was expressed and acted as the sole methane oxidizer at concentrations above 5 µM Cu. Using physical protein–protein interaction, we identified seven transporters, three cell wall biosynthesis or degradation proteins, Cu resistance operon proteins, and 18 hypothetical proteins that may be involved in Cu toxicity and homeostasis. These findings shed light on the key regulatory genes of the Cu switch that will have potential implications for bioremediation and biotechnology applications.

Effect of dissolved organic matter on copper bioavailability to a coastal dinoflagellate at environmentally relevant concentrations

The speciation and bioavailability of copper (Cu) in the marine environment are affected by the presence of dissolved organic matter (DOM). Previous studies conducted at dissolved Cu concentrations >100 nM confirmed that Cu bioavailability depends on the concentration of labile Cu, as measured by anodic stripping voltammetry (ASV), which aligns with the expectations of the biotic ligand model (BLM). However, ambient Cu concentrations in coastal waters are generally lower, ranging between 1 and 80 nM, and the effect of DOM on the bioavailability of Cu to marine organisms has not been tested within that range of Cu concentrations. The present study aims to assess the impact of two types of DOM, a commercially available fulvic acid, and marine DOM extracted by ultrafiltration, on Cu bioavailability to phytoplankton using short-term 65Cu internalisation by the marine dinoflagellate Prorocentrum micans. Results showed that Cu internalisation decreases with DOM additions as expected according to the BLM and in agreement with ASV measurements of labile Cu, at the highest tested Cu concentration (100 nM). On the contrary, at a lower Cu concentration (20 nM), organic complexes appear to be partially bioavailable, thereby challenging the general applicability of the BLM model at environmentally relevant concentrations in coastal areas.

Foliar enrichment of copper oxide nanoparticles promotes biomass, photosynthetic pigments, and commercially valuable secondary metabolites and essential oils in dragonhead (Dracocephalum moldavica L.) under semi-arid conditions

High alkaline and low organic carbon hinder micronutrients, such as copper (Cu), bioavailability in (semi-) arid soils, affecting plant nutrient quality and productivity. This study aimed at investigating the potential beneficial effects of foliar Cu oxide nanoparticles (CuONPs) and conventional chelated-Cu applications (0–400 mg Cu/L) on the biomass, physiological biomarkers of plant productivity and oxidative stress, Cu bioaugmentation, and essential oils and secondary metabolites in dragonhead (Dracocephalum moldavica [L.]) grown in Cu-limited alkaline soil in semi-arid condition. Employing a randomized complete block design with three replicates, two different sources of Cu (CuONPs and chelated-Cu), and a wide range of Cu concentrations (0, 40, 80, 160, and 400 mg Cu/L), plants were foliarly treated at day-60 and day-74. At day-120, plants were harvested at the end of the flowering stage. Results showed shoot Cu bioaccumulation, flavonoids and anthocyanin increased in a dose-dependent manner for both Cu compounds, but the beneficial effects were significantly higher with CuONPs compared to chelated-Cu treatments. Further, shoot biomass (23 %), photosynthetic pigments (chlorophyll-a and chlorophyll-b; 77 and 123 %, respectively), and essential oil content and yield (70 and 104 %, respectively) increased significantly with foliar application of 80 mg/L CuONPs compared to equivalent concentration of chelated-Cu, suggesting an optimal threshold beyond which toxicity was observed. Likewise, commercially important secondary metabolites' yield (such as geranyl acetate, geranial, neral, and geraniol) was higher with 80 mg/L CuONPs compared to 160 mg/L chelated-Cu (2.3, 0.5, 2.5, and 7.1 %, respectively). TEM analyses of leaf ultrastructure revealed altered cellular organelles for both compounds at 400 mg/L, corroborating the results of oxidative stress response (malondialdehyde and H2O2). In conclusion, these findings indicate significantly higher efficacy of CuONPs, with an optimal threshold of 80 mg/L, in promoting essential oil and bioactive compound yield in dragonhead and may pave a path for the use of nano-Cu as a sustainable fertilizer promoting agricultural production in semi-arid soils that are micronutrient Cu deficient.

Bioaccumulation and human health risk assessment of some heavy metals in sediments, Sparus aurata and Salicornia europaea in Güllük Lagoon, the south of Aegean Sea.

In the current study, it was aimed to determine the sediment in Sparus aurata (Linnaeus 1758) tissues and Salicornia europaea L. Cu, Zn Fe, Hg, Cd, Pb and Mn concentration distributions in Güllük Lagoon which is located in the south of Aegean Sea and under the influence of anthropogenic activities by means of metal pollution and health indices and to investigate the effects of these metals on public health. The concentration range of Cu, Zn, Fe, Hg, Cd, Pb and Mn in sediment samples was determined as 12.16-26.00, 1.62-2.03, 7.77-8.36, 7.52-16.15, 0.071-0.40, 7.99-13.74 and 12.11-12.63mgkg-1, respectively. Cu and Hg concentrations in sediment were found to be higher than sediment quality guidelines standards. In addition, according to the enrichment factor (EF), Hg, Cd and Cu were found to show above moderate enrichment. Cu, Zn, Fe, Hg, Cd and Mn concentrations in S. aurata muscle tissue were 1.31 ± 2.30, 1.01 ± 0.24, 3.43 ± 0.75, 2.79 ± 0.85, 0.01 ± 0.01 and 1.80 ± 1.12mgkg-1, respectively. S. europaea heavy metals (HMs) concentrations were determined as Cu = 10.97 ± 3.20, Zn = 0.74 ± 0.62, Fe = 5.69 ± 0.22, Hg = 9.62 ± 8.84, Cd = 0.53 ± 0.33, Pb = 0.22 ± 0.26 and Mn = 8.61 ± 0.14mgkg-1. It was seen that Hg in S. aurata muscle tissue and Hg in S. europaea concentrations exceeded the limit values determined for consumption purposes. Target hazard quotient (THQ) and total target hazard quotient (TTHQ) values were found to be < 1 for S. aurata and S. europaea. When all these results were considered, it was determined that the metals that could pose a potential ecological and health risk were Hg, Cd and Cu in the study area.

Investigating the mechanisms of dissolved organic matter protection against copper toxicity in fish of Amazon's black waters

We investigated how natural dissolved organic matter (DOM) of the Rio Negro (Amazon) affects acute copper (Cu) toxicity to local fish: the cardinal tetra (Paracheirodon axelrodi) and the dwarf cichlid (Apistogramma agassizii). It is established that Cu2+ complexation with DOM decreases Cu bioavailability (and thus toxicity) to aquatic organisms, as conceptualized by the Biotic Ligand Model (BLM). However, we also know that Rio Negro's DOM can interact with fish gills and have a beneficial effect on Na+ homeostasis, the main target of acute Cu toxicity in freshwater animals. We aimed to tease apart these potential protective effects of DOM against Cu-induced Na+ imbalances in fish. In the laboratory, we acclimated fish to Rio Negro water (10 mg L−1 DOC) and to a low-DOM water (1.4 mg L−1 DOC) with similar ion composition and pH (5.9). We measured 3-h Cu uptake in gills and unidirectional and net Na+ physiological fluxes across a range of Cu concentrations in both waters. Various DOM pre-acclimation times (0, 1 and 5 days) were evaluated in experiments with P. axelrodi. Copper exposure led to similar levels of net Na+ loss in the two fish, but with distinct effects on Na+ influx and efflux rates reflecting their different ionoregulation strategies. Rio Negro DOM protected against Cu uptake and toxicity in the two fish species. Both Cu uptake in fish gills and Na+ regulation disturbances were relatively well predicted by the modelled aqueous free Cu2+ ion concentration. These findings suggest that protection by DOM occurs mainly from Cu complexation under the tested conditions. The prevalence of this geochemical-type protection over a physiological-type protection agrees with the BLM conceptual framework, supporting the use of the BLM to assess the risk of Cu in these Amazonian waters.

Effects of divalent copper on tetracycline degradation and the proposed transformation pathway.

To reveal the characteristics of tetracycline (TC) photocatalytic degradation under Cu(II) coexistence, effects of Cu(II) on TC photocatalytic degradation by ZnO nanoparticles (ZnO NPs) as a function of pH, humic acid (HA), and initial Cu(II) concentration were investigated. Interaction of TC with Cu(II) in the treatment process was analyzed by circular dichroism (CD) spectroscopy, while TC degradation pathway was investigated by high-performance liquid chromatography-mass spectrometry. Sixty-five percent and ninety-one percent TC degradation within 60min in the absence and presence of Cu(II), respectively, was reported. Both adsorption and photocatalytic degradation of TC under Cu(II) coexistence increased with increasing pH from 3 to 6, while decreased with further increase in pH. HA inhibited the degradation of TC by ZnO NPs both in the presence as well absence of Cu(II), while TC degradation decreased from 91 to 73% and from 73 to 37% in the presence and absence of Cu(II), respectively. TC degradation by ZnO NPs first increased then decreased with increasing Cu(II). Maximum TC degradation (about 94%) was obtained in the optimum concentration range of Cu(II) (0.05-0.15mmol/L). In addition, there was a lag effect between TC adsorption and degradation on ZnO NPs. TC degradation was improved via Cu(II)-TC surface complexation and followed N-demethylation and hydroxylation routes. This study could be of potential importance in extrapolating the transformation of TC or other antibiotics under the coexistence of heavy metals in water.

Effect of Copper Ion Concentration on Bacteria and Cells.

In the oral cavity, dental implants—most often made of commercially pure titanium—come in contact with bacteria, and antibacterial management has been researched extensively to improve patient care. With antibiotic resistance becoming increasingly prevalent, this has resulted in copper being investigated as an antibacterial element in alloys. In this study, the objective was to investigate the copper ion concentrations at which cyto-toxicity is avoided while bacterial inhibition is ensured, by comparing Cu ion effects on selected eukaryotes and prokaryotes. To determine relevant copper ion concentrations, ion release rates from copper and a 10 wt. % Cu Ti-alloy were investigated. Survival studies were performed on MC3T3 cells and Staphylococcus epidermidis bacteria, after exposure to Cu ions concentrations ranging from 9 × 10−3 to 9 × 10−12 g/mL. Cell survival increased from <10% to >90% after 24 h of exposure, by reducing Cu concentrations from 9 × 10−5 to 9 × 10−6 g/mL. Survival of bacteria also increased in the same range of Cu concentrations. The maximum bacteria growth was found at 9 × 10−7 g/mL, probably due to stress response. In conclusion, the minimum inhibitory concentrations of Cu ions for these prokaryotes and eukaryotes were found in the range from 9 × 10−5 to 9 × 10−6 g/mL. Interestingly, the Cu ion concentration correlating to the release rate of the 10 wt. % Cu alloy (9 × 10−8 g/mL) did not kill the bacteria, although this alloy has previously been found to be antibacterial. Further studies should investigate in depth the bacteria-killing mechanism of copper.

Assessing relative sensitivity of marine and freshwater bivalves following exposure to copper: Application of classical and novel genotoxicological biomarkers

Determination of relative sensitivity of biota following exposures to contaminants including metals is important for environmental protection. Copper (Cu), although biologically essential can be highly toxic to biota if present at higher concentrations in the natural environment. Given its ubiquitous presence within coastal and inland water bodies, we compared Cu-induced genotoxicity in two ecologically important mussel species, the freshwater Dreissena polymorpha (DP) and marine Mytilus galloprovincialis (MG), along with its tissue specific accumulation. Novel biomarker in terms of induction of gamma H2AX (γ-H2AX) foci, along with comet assay and induction of micronuclei (MN) were used to determine DNA damage response (DDR) in these two species following exposure to a range of Cu concentrations (18, 32, 56 μg L−1) for 10 days. Concentration-dependent increases in Cu concentration in gill tissue, as determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), were paralleled by a greater degree of genotoxicity. An induction of γ-H2AX foci was present in all Cu exposure concentrations, proving this technique to be a sensitive and suitable biomarker of genotoxicity in bivalves. The multi-biomarker approach adopted here suggests firstly that in parallel with MG, which is widely used to assess the health of marine and coastal environment, DP is also suitable representative of inland water bodies, and that there is a similar mechanism of action for the induction of genotoxicity between the two species, following exposure to Cu. Secondly, for genotoxicity assessment a battery of responses could simultaneously be assessed in these two bivalve species. Finally, for adequate protection of the environment it is vital to adopt a multi-biomarker, multi-species approach to determine adverse biological effects to gain a holistic understanding of the real threat posed by contaminants to hydrosphere.

Copper toxicity in Bristol Bay headwaters: Part 2—Olfactory inhibition in low‐hardness water

We investigated the olfactory toxicity of copper (Cu) to rainbow trout in low-hardness (27 mg/L as CaCO3 ) water formulated in the laboratory over a 120-h period using a flow-through design. The fish's response to an alarm cue (e.g., reduction in activity) was recorded to determine the exposure concentrations and durations that inhibited olfactory detection of the cue after 3, 24, 48, and 96 h of Cu exposure and after 24 h of clean water recovery following the 96-h exposure period. Exposures were conducted with a range of Cu concentrations from 0.13 (control) to 7.14 μg Cu/L (dissolved Cu). We observed a dose-dependent response in olfactory inhibition with a 20% reduction in the probability of responding to the alarm cue, relative to controls, at 2.7 and 2.4 μg Cu/L after 24 or 96 h of exposure, respectively. Olfactory inhibition manifested between 3 and 24 h of exposure. Our 24- and 96-h 20% olfactory inhibition estimates fell between the criteria derived using the biotic ligand model (BLM; criterion maximum concentration [CMC] and criterion continuous concentration [CCC] values were 0.63 and 0.39 μg Cu/L, respectively) and water hardness-based criteria (CMC and CCC values were 3.9 and 2.9 μg Cu/L, respectively). Therefore, the hardness-based criteria do not appear to be protective and the BLM-derived criteria do appear to be protective against Cu-induced olfactory inhibition given our test water chemistry. Neither the hardness-based criteria nor the BLM-derived criteria appear to be protective against our estimated Cu behavioral avoidance response concentrations at 24- and 96-h exposures (0.54 and 0.50 μg Cu/L, respectively). Environ Toxicol Chem 2019;38:198-209. © 2018 SETAC.

Copper excess reduces nitrate uptake by Arabidopsis roots with specific effects on gene expression

Nitrate uptake by plants is mediated by specific transport proteins in roots (NRTs), which are also dependent on the activity of proton pumps that energize the reaction. Nitrogen (N) metabolism in plants is sensitive to copper (Cu) toxicity conditions. To understand how Cu affects the uptake and assimilation processes, this study assesses the inhibitory effects of elevated Cu levels on the expression of genes related to N absorption, transport and assimilation in roots of Arabidopsis. Plants were grown hydroponically for 45 days, being exposed to a range of Cu concentrations in the last 72 h or alternatively exposed to 5.0 μM Cu for the last 15 days. High Cu levels decreased the uptake and accumulation of N in plants. It down-regulated the expression of genes encoding nitrate reductase (NR1), low-affinity nitrate transporters (NRT1 family) and bZIP transcription factors (TGA1 and TGA4) that regulate the expression of nitrate transporters. Cu toxicity also specifically down-regulated the plasma membrane proton pump, AHA2, whilst having little effect on AHA1 and AHA5. In contrast, there was an up-regulation of high-affinity nitrate transporters from the NRT2 family when exposed to medium level of Cu excess, but this was insufficient for restoring N absorption by roots to control levels. These results demonstrate that plants display specific responses to Cu toxicity, modulating the expression of particular genes related to nitrate uptake, such as low-affinity nitrate transporters and proton pumps.

Sulfide enrichment at an oceanic crust-mantle transition zone: Kane Megamullion (23°N, MAR)

The Kane Megamullion oceanic core complex located along the Mid-Atlantic Ridge (23°30′N, 45°20′W) exposes lower crust and upper mantle directly on the ocean floor. We studied chalcophile elements and sulfides in the ultramafic and mafic rocks of the crust-mantle transition and the mantle underneath. We determined mineralogical and elemental composition and the Cu isotope composition of the respective sulfides along with the mineralogical and elemental composition of the respective serpentines. The rocks of the crust-mantle transition zone (i.e., plagioclase harzburgite, peridotite-gabbro contacts, and dunite) overlaid by troctolites are by one order of magnitude enriched in several chalcophile elements with respect to the spinel harzburgites of the mantle beneath. Whereas the range of Cu concentrations in spinel harzburgites is 7–69 ppm, the Cu concentrations are highly elevated in plagioclase harzburgites with a range of 90–209 ppm. The zones of the peridotite-gabbro contacts are even more enriched, exhibiting up to 305 ppm Cu and highly elevated concentrations of As, Zn, Ga, Sb and Tl. High Cu concentrations show pronounced correlation with bulk S concentrations at the crust-mantle transition zone implying an enrichment process in this horizon of the oceanic lithosphere. We interpret this enrichment as related to melt-mantle reaction, which is extensive in crust-mantle transition zones. In spite of the ubiquitous serpentinization of primary rocks, we found magmatic chalcopyrites [CuFeS2] as inclusions in plagioclase as well as associated with pentlandite [(Fe,Ni)9S8] and pyrrhotite [Fe1−xS] in polysulfide grains. These chalcopyrites show a primary magmatic δ65Cu signature ranging from −0.04 to +0.29 ‰. Other chalcopyrites have been dissolved during serpentinization. Due to the low temperature (<300 °C) of circulating fluids chalcophile metals from primary sulfides have not been mobilized and transported away but have been trapped in smaller secondary sulfides and hydroxides. Combined with the Cu deposits documented in the crust-mantle transition zones of various ophiolite complexes, our results indicate that the metal enrichment, increased sulfide modes, and potentially formation of small sulfide deposits could be expected globally along the petrological Moho.

Au–Cu Alloys Prepared by Pulse Electrodeposition toward Applications as Movable Micro-Components in Electronic Devices

Influences of pulse plating parameters on chemical composition, grain size, morphology, and mechanical property of Au–Cu alloy films electrodeposited with an Au-rich sulfite-based electrolyte were investigated. A wide range of Cu concentration (wCu) varied from 3.5 to 26.7 wt% was attained in the Au–Cu films. The galvanic displacement reaction occurred during the off-time period showed great influences on the composition and the grain size. Meanwhile, surface morphology of the Au–Cu films was interrelated with the alloy composition. An increase in the pulsed current density lead to roughening of the surface, and smoothening of the surface was achieved by promoting the displacement reaction. Micro-mechanical properties of the Au–Cu films were evaluated by micro-compression tests for applications as movable micro-components in electronic devices. A pronounced high yield strength at 1.38 GPa was achieved in the Au–Cu film with the smallest grain size at ca. 4.40 nm and the wCu at ca. 15 wt%, which is suggested to be a result of synergetic effects of grain boundary strengthening and solid solution strengthening mechanisms.

Dose-dependent sensitivity of Arabidopsis thaliana seedling root to copper is regulated by auxin homeostasis

Plants need copper for normal growth and development; however, excess Cu interfere numerous biological functions in plants. To investigate the complex network controlling Cu sensitivity and adaptability, a wide range of Cu concentrations were tested for their differential impacts on primary root length of Arabidopsis thaliana seedlings. Three CuSO4 concentrations (5, 20 and 60μm) were further analyzed in parallel due to their highly distinguishable impacts on multiple morphogenetic characteristics of Arabidopsis roots. Excessive (Cu60) but not moderate amount of Cu (Cu5 and Cu20) reduced the meristem size by affecting meristematic cell division potential and stem cell niche activity. As revealed by the DR5:GUS marker line, the Cu-triggered root phenotype was correlated well with the change in the levels of endogenous auxin in Arabidopsis. The role of auxin homeostasis in regulating the differential Cu responses was verified by exogenous NAA application and in Arabidopsis mutants defective in auxin production, signaling sensitivity or transport. This study also presents evidences for the involvement of an auxin/cytokinin crosstalk in regulating auxin biosynthesis upon Cu treatment and demonstrated that the decreased level of auxin is achieved by the repressed expression of TAA1 and YUCCA family members in the indole-3-pyruvic acid pathway.

Copper in the sediment: a major stressor for eelgrass, Zostera marina L.

Eelgrass (Zostera marina L.) is an important organism in coastal marine waters and is highly likely to encounter exposure to multiple stressors, both anthropogenic contaminants and natural stressors. Here, we exposed eelgrass to a range of Cu concentrations and salinities, and also varied exposure route between sediment and water. Measured endpoints were Cu accumulation in root and leaves, relative growth rate, leaf mortality, chlorophyll concentration, and maximum photosynthetic quantum yield. Cu accumulation from the sediment was translocated to all parts of the plant, while Cu taken up from the water showed a tendency to remain in leaves. Effects on relative growth rate and leaf mortality were found only following uptake of Cu from the sediment. We tested effects of different salinities, acting as multiple stressors, together with Cu, but found only weak effects with little interaction with Cu. Experiments with anthropogenic contaminants that marine plants are mainly exposed to through the sediment should be done using sediment exposure, as the common practice of using only water exposure will lead to underestimation of harmful effects. Future studies should take all relevant factors into consideration, as anthropogenic inputs and natural factors are prone to fluctuations due to e.g., climate change.

Novel blue-emitting gold nanoclusters confined in human hemoglobin, and their use as fluorescent probes for copper(II) and histidine

We report on the synthesis and characterization of highly fluorescent gold nanoclusters (Au-NCs) from a gold precursor using a single-step chemical reduction in the presence of human adult hemoglobin (Hb). The conformational changes of Hb before and after cluster formation were studied by various spectroscopic techniques. The Au-NCs in Hb display blue fluorescence with a peak centered at 450 nm (photoexcited at 365 nm) and a quantum yield of 2.8 %. The Au-NCs exhibit excellent photostability and long-term stability, and can be applied in the pH range 5–12 even in the presence of high electrolyte concentrations. The Au-NCs in Hb can act as a highly sensitive and selective fluorescent turn-off probe for Cu(II) ion. The observed reversible fluorescence recovery of Hb-AuNCs/Cu(II) aggregates was exploited to develop a selective and sensitive turn-on fluorescence assay for His. Under optimized conditions, the probe gives a fluorescent response that is linear in the 0.1 to 20.0 μM concentration range of Cu(II), with a limit of detection of 28 nM. The probe for His, in turn, has a linear range in the 1–21 μM concentration range, and the limit of detection is 0.6 μM.

Assembly processes under severe abiotic filtering: adaptation mechanisms of weed vegetation to the gradient of soil constraints.

QuestionsEffects of soil on vegetation patterns are commonly obscured by other environmental factors; clear and general relationships are difficult to find. How would community assembly processes be affected by a substantial change in soil characteristics when all other relevant factors are held constant? In particular, can we identify some functional adaptations which would underpin such soil-induced vegetation response?LocationEastern Serbia: fields partially damaged by long-term and large-scale fluvial deposition of sulphidic waste from a Cu mine; subcontinental/submediterranean climate.MethodsWe analysed the multivariate response of cereal weed assemblages (including biomass and foliar analyses) to a strong man-made soil gradient (from highly calcareous to highly acidic, nutrient-poor soils) over short distances (field scale).ResultsThe soil gradient favoured a substitution of calcicoles by calcifuges, and an increase in abundance of pseudometallophytes, with preferences for Atlantic climate, broad geographical distribution, hemicryptophytic life form, adapted to low-nutrient and acidic soils, with lower concentrations of Ca, and very narrow range of Cu concentrations in leaves. The trends of abundance of the different ecological groups of indicator species along the soil gradient were systematically reflected in the maintenance of leaf P concentrations, and strong homeostasis in biomass N:P ratio.ConclusionUsing annual weed vegetation at the field scale as a fairly simple model, we demonstrated links between gradients in soil properties (pH, nutrient availability) and floristic composition that are normally encountered over large geographic distances. We showed that leaf nutrient status, in particular the maintenance of leaf P concentrations and strong homeostasis of biomass N:P ratio, underpinned a clear functional response of vegetation to mineral stress. These findings can help to understand assembly processes leading to unusual, novel combinations of species which are typically observed as a consequence of strong environmental filtering, as for instance on sites affected by industrial activities.

Interactions of copper and thermal stress on mitochondrial bioenergetics in rainbow trout, Oncorhynchus mykiss

Thermal stress may influence how organisms respond to concurrent or subsequent chemical, physical and biotic stressors. To unveil the potential mechanisms via which thermal stress modulates metals-induced bioenergetic disturbances, the interacting effects of temperature and copper (Cu) were investigated in vitro. Mitochondria isolated from rainbow trout livers were exposed to a range of Cu concentrations at three temperatures (5, 15 and 25°C) with measurement of mitochondrial complex I (mtCI)-driven respiratory flux indices and uncoupler-stimulated respiration. Additional studies assessed effects of temperature and Cu on mtCI enzyme activity, induction of mitochondrial permeability transition pore (MPTP), swelling kinetics and mitochondrial membrane potential (MMP). Maximal and basal respiration rates, as well as the proton leak, increased with temperature with the Q10 effects being higher at lower temperatures. The effect of Cu depended on the mitochondrial functional state in that the maximal respiration was monotonically inhibited by Cu exposure while low and high Cu concentrations stimulated and inhibited the basal respiration/proton leak, respectively. Importantly, temperature exacerbated the effects of Cu by lowering the concentration of the metal required for toxicity and causing loss of thermal dependence of mitochondrial respiration. Mitochondrial complex I activity was inhibited by Cu but was not affected by incubation temperature. Compared with the calcium (Ca) positive control, Cu-imposed mitochondrial swelling exhibited variable kinetics depending on the inducing conditions, and was highly temperature-sensitive. A partial reversal of the Cu-induced swelling by cyclosporine A was observed suggesting that it is in part mediated by MPTP. Interestingly, the combination of high Cu and high temperature not only completely inhibited mitochondrial swelling but also greatly increased the respiratory control ratio (RCR) relative to the controls. Copper exposure also caused marked MMP dissipation which was reversed by N-acetyl cysteine and vitamin E suggesting a role of reactive oxygen species (ROS) in this response. Taken together, Cu impairs oxidative phosphorylation in part by inhibiting the electron transport chain (ETC), stimulating proton leak, inducing MPTP and dissipating MMP, with high temperature exacerbating these effects. Thus environmental temperature rise due to natural phenomenon or global climate change may sensitize fish to Cu toxicity by exacerbating mitochondrial dysfunction.

Effects of copper exposure on hatching success and early larval survival in marbled salamanders, Ambystoma opacum.

The creation of wetlands, such as urban and industrial ponds, has increased in recent decades, and these wetlands often become enriched in pollutants over time. One metal contaminant trapped in created wetlands is copper (Cu(2+)). Copper concentrations in sediments and overlying water may affect amphibian species that breed in created wetlands. The authors analyzed the Cu concentration in dried sediments from a contaminated wetland and the levels of aqueous Cu released after flooding the sediments with different volumes of water, mimicking low, medium, and high pond-filling events. Eggs and larvae of Ambystoma opacum Gravenhorst, a salamander that lays eggs on the sediments in dry pond beds that hatch on pond-filling, were exposed to a range of Cu concentrations that bracketed potential aqueous Cu levels in created wetlands. Embryo survival varied among clutches, but increased Cu levels did not affect embryo survival. At Cu concentrations of 500 µg/L or greater, however, embryos hatched earlier, and the aquatic larvae died shortly after hatching. Because Cu concentrations in sediments increase over time in created wetlands, even relatively tolerant species such as A. opacum may be affected by Cu levels in the posthatching environment.

Copper export contributes to low copper levels and copper tolerance in Emiliania huxleyi

Previous studies have documented that the coccolithophore Emiliania huxleyi maintains lower cellular copper concentrations and is more tolerant to high copper than are many other species of marine algae. We confirm that in comparison to the diatom Thalassiosira pseudonana, E. huxleyi maintains substantially lower Cu levels while maintaining near maximal growth rates. Using the stable isotope 65Cu as a tracer, we show that a constitutive or rapidly induced mechanism for Cu efflux is present in Cu‐replete E. huxleyi cells. This was revealed during uptake experiments in which the accumulation of the isotopically enriched spike, as measured by the isotope fractionation, exceeded the total accumulated copper. Evidence of efflux was not observed during Cu uptake in T. pseudonana cells. In contrast, copper‐limited E. huxleyi cells exhibited no observable efflux during uptake over the same time period, though net uptake rates were similar to replete cells, suggesting that efflux is modulated by cellular copper needs. This Cu‐efflux mechanism is effective over a wide range of Cu concentrations, as E. huxleyi is able to maintain much lower cellular Cu levels than T. pseudonana during 4 h uptake experiments. Direct measurement of export from copper‐loaded cells provided further evidence of a very effective efflux mechanism in E. huxleyi, while loss of copper from T. pseudonana was considerably less. Improvements to cell washes to remove surface‐bound Cu were implemented.