Toxicity Of Cu Research Articles

Elucidating the Differences in Metal Toxicity by Quantitative Adverse Outcome Pathways.

Numerous studies have reported that the toxicity differences among metals are widespread; however, little is known about the mechanism of differences in metal toxicity to aquatic organisms due to the lack of quantitative understanding of their adverse outcome pathway. Here, we investigated the effects of Cd and Cu on bioaccumulation, gene expression, physiological responses, and apical effects in zebrafish larvae. RNA sequencing was conducted to provide supplementary mechanistic information for the effects of Cd and Cu exposure. On this basis, we proposed a quantitative adverse outcome pathway (qAOP) suitable for metal risk assessment of aquatic organisms. Our work provides a mechanistic explanation for the differences in metal toxicity where the strong bioaccumulation of Cu enables the newly accumulated Cu to reach the threshold that causes different adverse effects faster than Cd in zebrafish larvae, resulting in a higher toxicity of Cu than that of Cd. Furthermore, we proposed a parameter CIT/BCF (the ratio of internal threshold concentration and bioaccumulation factor) that helps to understand the toxicity differences by combining the information of bioaccumulation and internal threshold of adverse effects. This work demonstrated that qAOP is an effective quantitative tool for understanding the toxicity mechanism and highlight the importance of toxicokinetics and toxicodynamics at different biological levels in determining the metal toxicity.

Biomolecular modifications in the sacfry of Mogurnda adspersa in response to copper stress

Copper (Cu) is one of the most harmful contaminants in fresh-water systems. Fish larvae such as sacfry are particularly vulnerable to metals such as copper (Cu) due to a less-developed excretory organ system and permeable skin that can absorb metals directly from the water. However, the sublethal effects of metals on this life stage are not well understood. This study assessed the sublethal toxicity of Cu on purple-spotted gudgeon sacfry (PSG, Mogurnda adspersa). For this purpose, 96h Cu toxicity bioassays were performed and toxic effects of Cu on PSG were measured at different levels of biological organization, from the individual (loss of equilibrium, wet weight), to tissue (chemical changes in retinal tissue composition) and molecular responses (whole body amino acid (AA) profiles). The EC10 and EC50 (ECx: effect concentration that affected X% of test organisms) were found to be 12 (9 - 15) µg Cu L-1 and 22 (19 - 24) µg Cu L-1, respectively. Copper stress caused a decrease in total amino acid content and changed the AA profile of PSG compared to the controls. Proton-induced X-ray emission (PIXE) mapping techniques showed accumulation of Cu in the retinal tissues disturbing the distribution of other elements such as zinc, sulfur, phosphorus and potassium. Fourier-transform infrared (FTIR) microspectroscopy of control and Cu treated eye tissues revealed a change in protein secondary structure in retinal tissues in response to Cu accumulation, as well as decreased levels of the molecular retinal, consistent with the degradation of rhodopsin, a key protein in the visual sensory system. This is the first study to demonstrate the multi-level responses of PSG arising from exposure to environmentally realistic Cu concentrations and suggests that AA profiling can serve as a useful tool to assess the impacts of metals on fresh-water organisms.

Toxicity mechanisms and bioavailability of copper to fish based on an adverse outcome pathway analysis

Copper (Cu) exists in a variety of forms in different aquatic environments, and affects their bioavailability. In this study we provide a systematic review on toxicity of Cu which focuses on identifying evidence in the mechanisms of Cu toxicity, and apply an adverse outcome pathway (AOP) analysis to identify multiple potential mechanisms and their interactions of Cu toxicity to fish. This analysis process included the mechanisms of behavior toxicant, oxidative toxicant, ion regulation disruption toxicity, as well as endocrine disruption toxicity. It was found that at low levels of Cu exposure, swimming, avoid predators, locating prey and other sensory functions will be impaired, and the organism will suffer from metabolic alkalosis and respiratory acidosis following the inhibition of the carbonic anhydrase active. The main pathway of acute toxicity of Cu to fish is the inhibition of the Na+/K+-ATPase enzyme, and lead to reduced intracellular sodium absorption, as well as Cu-induced increased cell permeability, in turn resulting in increased sodium ion loss, leading to cardiovascular collapse and respiratory insufficiency. The endocrine disruption toxicity of Cu to fish caused growth inhibition and reproductive reduction. In addition, there are several key pathways of Cu toxicity that are affected by hardness (e.g., Ca2+) and intracellular DOC concentrations, including inhibiting Cu-induction, improving branchial gas exchange, altering membrane transport functions, decreasing Na+ loss, and increasing Na+ uptake. The results of the AOP analysis will provide a robust framework for future directed research on the mechanisms of Cu toxicity.

Effects of Long-Term Exposure to Copper on Mitochondria-Mediated Apoptosis in Pig Liver.

Copper (Cu) is listed as one of the main heavy metal pollutants, which poses potential health risks to humans. Excessive intake of Cu has shown toxic effects on the organs of many animals, and the liver is one of the most important organs to metabolize it. In this study, pigs, the mammal with similar metabolic characteristics to humans, were selected to assess the effects of long-term exposure to Cu on mitochondria-mediated apoptosis, which are of great significance for studying the toxicity of Cu to humans. Pigs were fed a diet with different contents of Cu (10, 125, and 250mg/kg) for 80days. Samples of blood and liver tissue were collected on days 40 and 80. Experimental results demonstrated that the accumulation of Cu in the liver was increased in a dose-dependent and time-dependent manner. Meanwhile, the curve of pig's body weight showed that a 125mg/kg Cu diet promoted the growth of pigs during the first 40days and then inhibited it from 40 to 80days, while the 250mg/kg Cu diet inhibited the growth of pigs during 80days of feeding. Additionally, the genes and protein expression levels of Caspase-3, p53, Bax, Bak1, Bid, Bad, CytC, and Drp1 in the treatment group were higher than that in the control group, while Bcl-2, Bcl-xL, Opa1, Mfn1, and Mfn2 were decreased. In conclusion, these results indicated that long-term excessive intake of Cu could inhibit the growth of pigs and induced mitochondria-mediated apoptosis by breaking the mitochondrial dynamic balance. Synopsis: Long-term exposure to high doses of Cu could lead to mitochondrialdysfunction by breaking the mitochondrial dynamic balance, which ultimatelyinduced mitochondria-mediated apoptosis in the liver of pigs. This might be closelyrelated to the growth inhibition and liver damage in pigs.

Quantitative Estimation of Synergistic Toxicity of Cu and Zn on Growth of Arabidopsis thaliana by Isobolographic Method.

Heavy metal is one of the most frequent soil contaminants and contaminated soils generally include numerous metals. Although exposure to multiple metals may increase the toxicity to humans and ecosystems, only additive effects are considered in the risk assessment. In this study, the synergistic effect of heavy metals (Cu and Zn) on a model plant, Arabidopsis thaliana, was quantified by the isobolographic method. The plant was cultured via the growth assay method on a plant agar containing individual heavy metals or combinations of Cu + Zn in a growth chamber. The concentration of Cu varied by eight levels from 0 to 200 μM and the concentration of Zn also varied by eight levels from 0 to 400 μM. In the combination of metals, each of the three levels of Cu (25–75 μM) and Zn (20–100 μM) were applied. After 8 days, plants were harvested for root/shoot weight and measured for leaf chlorophyll and carotenoid content. The primary and secondary root elongation of A. thaliana was estimated using image analysis to calculate total root length. The EC50 values of Cu and Zn on A. thaliana, based on the total root length, were 40.0 and 76.4 μM, respectively. When two heavy metals were administered in combination, the EC values decreased less than those of the individual metals. The average value of the combination index was 0.6, proving the synergistic toxic effect on the root growth of A. Thaliana. As a result, the isobolograhic method is a useful tool for estimating the quantitative toxic effect of chemicals on plants.

DNA-based platform for efficient and precisely targeted bioorthogonal catalysis in living systems

As one of the typical bioorthogonal reactions, copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction holds great potential in organic synthesis, bioconjugation, and surface functionalization. However, the toxicity of Cu(I), inefficient catalytic activity, and the lack of cell specific targeting of the existing catalysts hampered their practical applications in living systems. Herein, we design and construct a DNA-based platform as a biocompatible, highly efficient, and precisely targeted bioorthogonal nanocatalyst. The nanocatalyst presents excellent catalytic efficiency in vitro, which is one order of magnitude higher than the commonly used catalyst CuSO4/sodium ascorbate. The theoretical calculation further supports the contribution of DNA structure and its interaction with substrates to the superior catalytic activity. More importantly, the system can achieve efficient prodrug activation in cancer cells through cell type-specific recognition and produce a 40-fold enhancement of transformation compared to the non-targeting nanocatalyst, resulting in enhanced antitumor efficacy and reduced adverse effects. In vivo tumor therapy demonstrates the safety and efficacy of the system in mammals.

Copper and Chromium toxicity is mediated by oxidative stress in Caenorhabditis elegans: The use of nanoparticles as an immobilization strategy

Environmental contamination by heavy metals (HMs) has impelled searching for stabilization strategies, where the use of zero-valent iron nanoparticles (nZVI) is considered a promising option. We have evaluated the combined effect of Cu(II)-Cr(VI) on two Caenorhabditis elegans strains (N2 and RB1072 sod-2 mutant) in aqueous solutions and in a standard soil, prior and after treatment with nZVI (5% w/w). The results showed that HMs aqueous solutions had an intense toxic effect on both strains. Production of reactive oxygen species and enhanced expression of the heat shock protein Hsp-16.2 was observed, indicating increased HM-mediated oxidative stress. Toxic effects of HM-polluted soil on worms were higher for sod-2 mutant than for N2 strain. However, nZVI treatment significantly diminished all these effects. Our findings highlighted C. elegans as a sensitive indicator for HMs pollution and its usefulness to assess the efficiency of the nanoremediation strategy to decrease the toxicity of Cu(II)-Cr(VI) polluted environments.

Growth Performance of Black poplar (Populus nigra L.) Under Drought Condition and Sewage Water Irrigation

The response of black poplar (Populus nigra L.) seedlings to the combination of both drought and sewage water irrigation have been investigated from 18-5-2019 to 18-8-2019. Two drought stress levels; D0; 90% or full water holding capacity (WHC) and D1; 30% WHC interacted with three sewage irrigation treatments; S0; 100 % of tap water as a control, S1; 50% of tap water and sewage water and S2; 100% of sewage water. The irrigation water quality index (IWQI) of S1 was 59.31 located under moderate restrictions class for irrigation purposes. Despite to its low pollution load index (PLI); 0.47. Fully irrigation to soil holding capacity with S1 (S1D0) led to a significant increase in seedling height, stem base diameter, fresh and dry biomass and moisture content in shoot system. In addition, the highest tolerance index (TI %) about 92.07 % of the seedlings recorded despite to the soil toxicity with Ni and Pb ions. Lowest accumulation and metal toxicity of Cu, Zn, Cd, Pb and Mn in the soil and leaves have been determined. Thus, diluting sewage water; 1:1 can be used to irrigate black poplar trees with high water demand and moderate to high tolerance to salts for reforestation and urban vegetation.

Nitric oxide overcomes copper and copper oxide nanoparticle-induced toxicity in Sorghum vulgare seedlings through regulation of ROS and proline metabolism.

This study aimed to investigate the phytotoxic effect of copper (Cu) and copper nanoparticles (CuONPs) and ameliorative potential of nitric oxide (NO) against these toxic materials in Sorghum vulgare Pers. seedlings. Data suggested that exposure of Cu and CuONPs significantly reduced growth, chlorophyll, carotenoids and protein in root and shoot, which coincided with increased Cu accumulation. However, addition of sodium nitroprusside (SNP, a donor of NO) lowered Cu and CuONPs mediated toxicity through restricting Cu accumulation and improving photosynthetic pigments and total soluble protein contents. Data further suggested that exposure of Cu and CuONPs significantly increased hydrogen peroxide (H2 O2 ), superoxide radicals (O2 •- ), and malondialdehyde (MDA) contents. Enhanced level of oxidative stress severely inhibited the enzymatic activities of glutathione reductase (GR), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and monodehydroascorbate reductase (MDHAR) but enhanced superoxide dismutase (SOD) and catalase (CAT) activity. However, addition of SNP positively regulated antioxidants enzymes activity, particularly the enzymes involved in the ascorbate-glutathione cycle to overcome Cu- and CuONPs-induced stress in Sorghum seedlings. Further, Cu and CuONPs enhanced accumulation of free proline through inducing Δ1 -pyrroline-5-carboxylate synthetase (P5CS) activity while lowering the proline dehydrogenase (PDH) activity. However, addition of SNP reversed these responses. Therefore, overall results revealed that SNP has enough potential of reducing the toxicity of Cu and CuONPs in Sorghum seedlings through regulation of proline metabolism and activity of enzymes of the ascorbate-glutathione cycle. These findings can be employed in developing new resistant varieties of Sorghum having enhanced tolerance against Cu or CuONP stress and improved productivity.

Understanding the bactericidal mechanism of Cu(OH)2 nanorods in water through Mg-substitution: high production of toxic hydroxyl radicals by non-soluble particles.

To date, there is still a lack of definite knowledge regarding the toxicity of Cu(OH)2 nanoparticles towards bacteria. This study was aimed at shedding light on the role played by released cupric ions in the toxicity of nanoparticles. To address this issue, the bactericidal activity of Cu(OH)2 was at first evaluated in sterile water, a medium in which particles are not soluble. In parallel, an isovalent substitution of cupric ions by Mg2+ was attempted in the crystal structure of Cu(OH)2 nanoparticles to increase their solubility and determine the impact on the bactericidal activity. For the first time, mixed Cu1-xMgx(OH)2 nanorods (x ≤ 0.1) of about 15 nm in diameter and a few hundred nanometers in length were successfully prepared by a simple co-precipitation at room temperature in mixed alkaline (NaOH/Na2CO3) medium. For E. coli, 100% reduction of one million CFU per mL (6 log10) occurs after only 180 min on contact with both Cu(OH)2 and Cu0.9Mg0.1(OH)2 nanorods. The entire initial inoculum of S. aureus is also killed by Cu(OH)2 after 180 min (100% or 6 log10 reduction), while 0.01% of these bacteria stay alive on contact with Cu0.9Mg0.1(OH)2 (99.99% or 4 log10 reduction). The bactericidal performances of Cu(OH)2 and the magnesium-substituted counterparts (i.e. Cu1-xMgx(OH)2) are not linked to cupric ions they release in water since their mass concentrations after 180 min are much lower than minimal concentrations inhibiting the growth of E. coli and S. aureus. Finally, an EPR spin trapping study reveals how these nanorods kill bacteria in water: only the presence of hydrogen peroxide, a by-product of the normal metabolism of oxygen in aerobic bacteria, allows the Cu(OH)2 and its magnesium-substituted counterparts to produce a lethal amount of free radicals, the majority of which are the highly toxic HO˙.

The ionophore thiomaltol induces rapid lysosomal accumulation of copper and apoptosis in melanoma.

In this report, we investigate the toxicity of the ionophore thiomaltol (Htma) and Cu salts to melanoma. Divalent metal complexes of thiomaltol display toxicity against A375 melanoma cell culture resulting in a distinct apoptotic response at submicromolar concentrations, with toxicity of Cu(tma)2>Zn(tma)2 >> Ni(tma)2. In metal-chelated media, Htma treatment shows little toxicity, but the combination with supplemental CuCl2, termed Cu/Htma treatment, results in toxicity that increases with suprastoichiometric concentrations of CuCl2 and correlates with the accumulation of intracellular copper. Electron microscopy and confocal laser scanning microscopy of Cu/Htma treated cells shows a rapid accumulation of copper within lysosomes over the course of hours, concurrent with the onset of apoptosis. A buildup of ubiquitinated proteins due to proteasome inhibition is seen on the same timescale and correlates with increases of copper without additional Htma.

Mineral Materials as a Neutralizing Agent Used on Soil Contaminated with Copper.

The aim of the investigation was to evaluate the response of plants, using black mustard (Brassica nigra L. Koch) as a model plant, to soil contamination with copper (0, 200, 400, 600 mg Cu kg−1 of soil), and to determine the effectiveness of the Cu immobilization with mineral neutralizing materials, such as lime, clay and zeolite. The plant yield depended on soil contamination and mineral amendments. In the series without neutralizing materials, the level of 600 mg Cu kg−1 reduced the yield and increased leaf greenness. Lime alleviated the toxicity of Cu in objects with 200 mg Cu kg−1. Zeolite slightly mitigated the harmful effects of Cu at the level of 400 and 600 mg kg−1. Zeolite lowered the SPAD index. In the chemical composition of plants, the content of Cu, K, Mg, Na and Ca in plants increased to 400 mg Cu kg−1, while the content of P decreased to 600 mg Cu kg−1. Among the materials, lime reduced the Cu accumulation in plants the most, followed by clay. Cu narrowed the majority of ratios and widened the Ca:P and K:Ca ratios in plants. The applied mineral materials, except lime, did not significantly affect the formation of these indicators.

Intracellular Biotransformation of Cu(II)/Cu(I) Explained High Cu Toxicity to Phytoplankton Chlamydomonas reinhardtii.

The toxicity of Cu is related to its redox species, but the differential toxicity of Cu(II) and Cu(I) remains unknown. In the present study, we developed a novel protocol to simultaneously detect the biologically produced extracellular Cu(I) and internalized Cu(II) in a freshwater phytoplankton Chlamydomonas reinhardtii. The intracellular Cu(I) was further imaged using a fluorometric probe. Combining these pieces of evidence, we demonstrated that Cu(I) dominated the Cu toxicity in algal cells under Fe-deficient conditions. Our results showed that the labile Cu(I) content increased significantly in the low Fe quota cells. Intracellular biotransformation from Cu(II) to Cu(I) rather than the direct uptake of Cu(I) was responsible for the high Cu toxicity. The abnormal biotransformation from Cu(II) to Cu(I) under Fe deficiency was not resulted from the increase of overall Cu bioaccumulation but was likely due to the change of Cu(II) metabolism. High contents of Cu(II) were accumulated in the normal cells and the low Zn quota cells upon Cu exposure but did not induce cell death, further suggesting that Cu(I) dominated the Cu toxicity to the algae. This is the first study to simultaneously consider the effect of Cu(I) and Cu(II) during Cu exposure in phytoplankton. The results uncovered the underlying mechanisms of high Cu toxicity under Fe deficiency and highlighted the critical role of modulation of Cu metabolism in phytoplankton.

Combined effects of copper and microplastics on physiological parameters of Tubastrea aurea corals.

Microplastics (MPs) have been a serious environmental problem because it can carry pollution like heavy metals and organic pollutants. However, the combined effect of MPs and bivalent copper ion (Cu(II)) on the coral azooxanthellate has been rarely studied. In the present study, the combined effects of PVC and Cu(II) on the physiological responses of Tubastrea aurea were studied. Our results showed that MPs alone enhanced the activity of catalase (CAT), superoxide dismutase (SOD), and reduced glutathione (GSH). The mixture groups had the same effects on the CAT and GSH, which enhanced CAT and GSH activity by 97% and 53% respectively. MPs alone and the combined treatment groups decreased the activity of lipid peroxide (LPO) and the content of metallothionein (MT) by 45% and 20% of the coral Tubastrea aurea. Cu(II) exposure always had negative effect on the physiological parameters of coral, and MPs decreased the toxicity of Cu(II) in the combined groups. This work is the first time to report the combined effects of Cu(II) and microplastics on azooxanthellate coral, which will provide important preliminary data for the following research.

Copper-binding ligands in deep-sea pore waters of the Pacific Ocean and potential impacts of polymetallic nodule mining on the copper cycle

The release of potentially toxic metals, such as copper (Cu), into the water column is of concern during polymetallic nodule mining. The bioavailability and thus toxicity of Cu is strongly influenced by its speciation which is dominated by organic ligand (L) complexation in seawater, with L-complexes being considered less bioavailable than free Cu2+. The presence of CuL-complexes in deep-sea sediments has, however, not been systematically studied in the context of deep-sea mining. We thus analyzed the Cu-binding L concentration ([L]) in deep-sea pore waters of two polymetallic nodule provinces in the Pacific Ocean, the Peru Basin and the Clarion-Clipperton-Zone, using competitive ligand equilibration–adsorptive stripping voltammetry. The pore-water dissolved Cu concentration ([dCu]) ranged from 3 to 96 nM, generally exceeding bottom water concentrations (4–44 nM). Based on fitting results from ProMCC and Excel, Cu was predominantly complexed by L (3–313 nM) in bottom waters and undisturbed pore waters. We conclude that processes like deep-sea mining are unlikely to cause a release of toxic Cu2+ concentrations ([Cu2+]) to the seawater as > 99% Cu was organically complexed in pore waters and the [Cu2+] was < 6 pM for 8 of 9 samples. Moreover, the excess of L found especially in shallow pore waters implied that even with a Cu release through mining activities, Cu2+ likely remains beneath toxic thresholds.

The effects of copper ions and copper nanomaterials on the output of amino acids from marine microalgae.

In this study, the marine microalgae Skeletonema costatum and Nitzschia closterium were exposed to different forms of copper, such as a metal salt (Cu2+), a nano-metal (nano-Cu), and nano-metal oxide (nano-CuO). During a 96-h exposure to nanoparticles (NPs) and salt, the cell number, Cu2+ concentration in the culture medium, morphology, and intracellular amino acids were measured to assess the toxicity of the copper materials and the toxicity mechanism of the NPs. As results, the toxicity of Cu2+, nano-Cu, and nano-CuO to marine phytoplankton decreased in order. The EC50 values of Cu2+ and nano-Cu for S. costatum and N. closterium ranged from 0.356 to 0.991 mg/L and 0.663 to 2.455 mg/L, respectively. Nano-Cu inhibits the growth of marine phytoplankton by releasing Cu2+; however, nano-CuO is harmful to microalgae because of the effect of NPs. The secretion of extracellular polymeric substances by microalgae could also affect the toxicity of nano-Cu and nano-CuO to microalgae. S. costatum was more sensitive to copper than N. closterium. Cu2+, nano-Cu, and nano-CuO all reduced per-cell amino acids and the total output of algae-derived amino acids by affecting the growth of the phytoplankton. This study helps to understand the risk assessment of nano-Cu and nano-CuO to marine microalgae.

Effect of site hydrological conditions and soil aggregate sizes on the stabilization of heavy metals (Cu, Ni, Pb, Zn) by biochar

Biochar is a popular material that would effectively immobilize heavy metals in soil, which can greatly decrease the health risk of heavy metals. Although many previous studies have studied the immobilization of heavy metals by biochar, the influence of hydrological conditions on the immobilization effect is still not clear. This paper carried out column experiments to study the effect of fluctuating groundwater table on Cu, Ni, Pb, Zn distribution and speciation with the addition of biochar from pyrolysis of swine manure. Experimental results showed that biochar could significantly decrease the leaching toxicity of Cu and Ni by 24.4% and 44.7% respectively, while the immobilization effect of Pb and Zn was relatively insignificant. The average reduction percentage of bioavailable Cu was 14.5%, 39.5% and 33.3% in the unsaturated zone, fluctuating zone and saturated zone respectively, showing the better immobilization effect in the fluctuating zone and saturated zone. The residual fraction of heavy metals increased significantly after the addition of biochar, and the increase of residual fraction was larger in small soil aggregates. This study helped illustrate the influence of hydrological conditions and soil aggregate sizes on the stabilization effect of heavy metals by biochar, which could be used to guide the remediation process of sites contaminated by heavy metals.

Strength of Cu-efflux response in Escherichia coli coordinates metal resistance in Caenorhabditis elegans and contributes to the severity of environmental toxicity

Without effective homeostatic systems in place, excess copper (Cu) is universally toxic to organisms. While increased utilization of anthropogenic Cu in the environment has driven the diversification of Cu-resistance systems within enterobacteria, little research has focused on how this change in bacterial architecture impacts host organisms that need to maintain their own Cu homeostasis. Therefore, we utilized a simplified host–microbe system to determine whether the efficiency of one bacterial Cu-resistance system, increasing Cu-efflux capacity via the ubiquitous CusRS two-component system, contributes to the availability and subsequent toxicity of Cu in host Caenorhabditis elegans nematode. We found that a fully functional Cu-efflux system in bacteria increased the severity of Cu toxicity in host nematodes without increasing the C. elegans Cu-body burden. Instead, increased Cu toxicity in the host was associated with reduced expression of a protective metal stress-response gene, numr-1, in the posterior pharynx of nematodes where pharyngeal grinding breaks apart ingested bacteria before passing into the digestive tract. The spatial localization of numr-1 transgene activation and loss of bacterially dependent Cu-resistance in nematodes without an effective numr-1 response support the hypothesis that numr-1 is responsive to the bacterial Cu-efflux capacity. We propose that the bacterial Cu-efflux capacity acts as a robust spatial determinant for a host’s response to chronic Cu stress.

Metabolomics combined with physiology and transcriptomics reveals how Citrus grandis leaves cope with copper-toxicity

Limited data are available on metabolic responses of plants to copper (Cu)-toxicity. Firstly, we investigated Cu-toxic effects on metabolomics, the levels of free amino acids, NH4+-N, NO3--N, total nitrogen, total soluble proteins, total phenolics, lignin, reduced glutathione (GSH) and malondialdehyde, and the activities of nitrogen-assimilatory enzymes in ‘Shatian’ pummelo (Citrus grandis) leaves. Then, a conjoint analysis of metabolomics, physiology and transcriptomics was performed. Herein, 59 upregulated [30 primary metabolites (PMs) and 29 secondary metabolites (SMs)] and 52 downregulated (31 PMs and 21 SMs) metabolites were identified in Cu-toxic leaves. The toxicity of Cu to leaves was related to the Cu-induced accumulation of NH4+ and decrease of nitrogen assimilation. Metabolomics combined with physiology and transcriptomics revealed some adaptive responses of C. grandis leaves to Cu-toxicity, including (a) enhancing tryptophan metabolism and the levels of some amino acids and derivatives (tryptophan, phenylalanine, 5-hydroxy-l-tryptophan, 5-oxoproline and GSH); (b) increasing the accumulation of carbohydrates and alcohols and upregulating tricarboxylic acid cycle and the levels of some organic acids and derivatives (chlorogenic acid, quinic acid, d-tartaric acid and gallic acid o-hexoside); (c) reducing phospholipid (lysophosphatidylcholine and lysophosphatidylethanolamine) levels, increasing non-phosphate containing lipid [monoacylglycerol ester (acyl 18:2) isomer 1] levels, and inducing low-phosphate-responsive gene expression; and (d) triggering the biosynthesis of some chelators (total phenolics, lignin, l-trytamine, indole, eriodictyol C-hexoside, quercetin 5-O-malonylhexosyl-hexoside, N-caffeoyl agmatine, N′-p-coumaroyl agmatine, hydroxy-methoxycinnamate and protocatechuic acid o-glucoside) and vitamins and derivatives (nicotinic acid-hexoside, B1 and methyl nicotinate). Cu-induced upregulation of many antioxidants could not protect Cu-toxic leaves from oxidative damage. To conclude, our findings corroborated the hypothesis that extensive reprogramming of metabolites was carried out in Cu-toxic C. grandis leaves in order to cope with Cu-toxicity.

Hydrogeochemical and ecological risk assessments of trace elements in the coastal surface water of the southern Caspian Sea.

This study investigates the occurrence, distribution, and potential ecological risk of trace elements (TEs) along with the hydrogeochemical characteristics of coastal surface water collected from the southern Caspian coasts in the Mazandaran province of Iran. Eighteen coastal water sites were sampled and analyzed using inductively coupled plasma-mass spectrometry and ion chromatography to determine concentrations of TEs and major ions, respectively. Mean concentrations (µg/l) of TEs in the water followed the order: Al (154.3) > Fe (73.6) > Zn (67.8) > Mn (29.9) > Cu (5.7) > Mo (3.7) > Cd (2.8) > Se (2.3) > V (1.9) > Co = As (1.8) > Sb (1.2) > Pb (0.6). TEs displayed high variations within samples, reflecting many sources that control their concentrations in the coastal water. Most TEs displayed elevated concentrations in the east and west of the study area. The Na-Cl water type in the majority of investigated sites indicates excess alkaline elements and strong acid anions relative to alkaline earth cations and weak acid anions. Considering the saturation states of minerals, carbonate and evaporite minerals are oversaturated and unsaturated in surface water, respectively. Compositional interrelations between aqueous species showed that reverse cation exchange may have occurred. The excess SO42- content, derived from irrigation return flow and domestic greywater, probably plays a crucial role in the mobilization and transport of Zn and Pb by binding to sulfate ligands and forming aqueous complexes. Ecological risk assessment of TEs revealed that water in the majority of sites is safe in terms of As, Se, Pb, and Cd content, and unsuitable with respect to Zn and Cu. Acute and chronic toxicities of Cu and Zn are reported in several sites, thus coastal water cannot be used for fishery and protecting "nature reserve" purposes. However, industrial activity and tourism are safe to be conducted in most coastal water sites.