Chronic Cu Toxicity Research Articles

Antagonistic Effect of Zinc Oxide Nanoparticles Dietary Supplementation Against Chronic Copper Waterborne Exposure on Growth, Behavioral, Biochemical, and Gene Expression Alterations of African Catfish, Clarias gariepinus (Burchell, 1822).

The harmful impact of waterborne copper (Cu) as a common abiotic stressor in aquatic environments has gained much more interest. The present study aimed to investigate the utilization of zinc oxide nanoparticles (ZnONPs) dietary supplementation to mitigate the chronic toxicity of Cu in African catfish (Clarias gariepinus). Two hundred and forty fish (92.94 ± 0.13 g) were assigned into six groups for 60 days. Control (C), ZnONPs20, and ZnONPs30 groups were fed on basal diets fortified with 0, 20, and 30 mg kg-1 ZnONPs without Cu exposure. Cu, Cu + ZnONPs20, and Cu + ZnONPs30 groups were exposed to Cu at a dose of 10mg L-1 and fed on basal diets fortified with 0, 20, and 30 mg kg-1 ZnONPs, respectively. The results revealed that the Cu-exposed fish experienced abnormal clinical signs and behavioral changes. The growth indices and acetylcholine esterase activity were significantly decreased (P < 0.05) in the Cu group. Meanwhile, hepatorenal and serum stress indices (P < 0.05) were significantly elevated with chronic Cu exposure. In addition, a higher expression of stress (P < 0.05) (heat shock protein 60 and hypoxia-inducible factor-1 alpha) and apoptotic-related genes (C/EBP homologous protein, caspase-3, and Bcl-2 Associated X-protein) with down-regulation (P < 0.05) of the anti-apoptotic-related genes (B-cell lymphoma 2 and proliferating cell nuclear antigen) was noticed in the Cu-exposed fish. Histopathological alterations in the gills, liver, kidney, and spleen were markedly reported in the Cu-exposed group. The dietary supplementation with ZnONPs significantly alleviated the negative impacts of chronic waterborne-Cu exposure on growth performance, physiological changes, gene expression, and tissue architecture, especially at 30 mg kg-1 diet level. In particular, the inclusion of ZnONPs at the 30 mg kg-1 diet level produced better outcomes than the 20 mg kg-1 diet. Overall, ZnONPs could be added as a feed supplement in the C. gariepinus diet to boost the fish's health and productivity and alleviate the stress condition brought on by Cu exposure.

Development of a Site-Specific Guideline Value for Copper and Aquatic Life in Tropical Freshwaters of Low Hardness.

Copper (Cu) is a contaminant of potential concern for a uranium mine whose receiving waters are in the World Heritage-listed Kakadu National Park in northern Australia. The physicochemical characteristics of the freshwaters in this region enhance metal bioavailability and toxicity. Seven tropical species were used to assess the chronic toxicity of Cu in extremely soft freshwater from a creek upstream of the mine. Sensitivity to Cu was as follows: Moinodaphnia macleayi > Chlorella sp. > Velesunio sp. > Hydra viridissima > Amerianna cumingi > Lemna aequinoctialis > Mogurnda mogurnda. The 10% effect concentrations (EC10s) ranged from 1.0 µg/L Cu for the cladoceran Moinodaphnia macleayi to 9.6 µg/L for the fish M. mogurnda. The EC50s ranged from 6.6 µg/L Cu for the mussel Velesunio sp. to 22.5 µg/L Cu for M. mogurnda. Geochemical modeling predicted Cu to be strongly bound to fulvic acid (80%-99%) and of low bioavailability (0.02%-11.5%) under these conditions. Protective concentrations (PCs) were derived from a species sensitivity distribution for the local biota. The 99% PC (PC99), PC95, PC90, and PC80 values were 0.5, 0.8, 1.0, and 1.5 µg/L Cu, respectively. These threshold values suggest that the current Australian and New Zealand default national 99% protection guideline value for Cu (1.0 µg/L) would not provide adequate protection in freshwaters of low hardness, particularly for this area of high conservation value. The continuous criterion concentration predicted by the Cu biotic ligand model for conditions of low pH (6.1), low dissolved organic carbon (2.5 mg/L), low hardness (3.3 mg/L), and 27 °C was 0.48 µg/L Cu, comparable with the PC99. Consideration of the natural water quality conditions of a site is paramount for protective water quality guidelines. Environ Toxicol Chem 2022;41:2808-2821. © 2022 Commonwealth of Australia. Environmental Toxicology and Chemistry © 2022 SETAC.

Chronic toxicity, bioavailability and bioaccumulation of Zn, Cu and Pb in Lactuca sativa exposed to waste from an abandoned gold mine

Abandoned mines with untreated waste cause environmental pollution. The complex mixture of mining waste includes high metal content, anthropogenic chemicals and sterile rocks. Adverse effects of contaminated soils have been widely assessed by the use of plants. The aim of this study was to assess the chronic toxicity of a contaminated soil by waste from an abandoned gold mine on Lactuca sativa and its relationship with the bioavailability and bioaccumulation of Zn, Cu and Pb. Soil samples were taken from the site of mining waste stacking and a reference site in La Planta (Argentina). Contamination indices were calculated and acute and chronic exposures on L. sativa were carried out. Phytotoxicity indices, morphological and biochemical parameters, and concentrations of Zn, Cu and Pb in pseudo total and bioavailable soil fractions and in plant tissue were determined. Concentration- and time-dependent toxicity effects were observed, especially on plant width, fresh aerial biomass, leaf area and percentage of plants with completely necrotic aerial biomass. High levels of Zn (1453.3 ± 220.3 μg g−1) were found in plant tissue compared to Pb (277.2 ± 18.0 μg g−1) and Cu (255.3 ± 25.6 μg g−1). Toxicological endpoints correlated with metal uptake and mining waste concentration. In addition, bioaccumulation factors correlated with mobilisable and water soluble fractions. The concentration of Pb in aerial biomass surpassed the permissible concentrations in leaf vegetables, even at the reference site, indicating that lettuce crop consumption could be risky for the local population's health. This study demonstrated a strong relationship between metal bioavailability, toxicity endpoints and bioaccumulation, contributing with novel information to future ecotoxicological risk assessments and remediation plans.

Acute and sub-chronic effects of copper on survival, respiratory metabolism, and metal accumulation in Cambaroides dauricus

Trace metal contamination in the aquatic ecosystem occurs worldwide: although copper is an essential trace metal, it is considered as a pollutant at certain levels in China. Freshwater crayfish Cambaroides dauricus is a commercially important wild species in northeastern China, in which is an important heavy industry area. The population of C. dauricus was decreasing sharply due to the environmental pollution and human intervention over the past 20 years. However, nothing is known regarding the responses of this species to trace metal toxicants. This study aimed to determine the acute and chronic toxicity of Cu and its toxicological effects on respiratory metabolism, as well as Cu accumulation in C. dauricus. The acute (96 h) median lethal concentration (LC50) value of 32.5 mg/L was detected in C. dauricus. Then, acute (96 h; 8.24, 16.48 mg/L) and sub-chronic (14 days; 2.06, 4.12 mg/L) exposure in Cu was investigated by estimating the oxygen consumption rate, ammonium excretion rate, and Cu accumulation. Both acute and sub-chronic Cu exposure induced an inhibition of the oxygen consumption rate and ammonium excretion rate, and thereby, an increased O:N ratio. The shift in O:N ratio indicated a metabolic substrate shift towards lipid and carbohydrate metabolism under Cu stress. Cu accumulation in the hepatopancreas and muscles throughout the study was found to be time-dependent and concentration-dependent. The maximum accumulation in the hepatopancreas and muscle were almost 31.6 folds of the control after 14 days’ exposure to 4.12 mg/L concentration. Based on the present work, we suggest that crayfish be considered a potential bioindicator of environmental pollution in freshwater systems. The study provides basic information for further understanding of the toxicological responses of this species to trace metals.

Redox dyshomeostasis in the experimental chronic hepatic overloads with iron or copper

Male rats of 80–90 g were overloaded with either Fe(II) or Cu(II) for 42 days by high concentrations of FeCl2 or CuSO4 in the drinking water. The animals were fed with a commercial rodent diet of 2780 kcal/100 g. Both metal treatments led to a liver redox imbalance and dyshomeostasis with oxidative stress and damage and the concomitant enhancement of oxidative processes as indicated by in vivo surface liver chemiluminescence, the sensitive and organ non-invasive assay for oxidative free radical reactions, and by ex vivo determined processes of phospholipid peroxidation and protein oxidation. In parallel, marked decreases in the antioxidant defense were observed. Liver reduced glutathione (GSH) content and the reduced/oxidized glutathione ratio (GSH/GSSG) were early indicators of oxidative metabolic disturbance upon the metal overloads. Thus, GSH plays a central role in the defense reactions involved in the chronic toxicity of Fe and Cu. Chronic overloads of Fe or Cu in rats afford an experimental animal model of hemochromatosis and of Wilson's disease, respectively. These two animal models could be useful in the study and development of the beneficial effects of pharmacological interventions in the two human diseases.

Experimentally derived acute and chronic copper Biotic Ligand Models for rainbow trout

We evaluated the effects of varying water chemistry ([Ca2+]=0.2–3mM, [Mg2+]=0.05–3mM, dissolved organic matter (DOM, natural, from maple leaves)=0.3–10mg of CL−1, pH=5.0–8.5) on the acute (96-h, unfed fish) and chronic (30-d, fed fish) toxicity of waterborne Cu to juvenile rainbow trout (Oncorhynchus mykiss) exposed in flow-through conditions. Acute and chronic Biotic Ligand Models (BLMs) were developed from the obtained toxicity data-sets, using the Visual MINTEQ software. Our results indicate that Cu is predominantly an acute toxicant to rainbow trout, as there were no observable growth effects and the 96-h and 30-d LC50 values were similar, with mortality mostly occurring within the first few days of exposure. Calcium and DOM were greatly protective against both acute and chronic Cu toxicity, but Mg seemed to only protect against chronic toxicity. Additional protection by pH 5.0 in acute exposure and by pH 8.5 in chronic exposure occurred. In the range of conditions tested, the observed 96-h LC50 and 30-d LC20 values varied by a factor of 39 and 27 respectively. The newly developed acute and chronic BLMs explained these variations reasonably well (i.e. within a 2-fold error), except at pH≥8 where the high observed acute toxicity could not be explained, even by considering an equal contribution of CuOH+ and Cu2+ to the overall Cu toxicity. The 96-h LC50 values of 59% of 90 toxicity tests from 19 independent studies in the literature were reasonably well predicted by the new acute BLM. The LC20 predictions from the new chronic BLM were reasonable for 7 out of 14 toxicity tests from 6 independent chronic studies (with variable exposure durations). The observed deviations from BLM predictions may be due to uncertainties in the water chemistry in these literature studies and/or to differences in fish sensitivity. A residual pH effect was also observed for both the acute and the chronic data-sets, as the ratio of predicted vs. observed LC values generally increased with the pH. Additional mechanistic studies are required to understand the influence of pH, Na, and Mg on Cu toxicity to trout. The present study presents the first experimentally developed chronic Cu BLM for the rainbow trout. To the best of our knowledge, it also presents the first acute Cu BLM that is based on a published data-set for trout. These newly developed BLMs should contribute to improving the risk assessment of Cu to fish in freshwater.

Weighted species sensitivity distribution method to derive site-specific quality criteria for copper in Tai Lake, China.

Tai Lake (Ch: Taihu), which is the largest lake in Jiangsu province, China, has been affected by human activities. As part of a concerted effort to improve water quality to protect the integrity of the Tai Lake ecosystem, a water quality criterion (WQC) was developed for copper (Cu) II. The acute WQC was based on 440 values for acute toxicity of Cu to 24 species from 6 phyla, 16 families, and 20 genera. In addition, 255 values for chronic toxicity of Cu to 10 species from 5 phyla, 8 families, and 9 genera were used to derive chronic WQC. Instead of using a traditional approach based species sensitivity distributions (SSD), a weighted species sensitivity distribution (WSSD) approach was used to calculate the cumulative probability based on endemic species to Tai Lake. Acute and chronic WQC developed by use of the WSSD were 5.3 and 3.7 μg Cu/L, respectively. While the WQC values were comparable to those of other countries, there were slight differences due to variability in species composition of different regions. The site-specific criteria indicated that the current standard set for surface water by the Chinese government might not be protective of aquatic organisms in Tai Lake.

Metal–metal interactions of dietary cadmium, copper and zinc in rainbow trout, Oncorhynchus mykiss

The influence of metal–metal interactions on uptake, accumulation, plasma transport and chronic toxicity of dietary Cu, Cd and Zn in rainbow trout ( Oncorhynchus mykiss) was explored. Juvenile rainbow trout were fed diets supplemented with (μg/g) 500 Cu, 1000 Zn and 500 Cd singly and as a ternary mixture at 2.5% body weight daily ration for 28 days. Complex interactions among the metals dependent on the tissue/organ, metals ratios and duration of exposure were observed. While Zn did not accumulate, whole-body Cd and Cu concentrations increased following linear and saturation patterns, respectively. Early enhanced whole-body Cu accumulation in fish exposed to the metals mixture was correlated with reduced Cd concentration whereas late enhancement of Cd accumulation corresponded with elevated Cd concentration. This suggests early mutual antagonism and late cooperation between Cd and Cu probably due to interactions at temporally variable metal accumulation sites. At the level of uptake, Cd and Cu were either antagonistic or mutually increased the concentrations of each other depending on the duration of exposure and section of the gut. At the level of transport, enhanced Cd accumulation in plasma was closely correlated with reduced concentrations of both Zn and Cu indicating competitive binding to plasma proteins and/or antagonism at uptake sites. Compared to the Cu alone exposure, Cu concentrations were either lower (gills and carcasses) or higher (liver and kidney) in fish exposed to the metals mixture. On the other hand, Cd accumulation was enhanced in livers and carcasses of fish exposed to the mixture compared to those exposed to Cd alone, while Zn stimulated Cu accumulation in gills. Chronic toxicity was demonstrated by elevated malondialdehyde levels in livers and reduced concentrations of Zn and Cu in plasma. Overall, interactions of Cd, Cu and Zn are not always consistent with the isomorphous competitive binding theory.

Bioavailability of metals in stream food webs and hazards to brook trout (Salvelinus fontinalis) in the upper Animas River watershed, Colorado.

The water quality, habitats, and biota of streams in the upper Animas River watershed of Colorado, USA, are affected by metal contamination associated with acid drainage. We determined metal concentrations in components of the food web of the Animas River and its tributaries-periphyton (aufwuchs), benthic invertebrates, and livers of brook trout (Salvelinus fontinalis)-and evaluated pathways of metal exposure and hazards of metal toxicity to stream biota. Concentrations of the toxic metals cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn) in periphyton, benthic invertebrates, and trout livers from one or more sites in the upper Animas River were significantly greater than those from reference sites. Periphyton from sites downstream from mixing zones of acid and neutral waters had elevated concentrations of aluminum (Al) and iron (Fe) reflecting deposition of colloidal Fe and Al oxides, and reduced algal biomass. Metal concentrations in benthic invertebrates reflected differences in feeding habits and body size among taxa, with greatest concentrations of Zn, Cu, and Cd in the small mayfly Rhithrogena, which feeds on periphyton, and greatest concentrations of Pb in the small stonefly Zapada, a detritivore. Concentrations of Zn and Pb decreased across each trophic linkage, whereas concentrations of Cu and Cd were similar across several trophic levels, suggesting that Cu and Cd were more efficiently transferred via dietary exposure. Concentrations of Cu in invertebrates and trout livers were more closely associated with impacts on trout populations and invertebrate communities than were concentrations of Zn, Cd, or Pb. Copper concentrations in livers of brook trout from the upper Animas River were substantially greater than background concentrations and approached levels associated with reduced brook trout populations in field studies and with toxic effects on other salmonids in laboratory studies. These results indicate that bioaccumulation and transfer of metals in stream food webs are significant components of metal exposure for stream biota of the upper Animas River watershed and suggest that chronic toxicity of Cu is an important factor limiting the distribution and abundance of brook trout populations in the watershed.

The effect of chronic toxicity of copper on the activity of Balbiani rings and nucleolar organizing region in the salivary gland chromosomes of Chironomus ninevah larvae

The effect of copper on the activity of Balbiani rings (BR1 and BR2) and nucleolar organizing region (NOR) in chromosome IV of the salivary gland of the 4th instar larvae of Chironomus ninevah has been investigated. Sublethal concentrations, i.e. 0·02, 0·1, 0·15 and 0·2 mg litre −1 suppress the activity of BR1, and are statistically significant. The same concentrations reduced the activity of BR2 and NOR, but without significant differences, except 0·1 mg litre −1 which was significant. The purpose of this work was to evaluate the effect of chronic toxicity of Cu 2+ on the expression of gene activity of midge salivary gland chromosomes Although there may be many more aquatic organisms that are more sensitive to copper, this midge was selected as a biological indicator species because of its association with benthic sediments.

Relationship between chronic toxicity and bioaccumulation of copper, cadmium and zinc as affected by water hardness and humic acid

Using daphnids, we have evaluated the effects of water hardness and humic acid (HA) on the chronic toxicity and bioaccumulation of Cu, Cd and Zn. Although changes in water hardness and HA concentration changed bioaccumulation or chronic toxicity, or both, of each of the three metals, there was no consistent relationship between changes in toxicity and changes in bioaccumulation. Only for Cu in hard water was there a positive correlation between toxicity and bioaccumulation. The uptake of 65Zn by molted exoskeletons suggests that changes in water chemistry do indeed modify the bioavailability of metals as would be expected. That is, an increase in the concentration of Ca 2+ or Mg 2+ or the increased chelation of metals by HA should decrease bioavailability. The complex storage, transformation and excretion processes in multicellular animals, however, result in there being a poor correlation among bioavailability, bioaccumulation and toxicity.

Effects of Available Dietary Carbohydrate and Water Temperature on the Chronic Toxicity of Waterborne Copper to Rainbow Trout (Salmo gairdneri)

We used a factorial design to determine the effects of dietary carbohydrate content (0.1 or 25.6%) and temperature (10 or 15 °C) on the chronic toxicity of waterborne copper (0 or 144 μg Cu∙L−1) to rainbow trout (Salmo gairdneri) over a 12-wk period. Relative to those on the low-carbohydrate (LC) diet, trout reared on the high-carbohydrate (HC) diet developed enlarged, glycogen-filled livers and attained reduced final mean wet body weights. This response was more pronounced at 10 than at 15 °C. Whereas Cu exposure had no significant effect on the growth or mortality of either LC or HC fish at 15 °C, or LC fish at 10 °C, HC fish at 10 °C showed significantly reduced final weight and increased mortality. Trends toward reduced liver glycogen and increased serum glucose, characteristic of chronic stress, were apparent in all Cu-exposed fish. In Cu-free water, mean Cu concentrations in liver tissue of HC fish were significantly lower than those in LC fish at both temperatures. Copper exposure resulted in significant increases in liver Cu concentration for all diet treatments except HC at 10 °C. Elevated serum sorbitol dehydrogenase levels, indicative of hepatic damage, were apparent in Cu-HC fish at 15 °C and in HC, Cu-HC, and Cu-LC fish at 10 °C. No treatment effects on either serum protein or hematocrit were evident. The treatments during the growth period affected subsequent lethal tolerance of Cu. While the 96-h Cu LC50 was not significantly altered by diet alone at 15 °C, it was elevated in HC fish, relative to LC fish at 10 °C. The Cu LC50 of Cu preexposed fish was significantly elevated, relative to parallel groups held in Cu-free water, for LC fish at both 10 and 15 °C. This acclimation to Cu was not apparent in HC fish at either temperature: no significant elevation in LC50 occurred as a result of preexposure. We conclude that increased dietary carbohydrate can significantly increase the chronic toxicity of Cu to trout, and that the impact is enhanced by reduced water temperature.

The toxicity and bioaccumulation of cadmium and copper as affected by humic acid

The toxicity of Cd and Cu to daphnids was evaluated over a 42-day exposure period at three humic acid concentrations. The addition of humic acid to test water decreased the acute and chronic toxicity of Cu but increased the acute and chronic toxicity of Cd. However, there was no effect of HA on the bioaccumulation of either metal. At the highest concentration, the presence of humic acid actually resulted in an increase in mean brood sizes of Cu-exposed animals and prevented a reduction in mean brood sizes of Cd-exposed animals.