Mosses are frequently used to monitor atmospheric metal contamination but few studies on metal adsorption under controlled conditions are available. Here, the accumulation of the heavy metals copper and zinc was studied in the acrocarp moss Atrichum undulatum. An in vitro culture of A. undulatum was established and the same line, size and equally old remets were exposed to six different treatments representing various metal exposure times and washing scenarios as rain simulation. The metal treatments were done in copper and zinc salts (Cu-acetate, CuSO4, ZnSO4 and ZnCl2, respectively). Energy-Dispersive X-ray microanalysis (EDX) was employed to detect bound heavy metals on the moss plantlets. Element distribution in stems and leaves was measured separately. The aqueous solution of metal salts facilitated an adsorption of both elements in the moss tissue as compared to solid medium. Furthermore, A. undulatum can tolerate pollution of zinc and copper in a distinctive extent; our data point towards a higher zinc tolerance whereas copper is rather harmful. However, semi-quantitatively, less zinc was detected within the moss tissue compared to copper. Interestingly, a strong positive correlation between the accumulation of copper/zinc and iron, and a strong negative correlation between copper/zinc and magnesium, respectively, was documented.

Gradual contamination of agricultural land with copper (Cu), due to the indiscriminate uses of fungicides and pesticides, and the discharge of industrial waste to the environment, poses a high threat for soil degradation and consequently food crop production. In this study, we combined morphological, physiological and biochemical assays to investigate the mechanisms underlying acetate-mediated Cu toxicity tolerance in lentil. Results demonstrated that high dose of Cu (3.0 mM CuSO₄. 5H₂O) reduced seedling growth and chlorophyll content, while augmenting Cu contents in both roots and shoots, and increasing oxidative damage in lentil plants through disruption of the antioxidant defense. Principle component analysis clearly indicated that Cu accumulation and increased oxidative damage were the key factors for Cu toxicity in lentil seedlings. However, acetate pretreatment reduced Cu accumulation in roots and shoots, increased proline content and improved the responses of antioxidant defense (e.g. increased catalase and glutathione-S-transferase activities, and improved action of glutathione-ascorbate metabolic pathway). As a result, excess Cu-induced oxidative damage was reduced, and seedling growth was improved under Cu stress conditions, indicating the role of acetate in alleviating Cu toxicity in lentil seedlings. Taken together, exogenous acetate application reduced Cu accumulation in lentil roots and shoots and mitigated oxidative damage by activating the antioxidant defense, which were the major determinants for alleviating Cu toxicity in lentil seedlings. Our findings provide mechanistic insights into the protective roles of acetate in mitigating Cu toxicity in lentil, and suggest that application of acetate could be a novel and economical strategy for the management of heavy metal toxicity and accumulation in crops.

The environmental consequences of nano-CuO particles have gained significant attention in recent decades. Identification of the mechanisms for soil and plant responses with respect to the chemical speciation of nano-CuO (mainly the exchangeable and reducible fractions) remains scarce. Here, we analyzed different chemical speciation of Cu and DTPA-extractable Cu over 42 days in (1) control soil without Cu addition; (2) soil treated with nano-CuO particles; and (3) soil treated with CuSO₄ solution. The applied dose was 500 mg Cu kg⁻¹ and maize was grown in these soils. Plant growth was inhibited, but the inhibition by nano-CuO was slightly weaker compared to CuSO₄. Cu accumulations were similar in the roots for CuSO₄ and nano-CuO treatments, but significantly higher in the shoots for CuSO₄ treatment. This indicates that Cu from nano-CuO-treated soils mainly accumulated in roots but rarely transferred to shoots. Enzyme activities on the rhizoplane visualized by zymography were strongly depressed by CuSO₄ but slightly inhibited by nano-CuO. Microbial community diversity measured by 16S rRNA was the lowest in CuSO₄-treated soils among three treatments. These results were explained by the following mechanisms: (1) Gradual increases of DTPA-extractable and exchangeable Cu were found in nano-CuO-treated soil, and the final concentrations at day 42 were only half of those in CuSO₄-treated soil; (2) Enzyme activities on the rhizoplane were positively related to soil pH and negatively correlated with DTPA-extractable and exchangeable Cu; (3) Even though reducible Cu in nano-CuO-treated soils was 1.3 times higher than in CuSO₄-treated soils, indicating stronger nano-accrued oxidative stress in nano-CuO-treated soils, the toxicity induced by nano-CuO particles was still weaker than CuSO₄. Nevertheless, the toxicity of Cu particles to plants and microbes mainly depends on the gradually-released bioavailable Cu. This demonstrates the greater importance of bioavailable Cu concentrations for toxicity modulation rather than the scale of Cu particles.

Pyrethroid-resistant Hyalella azteca with voltage-gated sodium channel mutations have been identified at multiple locations throughout California. In December 2013, H. azteca were collected from Mosher Slough in Stockton, CA, USA, a site with reported pyrethroid (primarily bifenthrin and cyfluthrin) sediment concentrations approximately twice the 10-d LC50 for laboratory-cultured H. azteca. These H. azteca were shipped to Southern Illinois University Carbondale and have been maintained in pyrethroid-free culture since collection. Even after 22 months in culture, resistant animals had approximately 53 times higher tolerance to permethrin than non-resistant laboratory-cultured H. azteca. Resistant animals held in culture also lacked the wild-type allele at the L925 locus, and had non-synonymous substitutions that resulted in either a leucine-isoleucine or leucine-valine substitution. Additionally, animals collected from the same site nearly three years later were again resistant to the pyrethroid permethrin. When resistant animals were compared to non-resistant animals, they showed lower reproductive capacity, lower upper thermal tolerance, and the data suggested greater sensitivity to, 4, 4′-dichlorodiphenyltrichloroethane (DDT), copper (II) sulfate, and sodium chloride. Further testing of the greater heat and sodium chloride sensitivity of the resistant animals showed these effects to be unrelated to clade association. Fitness costs associated with resistance to pyrethroids are well documented in pest species (including mosquitoes, peach-potato aphids, and codling moths) and we believe that H. azteca collected from Mosher Slough also have fitness costs associated with the developed resistance.

Copper nanoparticles (nCu) are widely used in industry and in daily life, due to their unique physical, chemical, and biological properties. Few studies have focused on nCu phytotoxicity, especially with regard to toxicity mechanisms in crop plants. The present study examined the effect of 15.6 μM nCu exposure on the root morphology, physiology, and gene transcription levels of wheat (Triticum aestivum L.), a major crop cultivated worldwide. The results obtained were compared with the effects of exposing wheat to an equivalent molar concentration of ionic Cu (Cu²⁺ released from CuSO₄) and to control plants. The relative growth rate of roots decreased to approximately 60% and the formation of lateral roots was stimulated under nCu exposure, possibly due to the enhancement of nitrogen uptake and accumulation of auxin in lateral roots. The expression of four of the genes involved in the positive regulation of cell proliferation and negative regulation of programmed cell death decreased to 50% in the Cu²⁺ treatment compared to that of the control, while only one gene was down-regulated to about half of the control in nCu treatment. This explained the decreased root cell proliferation and higher extent of induced cell death in Cu²⁺- than in nCu-exposed plants. The increased methane dicarboxylic aldehyde accumulation (2.17-fold increase compared with the control) and decreased antioxidant enzyme activities (more than 50% decrease compared with the control) observed in the Cu²⁺ treatment in relation to the nCu treatment indicated higher oxidative stress in Cu²⁺- than in nCu-exposed plants. Antioxidant (e.g., proline) synthesis was pronouncedly induced by nCu to scavenge excess reactive oxygen species, alleviating phytotoxicity to wheat exposed to this form of Cu. Overall, oxidative stress and root growth inhibition were the main causes of nCu toxicity.

Bulk Cu compounds such as Cu(OH)₂ are extensively used as pesticides in agriculture. Recent investigations suggest that Cu-based nanomaterials can replace bulk materials reducing the environmental impacts of Cu. In this study, stress responses of alfalfa (Medicago sativa L.) seedlings to Cu(OH)₂ nanoparticle or compounds were evaluated. Seeds were immersed in suspension/solutions of a Cu(OH)₂ nanoform, bulk Cu(OH)₂, CuSO₄, and Cu(NO₃)₂ at 25 and 75 mg/L. Six days later, the germination, seedling growth, and the physiological and biochemical responses of sprouts were evaluated. All Cu treatments significantly reduced root elongation (average = 63%). The ionic compounds at 25 and 75 mg/L caused a reduction in all elements analyzed (Ca, K, Mg, P, Zn, and Mn), excepting for S, Fe and Mo. The bulk-Cu(OH)₂ treatment reduced K (48%) and P (52%) at 75 mg/L, but increased Zn at 25 (18%) and 75 (21%) mg/L. The nano-Cu(OH)₂ reduced K (46%) and P (48%) at 75 mg/L, and also P (37%) at 25 mg/L, compared with control. Confocal microscopy images showed that all Cu compounds, at 75 mg/L, significantly reduced nitric oxide, concurring with the reduction in root growth. Nano Cu(OH)₂ at 25 mg/L upregulated the expression of the Cu/Zn superoxide dismutase gene (1.92-fold), while ionic treatments at 75 mg/L upregulated (∼10-fold) metallothionein (MT) transcripts. Results demonstrated that nano and bulk Cu(OH)₂ compounds caused less physiological impairments in comparison to the ionic ones in alfalfa seedlings.

The density-dependence of terrestrial plant–plant interactions in the presence of toxins has previously been explored using biodegradable compounds. We exposed barley and lettuce to four copper concentrations at four stand densities. We hypothesized that toxin effects would decrease and Cu uptake would increase at increasing plant densities. We analyzed toxin effects by (a) comparing plant biomasses and (b) using a recent regression model that has a separate parameter for the interaction of resource competition and toxin interference. Plant response to Cu was density-dependent in both experiments. Total Cu uptake by barley increased and the dose per plant decreased as plant density increased. This study is the first to demonstrate that plant density mediates plant response to metals in soil in a predictable way. This highlights the need to explore the mechanisms for and consequences of these effects, and to integrate the use of several plant densities into standard ecotoxicological testing.

In this report, we investigated how the presence of a polymer shell (poly(styrene-co-butyl acrylate) alters the toxicity of CuO NPs in Lemna gibba. Based on total Cu concentration, core–shell CuO NPs were 10 times more toxic than CuO NPs, inducing a 50% decrease of growth rate at 0.4 g l−1 after 48-h of exposure while a concentration of 4.5 g l−1 was required for CuO NPs for a similar effect. Toxicity of CuO NPs was mainly due to NPs solubilization in the media. Based on the accumulated copper content in the plants, core–shell CuO NPs induced 4 times more reactive oxygen species compared to CuO NPs and copper sulfate, indicating that the presence of the polymer shell changed the toxic effect induced in L. gibba. This effect could not be attributed to the polymer alone and reveals that surface modification may change the nature of NPs toxicity.

Cu nanoparticles (CuNPs) have been widely used in numerous products, and may become a potential threat to marine organisms, but their behavior in the marine environments and potential toxicity to marine organisms remain little known. In the present study, we investigated the behavior of CuNPs in seawater, as well as the toxicity and bioaccumulation of CuNPs and copper sulfate (CuSO4) in barnacle larvae (Balanus amphitrite), a dominant fouling invertebrate in marine environment. CuNPs tended to aggregate in natural seawater and released Cu ion rapidly into seawater. The aggregation and release were especially higher at a lower concentration of CuNPs, e.g., 94–96% of CuNPs were released as Cu ions at 20 μg/L after 24 h. The larger size of CuNPs (40 nm) tended to display a higher solubility than the 20 nm CuNPs did. Humic acids enhanced the aggregation and inhibited the dissolution of CuNPs, and had a protective effect on the survival of nauplii II at higher Cu concentrations (100–200 μg/L). Comparison of the lethal concentrations showed that CuNPs were generally less toxic to the two stages of barnacle larvae (nauplii II and VI) than the Cu ions. The calculated 48-h LC50 values for nauplii II were 189.5 μg/L, 123.2 μg/L, and 89.8 μg/L for 20 nm CuNPs, 40 nm CuNPs, and CuSO4, respectively. However, the lethal concentrations of Cu bioaccumulation in the barnacle larvae were comparable between CuNPs and Cu ions when expressed by the actual tissue Cu bioaccumulation. Barnacle larval settlement decreased with an increase of Cu concentrations of both CuNPs and CuSO4, and was significantly inhibited at 100 μg/L CuSO4 and 150 μg/L CuNPs. Our results indicated that the toxicity of CuNPs could not be solely explained by the released Cu ions, and both CuNPs and the released Cu ion contributed to their toxicity and bioaccumulation in barnacle larvae.

The feasibility of assessing copper accumulation in agricultural soils using avoidance tests with a Canadian strain of Folsomia candida was investigated under laboratory conditions. The avoidance response to nominal copper sulfate concentrations of 0, 200, 800, 1600 and 3200 mg kg⁻¹ in OECD soil was inconsistent between trials with the standard plastic cup or a modified Petri dish method requiring less soil. However, combined results from three Petri dish trials decreased variability and provided a 75% avoidance level, close to the 80% criterion proposed for avoidance tests. A Copper avoidance EC₅₀ₛ of 18 mg kg⁻¹was obtained using the Petri dish method whether tests were conducted with or without light. While Petri dish tests have potential as a cheap tool to distinguish metal contaminated soils from uncontaminated soils they would be unsuitable for tracking or quantifying changes in metal concentrations. throughout remediation. Advantages and limitations of the method have been presented.