Effect of Chloride Concentration on the Cytotoxicity, Bioavailability, and Bio-Reactivity of Copper and Silver in the Rainbow Trout Gut Cell Line (RTgutGC)

Chloride (Cl-) influences the bioavailability and toxicity of metals in fish, but the mechanisms by which it influences these processes is poorly understood. Here, we investigated the effect of chloride on the cytotoxicity, bioavailability (i.e., accumulation) and bio-reactivity (i.e., induction of mRNA levels of metal responsive genes) of copper (Cu) and silver (Ag) in the rainbow trout gut cell line (RTgutGC). Cells were exposed to metals in media with varying Cl- concentrations (0, 1, 5 and 146 mM). Metal speciation in exposure medium was analyzed using Visual MINTEQ software. Cytotoxicity of AgNO3 and CuSO4 was measured based on two endpoints: metabolic activity and membrane integrity. Cells were exposed to 500 nM of AgNO3 and CuSO4 for 24 hours in respective media to determine metal bioavailability and bioreactivity. Ag speciation changes from free ionic (Ag+) to neutral (AgCl), to negatively charged chloride complexes (AgCl2-, AgCl3-) with increasing Cl- concentration in exposure media whereas Cu speciation remains in two forms (Cu2+ and CuHPO4) across all media. Chloride does not affect Ag bioavailability but decreases metal toxicity and bio-reactivity. Cells exposed to Ag expressed significantly higher metallothionein mRNA levels in low Cl- media (0, 1, and 5 mM) than in high Cl- medium (146 mM). This suggests that chloride complexation reduces silver bio-reactivity and toxicity. Conversely, Cu bioavailability and toxicity were higher in the high chloride medium (146 mM) than in the low Cl- (0, 1, and 5 mM) media, supporting the hypothesis that Cu uptake may occur via a chloride dependent mechanism.

A Polymer Inclusion Membrane for Sensing Metal Complexation in Natural Waters

Metal speciation studies are of great importance in assessing metal bioavailability in aquatic environments. Functionalized membranes are a simple tool to perform metal chemical speciation. In this study, we have prepared and tested a polymer inclusion membrane (PIM) made of the polymer cellulose triacetate (CTA), the extractant di-(2-ethylhexyl) phosphoric acid (D2EHPA), and the plasticizer 2-nitrophenyloctyl ether (NPOE) as a sensor for Zn and Cu complexation studies. This PIM, incorporated in a device with an 0.01 M HNO3 receiving solution, is shown to effectively transport free metal ions, and it is demonstrated that the presence of ligands that form stable complexes with divalent metallic ions, such as ethylenediaminetetraacetic acid (EDTA) and humic acid (HA), greatly influences the accumulation of the metals in the receiving phase due to the increasing metal fraction complexed in the feed phase. Moreover, the effect of major ions found in natural waters has been investigated, and it is found that the presence of calcium did not decrease the accumulation of either Zn or Cu. Finally, the PIM sensor has been used successfully to evaluate metal complexation in a river water affected by Zn pollution.

The Use of Biochar of High Growth Rate Plants to Agriculturally Remediate Heavy Metal Polluted Acidic Mine Wastes

The chapter is meant to expose how a sound methodology can be instrumented to both, remediate acidic metal polluted mine wastes, taking advantage of the neutralizing power and high metal sorption affinity of biochar, and to utilize pyrolyzed material derived from high-rate growth plants (water hyacinth, Eichhornia crassipes Mart, and Eucalyptus, Eucalyptus globulus Labill), which have become of ecological relevance due to their unwanted proliferation over specific terrestrial, lacustrine or riverine environments. In addition, the proposal considers not only neutralizing the mine tailings and abating the toxic levels of specific heavy metals like Pb, Cd, Cu, Zn, etc., to fulfill the international and national standards and norms, but to conveniently combine biochar with widely used soil amendments to pass widely recognized biological tests of growth using heavy metal-sensitive plants. The approach addresses firstly: a) characterizing physiochemically mine tailings and biochar, in terms of their properties (metal speciation and contents, potential acidity and neutralization potential, chemical oxygen demand, heavy metal-biochar sorption-complexing affinities, among others), and secondly; b) creating a” fertile environment” by reconditioning, agriculturally, the heavy metal-polluted acidic mine waste to allow native vegetation, or other reforesting species, to regrow on the reclaimed site, based on the bioassay tests performances.

Impact of Zn Substitution on Fe(II)-induced Ferrihydrite 2 Transformation Pathways

Iron oxide minerals are ubiquitous in soils, sediments, and aquatic systems and influence the fate and availability of trace metals. Ferrihydrite is a common iron oxide of nanoparticulate size and poor crystallinity, serving as a thermodynamically unstable precursor to more crystalline phases. While aging induces such phase transformations, these are accelerated by the presence of dissolved Fe(II). However, the impact of trace metals on Fe(II)-catalyzed ferrihydrite phase transformations at ambient temperatures and the associated effects on trace metal speciation has seen limited study. In the present work, phase transformations of ferrihydrite that contains the trace metal zinc in its structure were investigated during aging at ambient temperature in the presence of two different Fe(II) concentrations at pH 7. X-ray diffraction reveals that low Fe(II) concentration (0.2 mM) generates hematite plus minor lepidocrocite, whereas high Fe(II) concentration (1.0 mM) promotes the production of a magnetite-lepidocrocite mixture. In both cases, a substantial fraction of ferrihydrite remains after 12 days. In contrast, Zn-free ferrihydrite forms primarily lepidocrocite and goethite in the presence of 0.2 mM Fe(II), with minor hematite and a trace of ferrihydrite remaining. For 1.0 mM Fe(II), magnetite, goethite, and lepidocrocite form when Zn is absent, leaving no residual ferrihydrite. Transformations of Zn-ferrihydrite produce a transient release of zinc to solution, but this is nearly quantitatively removed into the mineral products after 1 hour. Extended X-ray absorption fine structure spectroscopy suggests that zinc partitions into the newly formed phases, with a shift from tetrahedral to a mixture of tetrahedral and octahedral coordination in the 0.2 mM Fe(II) system and taking on a spinel-like local structure in the 1.0 mM Fe(II) reaction products. This work indicates that substituting elements in ferrihydrite may play a key role in promoting the formation of hematite in low temperature systems, such as soils or sediments. In addition, the retention of zinc in the products of ferrihydrite phase transformation shows that trace metal micronutrients and contaminants may not be mobilized under circumneutral conditions despite the formation of more crystalline iron oxides. Furthermore, mass balance requires that the abundance and isotopic composition of iron oxide-associated zinc, and possibly other trace metals, in the rock record may be retained during diagenetic phase transformations of ferrihydrite if catalyzed by dissolved Fe(II).

Metal Speciation in Sludges: a Tool to Evaluate Risks of Land Application and to Track Heavy Metals Contamination in Sewage Network

Agricultural spreading of dewatered sewage sludge from wastewater treatment plants is economically profitable but care must be taken to ensure that there is neither degradation of the agronomic quality of the soils nor contamination of them in the long term, particularly by accumulation of heavy metals. To evaluate the variability of the sludge in a given geographical area, we studied the sludge coming from five treatment plants in northern Algeria. We determined parameters that account for the agronomic quality of sludges and total content of Ag, Cd, Co, Cr, Cu, Ni, Pb, Ti and Zn. We evaluated the bioavailability of theses metals by determining their speciation by sequential extraction, each metal being distributed among five fractions: easily exchangeable, acido-soluble, bound to carbonates and partly to Fe-sulphides, bound to Fe- and Mn-oxides, bound to organic matter or sulphides, contained in poorly soluble minerals. We found that all the analysed sludges had satisfactory properties from an agronomic quality point of view. High total Ni content indicated that three sludges were not spreadable under French or Chinese regulations. Metal speciation, however, showed that Ni was contained in very poorly bioavailable fractions, and therefore presented a low risk in the soils concerned. In contrast, the total Cu was below the regulatory limit values, but contained in very bioavailable fractions with a risk of toxic effects by accumulation over less than 10 years. These results showed that regulations must take into account the bioavailability with regard to the characteristics of the soils on which sludge will be spread. Metal speciation in the sludge also made it possible to identify the zone of the sewerage network on which the sources of contamination must be sought and gave indications on the nature of these sources.