Development of a new tool for the long term in vitro ecotoxicity testing of nanomaterials using a rainbow-trout cell line (RTL-W1).

The current wide use of manufactured nanomaterials (MNs) is leading to the release of nanoparticles (NPs) to water bodies. Aquatic organisms, including fish, are exposed to low concentrations of NPs for long periods of time being necessary to develop laboratory toxicity tests reflecting realistic conditions. Additionally, today there is a demand of in vitro assays respecting the 3Rs principle. Thus, the main aim of this work was to stablish an in vitro tool for the assessment of long-term NPs ecotoxicity. Considering the key role of liver in detoxification, a rainbow trout liver cell line, RTL-W1, was used. CuO NPs were chosen to validate this tool taking into account their important production level. Cells were exposed for 21 days to 25 or 100 µg CuO NPs/ml. Every seven days cells were split and one fourth of them transferred to a new plate with appropriate concentrations of NPs in culture medium. Lower concentrations of CuO NPs did not cause any deleterious effect, whereas higher concentrations led to significant mortality after 14 days and to the intracellular accumulation of Cu particles. Identical results were observed in cells exposed to CuSO4 at the same Cu concentrations. Therefore, the observed toxic effects might be mainly due to Cu2+ ions.

Mercury availability by operationally defined fractionation in granulometric distributions of soils and mine wastes from an abandoned cinnabar mine.

Mercury contamination from historic cinnabar mines represents a potential risk to the environment. Asturias, in Northern Spain, was one of the largest metallurgic and mining producer areas of Hg in Europe during the 20th century until the end of activities in 1974. Mining operations have caused Hg release and dispersion throughout the area. In this study, soils collected from calcine piles and surrounding soils at an abandoned Hg mine and metallurgical plant in Mieres (Asturias, Spain) were distributed in different particle-size subsamples. Fractionation of Hg was performed by means of a Hg-specific sequential extraction procedure complemented with the selective determination of organic Hg fraction by a specific extraction method. Extremely high concentrations of total Hg were found in calcine piles. Concentrations and mobility of Hg decreased markedly with the distance in soils located 25 m both above and below the chimney of the metallurgical plant. The sequential extraction results indicated that Hg is primarily found as elemental Hg followed by sulfide Hg in the finest subsamples. However, this distribution is inverted in the coarser grain fractions where sulfide Hg prevails. Calcine piles exhibited exceptionally high values of mobile Hg (up to 5350 µg g(-1) in the finest subsample). Accumulation of Hg in the elemental Hg fraction was observed at decreasing grain size which is indicative of deposition of Hg vapors from the metallurgical plant. Enrichment of sulfide Hg was found in the finest subsamples of soils sampled below the chimney (up to 99 µg g(-1)). Significant organic Hg contents were observed in the soil samples (up to 2.8 µg g(-1)), higher than those found in other abandoned Hg mining sites. A strong correlation was observed between organic Hg and Hg humic and fulvic complexes, as well as with the elemental Hg fraction. This indicates that both humic and fulvic material and elemental Hg must be the primary variables controlling Hg methylation in these soils.

Arsenic input into the catchment of the River Caudal (Northwestern Spain) from abandoned Hg mining works: effect on water quality.

In Asturias (NW Spain) there are many abandoned mines, of which Hg mines are of particular significance from an environmental point of view, due to the presence of Hg and particularly As, which is found either in the form of specific (orpiment and realgar) or non-specific minerals (As-rich pyrite). The instability of these minerals leads to the presence of As-rich mine drainage and spoils heap leachates that enter surface waters or groundwaters. A study including the three most important Hg mines in the region (La Soterraña, Los Rueldos and El Terronal) has been conducted. Watercourses flowing through these mining areas are tributaries of the River Caudal, one of the most important rivers in the area. High concentrations of As were found in some of these waters, which were monitored over a period of three hydrological years and classified according to a water quality index. Those waters sampled close to the mines are generally of poor-to-bad quality, with low alkalinity and in some cases high metal content, but the quality of these waters improves with distance from the mines. The average mass load of As entering the River Caudal has been evaluated as: 200, 12 and 9,800 kg year(-1) from La Soterraña, Los Rueldos and El Terronal mine sites, respectively. Despite the constant input of about 10 tonnes of As per year, the total As concentration remains below analytical detection thresholds on account of the river's high water flow. Nevertheless, an important part of this As load is presumably retained in the river sediments, representing a potential risk of pollution of the aquatic ecosystems.