Toxicogenomics of natural and anthropogenic effectors in eleutheroembryos

The introduction of chemicals into the environment by human activities may represent a serious risk to environmental and human health. Environmental risk assessment requires the use of efficient and sensitive tools to determine the impact of contaminants on the ecosystems. The use of zebrafish for the toxicity assessment of pharmaceuticals, drugs, and pollutants, is becoming well accepted due to zebrafish unique advantages for the screening of compounds for hazard identification. The aim of the present work is to apply toxicogenomic approaches to identify novel biomarkers and uncovered potential modes of action of classic and emergent contaminants able to disrupt endocrine systems, such as the Retinoic Acid Receptor, Retinoid X Receptor and the Aryl Hydrocarbon Receptor. This study relies on different nuclear and cytosolic protein receptors and other conditional (ligand- or stress- activated) transcriptional factors that are intimately involved in the regulation of defensome genes and in mechanisms of chemical toxicity. The transcriptomic effects of organic compounds, endogenous compounds, and nanoparticles were analysed during the early stages of zebrafish development. Studying the gene expression profiles of exposed and unexposed organisms to pollutants using microarrays allowed the identification of specific gene markers and to establish a "genetic code" for the tested compounds. Changes in gene expression were observed at toxicant concentrations that did not cause morphological effects. Even at low toxicant concentrations, the observed changes in transcript levels were robust for some target genes. Microarray responses of selected genes were further complemented by the real time quantitative polymerase chain reaction (qRT-PCR) methodology. The combination of bio-informatic, toxicological analyses of differential gene expression profiles, and biochemical and phenotypic responses across the treatments allowed the identification of uncovered potential mechanisms of action. In addition, this work provides an integrated set of tools that can be used to aid management-decision making by improving the predictive capability to measure environmental stress of contaminants in freshwater ecosystems. This study also illustrates the potential of zebrafish embryos for the systematic, large-scale analysis of chemical effects on developing vertebrates.

Inputs from a mercury-contaminated lagoon: impact on the nearshore waters of the Atlantic Ocean

Ria de Aveiro, a Portuguese coastal lagoon that exchanges water with the Atlantic Ocean, received the effluent from a chlor-alkali industry for over 50 years; consequently several tons of mercury had been buried in the sediments of an inner basin. To assess the importance (and seasonal variation) of the lagoon waters as carriers of mercury to the nearby coastal area, we measured total mercury levels in several compartments: in surface sediments, in surface and deep waters (including dissolved and particulate matter!, and in biota. Dissolved (reactive and total) mercury concentrations both in surface and deep waters were low (<1 to 15 ng L '). Mean mercury values in suspended particulate matter varied hetween 0.2 and 0.6 jxg g ' and in sediments between 1 and 9 ng g '. Aquatic organisms displayed levels below regulatory limits but exhibited some bioaccumulation of mercury, with concentrations ranging from 0.05 to 0.8 ^ig g ' Idry weight (dw)|. No seasonal pattern was found in this study for mercury-related determinations. Levels found in the estuary mouth during ebb tide provide evidence for the transport of mercury to the coastal zone. No significant changes in the partition of mercury between dissolved and particulate phases were found in the coastal waters in comparison with the values found in the estuary mouth. In spite of the high levels of mercury found inside some areas of the lagoon, the wide web of islands and channels allows some spreading of contaminants before they reach the coastal waters. Moreover, the low efficiency of local marine sediments in trapping mercury contributes to a dilution of mercury transported in suspended particulate matter over a broader area, reducing the impact in the nearby manne coastal zone.