-Using several clonal lineages of Simocephalus vetulus (Cladocera, Daphniidae) as a random sample, we investigated the genetic component of the halotolerance of one brackish and two freshwater populations of this littoral filter feeder. We hypothesized that genotypes from the brackish population were more tolerant than freshwater ones, via adaptation to local environmental conditions. Clonal identity was established by a cost-effective molecular fingerprinting technique (microsatellite-primed polymerase chain reaction (MSP-PCR)). Two distinct methodologies were used to assess cladoceran sensitivity to syntheticgrade sodium chloride (NaCl): (i) standard 48-h acute assays and (ii) 12-h survival time (ST) trials. No correlation was found between acute EC 50 and ST values. The sensitivity of brackish and freshwater clones was comparable in terms of acute EC 50 (varied from 2.28 to 3.83 g.L x1). On the contrary, genetically determined differential tolerance to NaCl among populations was found for ST: all brackish genotypes, except one, were more resilient (ST > 120 min) than freshwater clones (ST < 120 min). Bearing in mind that these results were obtained with isolates from the extant population, it is surprising that the range of acute sensitivity of the freshwater and brackish genotypes was similar, and that the only difference between them was the ability of brackish clones to survive longer under high salinity stress (6 g.L x1, in ST trials). We must conclude that the effect of salinity (original environment context) on the selection of genotypes was weaker than we had expected and than other authors have shown for other stressors.

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Freshwaters commonly face human pressure, which produces multiple stressor scenarios that may threaten their ecological status. Biotic community assessments are useful tools to monitor such changes, and the European Water Framework Directive (WFD) has brought this into the legislative framework. The present study focused on a small mountain river facing multiple stresses, with the purpose of (1) evaluating the sensitivity of the macroinvertebrate community to the stresses, (2) assessing whether the ecological status was impaired in different years and (3) comparing the WFD approach with more refined community analyses. The river was generally in a very good ecological state. Despite seasonal and inter-annual fluctuations, no evident stressor-related effects were detected. We hypothesise that the observed resilience was the result of (1) the current dormant state of the abandoned mining areas (the most relevant potential source of contamination) and (2) the naturalness and strong current of the river, which buffered the impacts of contamination. Such a pattern may be generally valid for small mountain rivers, making them therefore more resilient to stresses. Nevertheless, small changes in community structure in the lowest reach of the river – undetected by the WFD assessment scheme – suggest that some disturbance was present, signalling the need for intervention by managers.

Site-specific risk assessment of contaminated areas indicates prior areas for intervention, and provides helpful information for risk managers. This study was conducted in the Ervedosa mine area (Bragança, Portugal), where both underground and open pit exploration of tin and arsenic minerals were performed for about one century (1857 – 1969). We aimed at obtaining ecotoxicological information with terrestrial and aquatic plant species to integrate in the risk assessment of this mine area. Further we also intended to evaluate if the assessment of other parameters, in standard assays with terrestrial plants, can improve the identification of phytotoxic soils. For this purpose, soil samples were collected on 16 sampling sites distributed along four transects, defined within the mine area, and in one reference site. General soil physical and chemical parameters, total and extractable metal contents were analyzed. Assays were performed for soil elutriates and for the whole soil matrix following standard guidelines for growth inhibition assay with Lemna minor and emergence and seedling growth assay with Zea mays. At the end of the Z. mays assay, relative water content, membrane permeability, leaf area, content of photosynthetic pigments (chlorophylls and carotenoids), malondialdehyde levels, proline content, and chlorophyll fluorescence (Fv/Fm and ΦPSII) parameters were evaluated. In general, the soils near the exploration area revealed high levels of Al, Mn, Fe and Cu. Almost all the soils from transepts C, D and F presented total concentrations of arsenic well above soils screening benchmark values available. Elutriates of several soils from sampling sites near the exploration and ore treatment areas were toxic to L. minor, suggesting that the retention function of these soils was seriously compromised. In Z. mays assay, plant performance parameters (other than those recommended by standard protocols), allowed the identification of more phytotoxic soils. The results suggest that these parameters could improve the sensitivity of the standard assays.

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