High-biomass crops can be considered as an alternative to hyperaccumulator plants to phytoremediate soils contaminated by heavy metals. In order to assess their practical capability for the absorption and accumulation of Hg in shoots, barley, white lupine, lentil, and chickpea were tested in pot experiments using several growth substrates. In the first experimental series, plants were grown in a mixture of vermiculite and perlite spiked with 8.35 microg g(-1) d.w. of soluble Hg. The mercury concentration of the plants' aerial tissues ranged from 1.51 to 5.13 microg g(-1) d.w. with lentil and lupine showing the highest values. In a second experiment carried out using a Hg-polluted soil (32.16 microg g(-1) d.w.) collected from a historical mining area (Almadén, Spain), the crop plants tested only reached shoot Hg concentration up to 1.13 microg g(-1) d.w. In the third experimental series, the Almadén soil was spiked with 1 microg g(-1) d.w. of soluble Hg; as a result, mercury concentrations in the plant shoots increased approximately 6 times for lupine, 5 times for chickpea, and 3.5 times for barley and lentil, with respect to those obtained with the original soil without Hg added. This marked difference was attributed to the low availability of Hg in the original Almadin soil and its subsequent increase in the Hg-spiked soil. The low mercury accumulation yields obtained for all plants do not make a successful decontamination of the Almadén soils possible byphytoremediation using crop plants. However, since the crops tested can effectively decrease the plant-available Hg level in this soil, their use could, to some extent, reduce the environmental risk of Hg pollution in the area.

Earthworms are standard test organisms in soil toxicity testing. They have been broadly used to assess environmental impact from heavy metal pollution; however, the knowledge on toxic effects from pesticides upon these organisms is still very limited. One of the ecotoxicological approaches to assess pollutant bioavailability and sublethal effects is the use of molecular and biochemical biomarkers. This review focuses on five issues that need further investigation: 1) field validation of earthworm biomarkers of pesticide exposure (e.g., cholinesterases) as well as testing and development in earthworms of those biomarkers of pesticide exposure currently used in other organisms (e.g., carboxylesterases), 2) the impact of environmental and biological interfering factors upon biomarker responses, 3) the development of biomarker-based approaches to assess long-term pesticide exposure, and 4) the need to develop biomarkers of behavioural and reproductive disruption with direct implications at individual and population levels.

An in vitro study was conducted to evaluate the ability of pyridine-2-aldoxime methochloride (2-PAM) to recover organophosphorus (OP)-inhibited cholinesterase (ChE) activity of two earthworm species (Eisenia fetida and Lumbricus terrestris). After inhibition of ChE activity by OP pesticides, an alkyl group may be released from the OP-ChE complex. This reaction is termed aging, and the esterase cannot be reactivated either spontaneously or by the action of reactivating agents, such as 2-PAM. We also examined the aging kinetics of OP-inhibited ChE activity to evaluate the suitability of 2-PAM reactivation methodology for field monitoring. A 2-PAM concentration of 5 x 10(-4) M was enough to reactivate the OP-inhibited ChE activity after 60 min of incubation at 25 degrees C. Chemical reactivation kinetics followed an exponential rise to a maximum of 70 to 80% of normal enzyme activity when ChEs were inhibited with methyl paraoxon or dichlorvos and up to 60% for the chlorpyrifos-inhibited ChE of E. fetida. The aging rates (ka) of the inhibited ChEs were strongly affected by the OP type, and these rates decreased for both earthworm species in the following order: Methyl paraoxon (ka = 0.023-0.033/h) > dichlorvos (ka = 0.008-0.009/h) > chlorpyrifos oxon (ka = 0.003-0.006/h). In particular, chlorpyrifos-inhibited ChE activity of L. terrestris aged slowly (median aging time, 190 h), which means that chemical reactivation of esterase activity with 2-PAM seems feasible one week after exposure to OP pesticides. We conclude that reactivation of earthworm ChE activity by treatment with 2-PAM is a complementary and specific methodology for assessing exposure to OP pesticides.

The efficiency of white lupine (Lupinus albus) to uptake and accumulate mercury from a soil polluted by mining activities was assessed in a pot experiment with chemically assisted phytoextraction. The mobilizing agents tested were ethylenediaminetetracetic acid (EDTA) and hydrochloric acid (HCl). Two doses of each amendment were used (0.5 and 1.0 g of amendment per kg of soil), and unamended pots were used as a control. Addition of HCl to the soil did not negatively affect plant biomass, while the use of EDTA led to a significant decrease in plant growth when compared to that found for non-treated pots, with plants visually showing symptoms of toxicity. The addition of hydrochloric acid increased root, shoot and total plant Hg uptake of white lupine by 3.7 times, 3.1 times and 3.5 times, respectively, in relation to non-amended plants. The greatest efficiency was obtained for the highest HCl dose. EDTA led to higher concentrations of total plant Hg than that found with the control, but, due to the aforementioned decrease in plant biomass, the Hg phytoextraction yield was not significantly increased. These results were attributed to the capability of both amendments to form stable Hg complexes. The concentration of Hg in the water of the soil pores after the phytoextraction experiment was very low for all treatments, showing that risks derived from metal leaching could be partially avoided by using doses and chemicals suitable to the concentration of metal in the soil and plant performance.

Single extraction methods have been extensively used to assess the availability of metals in polluted soils. This work focused on checking the feasibility of several chemicals, i.e. CaCl, EDTA, diethylenetriaminepentaacetic acid (DTPA) and a low-molecular-weight organic acid mixture (rhizosphere-based method), to be used as extractants for mercury (Hg) in a soil from the Almadén mining district (Spain). Moreover, the effect of several experimental parameters, i.e. extraction time (0.5, 1, 2, 5, 16 and 24 h), concentration of extractant (0.01, 0.05, 0.1 and 1 M) and soil/extractant ratio (1:2, 1:5 and 1:10), on the amount of Hg extracted was investigated. The Hg extraction ability followed the descending order EDTA > rhizosphere-based method > DTPA ≈ CaCl. This ranking was attributed to the higher complexation power of EDTA and organic acids. It was also found that extraction times between 2 and 5 h are required to avoid underestimation of mobile Hg and re-adsorption of the Hg previously extracted. Although some exceptions were found, Hg extraction efficiency was generally enhanced with higher extractant concentrations. Finally, the amount of Hg extracted by the four extractants increased with decreasing soil/extractant ratios.