The earthworm Lumbricus rubellus (Hoffmeister, 1843) is a terrestrial pollution sentinel. Enzyme activity and transcription of phase II detoxification superfamily glutathione transferases (GST) is known to respond in earthworms after soil toxin exposure, suggesting GST as a candidate molecular-based pollution biomarker. This study combined sub-proteomics, bioinformatics and biochemical assay to characterise the L. rubellus GST complement as pre-requisite to initialise assessment of the applicability of GST as a biomarker. L. rubellus possesses a range of GSTs related to known classes, with evidence of tissue-specific synthesis. Two affinity-purified GSTs dominating GST protein synthesis (Sigma and Pi class) were cloned, expressed and characterised for enzyme activity with various substrates. Electrospray ionisation mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) following SDS-PAGE were superior in retaining subunit stability relative to two-dimensional gel electrophoresis (2-DE). This study provides greater understanding of Phase II detoxification GST superfamily status of an important environmental pollution sentinel organism. This study currently provides the most comprehensive view of the Phase II detoxification enzyme superfamily of glutathione transferases within the important environmental pollution sentinel earthworm Lumbricus rubellus.

Atmospheric sampling of organochlorine pesticides (OCPs) was conducted using Semi Permeable Membrane Devices (SPMDs) deployed in the Alps at different altitudinal transects for two consecutive exposure periods of half a year and a third simultaneous year-long period. Along all the altitude profiles, the sequestered amounts of OCPs increased in general with altitude. SPMDs were still working as kinetic samplers after half a year for the majority of the OCPs. However, compounds with the lowest octanol-air partition coefficient (Koa), reached equilibrium within six months. This change in the SPMD uptake was determined for the temperature gradient along the altitude profile influencing Koa, OCPs availability in the gaseous phase, and SPMD performance. In sum, it seems two effects are working in parallel along the altitude profiles: the change in SPMD performance and the different availability of OCPs along the altitudinal transects determined by their compound properties and concentrations in air. SPMDs were in different uptake stages regarding OCPs, as they were influenced by the temperature (season, triolein state, and altitude) and Koa.

Guidelines provided by the OECD and EPPO allow the use of single-species tests performed in greenhouses to assess the risk of herbicides to non-target terrestrial plant communities in the field. The present study was undertaken to investigate the use of greenhouse data to determine effects of herbicides with a different mode of action on the biomass, seed production and emergence of field-grown plants. In addition, a single species approach was compared with a mixed species approach. Effects on the biomass of greenhouse and field-grown plants were found to be related at different effect levels, indicating that it might be possible to translate results from greenhouse studies to field situations. However, the use of single-species tests may not be valid. The response of a single plant species to sublethal herbicide dosages differed to the response of the same species grown in a mixture with other species. The use of single-species greenhouse tests in the ecological risk assessment of crop protection products may only be valid for single species in the field, not for vegetations. Keywords: Non-crop terrestrial plants; Tepraloxydim; Greenhouse; Field; Biomass

This paper evaluates approaches to calculate acceptable loads for metal deposition to forest ecosystems, distinguishing between critical loads, stand-still loads and target loads. We also evaluated the influence of including the biochemical metal cycle on the calculated loads. Differences are illustrated by examples of Cd, Cu, Pb and Zn for a deciduous forest on five major soil types in the Netherlands. Stand-still loads are generally lower than critical loads, which in turn are lower than the target loads indicating that present levels are below critical levels. Uncertainties in the calculated critical loads are mainly determined by the uncertainty in the critical limits and the chemical speciation model. Including the metal cycle has a small effect on the calculated critical loads. Results are discussed in view of the applicability of the critical load concept for metals in future protocols on the reduction in metal emissions. Critical load methods for metals can be used to assess future risks due to metal inputs.

Although ecotoxicological data on heavy metals are abundant, information on other potentially toxic elements with attributed deficiency and/or toxic disturbances is scarce. Here we quantify zinc, copper, iron, manganese, chromium, molybdenum, strontium, barium, and boron in bones of greater white-toothed shrews, Crocidura russula, inhabiting two protected Mediterranean coastal sites: the Ebro Delta, a wetland impacted by human activities, and the Medas Islands, a reference site. Natural and anthropogenic inputs significantly increase Fe, Mn, Mo, Sr, Ba, and B in specimens from the Ebro Delta, whereas Cu and Cr were higher in Medas' shrews. Principal component analysis allowed complete separation between sites along the first two axes in particular due to B, Sr, and Cu. This study provides metal reference values in bones of insectivores, explores their variability and bioaccumulation patterns in depth, and assesses the potential environmental risk and toxicity for biota exposed to the above elements. Sr, Ba, Fe, Mn, Mo and B were bioaccumulated in bones of shrews from the Ebro Delta area and Cu in Medas Islands, whereas Cr and Zn showed similar levels at both sites.

Molluscs are sensitive species to the toxic effects of organotin compounds, particularly to masculinisation. Both tributyltin (TBT) and triphenyltin (TPT) have been recently shown to bind to mollusc retinoid X receptor (RXR). If RXR is involved in lipid homeostasis, exposure to TPT would have an immediate effect on endogenous lipids. To test this hypothesis, the ramshorn snail Marisa cornuarietis was exposed to environmentally relevant concentrations of TPT (30, 125, 500 ng/L as Sn) in a semi-static water regime for 7 days. Percentage of lipids and total fatty acid content decreased significantly in TPT-exposed females while the activity of peroxisomal acyl-CoA oxidase, involved in fatty acid catabolism, increased. In addition, fatty acid profiles (carbon chain length and unsaturation degree) were significantly altered in exposed females but not in males. This work highlights the ability of TPT to disrupt lipid metabolism in M. cornuarietis at environmentally realistic concentrations and the higher susceptibility of females in comparison to males. Short-term exposure to the fungicide TPT disrupts lipid metabolism in M. cornuarietis at environmentally realistic concentrations.

The troposphere is subject to continuous inputs, production and removal processes of ozone and its precursors from natural processes and human activities acting together within a very complex system. In order to assess the behaviour of background ozone in the Mediterranean area, a description of trends, seasonal and diurnal behaviours of free tropospheric ozone is provided. In the Mediterranean area and southern Europe the background tropospheric ozone concentration appears significantly affected by three main air mass transport processes: (i) transport of polluted air masses on regional and long-range scales, (ii) downward transport of stratospheric air masses, and (iii) transport of mineral dust from the Sahara desert. In this review of the literature of the last two decades, we present an overview of these phenomena, mainly monitored at high baseline mountain stations representative of background atmospheric conditions. How background ozone is influenced by vertical and horizontal transport processes in the southern Europe and the Mediterranean area.

Phenanthrene uptake by Medicago sativa L. was investigated under the influence of an arbuscular mycorrhizal fungus. Inoculation of lucerne with the arbuscular mycorrhizal fungus Glomus etunicatum L. resulted in higher phenanthrene accumulation in the roots and lower accumulation in the shoots compared to non-mycorrhizal controls. Studies on sorption and desorption of phenanthrene by roots and characterization of heterogeneity of mycorrhizal and non-mycorrhizal roots using solid-state 13C nuclear magnetic resonance spectroscopy (13C NMR) demonstrated that increased aromatic components due to mycorrhizal inoculation resulted in enhanced phenanthrene uptake by the roots but lower translocation to the shoots. Direct visualization using two-photon excitation microscopy (TPEM) revealed higher phenanthrene accumulation in epidermal cells of roots and lower transport into the root interior and stem in mycorrhizal plants than in non-mycorrhizal controls. These results provide some insight into the mechanisms by which arbuscular mycorrhizal inoculation may influence the uptake of organic contaminants by plants. Colonization by an arbuscular mycorrhizal fungus promoted root uptake and decreased shoot uptake of phenanthrene by Medicago sativa L.

Soil metal pollution can trigger evolutionary adaptation in soil-borne organisms. An in vitro screening test showed cadmium adaptation in populations of Suillus luteus (L.: Fr.) Roussel, an ectomycorrhizal fungus of pine trees. Cadmium stress was subsequently investigated in Scots pine (Pinus sylvestris L.) seedlings inoculated with a Cd-tolerant S. luteus, isolated from a heavy metal contaminated site, and compared to plants inoculated with a Cd-sensitive isolate from a non-polluted area. A dose-response experiment with mycorrhizal pines showed better plant protection by a Cd-adapted fungus: more fungal biomass and a higher nutrient uptake at high Cd exposure. In addition, less Cd was transferred to aboveground plant parts. Because of the key role of the ectomycorrhizal symbiosis for tree fitness, the evolution of Cd tolerance in an ectomycorrhizal partner such as S. luteus can be of major importance for the establishment of pine forests on Cd-contaminated soils. The evolutionary adaptation for higher Cd tolerance in Suillus luteus, an ectomycorrhizal fungus, is of major importance for the amelioration of Cd toxicity in pine trees exposed to high Cd concentrations.

Plants react to O3 threat by setting up a variety of defensive strategies involving the co-ordinated modulation of stress perception, signalling and metabolic responses. Although stomata largely controls O3 uptake, differences in O3 tolerance cannot always be ascribed to changes in stomatal conductance but cell protective and repair processes should be taken into account. O3-driven ROS production in the apoplast induces a secondary, active, self-propagating generation of ROS, whose levels must be finely tuned, by many enzymatic and non-enzymatic antioxidant systems, to induce gene activation without determining uncontrolled cell death. Additional signalling molecules, as ethylene, jasmonic and salicylic acid are also crucial to determine the spreading and the containment of leaf lesions. The main recent results obtained on O3 sensing, signal transduction, ROS formation and detoxification mechanisms are here discussed. A dissection of the complex network of interacting mechanisms which determine the cell fate under ozone stress.