As part of the Endis-Risks project, the current study describes the occurrence of the chlorotriazine pesticides atrazine, simazine and terbutylazine in water, sediment and suspended matter in the Scheldt estuary (B-Nl) from 2002 to 2005 (3 samplings a year, 8 sampling points). Atrazine was found at the highest concentrations, varying from 10 to 736 ng/l in water and from 5 up to 10 ng/g in suspended matter. Simazine and terbutylazine were detected at lower concentrations. Traces of the targeted pesticides were also detected in sediments, but these were below the limit of quantification. As part of an ecotoxicological assessment, we studied the potential effect of atrazine on molting of Neomysis integer (Crustacea:Mysidacea), a resident invertebrate of the Scheldt Estuary and a proposed test organism for the evaluation of endocrine disruption. Following chronic exposure (3 weeks), atrazine did not significantly affect mysid molting at environmentally relevant concentrations (up to 1 μg/l). The water of the Scheldt estuary and its associated suspended solids are contaminated with chlorotriazines at concentrations that do not affect mysid molting.

Photosynthetic stimulation and stomatal conductance (Gs) depression in Quercus ilex leaves at a CO2 spring suggested no down-regulation. The insensitivity of Gs to a CO2 increase (from ambient 1500 to 2000 μmol mol-1) suggested stomatal acclimation. Both responses are likely adaptations to the special environment of CO2 springs. At the CO2-enriched site, not at the control site, photosynthesis decreased 9% in leaves exposed to 2x ambient O3 concentrations in branch enclosures, compared to controls in charcoal-filtered air. The stomatal density reduction at high CO2 was one-third lower than the concomitant Gs reduction, so that the O3 uptake per single stoma was lower than at ambient CO2. No significant variation in monoterpene emission was measured. Higher trichome and mesophyll density were recorded at the CO2-enriched site, accounting for lower O3 sensitivity. A long-term exposure to H2S, reflected by higher foliar S-content, and CO2 might depress the antioxidant capacity of leaves close to the vent and increase their O3 sensitivity. Very high CO2 concentrations did not compensate for the effects of O3 on holm oak photosynthesis.

The growth of transgenic canola (Brassica napus) expressing a gene for the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase was compared to non-transformed canola exposed to flooding and elevated soil Ni concentration, in situ. In addition, the ability of the plant growth-promoting bacterium Pseudomonas putida UW4, which also expresses ACC deaminase, to facilitate the growth of non-transformed and transgenic canola under the above mentioned conditions was examined. Transgenic canola and/or canola treated with P. putida UW4 had greater shoot biomass compared to non-transformed canola under low flood-stress conditions. Under high flood-stress conditions, shoot biomass was reduced and Ni accumulation was increased in all instances relative to low flood-stress conditions. This is the first field study to document the increase in plant tolerance utilizing transgenic plants and plant growth-promoting bacteria exposed to multiple stressors. Using transgenic plants and plant growth-promoting bacteria as phytoremediation methods increased plant tolerance at a metal-contaminated field site under low flood conditions.

The purpose of this study is to develop a mechanistic model to describe a conceptually new “flux-biological response” approach based on biotic ligand model (BLM) and Michaelis-Menten (M-M) kinetics to allow the linkage between valve closure behavior and sodium (Na) transport mechanism in freshwater clam Corbicula fluminea in response to waterborne copper (Cu). We test the proposed model against published data regarding Na uptake kinetics in rainbow trout and Na uptake profile in C. fluminea, confirming that the predictive model is robust. Here, we show that the predicted M-M maximum Cu internalization flux in C. fluminea is 0.369 μmol g-1 h-1 with a half-saturation affinity constant of 7.87 x 10-3 μM. Dynamics of Na uptake and valve closure daily rhythm driven by external Cu can also be predicted simultaneously. We suggest that this “Na transport-valve closure behavior” approach might provide the basis of a future design of biomonitoring tool. A new flux-biological response model can link valve closure and sodium transport mechanisms in freshwater clam in response to copper.

The use of a vegetation cover for the management of heavy metal contaminated soils needs prior investigations on the plant species the best sustainable. In this work, behaviors of Trifolium repens and Lolium perenne, growing in a metal-polluted field located near a closed lead smelter, were investigated through Cd, Pb and Zn-plant metal concentrations and their phytotoxicity. In these plant species, metals were preferentially accumulated in roots than in shoots, as follow: Cd > Zn > Pb. Plant exposure to such metals induced oxidative stress in the considered organs as revealed by the variations in malondialdehyde levels and superoxide dismutase activities. These oxidative changes were closely related to metal levels, plant species and organs. Accordingly, L. perenne seemed to be more affected by metal-induced oxidative stress than T. repens. Taken together, these findings allow us to conclude that both the plant species could be suitable for the phytomanagement of metal-polluted soils. Usefulness of Trifolium repens and Lolium perenne for the phytomanagement of heavy metal-contaminated soils.

Boron (B) is a widespread environmental contaminant that is mobile relative to other trace elements. We investigated the potential of hybrid poplar (Populus sp.) for B phytomanagement using a lysimeter experiment and a field trial on B-contaminated wood-waste. In both studies, poplars enhanced evapotranspiration from the wood-waste, reduced B leaching, and accumulated B in the aerial portions of the tree. When grown in a substrate containing 30 mg/kg B, poplar leaves had an average B concentration of 845 mg/kg, while the stems contained 21 mg/kg B. Leaf B concentrations increased linearly with leaf age. A decomposition experiment revealed that abscised leaves released 14% of their B during the winter months. Fertiliser application enhanced tree growth without decreasing the leaf B concentrations. Harvesting alternate rows of trees on a contaminated site would reduce leaching from the site while removing B. Harvested plant material may provide bioenergy, stock fodder, or an amendment for B-deficient soils. Poplars reduce boron leaching from contaminated sites.

Tobacco plants transformed with TaLCT1 were cultured on Knop's medium with modified calcium concentrations (0.01-3 mM) in the presence of Pb2+, and in soil contaminated by lead. A 4-5 μM Pb2+ administered in the presence of 1 mM Ca2+ inhibited the root growth of transgenic plants to much lesser degree than of control plants, whereas in the presence of 3 mM Ca2+ no differences were found between the studied lines. The reduction of Pb2+ toxicity in the presence of 1 mM Ca2+ was not accompanied by a change in the lead tissue concentration. However, when Ca2+ level in the medium was lowered to 0.01 mM, several fold higher root/shoot Pb ratio in transgenic plants was observed, twofold increase in the total amount of metal accumulated, and lower concentration of Pb in the xylem sap. Results suggest the involvement of TaLCT1 in the regulation of Ca-dependent Pb-detoxification, and under conditions of low calcium in lead uptake and distribution. Ca2+-dependent Pb2+ detoxification and uptake was regulated by TaLCT1.

Visible ozone symptoms on leaves are expressions of physiological mechanisms to cope with oxidative stresses. Often, the symptoms consist of stippling, which corresponds to localized cell death (hypersensitive response, HR), separated from healthy cells by a layer of callose. The HR strategy tends to protect the healthy cells and in most cases the efficiency of chlorophyll to trap energy is not affected. In other cases, the efficiency of leaves to produce biomass declines and the plant loses its photosynthetic apparatus replacing it with a new, more efficient one. Another strategy consists of the production of pigments (anthocyanins), and leaves become reddish. In these cases, the most significant physiological manifestation consists of the enhanced dissipation of energy. These different behavior patterns are reflected in the initial events of photosynthetic activity, and can be monitored with techniques based on the direct fluorescence of chlorophyll a in photosystem II, applying the JIP-test. Analytical techniques based on the direct fluorescence of chlorophyll a, allow us to discriminate species-specific physiological behavior in relation to ozone air pollution.

An open top chamber experiment was carried out in the summer of 2003 to examine the effect of nitrous acid (HONO) gas on the physiological status of Scots pine saplings (Pinus sylvestris). Four-year-old pine trees were exposed to two different levels of HONO gas (at ca. 2.5 ppb and 5.0 ppb) and a control (filtered air) from early evening to early morning (18:00-6:00), in duplicate open top chambers. Significant decreases in the ratios of chlorophylls a to b, an increase in the carbon to nitrogen (C/N) ratio, and a reduction of maximum yield of PS II (Fv/Fm) in pine needles were also observed after the 2 months' fumigation. Cation contents of pine needles were also decreased by the fumigation with HONO gas. The results could be explained by the harmful effects of OH radicals, generated from photolysis of HONO gas, and/or aqueous phase HONO (NO2-/HONO), on the photosynthetic capacity of pine needles. Exposure to HONO affects photosynthesis and nutrient status of pine trees.

Berkheya coddii Roessler (Asteraceae) is a hyper-accumulator of nickel, which can be used in phytomining and phytoremediation. Chrysolina pardalina Fabricius (Chrysomelidae) is a phytophagous leaf beetle, which may be useful in controlling population levels of B. coddii after it has been introduced into a new habitat. The aim of this study was to investigate the response of C. pardalina to topical application of dimethoate. Data recorded included the activity of acetylcholinesterase (AChE), the concentration of glutathione (GSH), and the activity of selected enzymes connected with GSH metabolism. Assays were carried out several times during the first 24 h after exposure to dimethoate. At the dosages used in this study, dimethoate was not as toxic as expected. AChE activity was significantly decreased 14 and 24 h after application. GST activity was significantly decreased 24 h after application. GSTPx activity was significantly decreased 2, 14 and 24 h after application. GR activity was significantly increased 4 h after application. GSH concentration was significantly increased 24 h after application. Long-term exposure to high levels of nickel may have caused adaptive changes in the enzymes that enable C. pardalina to deal with other stressors, including organophosphate pesticides. Long-term exposure to high levels of nickel may have caused adaptive changes in the enzymes that enable Chrysolina pardalina to deal with other stressors, including organophosphate pesticides.