Microcystins are cyanobacterial hepatotoxins capable of accumulation into animal tissues. The toxins act by inhibiting specific protein phosphatases and both non-covalent and covalent interactions occur. The 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB) method determines the total, i.e. the sum of free and protein-bound microcystin in tissues. The aim of the method development in this paper was to tackle the problems with the MMPB methodology: the rather laborious workflow and the loss of material during different steps of the method. In the optimised workflow the oxidation recovery was of acceptable level (29–40%), the extraction efficiency good (62–97%), but the signal suppression effect from the matrix remained severe in our system (16–37% signal left). The extraction efficiency for the determination of the free, extractable microcystins, was found to be good, 52–100%, depending on the sample and the toxin variant and concentration. The study concerns method development for the LC–MS–MS analysis of both free and protein-bound microcystin in tissue materials.

International audience | Microcystins are cyanobacterial hepatotoxins capable of accumulation into animal tissues. The toxins act by inhibiting specific protein phosphatases and both non-covalent and covalent interactions occur. The 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB) method determines the total, i.e. the sum of free and protein-bound microcystin in tissues. The aim of the method development in this paper was to tackle the problems with the MMPB methodology: the rather laborious workflow and the loss of material during different steps of the method. In the optimised workflow the oxidation recovery was of acceptable level (29-40%), the extraction efficiency good (62-97%), but the signal suppression effect from the matrix remained severe in our system (16-37% signal left). The extraction efficiency for the determination of the free, extractable microcystins, was found to be good, 52-100%, depending on the sample and the toxin variant and concentration.

Benthic invertebrates can uptake metals through diffusion of free ion solutes, or ingestion of sediment-bound forms. This study investigated the efficacy of the metal chelating resin SIR 300™ in adsorbing porewater metals and isolating pathways of metal exposure. A field experiment (Botany Bay, Sydney, Australia) and a laboratory toxicity test each manipulated the availability of porewater metals within contaminated and uncontaminated sediments. It was predicted that within contaminated sediments, the resin would adsorb porewater metals and reduce toxicity to invertebrates, but in uncontaminated sediments, the resin would not significantly affect these variables. Whereas in the laboratory, the resin produced the predicted results, in the field the resin increased porewater metal concentrations of contaminated sediments for at least 34 days and decreased abundances of four macroinvertebrate groups, and richness in all sediments. These contrasting findings highlight the limits of extrapolating the results of laboratory experiments to the field environment.

Benthic invertebrates can uptake metals through diffusion of free ion solutes, or ingestion of sediment-bound forms. This study investigated the efficacy of the metal chelating resin SIR 300 in adsorbing porewater metals and isolating pathways of metal exposure. A field experiment (Botany Bay, Sydney, Australia) and a laboratory toxicity test each manipulated the availability of porewater metals within contaminated and uncontaminated sediments. It was predicted that within contaminated sediments, the resin would adsorb porewater metals and reduce toxicity to invertebrates, but in uncontaminated sediments, the resin would not significantly affect these variables. Whereas in the laboratory, the resin produced the predicted results, in the field the resin increased porewater metal concentrations of contaminated sediments for at least 34 days and decreased abundances of four macroinvertebrate groups, and richness in all sediments. These contrasting findings highlight the limits of extrapolating the results of laboratory experiments to the field environment.

Gene expression responses of paper birch (Betula papyrifera) leaves to elevated concentrations of CO2 and O3 were studied with microarray analyses from three time points during the summer of 2004 at Aspen FACE. Microarray data were analyzed with clustering techniques, self-organizing maps, K-means clustering and Sammon's mappings, to detect similar gene expression patterns within sampling times and treatments. Most of the alterations in gene expression were caused by O3, alone or in combination with CO2. O3 induced defensive reactions to oxidative stress and earlier leaf senescence, seen as decreased expression of photosynthesis- and carbon fixation-related genes, and increased expression of senescence-associated genes. The effects of elevated CO2 reflected surplus of carbon that was directed to synthesis of secondary compounds. The combined CO2 + O3 treatment resulted in differential gene expression than with individual gas treatments or in changes similar to O3 treatment, indicating that CO2 cannot totally alleviate the harmful effects of O3. Clustering analysis of birch leaf gene expression data reveals differential responses to O3 and CO2.

This study examines the food-chain transfer of Zn from two plant species, Urtica dioica (stinging nettle) and Acer pseudoplatanus (sycamore maple), into their corresponding aphid species, Microlophium carnosum and Drepanosiphum platanoidis. The plants were grown in a hydroponic system using solutions with increasing concentrations of Zn from 0.02 to 41.9 mg Zn/l. Above-ground tissue concentrations in U. dioica and M. carnosum increased with increasing Zn exposure (p < 0.001). Zn concentrations in A. pseudoplatanus also increased with solution concentration from the control to the 9.8 mg Zn/l solution, above which concentrations remained constant. Zn concentrations in both D. platanoidis and the phloem tissue of A. pseudoplatanus were not affected by the Zn concentration in the watering solution. It appears that A. pseudoplatanus was able to limit Zn transport in the phloem, resulting in constant Zn exposure to the aphids. Zn concentrations in D. platanoidis were around three times those in M. carnosum. Concentrations of Zn in two aphid species are dependant on species and exposure.

This study examines the food-chain transfer of Zn from two plant species, Urtica dioica (stinging nettle) and Acer pseudoplatanus (sycamore maple), into their corresponding aphid species, Microlophium carnosum and Drepanosiphum platanoidis. The plants were grown in a hydroponic system using solutions with increasing concentrations of Zn from 0.02 to 41.9 mg Zn/l. Above-ground tissue concentrations in U. dioica and M. carnosum increased with increasing Zn exposure (p < 0.001). Zn concentrations in A. pseudoplatanus also increased with solution concentration from the control to the 9.8 mg Zn/l solution, above which concentrations remained constant. Zn concentrations in both D. platanoidis and the phloem tissue of A. pseudoplatanus were not affected by the Zn concentration in the watering solution. It appears that A. pseudoplatanus was able to limit Zn transport in the phloem, resulting in constant Zn exposure to the aphids. Zn concentrations in D. platanoidis were around three times those in M. carnosum. Concentrations of Zn in two aphid species are dependant on species and exposure.

Photosynthetic acclimation under elevated carbon dioxide (CO2) and/or ozone (O3) has been the topic of discussion in many papers recently. We examined whether or not aspen plants grown under elevated CO2 and/or O3 will acclimate after 11 years of exposure at the Aspen Face site in Rhinelander, WI, USA. We studied diurnal patterns of instantaneous photosynthetic measurements as well as A/Ci measurements monthly during the 2004-2008 growing seasons. Our results suggest that the responses of two aspen clones differing in O3 sensitivity showed no evidence of photosynthetic and stomatal acclimation under either elevated CO2, O3 or CO2 + O3. Both clones 42E and 271 did not show photosynthetic nor stomatal acclimation under elevated CO2 and O3 after a decade of exposure. We found that the degree of increase or decrease in the photosynthesis and stomatal conductance varied significantly from day to day and from one season to another. We report of no evidence of photosynthetic and stomatal acclimation in aspen trees grown under elevated CO2 and O3 after over a decade of exposure.

Linear Alkylbenzene Sulfonates (LAS) effects (mortality, egestion rate, behaviour) on the marine deposit feeder Hydrobia ulvae were assessed in whole-sediment and water-only systems. The results were combined with a bioenergetic-based kinetic model of exposure pathways to account for the observed toxicity. The 10-d LC50 value based on the freely dissolved fraction was 9.3 times lower in spiked sediment (0.152 ± 0.001 (95% CI) mg/L) than in water-only (1.390 ± 0.020 (95% CI) mg/L). Consequently, the actual 10-d LC50 value (208 mg/kg) was overestimated by the Equilibrium Partitioning calculation (1629 mg/kg). This suggests that the sediment associated LAS fraction was bioavailable to the snails. It could also be due to modifications in physiological parameters in absence of sediment, the organism natural substrate. Lethality of the marine gastropod deposit feeder Hydrobia ulvae exposed to LAS in water-only system was inappropriate to predict LAS toxicity in sediment system.

7 páginas, 4 figuras, 4 tablas. | Linear Alkylbenzene Sulfonates (LAS) effects (mortality, egestion rate, behaviour) on the marine deposit feeder Hydrobia ulvae were assessed in whole-sediment and water-only systems. The results were combined with a bioenergetic-based kinetic model of exposure pathways to account for the observed toxicity. The 10-d LC50 value based on the freely dissolved fraction was 9.3 times lower in spiked sediment (0.152 ± 0.001 (95% CI) mg/L) than in water-only (1.390 ± 0.020 (95% CI) mg/L). Consequently, the actual 10-d LC50 value (208 mg/kg) was overestimated by the Equilibrium Partitioning calculation (1629 mg/kg). This suggests that the sediment associated LAS fraction was bioavailable to the snails. It could also be due to modifications in physiological parameters in absence of sediment, the organism natural substrate. | This research was supported by the European Commission through the FP6 program (EST Marie Curie Fellowship) and it was performed in the framework of the RISICO project (MEST-CT-2004-505182). We also acknowledge the financial support of ERASM, ECOSOL and PETRESA. | Peer reviewed

The purpose of this work is to assess the effectiveness of two grass covers (buffer zone and grass-covered inter-row), to reduce pesticide leaching, and subsequently to preserve groundwater quality. Lower amounts of pesticides leached through grass-cover soil columns (2.7-24.3% of the initial amount) than the bare soil columns (8.0-55.1%), in correspondence with their sorption coefficients. Diuron was recovered in higher amounts in leachates (8.9-32.2%) than tebuconazole (2.7-12.9%), in agreement with their sorption coefficients. However, despite having a sorption coefficient similar to that of diuron, more procymidone was recovered in the leachates (10.2-55.1%), probably due to its facilitated transport by dissolved organic matter. Thus even in this very permeable soil, higher organic matter contents associated with grass-cover reduce the amount of pesticide leaching and limit the risk of groundwater contamination by the pesticides. The results of diuron and tebuconazole transfer through undisturbed buffer zone soil columns are in agreement with field observations on the buffer zone.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]ARCEAU | The purpose of this work is to assess the effectiveness of two grass covers (buffer zone and grass-covered inter-row), to reduce pesticide leaching, and subsequently to preserve groundwater quality. Lower amounts of pesticides leached through grass-cover soil columns (2.7e24.3% of the initial amount) than the bare soil columns (8.0e55.1%), in correspondence with their sorption coefficients. Diuron was recovered in higher amounts in leachates (8.9e32.2%) than tebuconazole (2.7e12.9%), in agreement with their sorption coefficients. However, despite having a sorption coefficient similar to that of diuron, more procymidone was recovered in the leachates (10.2e55.1%), probably due to its facilitated transport by dissolved organic matter. Thus even in this very permeable soil, higher organic matter contents associated with grass-cover reduce the amount of pesticide leaching and limit the risk of groundwater contamination by the pesticides. The results of diuron and tebuconazole transfer through undisturbed buffer zone soil columns are in agreement with field observations on the buffer zone.

Betula papyrifera trees were exposed to elevated concentrations of CO2 (1.4 × ambient), O3 (1.2 × ambient) or CO2 + O3 at the Aspen Free-air CO2 Enrichment Experiment. The treatment effects on leaf surface characteristics were studied after nine years of tree exposure. CO2 and O3 increased epidermal cell size and reduced epidermal cell density but leaf size was not altered. Stomatal density remained unaffected, but stomatal index increased under elevated CO2. Cuticular ridges and epicuticular wax crystallites were less evident under CO2 and CO2 + O3. The increase in amorphous deposits, particularly under CO2 + O3, was associated with the appearance of elongated plate crystallites in stomatal chambers. Increased proportions of alkyl esters resulted from increased esterification of fatty acids and alcohols under elevated CO2 + O3. The combination of elevated CO2 and O3 resulted in different responses than expected under exposure to CO2 or O3 alone. The combined effects of CO2 and O3 on birch leaf surface characteristics cannot be predicted on the basis of studies examining each of these gases separately.