Maria de Lurdes Simões Gonçalves

Metal ion binding of copper(II), zinc(II) and lead(II) by the alga Selenastrum capricornutum printz

Abstract From titration curves of formaldehyde-treated cells of the alga Selenastrum capricornutum with metals, specific adsorption was determined and quantified in terms of average surface complex formation constants and differential equilibrium functions. The values obtained are compared with those obtained in the same conditions for the bacterium Klebsiella pneumoniae. The same analytical experiment allowed the determination of Cu(II) bound to the solute exudate ligands.

Cadmium accumulation and antioxidative defences in Brassica juncea L. Czern, Nicotiana tabacum L. and Solanum nigrum L.

Remediation of sites contaminated with heavy metals using hyperaccumulators seems a promising alternative to engineering approaches. In this work, we compared cadmium (Cd) accumulation and tolerance (based on responses to oxidative stress) in three different species, Brassica juncea (L.) Czern, Nicotiana tabacum L. and Solanum nigrum L., described in the literature as very tolerant or even as hyperaccumulators. The plants were grown in soil spiked with different Cd concentrations (0 – 35 mg kg−1) over a period of 90 days. The translocation factor (TF), used to measure the effectiveness to translocate Cd from roots to shoots, depended greatly on the species. N. tabacum was the plant which exhibited the highest TF values. It was the only plant under study that fulfilled the conditions of a hyperaccumulator for all levels of soil contamination. On the other hand, S. nigrum presented the highest Cd concentration in plant tissues, with TF >1 in the presence of 5 mg Cd kg−1 of soil. Although B. juncea had presented the lowest TF and Cd concentrations, it was the only plant with TF values increasing with the level of cadmium. Oxidative stress in plants was evaluated by lipid peroxidation and activities of catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and superoxide dismutase (SOD), both in roots and shoots. A significant enhancement (versus control) on lipid peroxidation and enzymatic activity of CAT and APX in shoots of B. juncea, N. tabacum and S. nigrum was observed for the highest levels of Cd in soil, 15 and/or 35 mg Cd kg−1. B. juncea presented the most sensitive response of GPX, for all levels of Cd in soil. Lipid peroxidation and CAT activity were greater in shoots than in roots for all plants and soil Cd concentrations. SOD activity did not present consistent trends for any plant.

Depuration kinetics of paralytic shellfish toxins in Mytilus galloprovincialis exposed to Gymnodinium catenatum: laboratory and field experiments.

The kinetics of paralytic shellfish toxins in Mytilus galloprovincialis, previously exposed to Gymnodinium catenatum, was studied under depuration laboratory conditions and over a declining bloom of the dinoflagellate in the field. The variation of the levels observed throughout the laboratory experiment was characterized by a fast depuration of B1, C1 + 2, dcSTX and dcGTX2 + 3, possibly due to the gut evacuation of unassimilated toxins or microalgae cells, or loss during digestive mechanisms. Subsequent enhancements were observed for all compounds with emphasis to dcSTX and dcGTX2 + 3, pointing to biotransformation of the assimilated toxins. Then levels decreased gradually. A first-order depuration kinetic model fitted well to the decrease of B1, C1 + 2 and dcGTX2 + 3 concentrations, but not for dcSTX. Mussels exposed to a declining bloom of Gymnodinium catenatum exhibited a loss of toxins following the same pattern. Despite the low abundance of this dinoflagellate, a similar kinetic model was applied to the field data. The depuration rate of dcGTX2 + 3 in the field experiment (0.153 ± 0.03 day(-1)) significantly exceeded the value calculated in the laboratory (0.053 ± 0.01 day(-1)), while smaller differences were obtained for B1 (0.071 ± 0.02 and 0.048 ± 0.01 day(-1)) and similar values for C1 + 2 (0.082 ± 0.03 and 0.080 ± 0.03 day(-1)). The slower depuration rate of dcGTX2 + 3 in the heavily contaminated mussels at the laboratory may be related to a more effective contribution of C1 + 2 biotransformation.

Comparison of cadmium-induced oxidative stress in Brassica juncea in soil and hydroponic cultures

AimsThe objective of this study was to investigate the response of Brassica juncea in the presence of Cd, in hydroponic and soil experiments, and to conclude about common and divergent trends in both cultures.MethodsWe studied the effect of Cd on growth, oxidative damage and antioxidant responses in roots and shoots of B. juncea grown in soil and hydroponic cultures, using typical time-scales for each one. Major ROS-scavenging enzymes such as catalase, ascorbate peroxidase and guaiacol peroxidase were evaluated, as well as lipid peroxidation.ResultsSmall Cd concentrations in the plant led to enhanced plant growth, while large Cd concentrations impaired growth. The increase in lipid peroxidation observed in the presence of Cd was always greater in shoots than in roots. The physiological response to enhanced levels of reactive oxygen species in the presence of Cd included an increase in guaiacol peroxidase, ascorbate peroxidase and catalase activities, but those enzymes were not always enhanced in a similar manner in both cultures.ConclusionsThe main factors responsible for the differences between the experiments in soil and nutrient solution, were the different Cd content in the plant tissues and the different time-scale of the experiments.

Variations of Mn, Fe and S concentrations in sediment pore waters of Ria Formosa at different time scales

Intertidal sediments of Ria Formosa have been surveyed at different time scales. Pore water of short sediment cores was collected bimonthly at five stations, from May 1993 to June 1994, around low-tide. One location was studied intensively every 1.5 hour, during a 6-hour air-exposure period, and during 20 minutes (1, 5, 10 and 20 minutes) immediately after tidal water had inundated the sampling site. Determinations of vertical profiles of pH, Eh, water content, O2, total dissolved Mn (Mndiss), total dissolved Fe (Fediss) and inorganic sulphur [(HS−)t] were carried out in all samples. The results obtained indicate that daily variations of Mndiss, Fediss and (HS−)t concentrations appear to be superimposed to the seasonal ones. Probably, the renewal of the pore water at periods of tidal inundation causes advective transport of manganese and exchange of iron, between the pore water and the solid fraction of the sediment that masks seasonal fluctuations induced by temperature and organic matter input variations.

Sorghum detoxification mechanisms

ABSTRACT The influence of cadmium (Cd) on internal and external defense mechanisms of Sorghum bicolor (L.) Moench. x Sorghum sudanense was studied by electrochemical methods to infer the type of detoxification processes developed by the plant under environmental stress conditions. Hydroponic experiments with sorghum were conducted in nutrient solutions in the absence and presence of 0.1 mg Cd L−1. Plant exposure to Cd stimulated the release of root exudates with metal-binding affinity. However, their presence should not affect significantly the bioavailability of Cd, since the complex was dissociated within a very short period (≤ 50 milliseconds) in response to the consumption of the free metal ion at the root interface. The presence of Cd in the solution also stimulated a very significant increase of thiolic groups inside the plant, even at the very low concentration of 0.1 mg Cd L−1. These results suggest that the main defense mechanism developed by sorghum against metal toxicity is an internal process, i.e., the synthesis of phytochelatins.

Comparative study between probe focussed sonication and conventional stirring in the evaluation of cadmium and copper in plants.

AbstractUltrasound (US)-assisted extraction has been widely used for metal ion extraction in plants due to its unique properties of decreased extraction time, minimal contamination, low reagent consumption and low cost. However, very few papers present a sound comparison between probe-focussed sonication and conventional stirring in the evaluation of metal ion extraction in plants. In this study, ultrasonic-assisted digestion has been evaluated and compared to magnetic stirring for total copper and cadmium determination by atomic absorption spectrometry in biological samples (plants, plankton and mussels). The same experimental conditions of sample amount and particle size, extractant solution and extraction time were applied for both ultrasound and magnetic stirring-assisted extraction methods in order to truly compare their effect on metal ion solubilisation. To gain further insight in this issue, dried and fresh plants were tested. The results obtained indicated that osmotic tension in cell walls, produced when dried and powdered samples were immersed in the extractant solution, had an important contribution to metal ion solubilisation, the enhancement due to US for the same purpose being negligible. FigureSorghum in soil experiments with different levels of cadmium.

Improvement in soil and sorghum health following the application of polyacrylate polymers to a Cd-contaminated soil

Contamination of soils with cadmium (Cd) is a serious global issue due to its high mobility and toxicity. We investigated the application of insoluble polyacrylate polymers to improve soil and plant health. Sorghum was grown in a Cd-contaminated sandy soil. Polyacrylate polymers at 0.2% (w/w) were added to half of the soil. Control soil without plants was also included in the experiment. Growth of sorghum was stimulated in the polymer-amended soil. The concentration of Cd in the shoots, and the activities of catalase and ascorbate peroxidase decreased in plants from polymer-amended soil compared with unamended control. The amount of CaCl(2)-extractable Cd in the polymer-amended soil was 55% of that in the unamended soil. The Cd extracted in sorghum shoots was 0.19 mg per plant grown on soil without polymer and 0.41 mg per plant grown on polymer-amended soil. The total amount of Cd removed from each pot corresponded to 1.5 and more than 6% of soil CaCl(2)-extractable Cd in unamended and polymer-amended soil, respectively. The activities of soil acid phosphatase, beta-glucosidase, urease, protease and cellulase were greatest in polymer-amended soil with sorghum. In conclusion, the application of polyacrylate polymers to reduce the bioavailable Cd pool seems a promising method to enhance productivity and health of plants grown on Cd-contaminated soils.

Determination of nickel, calcium and magnesium in xylem sap by flame atomic absorption spectrometry using a microsampling technique.

INTRODUCTION Knowledge of xylem sap chemical composition is important to the understanding of translocation, detoxification and tolerance mechanisms. However, the small amount of sample available often hampers its characterisation. Hence, low volume consumption techniques are needed for xylem sap analysis. OBJECTIVE To develop a microsampling technique for the determination of elements in xylem sap from different plants by flame atomic absorption spectrometry (FAAS). METHODOLOGY The microsampling device was optimised in terms of sample volume and integration time. The analytical characteristics of the microsampling technique (micro-FAAS) were established and compared with those of FAAS with traditional continuous nebulisation. The method was validated by means of an independent technique. RESULTS Ca, Mg and Ni were determined in a 50 microL aliquot of xylem sap solution/element that was introduced directly into the flame via the microsampling accessory. Good precision was obtained with relative standard deviations of 1.1, 0.6 and 2.3% for Ca, Mg and Ni, respectively. Matrix effects resulting from the physical characteristics of the samples and possible chemical interferences caused by phosphate and/or sulphate were ruled out. CONCLUSION A simple, rapid and reproducible microsampling technique coupled to FAAS was developed and successfully applied in the determination of Ca, Mg and Ni in xylem sap.

Competition of copper and/or calcium in nickel-pectin interaction by potentiometric and voltammetric techniques.

The interaction between nickel and pectin extracted from citrus fruit was studied in 0.10 M KNO(3), at pH 5.5 and 25 degrees C. Differential pulse and/or square wave polarography were used to determine free nickel. For a high coverage degree (theta) of the pectin by the metal ion a good fitting was observed between experimental results and the model that includes both complex species, ML and ML(2) (M for the metal ion and L for the ligand). In the ML(2) species, Ni(II) interacts with two carboxylate groups of different chains, resulting in an inter-chain association. For low theta values, the formation of ML(2) is hindered due to the repulsion between the negative charges of carboxylic groups in two independent segments of pectin. The influence of calcium or copper ions on the free nickel concentration, in the presence of pectin, may lead to a decrease in free nickel concentration, contrary to what would be expected from direct competition between Ca(II) or Cu(II) and Ni(II) for the pectin binding sites. This is due to the partial neutralisation of the negative carboxylic charges by the positive charges of the divalent cations, which favours NiL(2) formation through the association of independent chains.