Does anoxia promote the mobilization of P-bearing colloids from dam reservoir sediment?

In the context of a reservoir, the anoxia that develops in the bottom sediment induces the release of phosphorus (P) into the overlying water, thus supporting eutrophication. Most studies focusing on P dynamic in an aquatic environment fail to consider the "truly" dissolved and colloidal fractions, hence the colloidal P has gone largely unexplored. The aim of this study was to investigate the release of sedimentary P under oscillating aerobic, anoxic and aerobic conditions, in taking into account the colloidal (10 kDa-1 µm) and truly dissolved (< 10 kDa) fractions. Laboratory incubations of wet sediment originating from a dam reservoir were performed over 63 days, consisting of 25 days of aerobic conditioning (lasting 2 periods) and 38 days of anoxia. Results showed that oxic conditions induced a very limited release of phosphorus, both in truly dissolved and colloidal forms. In turn, the development of anoxic conditions caused a large release of P, mainly in the colloidal fraction, representing about 90 % of the total water-mobilizable P (PWM < 1 µm). The initial release of truly dissolved P during the anoxic stage gradually diminished over time, possibly due to the formation of secondary minerals or re-adsorption processes. Approximately half of the PWM released during anoxia persisted under subsequent oxic conditions and consisted solely of colloidal P. The dynamics of PWM were primarily influenced by two main factors: (i) the reductive dissolution of iron, which released both dissolved and colloidal P, and (ii) the release of indigenous organic matter, which impacted the stability of the released colloids through bridging mechanisms.

Are cysteine, glutathione and phytochelatins responses of Myriophyllum alterniflorum to copper and arsenic stress affected by trophic conditions?

The aim of this article is to study the impact of both copper (Cu2+) and arsenic (As (V)) at 100 µg/L, with each element being combined with trophic conditions at the level of glutathione, cysteine and phytochelatins in the aquatic macrophyte Myriophyllum alterniflorum, whose potential for bioindication and phytoremediation of metal/metalloid pollution has already been demonstrated. To achieve this goal, a synthetic medium, of a composition similar to the water found in the Vienne River in France's Limousin Region and modified for eutrophic or oligotrophic conditions, is prepared. The analysis of cysteine, glutathione and phytochelatins is performed at 0, 3, 7, 14 and 21 days. Our results indicate that the eutrophic medium without contaminant only induces a significant increase in the glutathione level when compared to the oligotrophic medium. However, the joint presence of As (V) and Cu is able to increase the synthesis of cysteine, glutathione and phytochelatins (PC2 and PC3) under both eutrophic and oligotrophic conditions, with a significant increase in the eutrophic medium compared to the oligotrophic one. Phytochelatins (PC2 and PC3) are induced after as little as 3 days of exposure to copper and arsenic under both trophic conditions. Copper induces the synthesis of more PC3 than PC2, unlike arsenic. Our results confirm the potential use of phytochelatins as a specific biochemical biomarker for metal/metalloid stress. In conclusion, the eutrophic condition combined with copper or arsenic does change the response of Myriophyllum alterniflorum by enhancing its antioxidative defense. Thus, M. alterniflorum phytochelatins represent a potential dedicated biomarker to monitor water quality in terms of metal/metalloid stress regardless of the trophic level.

In search for potential biomarkers of copper stress in aquatic plants.

Over the last few decades, the use of pesticides and discharge of industrial and domestic wastewater on water surfaces have increased. Especially, Copper (Cu) pollution in aquatic ecosystems could constitute a major health problem, not only for flora and fauna but also for humans. To cope with this challenge, environmental monitoring studies have sought to find Cu-specific biomarkers in terrestrial and aquatic flora and/or fauna. This review discusses the toxic effects caused by Cu on the growth and development of plants, with a special focus on aquatic plants. While copper is considered as an essential metal involved in vital mechanisms for plants, when in excess it becomes toxic and causes alterations on biomarkers: biochemical (oxidative stress, pigment content, phytochelatins, polyamines), physiological (photosynthesis, respiration, osmotic potential), and morphological. In addition, Cu has a detrimental effect on DNA and hormonal balance. An overview of Cu toxicity and detoxification in plants is provided, along with information regarding Cu bioaccumulation and transport. Awareness of the potential use of these reactions as specific biomarkers for copper contamination has indeed become essential.

Effect of extraction methods on mobilizable colloids and associated phosphorus from reservoir sediment.

Mobilizable colloids from reservoir sediment contain nutrients and contaminants, thus may affect water quality once being released. A major obstacle to evaluate the quantity and quality of mobilizable colloids in natural system is the using of appropriate method for colloid extraction from sediment and their separation from dissolved and particulate phases. This work evaluates the role of different extraction methods (agitation, sonication at sediment pH, and sonication at alkaline pH) on the characteristics (mass, size, shape and composition) of water-mobilizable colloids from sediment of Champsanglard dam reservoir (France). Attention has been paid to phosphorus (P), an important element in controlling eutrophication. Recovered colloids were highly affected on both quantity and quality according to the different applied protocols. The less aggressive agitation liberated low-energy water-dispersible colloids without physical damage and with less modification in colloidal chemical composition and shape, whereas sonication released 10-20 times higher colloid quantity but in lower size, due to physically disruption of fragile sediment structure or aggregated/chained colloids. In contrast, alkaline pH intensified colloid release by fortified repulsive forces between colloids and dissolution of organic coat. Concerning phosphorus, competition with hydroxide ions for sorption site or dissolution of phosphate minerals in alkaline pH caused release of dissolved P to solution and decrease of P content in recovered colloids. A special care should be paid to method selection according to the aim of the study and when comparing data from experiments conducted with different colloid extraction methods.

Assessment of metal/metalloid occurrence in rivers with their accumulation in macrophyte case study with Myriophyllum alterniflorum.

Water quality monitoring with integrative tools is a main issue of concern for environment assessment. Submerged aquatic macrophyte can be a good candidate for the evaluation of contaminant content in rivers. Indeed, owing to their habitat, aquatic macrophytes interact directly with surface water; they can absorb contaminants and thus allow to detect their presence in water. In situ studies were conducted over 28 days in five aquatic environments, affected by different levels of anthropogenic pressure (domestic wastewater plant, industrial activities), during two field campaigns. We have investigated whether the accumulation of some metals and a metalloid (As) in Myriophyllum alterniflorum could be used to detect their occurrence in river waters. Our results demonstrated that long time bioaccumulation was correlated with the contaminant levels in water. However, the water composition and the duration of exposure affected the studied pollutants' absorption. On an operational point of view, the optimal duration of exposure of Myriophyllum alterniflorum to assess the water quality is conditioned by the contaminant contents in waters that can induce different defense mechanisms as the reduction of pollutant absorption and their efflux. In addition, the nutrient concentration influenced the accumulation of pollutants since the higher the nutrient level, the higher the essential metal accumulation was observed.

Correction to: Sedimentary phosphorus accumulation and distribution in the continuum of three cascade dams (Creuse River, France).

The missing Electronic Supplementary Material in the original paper is included in this paper.

Sedimentary phosphorus accumulation and distribution in the continuum of three cascade dams (Creuse River, France).

Dam construction leads to both sediment discontinuities and the creation of internal phosphorus (P) loads in reservoirs capable of supporting eutrophication. Today, majority of large rivers are dammed and numerous of these infrastructures are constructed in cascade. However, few studies focus on the cumulative effect of the presence of dam on sediment P mobility and bioavailability in downstream reservoirs and rivers parts or throughout the continuum. The influence of three cascade dams has been studied herein on the sedimentary P distribution in surface bed sediments along a 17-km fluvial continuum of the Creuse River (Massif Central, France). The sediments (17 samples) were analyzed for their physical (grain size, specific surface area) and chemical (pH, contents of P, Fe, Al, Ca, Mn, organic matter (OM), and P fractionation) characteristics. Results indicated an amount of P 3 to 7 times higher in dam sediments (1.59 ± 0.51 mgP/g DW) than in free-flowing river sections (0.27 ± 0.11 mgP/g DW). Unexpectedly, sedimentary TP content did not decrease from the first to the third reservoir. The spatial variations of sediment characteristics between river and reservoirs were correlated with the retention of particles sized under 200 µm within the reservoirs. In reservoir sediment, P was mainly associated with the ascorbate fraction (P associated with the redox-sensitive Fe/Mn precipitates). Inside each dam reservoir, longitudinal variations of the sedimentary P distribution were mainly due to the increase of amorphous Fe precipitate content accumulated in fine sediments toward the dam, as characterized by a low Fe-Asc/P-Asc molar ratio. In the river sections, P distribution (mainly associated with HCl and ascorbate fractions) was not significantly influenced by cascade dams.

Are Myriophyllum alterniflorum biomarker responses to arsenic stress differentially affected by hydrodynamic conditions?

Arsenic (As) is a significant contaminant in the environment and its detection through macrophytes can provide a powerful tool. Myriophyllum alterniflorum constitutes a good candidate by virtue of its ability to accumulate contaminants, and moreover its biomarkers can respond to the presence of trace metals and metalloids. The objective of this study therefore is to evaluate the watermilfoil response to As exposure under several hydrodynamic conditions since it is well known that hydrodynamics affect plant functioning. For this purpose, fresh watermilfoil plants are subjected to three hydrodynamic conditions, namely laminar, turbulent and calm, in a synthetic medium either enriched or not by 100 µg.L-1 arsenic for 21 days. Growth, pigment content (chlorophyll a, b and carotenoids), respiratory and photosynthetic activities, osmotic potential and hydrogen peroxide concentration are all monitored. Arsenic accumulation is measured separately in the roots and shoots of Myriophyllum alterniflorum. On the one hand, it should be noted that arsenic induces: (i) a significant increase in H2O2 content; (ii) a decrease in osmotic potential, pigment content, photosynthesis and respiration rates, shoot and root growth; and (iii) an inhibition of shoot branching. Moreover, a higher accumulation of this metalloid in roots than in shoots, regardless of the hydrodynamic condition, is witnessed. While on the other hand, hydrodynamic conditions only affect watermilfoil morphology and arsenic accumulation. Also, the younger and older parts have experienced differential toxic effects. Overall, our results suggest the effective use of M. alterniflorum in both water quality biomonitoring and phytoremediation studies.

Phosphorus mobility in dam reservoir affected by redox oscillations: An experimental study.

The internal sedimentary phosphorus (P) load of aquatic systems is able to support eutrophication, especially in dam-reservoir systems where sedimentary P stock is high and where temporary anaerobic conditions occur. The aim of this study therefore is to examine the response of sedimentary P exposed to redox oscillations. Surface sediments collected in the Champsanglard dam-reservoir (on the Creuse River, France) were subjected to two aerobic phases (10 and 12 days) alternated with two anaerobic periods (21 and 27 days) through batch incubations. The studied sediment contained 77 ± 3 µmol/g DW of P, mainly associated with the ascorbate fraction (amorphous Fe/Mn oxyhydroxides). The used sediment was rich in organic matter (OM) (21% ± 1%) with primarily allochthone signature. Our results showed that redox oscillations enhance dissolved inorganic phosphorus release at sediment/water interface. During the first anaerobic stage, the P release was mainly controlled by the dissolution/precipitation of iron minerals. The more pronounced increase of P release during the second anaerobic stage (44%) was due to various mechanisms related to the change in quality of dissolved organic matter (DOM), namely a higher SUVA254 and humification indices. The release of more refractory DOM (rDOM) served to lower the microbial metabolism activity, possibly favored iron oxyhydroxide aggregation and thus limiting iron reduction. In addition, rDOM is able to compete for mineral P sorption sites, leading to a greater P release. In reservoir with predominant allochthone OM input, the release of more aromatic DOM therefore plays an important role in P mobility.

Combined effect of copper and hydrodynamic conditions on Myriophyllum alterniflorum biomarkers.

The aim of this study is to determine the combined effect of copper and hydrodynamic conditions on the response of certain biomarkers of an aquatic macrophyte, namely Myriophyllum alterniflorum. Watermilfoil biomarkers are monitored in a synthetic medium enriched or not with copper (100 µg.L-1) for 21 days in aquarium systems (150 L), under three hydrodynamic conditions: laminar, turbulent, and calm. The studied biomarkers are: respiratory and photosynthetic activities; concentrations of chlorophyll a, b and carotenoids; osmotic potential; hydrogen peroxide content; and growth. In addition, Cu contents in water and in Myriophyllum alterniflorum (roots and shoots) are investigated. The hydrodynamic conditions only affect watermilfoil morphology. Copper accumulates less in turbulent zones; moreover, it is more likely to accumulate in shoots than in roots, except within the calm zone. Cu leads to: i) a significant increase in H2O2 content, ii) a decrease in root growth, pigment content, osmotic potential, photosynthesis and respiration rates, and iii) an inhibition of shoot branching. Differential effects are also observed between younger and older parts, thus indicating the benefit of considering these two plant parts separately in water quality biomonitoring.

Phosphate removal from aqueous solution using ZVI/sand bed reactor: Behavior and mechanism.

This research reports on phosphate removal from aqueous solution using ZVI/sand packed columns. The influence of column preconditioning, consisting of ZVI pre-oxidation before feeding the columns with phosphate solution, revealed that a column aged for 1 day was more efficient than un-conditioned column, 5-days and 10-days preconditioned columns. The distribution of phosphate trapped inside the columns was evaluated by measuring phosphate concentration in the solids at different levels (P1, P2 and P3) along the depth of the columns. The distribution of phosphate inside the columns was determined for a time period up to 46 days, corresponding to column saturation. Results showed heterogeneous trapping along the column before saturation and homogeneous distribution upon saturation. The maximum cumulative trapped phosphate after column dismantling was determined before saturation (after 17 days running) at 130, 68 and 31 mgP/gFe at the inlet-P1, P1-P2 and P2-P3 layers, respectively, whereas the homogeneous distribution of phosphate upon saturation was determined at 132 mgP/gFe throughout the column. Solid supports were characterized using SEM, XRD and XPS. Lepidocrocite and maghemite/magnetite were the only iron oxidation products identified at the different layers inside the columns. XPS results confirmed the sorption of phosphate at the surface of ZVI and its oxidation products and highlighted the formation of an iron phosphate complex.

Absorption and translocation of copper and arsenic in an aquatic macrophyte Myriophyllum alterniflorum DC. in oligotrophic and eutrophic conditions.

The aim of this study is to evaluate copper and arsenic accumulation and translocation at a concentration of 100 µg/L of a submersed macrophyte Myriophyllum alterniflorum. The trophic level (eutrophic and oligotrophic conditions) of the medium was also considered. To achieve this goal, plants were incubated for 21 days in the presence of 100 µg/L of Cu or AsV. The heavy metal transfers from the contaminated medium to plants and into plant tissues was discussed in terms of the bioconcentration factor (BCF) and the translocation factor (TF). Malondialdehyde (MDA) content in tissues was analyzed in order to study the toxicity of these two contaminants. Our results show that copper was more accumulated in shoots, than roots, whereas the opposite trend was observed for arsenic. In addition, the two contaminants were more accumulated in oligotrophic than eutrophic medium. The BCF of copper in shoots was 1356 in oligotrophic condition, while that of arsenic was higher in roots about 620 in the same condition. The TF was less than 1 for arsenic, and higher than 1 for copper, indicating that watermilfoil restrains the translocation of arsenic to shoots, while it has a low capacity to control the translocation of an essential micronutrient like copper. An increase in MDA content was observed under Cu and As stress. On the basis of this experiment, M. alterniflorum has a higher accumulation potential of copper and arsenic, and therefore, it can be a good candidate for the phytofiltration of these two contaminants from water.

Adsorption of As(III), As(V) and dimethylarsinic acid onto synthesized lepidocrocite.

The trapping of arsenic by zero valent iron is strongly dependant on iron by-products. Among these, lepidocrite has been scarcely studied. In this work, we studied the adsorption of two inorganic (As(III), As(V)) and one organic (dimethylarsinic acid, DMA) arsenic species onto lepidocrocite. pH influence was considered in the range 5 to 9, which corresponds to natural water pH. Langmuir model was used to simulate As adsorption isotherms. Our results showed that lepidocrocite offers high adsorption capacities: up to 0,25, 0,41 and 1 mol for As(V), DMA and As(III) could be respectively trapped per kilogram of zero-valent iron. pH influence varied from one arsenic species to another: increasing pH improve As(III) and DMA sorption whereas it has a very low effect on As(V) sorption.

AsIII oxidation by Thiomonas arsenivorans in up-flow fixed-bed reactors coupled to As sequestration onto zero-valent iron-coated sand.

The combined processes of biological As(III) oxidation and removal of As(III) and As(V) by zero-valent iron were investigated with synthetic water containing high As(III) concentration (10 mg L(-1)). Two up-flow fixed-bed reactors (R1 and R2) were filled with 2 L of sieved sand (d = 3 ± 1 mm) while zero-valent iron powder (d = 76 µm; 1% (w/w) of sand) was mixed evenly with sand in R2. Thiomonas arsenivorans was inoculated in the two reactors. The pilot unit was studied for 33 days, with HRT of 4 and 1 h. The maximal As(III) oxidation rate was 8.36 mg h(-1) L(-1) in R1 and about 45% of total As was removed in R2 for an HRT of 1 h. A first order model fitted well with the As(III) concentration evolution at the different levels in R1. At the end of the pilot monitoring, batch tests were conducted with support collected at different levels in R1. They showed that bacterial As(III) oxidation rate was correlated with the axial length of reactor, which could be explained by biomass distribution in reactor or by bacterial activity. In opposition, As(III) oxidation rate was not stable in R2 due to the simultaneous bacterial As(III) oxidation and chemical removal by zero-valent iron and its oxidant products. However, a durable removal of total As was realized and zero-valent iron was not saturated by As over 33 days in R2. Furthermore, the influence of zero-valent iron and its oxidant corrosion products on the evolution of As(III)-oxidizing bacteria diversity was highlighted by the molecular fingerprinting method of PCR-DGGE using aoxB gene as a functional marker of aerobic As(III) oxidizers.

Short term copper toxicity on Microcystis aeruginosa and Chlorella vulgaris using flow cytometry.

Copper sulphate is a common algaecide applied to ponds in order to keep phytoplanktonic blooms under control, especially those prone to cyanobacterial development. The copper toxicity was evaluated for two phytoplanktonic species with the same size and shape: one cyanobacteria Microcystis aeruginosa and one chlorophyceae Chlorella vulgaris. Under controlled conditions in order to keep free copper concentration constant, three bioassays were carried out: growth inhibition tests and two sets of flow cytometric analysis (autofluorescence and esterase activity inhibition). The results showed: (1) that regardless of the cell type, as copper concentrations increased, cell division rate and autofluorescence decreased, (2) a difference in sensitivity according to the length of exposure to copper: 24 or 48 h, and (3) that M. aeruginosa showed a greater sensitivity than C. vulgaris in the 24h copper exposure assay according to esterase activity data.

Planktonic microbial community responses to added copper.

It is generally agreed that autotrophic organisms and especially phytoplanktonic species can be harmed by copper through its effect on photosystem. However, the impact of copper on other components of the pelagic food web, such as the microbial loop (autotrophic and heterotrophic picoplankton, pigmented and non-pigmented flagellates and ciliates) has received little attention. Indoor experiments were conducted to evaluate the direct and indirect effects of copper, supplied in the range of concentrations used to control cyanobacteria growth in ponds, on non-targeted organisms of natural microbial loop communities sampled in spring and summer. Two copper concentrations were tested (80microgL(-1) and 160microgL(-1) final concentrations), set, respectively, below and above the ligand binding capacity of the water samples. Both caused a significant decrease in the biomass and diversity of pigmented organisms (picophytoplankton and pigmented flagellates). Conversely, the heterotrophic bacterioplankton and the heterotrophic flagellates did not seem to be directly affected by either copper treatment in terms of biomass or diversity, according to the descriptor chosen. The ciliate biomass was significantly reduced with increasing copper concentrations, but differences in sensitivity appeared between spring and summer communities. Potential mixotrophic and nanoplanktorivorous ciliates appeared to be more sensitive to copper treatments than bacterivorous ciliates, suggesting a stronger direct and (or) indirect effect of copper on the former. Copper sulphate treatments had a significant restructuring effect on the microbial loop communities, resulting in a dominance of heterotrophic bacterioplankton among microbial microorganisms 27 days after the beginning of the treatment. The spring microbial communities exhibited a greater sensitivity than the summer communities with respect to their initial compositions.

Optimization of a GFAAS method for determination of total inorganic arsenic in drinking water.

The new 10mugl(-1) arsenic standard in drinking water has been a spur to the search for reliable routine analytical methods with a limit of detection at the mugl(-1) level. These methods also need to be easy to handle due to the routine analyses that are required in drinking water monitoring. Graphite furnace atomic absorption spectrometry (GFAAS) meets these requirements, but the limit of detection is generally too high except for methods using a pre-concentration or separation step. The use of a high-intensity boosted discharge hollow-cathode lamp decreases the baseline noise level and therefore allows a lower limit of detection. The temperature program, chemical matrix modifier and thermal stabilizer additives were optimized for total inorganic arsenic determination with GFAAS, without preliminary treatment. The optimal furnace program was validated with a proprietary software. The limit of detection was 0.26mugAsl(-1) for a sample volume of 16mul corresponding to 4.2pgAs. This attractive technique is rapid as 20 samples can be analysed per hour. This method was validated with arsenic reference solutions. Its applicability was verified with artificial and natural groundwaters. Recoveries from 91 to 105% with relative standard deviation <5% can be easily achieved. The effect of interfering anions and cations commonly found in groundwater was studied. Only phosphates and silicates (respectively at 4 and 20mgl(-1)) lead to significant interferences in the determination of total inorganic arsenic at 4mugl(-1).

Effect of copper sulphate treatment on natural phytoplanktonic communities.

Copper sulphate treatment is widely used as a global and empirical method to remove or control phytoplankton blooms without precise description of the impact on phytoplanktonic populations. The effects of two copper sulphate treatments on natural phytoplanktonic communities sampled in the spring and summer seasons, were assessed by indoor mesocosm experiments. The initial copper-complexing capacity of each water sample was evaluated before each treatment. The copper concentrations applied were 80 microg l(-1) and 160 microg l(-1) of copper, below and above the water complexation capacity, respectively. The phytoplanktonic biomass recovered within a few days after treatment. The highest copper concentration, which generated a highly toxic environment, caused a global decrease in phytoplankton diversity, and led to the development and dominance of nanophytoplanktonic Chlorophyceae. In mesocosms treated with 80 microg l(-1) of copper, the effect on phytoplanktonic community size-class structure and composition was dependent on seasonal variation. This could be related to differences in community composition, and thus to species sensitivity to copper and to differences in copper bioavailability between spring and summer. Both treatments significantly affected cyanobacterial biomass and caused changes in the size-class structure and composition of phytoplanktonic communities which may imply modifications of the ecosystem structure and function.

Laboratory investigation of the phosphorus removal (SRP and TP) from eutrophic lake water treated with aluminium.

Mechanisms involved in phosphorus (P) removal from eutrophic lake water with aluminium (Al) were assessed by jar tests. For this purpose, eutrophic lake water enriched with soluble reactive phosphorus (SRP), algae or sediments in order to mimic the various conditions found in shallow eutrophic lakes was studied. Total phosphorus (TP) removal was reached after floc settling, the maximal TP removal efficiency (90-95%) was obtained for an Al concentration ranging from 2 to 5 mg L(-1), depending on the organic matter (OM) origin (algae or sediments). Algae appeared to limit macro-floc formation (those able to settle). In contrast, in the presence of sediments, macro-floc formation was favoured at low Al dose (2-3 mg L(-1) Al). High SRP removal was obtained with the lowest Al dose (1 mg L(-1) Al) and remained greater than 60% for an SRP concentration up to 350 microg L(-1). SRP removal was not influenced by the OM origin. The experimental data and literature were used to suggest a hypothetical model for floc formation and P removal with Al under the conditions observed in treated lakes.

Toxicity of copper excess on the lichen Dermatocarpon luridum: antioxidant enzyme activities.

The aim of this study was to investigate the toxicity of copper on the aquatic lichen Dermatocarpon luridum focusing on the activities of some antioxidant enzymes. Investigations were conducted using increasing copper concentrations (0.00, 0.25, 0.50, 0.75 and 1.00 mM CuSO(4) x 5H(2)O) in synthetic freshwater that emulated the major ion compositions of its natural water biota; time course measurement was 0, 3, 6, 12, 24 and 48 h. The copper concentration in thalli increased with its increase in the medium and the duration of treatment. Copper induced lipid peroxidation, measured using the hydroperoxi-conjugated dienes (HPCD) concentration. The decrease in the protein concentrations was similar in thalli exposed to copper concentrations above 0.50 mM and the decrease was twice lower in thalli exposed to 0.25 mM copper. The activities of antioxidant enzymes measured were differently affected by copper excess. For 0.25 mM copper, the activities of SOD (superoxide dismutase) and APX (ascorbate peroxidase) were unchanged when compared with unstressed thalli whereas the CAT (catalase) activity increased and the GR (glutathione reductase) activity decreased. The activities of SOD and APX increased in thalli exposed to concentrations above 0.50mM copper. The CAT activity increased after the first 3h of experiments at these concentrations and then decreased with the duration of treatment at an activity lower than in the unstressed plant. Whereas the APX activity increased, the GR activity similarly decreased for the copper concentration tested whatever the duration of the experiment.