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Renata Behra

Publications and source records attributed to Renata Behra.

13 recordsLinked to original sources
PubMed2006

Thiols in Scenedesmus vacuolatus upon exposure to metals and metalloids.

Phytochelatins are intracellular metal ligands produced by algae when exposed to elevated metal concentrations. In freshwater ecosystems, algae are exposed to a wide range of metals and metalloids. The aim of this study was thus to investigate phytochelatin induction in freshwater algae upon metal and metalloid exposure. To that purpose, the unicellular green alga Scenedesmus vacuolatus, was exposed to Cu, Zn, Ni, Pb and Ag, as well as to As(III), As(V), Sb(III) and Sb(V), and examined for its thiol content (gamma-glutamylcysteine, glutathione and phytochelatins). Glutathione content was found to decrease upon the exposure to Zn and to increase upon the exposure to Pb and Ag. Phytochelatins were only induced by Cu (at [Cu2+] = 8x10(-11) M) and Pb (at [Pb2+] = 8x10(-11) to 8x10(-10) M), where [Cu2+] and [Pb2+] are computed free metal ion concentrations. Glutathione content also decreased upon the exposure to Sb(V) whereas an increase was observed as a result as the exposure to As(III) and As(V). The metalloids As(III), As(V) and Sb(III) in the concentration range from 8x10(-6) to 2x10(-4) M (total concentrations of oxyanions) were inducing phytochelatins. Glutathione and phytochelatin content in S. vacuolatus do thus sensitively respond to exposure to a number of metals and metalloids.

Arsenic↗
PubMed2005

Thiol and metal contents in periphyton exposed to elevated copper and zinc concentrations: a field and microcosm study.

Phytochelatins are metal-binding polypeptides produced by algae under metal exposure. The aim of this study was to investigate the effects of metal concentration variations in natural systems on periphyton at the biochemical level by analyzing its intracellular thiol content, in particular phytochelatins. To that purpose, two field campaigns were conducted in a stream subject to an increase of dissolved metal concentrations (particularly Cu and Zn) during rain events, which results in an increase of their accumulation in periphyton. At background metal concentrations, several thiols were detectable in periphyton, namely, glutathione (GSH), gamma-glutamylcysteine (gammaGluCys), phytochelatins (PC2), and some unidentified thiols, U1 and U2. Glutathione and gammaGluCys contents were found to vary independently of the rain, as well as U1 and U2, whereas the phytochelatin content increased during the rain events. To investigate whether Cu or Zn may be responsible for this increase, microcosm experiments were carried out with natural water enriched with Cu, Zn, and Cd separately, and Cu and Zn in combination. In this study, GSH, PC2, and U1 were also detected, but not gammaGluCys. An increase in accumulated Cu content did not induce any changes in thiol content, whereas an increase of the Zn content induced a decrease in GSH content and an increase in phytochelatin content. Zinc rather than Cu may thus induce a phytochelatin content increase in periphyton in the field studies. Addition of Cu and Zn in combination also induced an increase in phytochelatin content. Cadmium was found to be the most effective inducer, with the production of larger phytochelatins (PC3-4). This study is the first one to report changes in thiol content in periphyton in response to an increase of the metal concentration in natural freshwaters.

Analysis of Variance↗
PubMed2005

Impact of zinc acclimation on bioaccumulation and homeostasis in Chlorella kesslerii.

Growth curves, cellular Zn contents and cellular protein expression were examined for the green alga, Chlorella kesslerii, as a function of different Zn growth regimes (growth in 16 pM, 1.7 nM or 1.6 microM calculated Zn2+). Zn homeostasis was responsible for observed differences in the capacity of the organism to accumulate Zn. The rapid acclimation that occurred in response to a Zn deficiency was likely due to the production of Zn transport sites. No differences were observed among cellular phytochelatin contents or efflux rate constants, although efflux did play an important role in regulating Zn cellular content. A long-term adaptation to Zn was not thought to occur since bioaccumulation and biological responses were similar for four successive cultures (30-40 days, 16-19 cell cycles) at different [Zn2+]. Among proteins that were influenced by the Zn growth regime, the Rubisco and histone H3 proteins were identified as being induced in the presence of 1.6 microM Zn2+ as compared with 1.7 nM Zn2+. The impact of the Zn preconditioning demonstrated that the concentrations of essential metals in the algal growth media would have an important, if not predominant effect on toxicity or bioaccumulation assessments. Furthermore, the high regulation of Zn transport and intracellular events by the microorganisms will likely preclude the use of simple metal uptake models including the free ion activity model and the biotic ligand model to predict either bioaccumulation or toxicological effects of Zn and perhaps other essential metals.

Blotting, Western↗
PubMed2005

Tolerance of Oocystis nephrocytioides to copper: intracellular distribution and extracellular complexation of copper.

Several mechanisms have been proposed to explain how algae can tolerate heavy metals. In order to better understand the mechanisms determining metal tolerance, we examined the interaction of copper with two strains of the copper-tolerant green algae Oocystis nephrocytioides, isolated from algal communities differing only in copper exposure. The strains were cultured in chemically-defined media containing 0.04 microM Cu(total) (pCu 12.4) or 2 microM Cu(total) (pCu 10.6). Growth, photosynthesis rate, content of chlorophyll a and b, copper accumulation, its cellular distribution and ultrastructural localization, as well as the influence of algal growth on extracellular copper complexation were determined. Both strains had comparable growth and photosynthesis rates. The cellular content of both chlorophyll a and b was reduced, by roughly the same extent, at pCu 10.6 compared to pCu 12.4. Copper titration of the media indicated the production of copper-complexing ligands by O. nephrocytioides cultured at pCu 12.4 that increased with increased algal density during cell growth. No additional ligands were detected at pCu 10.6. Copper-complexing ligands had a conditional stability constant of K = 10(13) at pH 7.3. The intracellular concentration of copper in O. nephrocytioides was 80 microM at pCu 12.4 and increased to 7.5mM at pCu 10.6. The proportion of intracellular Cu accumulated increased from 8% of total Cu content at pCu 12.4 to 60% at pCu 10.6. By electron spectroscopic imaging, intracellular Cu was detected in the thylakoids and the pyrenoid of O. nephrocytioides cells. The results indicate that the tolerance of O. nephrocytioides to Cu is constitutive and does not need to be induced by previous exposure to Cu. We propose that accumulation and sequestration of Cu in thylakoids and, to a lesser extent, adsorption of copper to the algal cell surface represent the most important tolerance mechanism, for O. nephrocytioides.

Chlorophyll↗
PubMed2005

Modeling the concentration-response function of the herbicide dinoseb on Daphnia magna (survival time, reproduction) and Pseudokirchneriella subcapitata (growth rate).

Models describing dose-response relationships are becoming increasingly popular in ecotoxicology. They allow simple and thorough evaluations of toxicity test results, including inter- and extrapolations to concentrations or exposure times other than those tested. Simple parametric regression models are of particular interest because their parameters may be attributed mechanistic meanings and they can be applied without sophisticated mathematical and computational support. We recently proposed a four-parameter logistic regression model to fit the survival data of Daphnia magna under dinoseb stress. The model parameters are the maximum survival time, the minimum time required for an individual to die, effect concentration, EC(50), and a curve shape parameter. This model has now been applied to compare the lethality and reproduction toxicity of D. magna and the growth inhibition of Pseudokirchneriella subcapitata under dinoseb stress. It can be fitted adequately to all the measured data and the parameters can be attributed biological meanings in any of the three endpoints. A comparison of the modeled concentration-response functions of all three endpoints for dinoseb toxicity shows that the range of ECs with respect to both D. magna and algae is steep (a decrease of between 0.1 and 0.6 mg/L). The survival and reproduction of D. magna exhibit similar characteristic concentration-response functions and toxicities. The statistical no-effect concentration (SNEC) is 0.14 (survival) and 0.11 (reproduction)mg/L, respectively. On the other hand, algae seem to be less sensitive to dinoseb than D. magna (SNEC: 0.48 mg/L). However, further investigations of individual algae may lead to a more suitable comparison. We speculate that the four parameters of the model function can be related to specific properties of chemicals and organisms. Characterization of these properties would allow simple and appropriate estimation of the toxic effects of these chemicals.

2,4-Dinitrophenol↗
PubMed2005

Phytochelatin induction, cadmium accumulation, and algal sensitivity to free cadmium ion in Scenedesmus vacuolatus.

Phytochelatins are small, intracellular, metal-binding polypeptides produced by algae on exposure to increased metal concentration in their environment. The aim of the present study was to examine the relationship between phytochelatin concentration, bioaccumulated metal, and sensitivity of algal growth on cadmium exposure. For that purpose, intracellular cadmium concentration and thiol (glutathione, gamma-glutamylcysteine [gammaGluCys], and phytochelatins [PCn]) content were determined in the freshwater green alga Scenedesmus vacuolatus exposed to growth-inhibitory and noninhibitory concentrations of free Cd2+ in the range from 10(-14) to 10(-7) M. The algal growth rate was optimal up to a free Cd2+ concentration of 10(-9) M and then decreased by 40% at higher concentrations. The intracellular cadmium content increased sharply from 0.22 to 746 amol/cell over this free Cd2+ range. At the lowest Cd2+ concentration (control), glutathione was the only detectable thiol (127 amol/cell). With increasing Cd2+, formation of gammaGluCys and phytochelatins from PC2 to PC6 were observed. The predominant oligomer was PC3, with 42 amol/cell at the highest Cd2+ concentration (10(-7) M). The ratio of the concentration of total thiol groups to intracellular cadmium was important for maintaining optimal growth. In contrast, thiol groups from phytochelatins were never measured in excess to intracellular cadmium content.

Cadmium↗
PubMed2004

Influence of metal speciation in natural freshwater on bioaccumulation of copper and zinc in periphyton: a microcosm study.

The free ion activity model (FIAM) has already been confirmed under laboratory conditions for many trace metals but has still to be validated under natural conditions where the presence of natural organic ligands influences metal speciation and bioavailability. The goal of this study was to test if the FIAM is followed under natural conditions by measuring copper and zinc speciation as well as metal accumulation in periphyton. Periphyton was exposed in microcosms to natural river water with different added concentrations of copper (25-258 nM) or zinc (18-501 nM) and additions of a synthetic ligand (NTA). Free Cu2+ was in the range of 10(-16.5)-10(-14.5) M and Zn(2+) was in the range of 0.7-8.7 nM, as measured by competitive ligand exchange coupled with cathodic/anodic stripping voltammetry. Other metal complexes were either measured or computed. Bioaccumulation of zinc in periphyton appeared to be controlled by the free zinc ion concentration, confirming the FIAM. In contrast, bioaccumulation of copper was controlled by weakly complexed copper (including Cu2+ plus inorganic and weak organic complexes), which is in disagreement with the FIAM, and appears to be caused by limitation of copper diffusion due to very low free Cu2+ occurring in natural environments.

Biofilms↗
PubMed2004

Mechanistic approaches for evaluating the toxicity of reactive organochlorines and epoxides in green algae.

Reactive electrophilic chemicals, such as reactive organochlorine compounds or epoxides, react specifically with a broad spectrum of nucleophilic biomolecules, including proteins and DNA. Conventional toxicity tests for algae, involving the observation of growth inhibition, i.e., the inhibition of cell multiplication, after several days, yield unreliable information for risk assessment because reactive compounds hydrolyze to different extents during the exposure period. The diversity of their modes of toxic action further complicates effect assessment and calls for methods yielding additional information on the mechanisms of toxicity. One of the primary targets of reactive chemicals in cells is the tripeptide glutathione (GSH), which is important for detoxification but can also be regarded as a toxicity sensor because changes in glutathione levels indicate stress. A vital system for algae is the photosynthetic system, which is indirectly affected by reactive chemicals. The test systems developed in this study for the assessment of reactive toxicity toward algae were therefore based not only on nonspecific toxicity indicators like growth inhibition but also on indicators for disturbance of photosynthesis (inhibition of photosystem II quantum yield) and glutathione metabolism. The application of the developed test systems on Scenedesmus vacuolatus after short-term exposure of 2 h showed that these tests can be used as fast screening tests for algal toxicity and in mode-of-action-based test batteries.

Chlorophyta↗
PubMed2003

Accumulation of copper and zinc in periphyton in response to dynamic variations of metal speciation in freshwater.

Although the free ion activity model (FIAM) has been well-established in laboratory studies, there remains the need for field data in order to validate the applicability of this model in natural systems. The objective of this study was to investigate the response of copper and zinc accumulation in periphyton to short-term variations in metal concentration and speciation in freshwater. During heavy rain events, dissolved Cu in the Furtbach stream increased from 40 to 118 nM, while dissolved Zn increased from 45 to 147 nM due to the release of metals from contaminated sediments. Increases in free copper and free zinc ions in the water (from 10(-14) to 10(-11.5) M for Cu2+; from 1 to 15 nM for Zn2+) were observed during the onset of heavy rain events. Periphytic algae collected from artificial substrates had an intracellular copper content (0.2-2.8 micromol/g dry weight (dw)) that varied as a function of the exchangeable copper in the water (labile form) rather than the free Cu2+. Intracellular zinc content (1.5-8.0 micromol/g dw) was found to follow the same trend as the free zinc ion concentration. Adsorbed Cu and Zn on periphyton showed a very dynamic response to variations in dissolved metal concentration. Different concentrations of dissolved manganese during the two time periods may affect the accumulation of zinc and copper by competition for metal uptake.

Copper↗
PubMed2002

Effects of dinoseb on the life cycle of Daphnia magna: modeling survival time and a proposal for an alternative to the no-observed-effect concentration.

Risk assessment is in urgent need of more accurate toxic effect endpoints than those currently in use, especially for low concentrations. Often such endpoints are estimated by analysis of variance, linear interpolation, or smoothing. As these statistical methods are not always satisfactory, some authors have proposed to describe the entire dose-response curves by fully formalized parametric regression models whose parameters have toxicological meaning. These models allow a better evaluation of pollutant effects, including inter- and extrapolation to any other than the measured effect values. Following this line, a four-parameter logistic regression model (standard model) was fitted to survival data of Daphnia magna under pesticide (dinoseb) stress. The heterogeneity of the variance was taken into account with a both-sides logarithmic transformation. Besides the standard model, a hormesis and a threshold model were tested too. These two others models have been described in the literature and might better represent the dose-response function we are looking for. All three models showed a good fit to our data, and the statistics gave no hints as to which model is the most appropriate. As no evidence was seen for hormesis or for the existence of a threshold concentration, we used the simplest, namely, the standard model, for most of our calculations. Model calculations allow the quantification of the effects on individuals' longevity as well as on mean survival time of the population. We used them to define a no-effect value, the statistical-no-effect concentration (SNEC). The SNEC is based on the confidence bands of the modeled regression and represents the highest value for which an effect is statistically not different from the control. The SNEC is an alternative to classical endpoints, like the no-observed-effect concentration (NOEC) or the low-effect concentrations (e.g., EC10, EC5, EC1).

2,4-Dinitrophenol↗
PubMed2002

Ecotoxicological assessment of surface waters: a modular approach integrating in vitro methods.

Today ecotoxicological evaluations of surface water quality are either based on field surveys or online biomonitoring, whereas the ecotoxicological quality of wastewater is mostly determined with standardised acute toxicity tests. In this paper we present a concept for the ecotoxicological evaluation of surface waters, where mainly in vitro tests are used for the screening of water samples, presenting the first tier of a two-tiered approach. In this first tier a battery of fast and cost-efficient test-systems are used as an early warning system. Thereby, the toxic potential of water samples will be identified. This modular approach allows the exchange or addition of test-systems if necessary. If a toxic potential is identified in a water sample, this sample can be investigated more thoroughly in a second tier where organisms are used. In this paper we focus mainly on the general approach and the description of the first tier.

Ecosystem↗