Poikilothermic organisms experience trade-offs by differential physiological demands generated by temperature extremes. Many such organisms exhibit acclimatory effects, adjusting their metabolism and physiology to recently experienced temperatures. One such acclimatory effect is metabolic compensation, the deceleration of biological rates below Arrhenius expectations. Daphnia magna is eurythermal, and if acclimated to mildly stressful temperatures first, survives longer in lethal temperatures. This study examined the effect of ambient temperature (5°C-37°C) and acclimation history (lifetime at 10°C or 25°C) on the oxygen consumption rates of 8 genotypes of Daphnia with high or low acute temperature tolerance. There were decelerations of respiratory rates across a temperature gradient when acclimated to 25°C or following short 8- hour acclimation to measurement temperatures. Daphnia exposed to a near-lethal temperature (35°C) with a 24-hour recovery period at 25°C-acclimation temperature showed no difference in respiratory control compared to unexposed 25°C-acclimated Daphnia. Genotypes showed no difference in potential compensatory ability.

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<p> In this study the influence of several water chemistry parameters on the toxicity of Ni to Daphnia pulex in soft water were tested. A reconstituted soft water (pH 7.8, hardness 31.5 mg/L CaCO3) was used as the basis for culture and testing. Daphnia pulex was chosen as a typical cladoceran, one which can be acclimated to very soft water. An understanding of the influence of water chemistry on Ni toxicity in soft water is relevant because metals have higher bioavailability in soft water. The 48h EC50 in the reconstituted soft water (RSW) was 974 μg/L (16 μM) dissolved Ni (95%CI 830- 1081 μg/L). The following factors were examined for their potential for modifying Ni toxicity: Ca, Mg, Na, K, Cl, pH (3 different approaches used) and natural organic matter (NOM, 2 sources tested). Both Ca and Mg protected against Ni toxicity and the relative effect was greater for Ca. Varying the concentrations of Na, Cl or K did not alter the toxicity of Ni. Tests at different pH showed that as pH increased, Ni toxicity decreased. When the test solution pH was adjusted with the organic buffer 3-morpholinepropanesulfonic acid, there was a clear correlation between increasing pH and increasing EC50. The pH tests using bicarbonate to adjust pH did not show this relationship as clearly. Both types of NOM showed a protective effect on Ni toxicity with Nordic Reservoir NOM having a 4 fold greater effect than that of Suwannee River NOM. This research illustrated that the effect of alterations in water chemistry were generally as predicted within the context of the biotic ligand model (BLM) approach. The data provide the information required to develop a BLM for the acute effects of Ni in soft water.</p> / Thesis / Master of Science (MSc)

Polyaromatic hydrocarbons (PAHs) are contaminants synthesized through incomplete combustion of carbon based substances. PAHs are known to be photodynamic and toxicity increases exponentially when in contact with ultraviolet radiation (UV). The effect of UV absent recovery periods and potential for latent toxicity during photo-induced toxicity are previously unknown and are not included within the toxicity model. Results of equal interval tests further support the current reciprocity model as a good indicator of PAH photo-induced toxicity. Interval test results also indicate a possible presence of time-dependent toxicity and recovery thresholds and should be included into toxicity risk assessments. Moreover, results of latent effects assays show that latent mortality is a significant response to PAH photo-induced toxicity and should be included into toxicity risk assessments. The present research demonstrates that UV exposure time rate is a significant driving force of PAH photo-induced toxicity.

Xenohormesis suggests that consuming stressed food provides physiological adaptation to the consumer for similar stresses. To evaluate this, Daphnia magna was provided with heat-stressed Nannochloropsis limnetica grown at 25℃ whereas the control group was provided Nannochloropsis limnetica grown at 12℃. In the heat tolerance experiment, Daphnia kept at 28℃ and 20℃ while consuming 25℃ algae had significantly higher heat tolerance compared to the control. Additionally, in stressed conditions, Daphnia consuming 25℃-grown food had significantly higher lifespan compared to the control. Next, Daphnia consuming 25℃-grown algae had significantly higher levels of antioxidants which could explain the lifespan extension. Finally, in fatty acid analysis, eicosatetraenoic acid (EPA) was found lower in Daphnia consuming 25℃ cultures compared to Daphnia consuming 20℃ cultures. We speculate that this lower EPA level is increasing membrane rigidity and hence higher heat tolerance is observed. These findings indicate consumption of stressed algae might provide physiological benefits in Daphnia.

Ce doctorat s’intègre dans le projet ANR BIOMET (JC05_59809) et a eu pour vocation d’améliorer les connaissances actuelles sur l’influence de la matière organique dissoute sur la spéciation et la biodisponibilité des métaux particulièrement dans le cas de système sous forte pression urbaine tel que la Seine. La compréhension de l’influence de la MOD sur la spéciation et la biodisponibilité des métaux suppose en outre que l’on connaisse précisément sa composition chimique ou plus exactement ses groupements fonctionnels. C’est pourquoi une partie de ces travaux est consacrée à l’extraction et la caractérisation de la matière organique dissoutes d’origine naturelle et urbaine. La caractérisation des fractions de MOD a été réalisée de manière originale par une approche multidimensionnelle à l’aide d’un véritable éventail de techniques analytiques (analyses élémentaires; isotopiques; fonctionnelles et moléculaires) permettant ainsi de mieux aborder l’influence de la MOD sur la spéciation et la biodisponibilité du cuivre et du plomb. Parallèlement à l’étude de caractérisation de la MOD, ce travail s’attache à mieux cerner le rôle de la MOD d’origine urbaine en particulier de la MOD hydrophile sur la spéciation et la biodisponibilité du cuivre et du plomb. Dans cet objectif, les techniques : potentiométrique à l’aide d’électrode ionique spécifique et la récente technique électrochimique AGNES ainsi que des biotests de toxicité aiguë (Daphnia magna) et de bioaccumulation (Fontinalis antipyretica) ont permis : - d’étudier la complexation du cuivre et du plomb par les fractions de MOD obtenues afin notamment de fournir des paramètres de complexations des MOD hydrophiles peu connues jusqu’à présent et ainsi mieux prédire le transport des métaux en milieu urbain. - d’évaluer le rôle protecteur de la MOD d’origine urbaine sur la biodisponibilité du cuivre. Les résultats ont montré que la MOD issue des effluents de station d’épuration et rejetée dans le milieu récepteur présente des caractéristiques originales à savoir un fort pourcentage de MOD hydrophile, c’est à dire une faible hydrophobicité et un degré d’aromaticité peu marqué soulignant le caractère peu réfractaire de cette MOD. En revanche une plus grande diversité fonctionnelle de la MOD d’origine urbaine par rapport à la MOD naturelle a été mise en évidence notamment par un taux de structures protéiques très important. Les résultats obtenus ici pour la première fois vis-à-vis des MOD hydrophiles d’origine urbaine ont montré une réactivité particulière vis-à-vis du cuivre. En effet les fractions HPI et plus généralement les fractions issues des effluents de station d’épuration ont montré une teneur en sites complexants plus élevée que ce qu’il avait été déjà observé pour des matières organiques naturelles. Les valeurs des constantes ne semblent pas varier d’une fraction à l’autre et assez peu suivant l’origine de la MOD. Le rôle particulier de l’azote dans la complexation du cuivre notamment sous la forme de groupements amines a été mis en évidence. Les approches utilisées pour évaluer la biodisponibilité du cuivre ont révélé pour l’ensemble des fractions de MOD un effet protecteur vis-à-vis du cuivre sur les organismes vivants. Néanmoins cet effet protecteur ne se trouve pas à la hauteur de celui pouvant être prédit par le modèle de l’ion libre principalement dû à une biodisponibilité de certains complexes organiques. Ceci peut être expliqué par l’important taux de structures protéiques dans ces MOD, les protéines étant connues pour jouer un rôle important dans les mécanismes de transports des métaux au sein des organismes. Par ailleurs, l’outil DGT (Diffusive gradient in thin films) a montré son efficacité pour évaluer la fraction biodisponible du cuivre / This thesis was carried out on the framework of the ANR BIOMET research project (JC05_59809). The main objectives were to improve the current knowledge on the influence of dissolved organic matter (DOM) on metal speciation and bioavailability in urban aquatic system. The understanding of the influence of DOM on trace metal speciation and bioavailability required that the chemical composition of DOM, more exactly its binding sites should be well understood. Therefore, one part of this research focused on the isolation and characterization of DOM. The characterization of DOM, with a multidimensional approach was realized with a lot of various techniques (such as elemental, isotopic, functional and molecular) that were of prime importance to better understand the influence of DOM on trace metal speciation and bioavailability. To accomplish these objectives, the techniques such as potentiometry with ionic selective electrode and the recent electroanalytical technique AGNES and bioassays such as a bioaccumulation test (Fontinalis antipyretica) and an acute toxicity test (Daphnia magna) were performed in order to: - study the copper and lead binding by isolated DOM fractions in order to give some binding parameters of hydrophilic DOM and to better predict the fate of the trace metal. - evaluate the protective role of urban DOM on copper bioavailability. Results showed some particular characteristics of DOM from wastewater effluent such as a high proportion of hydrophilic DOM, i.e. a low hydrophobicity and a low degree of aromaticity underlying the low refractory character of urban DOM. Nevertheless a higher content of various functional groups was determined in urban DOM than in natural DOM. A very high content of proteinaceous structures was particularly identified in urban DOM. Binding experiments revealed for the first time on urban hydrophilic DOM, a higher content of binding sites than in natural DOM and these sites are strongly correlated to amino groups. The values of binding constant seem to not vary according to the nature of the fraction or the origin. The different approaches used to evaluate copper bioavailability depicted a protective role of DOM to the organisms Daphnia magna and Fontinalis antipyretica. Nevertheless this protective effect is hardly explained by the free ion concentration due to a bioavailability of some organic complexes. This feature could be explain with the high content of proteinaceous structures in these DOM fractions, since proteins are well know to play an important role in the transport mechanisms of trace metals into the organisms. Moreover, the DGT device (diffusive gradient in thin films) shows a good efficiency to assess the bioavailable copper