L’étude des effets des radionucléides et des métaux sur les organismes vivants est nécessaire pour évaluer leur toxicité et leur risque écologique. Notre approche a visé dans un premier temps à étudier les impacts du cadmium (Cd) et de l’Uranium (U) sur différents niveaux biologiques de l’écrevisse Procambarus clarkii après exposition aigue et chronique. Nous avons évalué leurs impacts sur les mitochondries, les réponses face à un stress oxydant, les structures histologiques et la survie. Nous avons tenté de lier ces effets entre eux et à la bioaccumulation dans les branchies et l’hépatopancréas. Nous avons aussi essayé de différencier la chimio- et la radiotoxicité de l’U en exposant des écrevisses soit à l’U appauvri soit au 233U (présentant une activité spécifique plus élevée) au travers des mêmes critères d’effets. Nous avons démontré que le gène mt codant pour la métallothionéine était toujours surexprimé en présence du Cd. De ce fait, il semble être un bon biomarqueur de toxicité du Cd chez P. clarkii. Nous avons mis en évidence que les deux métaux affectent les mitochondries grâce au suivi des niveaux d’expressions des gènes mitochondriaux (12s, atp6 et cox1), et que leur mécanismes d’action ne semblent pas être toujours les mêmes. Nous avons aussi prouvé que l’U génère plus de stress oxydant que le Cd grâce à la comparaison des niveaux d’expressions de gènes qui codent pour des antioxidants (sod(Mn) et mt) et des réponses enzymatiques de la superoxyde dismutase, la catalase, la glutathion peroxydase et la glutathion S transférase. Toutefois, les symptômes des atteintes histo-pathologiques semblent être les mêmes pour les deux métaux. En comparant leurs effets sur la survie des écrevisses, nous avons conclu que le Cd était plus toxique que le radioélément. D’autre part, nous avons démontré que les effets toxiques de l’U aux concentrations rencontrées dans l’environnement, sont plus liés à la chimiotoxicité qu’à la radiotoxicité de cet élément. Nous avons démontré que, les réponses moléculaires varient en fonction de l’intensité et la durée du stress chimique imposé aux organismes. Nous avons proposé d’utiliser les expressions de l’ensemble des gènes étudiés en tant que biomarqueurs de toxicité de l’U plutôt que les activités enzymatiques à cause de leur sensibilité. Ce travail de thèse propose des mécanismes d’actions de l’U basé surtout sur les réponses moléculaires et confirme la nécessité d’approfondir l’étude du profil écotoxicologique de ce radioélément. / The study of the effects of radionuclides and metals on organisms is necessary for the evaluation of their toxicity and their ecological threats. We first aimed to study the impacts of cadmium (Cd) and Uranium (U) on different biological levels of the crayfish Procambarus clarkii after acute and chronic exposures. We evaluated their impacts on mitochondria, oxidative stress responses, on histological structures, and the survival rates. We tried to connect these effects between them and to the bioaccumulation in the gills and the hepatopancreas. We also tried to discriminate the chemo and the radiotoxicity of U by exposing crayfish to either depleted or enriched U (233U: presenting a higher specific activity) using the same criteria of effects. We demonstrated that the gene mt encoding for the metallothionein was always over-expressed in the presence of Cd. Therefore, it seems to be a good biomarker of Cd toxicity in P. clarkii. The follow up of mitochondrial genes expressions (12s, atp6 and cox1), showed that both metals affect mitochondria and that their mechanisms of action do not seem to be always the same. We also observed that U generates more oxidative stress than Cd when comparing the expression levels of genes encoding for antioxidants (sod (Mn) and mt) and the enzymatic activities of superoxide dismutase, the catalase, the glutathione peroxidase and the glutathione S transferase. However, the symptoms of histo-pathological damages after Cd and U contamination were similar in both conditions. After comparing the survival rates of the crayfish, we concluded that Cd was more toxic than the radioelement. Moreover, we demonstrated that the toxic effect of U on P. clarkii exposed to a low environmental concentration is mainly due to its chemotoxicity rather than to its radiotoxicity. We established that, the molecular answers vary according to the intensity and the duration of the chemical stress applied to the organisms. We suggested the use of the expressions of all the studied genes as biomarkers of toxicity of U rather than the enzymatic activities because of their sensitivity. This work proposes mechanisms of actions of U based especially on the molecular responses and confirms the necessity of studying the ecotoxicological profile of this radioelement.

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The thesis is focused on anatomy and implementation of dissections of 4 selected model invertebrates in school practice. The selected model species include swan mussel (Anodonta cygnea), stick insect (Medauroidea extradentata), cockchafer (Melolontha melolontha) and sea urchin (Echinus esculentus). In case of the cockchafer, the text is based on this model species, but the supporting photographs of dissection are taken from the tropical cetoniid beetle Pachnoda, which has very similar internal anatomy with the cockchafer. For all selected animals, I describe external and internal morphology as well as recommended methods of obtaining the material, proper killing of specimens, and dissection techniques. The text is supplemented with original digital macrophotographs. The thesis includes also an overview of primary and secondary school textbooks with focus on anatomy of the selected animals. The text is supplemented by a glossary of morphological terminology used in this thesis. Attached is a photographic atlas of dissections, which is a combination of this master thesis and my bachelor thesis (defended in 2010), in which I dealt with five model invertebrates in the same way. This is Ascaris suum, Lumbricus terrestris, Helix pomatia, Procambarus aff. fallax and Archimandrita tesselata. The complete...

Environmental toxicologists and public health officials are responsible for assisting in the identification, management, and mitigation of public health hazards. As a result, there is a continued need for robust analytical tools that can aid in the rapid quantification and characterization of chemical exposure. In the first research phase, we demonstrated that a current tool for estimating human organophosphate pesticide exposure, measuring dialkyl phosphate (DAPs) metabolites in urine as chemical biomarkers of pesticide exposure, could represent exposure to DAPs themselves and not to pesticides. We showed that DAPs are metabolically stable, have high oral bioavailability, and are rapidly excreted in the urine following oral exposure. Results suggest that DAP measurements may lead to overestimates of human organophosphate pesticide exposure. In the second phase of research, a quick, easy, cheap, effective, rugged, and safe (QuEChERS) based analytical method was developed and validated for quantifying polycyclic aromatic hydrocarbons (PAHs) in biotic matrices with fat contents that ranged from 3 to 11%. Our method improved PAH recoveries 50 to 200% compared to traditional QuEChERS methods, performed as well or better than state of the art Soxhlet and accelerated solvent extraction methods, had sensitivity useful for chemical exposure assessments, and reduced sample preparation costs by 10 fold. The validated QuEChERS method was subsequently employed in a human exposure assessment. Little is known about how traditional Native American fish smoke-preserving methods impact PAH loads in smoked foods, Tribal PAH exposure, or health risks. Differences in smoked salmon PAH loads were not observed between Tribal smoking methods, where smoking methods were controlled for smoking structure and smoke source. PAH loads in Tribally smoked fish were up to 430 times greater than those measured in commercially available smoked fish. It is not likely that dietary exposure to non-carcinogenic PAHs at heritage ingestion rates of 300 grams per day poses an appreciable risk to human health. However, levels of PAHs in traditionally smoked fish may pose and elevated of risk of cancer if consumed at high rates over a life time. Accurately estimating PAH exposure in cases where aquatic foods become contaminated is often hindered by sample availability. To overcome this challenge, we developed a novel analytical approach to predict PAH loads in resident crustacean tissues based on passive sampling device (PSD) PAH measurements and partial least squares regression. PSDs and crayfish collected from 9 sites within, and outside of, the Portland Harbor Superfund site captured a wide range of PAH concentrations in a matrix specific manner. Partial least squares regression of crayfish PAH concentrations on freely dissolved PAH concentrations measured by PSDs lead to predictions that generally differed by less than 12 parts per billion from measured values. Additionally, most predictions (> 90%) were within 3-fold of measured values, while state of the art bioaccumulation factor approaches typically differ by 5 to 15-fold compared to measured values. In order to accurately characterize chemical exposure, new analytical approaches are needed that can simulate chemical changes in bioavailable PAH mixtures resulting from natural and/or remediation processes. An approach based on environmental passive sampling and in-laboratory UVB irradiation was developed to meet this need. Standard PAH mixtures prepared in-lab and passive sampling device extracts collected from PAH contaminated environments were used as model test solutions. UV irradiation of solutions reduced PAH levels 20 to 100% and lead to the formation of several toxic oxygenated-PAHs that have been previously measured in the environment. Site specific differences in oxygenated-PAH formation were also observed. The research presented in this dissertation can be used to advance chemical exposure estimation techniques, rapidly and cost-effectively quantify a suite of PAHs in biotic tissues, and simulate the effect of abiotic transformation processes on the bioavailable fraction of environmental contaminants. / Graduation date: 2013 / Access restricted to the OSU Community at author's request from April 3, 2013 - April 3, 2014

Invasive exotic species threaten native biodiversity, alter ecosystem structure and function, and annually cost over $100 billion in the US alone. Determining the ecological traits and interactions that affect invasion success are thus critical for predicting, preventing, and mitigating the negative effects of biological invasions. Native herbivores are widely assumed to facilitate exotic plant invasions by preferentially consuming native plants and avoiding exotic plants. Here, I use freshwater plant communities scattered broadly across the Southeastern U.S. to show that herbivory is an important force driving the ecology and evolution of freshwater systems. However, native consumers often preferentially consume rather than avoid exotic over native plants. Analyses of 3 terrestrial datasets showed similar patterns, with native herbivores generally preferring exotic plants. Thus, exotic plants appear defensively nave against these evolutionarily novel consumers, and exotic plants may escape their coevolved, specialist herbivores only to be preferentially consumed by the native generalist herbivores in their new ranges. In further support of this hypothesis, a meta-analysis of 71 manipulative field studies including over 100 exotic plant species and 400 native plant species from terrestrial, aquatic, and marine systems revealed that native herbivores strongly suppressed exotic plants, while exotic herbivores enhanced the abundance and species richness of exotic plants by suppressing native plants. Both outcomes are consistent with the hypothesis that prey are susceptible to evolutionarily novel consumers. Thus, native herbivores provide biotic resistance to plant invasions, but the widespread replacement of native with exotic herbivores eliminates this ecosystem service, facilitates plant invasions, and triggers an invasional meltdown. Consequently, rather than thriving because they escape their co-evolved specialist herbivores, exotic plants may thrive because their co-evolved generalist herbivores have stronger negative effects on evolutionarily nave, native plants.

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The most abundant organic compound produced by plants is cellulose, however it has long been accepted that animals do not secrete the hydrolytic enzymes required for its degradation, but rely instead on cellulases produced by symbiotic microbes. The recent discovery of an endogenous cDNA transcript encoding a putative GHF9 endoglucanase in the parastacid crayfish Cherax quadricarinatus (Byrne et al., 1999) suggests that similar cellulase genes may have been inherited by a range of crustacean taxa. In this study, the evolutionary history of the C. quadricarinatus endoglucanase gene and the presence of additional GHF9 genes in other decapod species were investigated. The activity of endoglucanase and endoxylanase enzymes within several cultured decapod species were also compared. The evolutionary history of the C. quadricarinatus endoglucanase gene was assessed by comparing intron/exon structure with that of other invertebrate and plant GHF9 genes. The coding region of the gene was found to be interrupted by eleven introns ranging in size from 102-902 bp, the position of which was largely conserved in both termite and abalone GHF9 genes. These structural similarities suggest GHF9 genes in crustaceans and other invertebrate taxa share a common ancestry. In addition, two introns were observed to share similar positions in plant GHF9 genes, which indicates this enzyme class may have been present in ancient eukaryote organisms. The presence of GHF9 genes in C. quadricarinatus and various other decapod species was then explored via degenerate primer PCR. Two distinct GHF9 gene fragments were determined for C. quadricarinatus and several other Cherax and Euastacus parastacid freshwater crayfish species, and a single GHF9 gene fragment was also determined for the palaemonid freshwater prawn Macrobrachium lar. Phylogenetic analyses of these fragments confirmed the presence of two endoglucanase genes within the Parastacidae, termed EG-1 and EG-2. The duplication event that produced these two genes appears to have occurred prior to the evolution of freshwater crayfish. In addition, EG-2 genes appear to have duplicated more recently within the Cherax lineage. The presence of multiple GHF9 endoglucanase enzymes within the digestive tract of some decapod species may enable more efficient processing of cellulose substrates present in dietary plant material. Endoglucanase and endoxylanase enzyme activities were compared in several parastacid crayfish and penaeid prawn species using dye-linked substrates. Endoglucanase activity levels were higher in crayfish compared with prawn species, which corresponds with the known dietary preferences of these taxa. Endoglucanase temperature and pH profiles were found to be very similar for all species examined, with optimum activity occurring at 60°C and pH 5.0. These results suggest endoglucanase activity in penaeid prawns may also be derived from endogenous sources. Additional in vitro studies further demonstrated crayfish and prawn species liberate comparable amounts of glucose from carboxymethyl-cellulose, which indicates both taxa may utilise cellulose substrates as a source of energy. Endoxylanase temperature and pH profiles were also similar for all crayfish species examined, with optimal activity occurring at 50°C and pH 5.0. These results suggest xylanase activity in crayfish may originate from endogenous enzymes, although it is unclear whether this activity is derived from GHF9 enzymes or a different xylanase enzyme class. In contrast, no endoxylanase activity was detected in the three prawn species examined. Together, these findings suggest a wide range of decapod crustacean species may possess endogenous GHF9 endoglucanase genes and enzymes. Endoglucanases may be secreted by various decapod species in order to digest soluble or amorphous cellulose substrates present in consumed plant material. Further biochemical studies may confirm the presence and functional attributes of additional endoglucanase genes and enzymes in decapods, which may ultimately assist in the design of optimal plant based crustacean aquaculture feeds.

La séquence sédimentaire du Dur At-Talah, située dans la partie sud du bassin de Syrte au centre de la Libye, est composée de 150 m d'épaisseur de roches principalement siliciclastiques (grès, siltites et argiles). L'importance de cette séquence est liée en partie à l'importance du bassin de Sirt qui est un des plus grands réservoirs d'hydrocarbures de Libye. La séquence est également un des plus important site de fossiles de vertébrés de l’Eocène supérieur, l'âge de la séquence. Les études antérieures, bien que très limitées par rapport à l'importance de cette zone, se sont concentrées principalement sur le contenu paléontologique de la séquence. L’étude sédimentologique n’avait été jusqu’à ce jour que très peu abordée. Cette thèse est un travail basé sur l’examen des affleurements et où l'accent est mis sur les structures sédimentaires et biogéniques (traces fossiles), visant à définir et à interpréter les faciès sédimentaires qui ont construit la séquence du Dur At Talah. L’ étude de la séquence a conduit à diviser la totalité de l’affleurement en trois intervalles génétiquement liés. Le plus ancien que nous avons appelé la « New Idam Unit » (environ 80 m d’épaisseur), est composé de grès très fins, de siltites et d’argiles. Cette unité (New Idam Unit) est recouverte en discordance par la « Sarir Unit » (environ 50 m d’épaisseur), composée de grès fins à moyens à faisceaux de litages entrecroisées dans sa partie inférieure (environ 25-30 m d’épaisseur) et de grès grossiers à microconglomératiques dans sa partie supérieure (environ 20-30 m d’épaisseur). La « Sarir Unit » est ainsi divisée en « Sarir inférieur » et « Sarir supérieur ». La « New Idam Unit » présente des dépôts qui sont attribués à un milieu main estuarien. La série débute par des dépôts d’environ 35 m d’épaisseur typiques d’un estuaire externe. Les 45 mètres qui lui font suite passent progressivement à des dépôts tidaux caractéristiques d’un estuaire interne. La Surface d'inondation maximale se situe entre les deux intervalles. Au-dessus de cette surface les indicateurs fluviaux augmentent et les indicateurs de marées diminuent progressivement, offrant ainsi un indice pour la migration de la rive de rivage vers le bassin (vers le Nord). La sous-unité du Sarir inférieur qui avait été interprétée avant ce travail comme dépôts fluviatiles, préserve pourtant des structures sédimentaires multi-échelles qui sont le résultat incontestable de processus de marée. Ceci est particulièrement évident dans la partie inférieure de l’unité « Sarir inférieur » (la Lower Lower Sarir Unit ; LLS). Dans la partie supérieure de l’unité Sarir inférieur (Upper Lower Sarir; ULS), les indicateurs de dynamique fluviale dominent largement sur ceux de la marée. Le Sarir inférieur est donc interprété comme un système deltaique mis en place lors d’une régression normale. Cette fois-ci, la surface maximale d'inondation se situe entre le LLS et ULS. Le « Upper Lower Sarir » (ULS) se terminée par une discordance subaérienne, avec de nombreuses traces d’émersion conservées au sommet de l'ULS. Celles-ci sont recoupées par le « Upper Sarir » qui montre des marqueurs fiables d'environnement de dépôt strictement fluvials. Les dépôts du « Upper Sarir » enregistre la séquence de bas niveau marin. Cette étude fournit des informations précieuses concernant le dépôt des séquences dans le bassin de Syrte au cours de la fin de l'Éocène. Elle fournit également une étude originale sur la dynamique tidale dans des milieux marins margino-littoraux. / Dur At Talah sedimentary sequence, located at the southern side of the Sirt Basin in central Libya, is composed of 150 m thick of mainly siliciclastic rocks. The importance of this sequence is linked to the importance of the Sirt Basin as one of large hydrocarbon reservoirs in Libya. The sequence is also an excellent site for vertebrate fossils of Late Eocene, the age of the sequence. Previous studies, though very limited compared to the importance of this area, are focused on its paleontological content. Sedimentology received only scant attention before this project. This thesis is an outcrop based study in which the focus is given to the sedimentary and biogenic (trace fossils) structures, aiming at defining and interpreting depositional facies which building up the sequence. The study is mainly based on field data which are analyzed on the light of related published literature and on the comparison with modern sedimentary environments. Results of facies analysis have led to splitting the entire sequence into three genetically related intervals. The oldest, we called the New Idam Unit (around 80m), is composed of very fine sandstones to mudstones. New Idam Unit is unconformably overlain by the Sarir Unit (around 50m), composed of medium grained cross bedded sandstones (the lower 25-30 m) changes up to very coarse and microconglomeratic sandstone (the upper 20-30 m). Thus, the Sarir Unit is split into the lower Sarir Subunit and upper Sarir subunit. The New Idam Unit presents both classical and unusual sedimentary and biogenic indicators that attribute this unit to estuarine depositional environment. It starts with outer estuarine (the lower 35 m) and ends up with inner estuarine (the upper 45 m). Maximum flooding surface is located in between. Above this surface the fluvial indicators increase and tidal indicators decrease, thus providing clue for basinward (North) migration of the shoreline. The lower Sarir subunit which was previously interpreted as fluvial deposits, preserves multi-scale sedimentary structures that undoubtedly belong to tidal processes. This is especially evidenced at the lower part of the lower Sarir Subunit (LLS). Fluvial indications over dominates the tidal ones in the upper part of the lower Sarir (ULS). Due to this configuration the whole lower Sarir subunit is interpreted as shallow marine, deltaic, depositional system, occurred during sea level ¨normal¨ regression. This time, maximum flooding surface is located between the LLS and ULS. The lower Sarir subunit is terminated by subaerial unconformity, with evidences of subaerial exposure preserved at the top of the ULS. These are intruded by the upper Sarir subunit which presents clear evidences of strictly fluvial environment of deposition. The deposits of the upper Sarir subunit record the low stand system tract part of the Dur At Talah sequence. In addition to the outlined results, the sequential pattern of the depositional events is suggested for the entire sequence of Dur At Talah. This study provides a valuable information regarding the depositional and sequential aspects of the Sirt Basin during the late Eocene, it also provide an unique case study for the better understanding of the shallow marine tidal deposits.