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Moluscos bivalves são organismos filtradores capazes de concentrar susbtâncias produzidas por microalgas tóxicas. No estado de Santa Catarina, líder na produção nacional, os cultivos têm sido oficialmente monitorados para a detecção de ficotoxinas causadoras dos Envenenamentos Diarreico (DSP), Amnésico (ASP) e Paralisante (PSP) por Consumo de Moluscos nas partes comestíveis. Amostras de água também são coletadas para a quantificação de algas nocivas. O objetivo deste trabalho foi sugerir o uso de áreas amostrais para coletas semanais, bem como calcular o tamanho das amostras agrupadas e analisar os dados de ocorrência. Diferentes cenários foram desenvolvidos para simular a variação dos tamanhos amostrais, utilizando-se o EpiTools®. Considerando-se uma alta prevalência e altas sensibilidades dos testes, é possível sugerir dois pools amostrais para a detecção de Toxinas Lipofíficas (2x30), duas para detectar PSP (2x15) e uma para detectar ASP (1x20) em cada uma das 24 áreas amostrais sugeridas. Se o teste de Cromatografia Líquida com Espectrometria de Massa (LC-MS/MS) for validado para todas as biotoxinas, apenas um pool amostral seria suficiente (1x15). Informações espaçotemporais de ocorrência também foram analisadas e apenas ficotoxinas causadoras de DSP foram encontradas. Utilizando-se os softwares SaTScan® e QGIS 2.12.2- Lyon®, foram desenvolvidos mapas de calor com os dois clusters espaciais encontrados para as detecções de DSP em moluscos e os quatro para Dinophysis acuminata (≥100cels/L) em amostras de água. Os resultados com maiores riscos relativos corresponderam ao cluster temporal do segundo semestre de 2014, os clusters espaciais das áreas 7 a 11 para DSP e áreas de 7 a 9 para D. acuminata. Esses resultados poderão contribuir para o planejamento de estratégias a serem incorporadas num futuro sistema de vigilância de moluscos bivalves do estado. / Bivalve molluscs are filtering organisms capable to concentrate substances produced by toxic microalgae. In Santa Catarina state, main Brazilian producer, the crops have been officially monitored for the detection of phycotoxins that cause Diarrheic (DSP), Amnesic (ASP), and Paralyzing (PSP) Shellfish Poisonings in edible parts. Water samples are also collected for harmful algae quantification. The goal of this study was to suggest the use of areas to be weekly sampled, as well as to calculate pooled sample sizes and to analyze the occurrence data. Different scenarios were developed to simulate the variation of sample sizes in EpiTools®. Considering a high prevalence and high tests sensitivities, we can suggest two pools to detect Lipophilic Toxins (2x30), two to detect PSP (2x15), and one to detect ASP (1x20) in each of the 24 suggested sampling areas. If the test of Liquid Chromatography with Mass Spectrometry (LC-MS/MS) becomes validated for all biotoxins, only one pool would be enough (1x15). Space-time occurence information was also analyzed and only phycotoxins causing DSP were found. Using SaTScan® and QGIS 2.12.2-Lyon® softwares, we developed heatmaps with two clusters found for DSP detection in shellfish and the four found for Dinophysis acuminate (≥100cels/L) in water samples. The results with higher relative risk values corresponded to the time cluster of the second semester of 2014, spatial cluster of the areas 7 to 11 for DSP, and areas 7 to 9 for D. acuminate. These results can contribute for the strategic plans to be incorporated in a future bivalve molluscs surveillance system of the state.

Ce travail a pour objectif d’étudier la biodiversité et la dynamique du zooplancton en relation avec les paramètres environnementaux dans la lagune Ghar El Melh au niveau de 5 stations à travers un suivi bimensuel allant de février 2011 à janvier 2012. Cette étude vise également à déterminer l’impact des régimes alimentaires à base de microalgues nuisibles, en conditions expérimentales, sur la survie, le comportement et la reproduction du copépode Calanoïde Acartia clausi collecté dans la lagune Ghar El Melh pendant la saison printanière.L’analyse hydrologique du milieu a mis en évidence une variation saisonnière de la totalité des paramètres avec des valeurs importantes de la température, de la salinité, du pH et de la Chl a en été et des teneurs élevées en nutriments en automne/hiver. L’étude spatio-temporelle du zooplancton a révélé la présence de 23 groupes zooplanctoniques dominés par les copépodes (37,25%) et les ciliés (21,09%). La lagune représente également un lieu de reproduction pour plusieurs espèces compte tenu de l’abondance du méroplancton tels que les larves de lamellibranches (14,9%) et les véligères de gastéropodes (12,5%). La température, la salinité, les caractéristiques hydrodynamiques, la teneur en ammonium et la richesse trophique ont été les facteurs déterminants pour le développement du zooplancton. Les relations d’ordre trophique semblent avoir un effet important sur la distribution des copépodes, ciliés et rotifères. La communauté des copépodes a été composée par 13 espèces dominées par les espèces opportunistes Oithona nana (59,5%), Acartia clausi (21%) et Euterpina acutifrons (8,87%) dont les abondances et la saisonnalité semblent être tributaires, en plus de l’abondance des proies, des caractéristiques hydrologiques de l’eau notamment de la température et la salinité.Les essais portant sur l’impact des microalgues nuisibles sur Acartia clausi ont mis en évidence i) une réduction du taux de survie (55%) et du nombre d’œufs produits (3,9 ± 3,2 œufs femelle-1 j-1) due à Prorocntrum micans malgré l’importance de l’activité alimentaire et du taux d’éclosion des œufs ii) Une altération importante de l’activité alimentaire, de la reproduction et de la survie due à Prorocentrum lima dès le premier jour iii) une létalitè importante de la souche Ostreopsis ovata se caractérisant par la mortalité de tous les individus après 24 heures iv) une altération importante du mouvement (33,6%) observée dès le 8ième jour, accompagnée d’une production réduite d’œufs (2,8 ± 3,9 œufs femelle-1 j-1), un faible taux d’éclosion (27,9 ± 30,6%) et d’une réduction importante de la survie due à Alexandrium catenella. / The aim of this work is to study the biodiversity and seasonal distribution of zooplankton community along with environmental factors in Ghar El Melh lagoon at 5 stations by twice monthly sampling from February 2011 to January 2012. This study also focused on the impact of several diets based on harmful microalgae on the survival, behavior and reproduction of the Calanoïd copepod Acartia clausi collected from Ghar El Melh lagoon during spring under laboratory control.The hydrological caracteristics revealed a seasonal variation of all parameters with high water temperature, salinity, pH and Chlorophyll a in summer and high nutrient concentrations in autumn/winter. The study of spatial and temporal variation of zooplankton revealed the presence of 23 groups mainly dominated by copepods (37.25%) and ciliates (21.09%). High abundance of meroplankton such as bivalve larvae (14.9%) and gastropod veligers (12.5%) showed that the lagoon can be considered as a breeding place for several marines species. Temperature, salinity, hydrodynamic characteristics, ammonium concentration and abundance of prey were the determining factors for the development of zooplankton. Trophic relationships seem to have an important effect on the distribution of copepods, ciliates and rotifers. The copepod community was composed of 13 species chiefly represented by opportunistic species Oithona nana (59.5%), Acartia clausi (21%) and Euterpina acutifrons (8.87%). Abundance and seasonality of this opportunistic community seem to be related to the abundance of prey and hydrological characteristics of water such as temperature and salinity.Bioassay on Acartia clausi using several harmful microalgae species showed (i) reduction of survival rate (55%) and egg production (3.9 ± 3.2 eggs female-1 day-1) due to Prorocntrum micans despite the high feeding rate and egg hatching success (80%) ii) a high decrease of feeding, reproduction and survival rates observed after 1 day of exposure to the toxic Prorocentrum lima iii) a very high lethality of used Ostreopsis ovata strain causing the mortality of all copepods after only 24 hours iv) high movement alteration rate (33.6%) observed after 8 days of exposure to Alexandrium catenella, associated to a reduced egg production rate (2.8 ± 3.9 eggs female-1 day-1), a low hatching success (27.9 ± 30.6%) and a reduced survival rate.

Moluscos bivalves são organismos filtradores capazes de concentrar susbtâncias produzidas por microalgas tóxicas. No estado de Santa Catarina, líder na produção nacional, os cultivos têm sido oficialmente monitorados para a detecção de ficotoxinas causadoras dos Envenenamentos Diarreico (DSP), Amnésico (ASP) e Paralisante (PSP) por Consumo de Moluscos nas partes comestíveis. Amostras de água também são coletadas para a quantificação de algas nocivas. O objetivo deste trabalho foi sugerir o uso de áreas amostrais para coletas semanais, bem como calcular o tamanho das amostras agrupadas e analisar os dados de ocorrência. Diferentes cenários foram desenvolvidos para simular a variação dos tamanhos amostrais, utilizando-se o EpiTools®. Considerando-se uma alta prevalência e altas sensibilidades dos testes, é possível sugerir dois pools amostrais para a detecção de Toxinas Lipofíficas (2x30), duas para detectar PSP (2x15) e uma para detectar ASP (1x20) em cada uma das 24 áreas amostrais sugeridas. Se o teste de Cromatografia Líquida com Espectrometria de Massa (LC-MS/MS) for validado para todas as biotoxinas, apenas um pool amostral seria suficiente (1x15). Informações espaçotemporais de ocorrência também foram analisadas e apenas ficotoxinas causadoras de DSP foram encontradas. Utilizando-se os softwares SaTScan® e QGIS 2.12.2- Lyon®, foram desenvolvidos mapas de calor com os dois clusters espaciais encontrados para as detecções de DSP em moluscos e os quatro para Dinophysis acuminata (≥100cels/L) em amostras de água. Os resultados com maiores riscos relativos corresponderam ao cluster temporal do segundo semestre de 2014, os clusters espaciais das áreas 7 a 11 para DSP e áreas de 7 a 9 para D. acuminata. Esses resultados poderão contribuir para o planejamento de estratégias a serem incorporadas num futuro sistema de vigilância de moluscos bivalves do estado. / Bivalve molluscs are filtering organisms capable to concentrate substances produced by toxic microalgae. In Santa Catarina state, main Brazilian producer, the crops have been officially monitored for the detection of phycotoxins that cause Diarrheic (DSP), Amnesic (ASP), and Paralyzing (PSP) Shellfish Poisonings in edible parts. Water samples are also collected for harmful algae quantification. The goal of this study was to suggest the use of areas to be weekly sampled, as well as to calculate pooled sample sizes and to analyze the occurrence data. Different scenarios were developed to simulate the variation of sample sizes in EpiTools®. Considering a high prevalence and high tests sensitivities, we can suggest two pools to detect Lipophilic Toxins (2x30), two to detect PSP (2x15), and one to detect ASP (1x20) in each of the 24 suggested sampling areas. If the test of Liquid Chromatography with Mass Spectrometry (LC-MS/MS) becomes validated for all biotoxins, only one pool would be enough (1x15). Space-time occurence information was also analyzed and only phycotoxins causing DSP were found. Using SaTScan® and QGIS 2.12.2-Lyon® softwares, we developed heatmaps with two clusters found for DSP detection in shellfish and the four found for Dinophysis acuminate (≥100cels/L) in water samples. The results with higher relative risk values corresponded to the time cluster of the second semester of 2014, spatial cluster of the areas 7 to 11 for DSP, and areas 7 to 9 for D. acuminate. These results can contribute for the strategic plans to be incorporated in a future bivalve molluscs surveillance system of the state.

During summer, the Baltic Sea is subjected to the world’s largest cyanobacterial blooms. These blooms are linked to eutrophication and raise many questions concerning their effects on the ecosystem. To understand their impacts on the food web dynamics, it is essential to assess growth responses of grazers to these cyanobacteria. In the northern Baltic proper, copepods are the most important herbivores providing an essential link between the primary producers and higher trophic levels. In this Thesis, Papers I &amp; II evaluate methods to estimate copepod growth in response to feeding conditions in situ. The most conspicuous diazotrophic filamentous cyanobacterium in the Baltic Sea is Nodularia spumigena, a producer of nodularin which is highly toxic to vertebrates, yet its ecological role is largely unknown. In Paper III, reciprocal interactions between cyanobacteria, sympatric algae and copepods are studied. The results suggest that nodularin is likely involved in allelopathic interactions, but it is not an inducible defense against grazers. Furthermore, the results of Papers IV &amp; V, indicate that natural assemblages of N. spumigena and Anabaena spp. may support copepod reproduction and that total diazotrophic filamentous cyanobacteria appear to provide a beneficial feeding environment for the feeding stages of copepod nauplii, most probably by stimulating the microbial communities that nauplii feed upon. Since cyanobacterial blooms are projected to increase due to global climate change, the combined effects of toxic cyanobacteria, ocean acidification and global warming predicted for year 2100 are further investigated on copepods in Paper IV. Taken together, these studies indicate that filamentous diazotrophic cyanobacteria contribute to sustaining secondary productivity and have potential implications of management practices with respect to combating eutrophication, global climate change and sustaining fish feeding conditions. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 4: Manuscript. Paper 5: Manuscript.</p>

Les rythmes biologiques constituent une propriété fondamentale de la vie, permettant aux organismes d’appréhender leur environnement et d’en anticiper les changements. Ces rythmes possèdent une double origine : une horloge moléculaire génère ces rythmes, qui sont ensuite synchronisés par des facteurs environnementaux. Si les organismes terrestres sont essentiellement soumis au rythme d’alternance jour/nuit, les espèces marines côtières et estuariennes occupent un biotope plus changeant encore : ils sont en effet également confrontés au rythme des marées. Pourtant, leurs rythmes biologiques sont à ce jour encore mal connus et les mécanismes moléculaires de(s) (l’) horloge(s) sous-jacente(s) ne sont pas caractérisés. Parallèlement, les efflorescences d’algues toxiques, en constante augmentation depuis 1970, constituent un problème écologique majeur, tant local qu’international. Les objectifs du présent travail consistaient à caractériser les rythmes de référence de l’huître Crassostrea gigas et d’en déterminer l’origine (moléculaire, zeitgebers). Il s’agissait ensuite d’étudier les perturbations potentielles de ces rythmes par l’algue toxique Alexandrium minutum, qui produit des toxines paralytiques et est régulièrement présente dans de nombreuses mers du globe. Les travaux réalisés ont permis de mettre en évidence l’existence d’un rythme d’activité valvaire circadien, faible et dual, et n’a pas permis de supporter l’hypothèse de l’existence d’une horloge circatidale. Nous avons formulé l’hypothèse que, chez C. gigas, le rythme tidal est soit d’origine exogène, soit produit par l’horloge circadienne synchronisée par les marées. Les analyses moléculaires réalisées sur le gène circadien cryptochrome dans le muscle adducteur - effecteur du mouvement des valves - ont montré que ce gène oscille à une fréquence tidale dans le muscle strié, favorisant notre seconde hypothèse. Par ailleurs, au-delà des gènes de l’horloge, l’algue A. minutum réprime l’expression de gènes impliqués dans différentes voies métaboliques importantes : la lutte contre le stress oxydant (cat, gpx), la respiration mitochondriale (cox1), l’immunité (ilk), la détoxification (mdr). Finalement nos analyses ont permis de mettre en évidence un impact génotoxique d’A. minutum chez C. gigas. / Biological rhythms constitute a fundamental property of life, allowing organisms to anticipate and adapt to their changing environment. These rhythms present a double origin: they are generated by a molecular clock and synchronized by environmental cues. Whereas terrestrial organisms are mainly subjected to day/night alternation, coastal and estuarine marine species inhabit an even more cycling biotope. They are indeed also submitted to tides. Nevertheless, biological rhythms in marine species are still unrecognized and molecular mechanisms of the underlying oscillator(s) are to date not determined. At the same time, toxic algae blooms are increasing since the 1970s and represent a major ecological concern, both at local and international levels. An objective of the present work was the characterization of biological rhythms in the oyster Crassotrea gigas and of their origin (molecular mechanism, zeitgebers). Furthermore, the work was designed to study the potential disruption of these rhythms by the toxic algal of worldwide distribution Alexandrium minutum, which produces paralytic toxins. The present results show the existence of a weak and dual circadian rhythm of valve activity in the oyster, and do not provide evidence for the existence of any circatidal clock. We suggested that, in the oyster C. gigas, the tidal rhythm could either be generated exogenously or endogenously by the tidally-synchronized circadian clock. Molecular analyses performed on the circadian gene cryptochrome in the adductor muscle of oyster, the effector of valve movements, revealed that Cgcry oscillates at tidal frequency in the striated muscle. This result supports our second hypothesis. Furthermore, A. minutum represses the expression of genes involved in key metabolic pathways: struggle against oxidative stress (cat, gpx), mitochondrial respiration (cox1), immunity (ilk), detoxification (mdr). Moreover, A. minutum impacts C. gigas at DNA level, being thus genotoxic.

Se ha estudiado la dinámica del fitoplancton en las lagunas costeras de Aiguamolls de l'Empordà. El fitoplancton esta sujeto principalmente al control "bottom-up", la variabilidad hidrológica y la disponibilidad de nutrientes tienen una mayor influencia en la composición y distribución de tamaños del fitoplancton, que el zooplancton. La concentración de materia orgánica disuelta es el factor ambiental más correlacionado con el crecimiento de la biomasa fitoplanctónica. Dada la proximidad entre las lagunas costeras y el mar, donde la ocurrencia de Proliferaciones de Algas Nocivas es cada vez más frecuente, se realizan un inventario general de las especies más abundantes del fitoplancton y se llevan a cabo análisis extensivos de la toxicidad. La mayoría de especies de dinoflagelados encontradas son potencialmente nocivas. Hay pocas especies en común entre el mar y las lagunas, sin embargo, existen especies productoras de PANs características de los ambientes lagunares. / The phytoplankton dynamics in the coastal lagoons of Empordà salt marshes was studied. The phytoplankton was mainly subject to the "bottom-up" control, the hydrologic variability and the nutrients availability have a greater influence in the composition and size distribution of the phytoplankton, than zooplankton. The concentration of dissolved organic matter was the environmental factor more correlated with the phytoplanktonic biomass growth. Due to the proximity between the coastal lagoons and the sea environment, where the occurrence of Harmful Algae Blooms is frequent, a general inventory of the most abundant species of the phytoplankton was made and an extensive monitoring of the toxicity was carried out. Most of dinoflagellates species found were potentially harmful. There were few species in common between the sea and the lagoons, nevertheless, some HABs species, characteristic of coastal lagoons, were found.