Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2014-09-05T14:43:24Z No. of bitstreams: 1 Tese de Doutorado Gabriel Luiz dos Santos.pdf: 1820652 bytes, checksum: 813a41cbb9ae01d5c374c54cce45816c (MD5) / Approved for entry into archive by Fatima Cleômenis Botelho Maria (botelho@ufba.br) on 2014-09-05T16:01:28Z (GMT) No. of bitstreams: 1 Tese de Doutorado Gabriel Luiz dos Santos.pdf: 1820652 bytes, checksum: 813a41cbb9ae01d5c374c54cce45816c (MD5) / Made available in DSpace on 2014-09-05T16:01:28Z (GMT). No. of bitstreams: 1 Tese de Doutorado Gabriel Luiz dos Santos.pdf: 1820652 bytes, checksum: 813a41cbb9ae01d5c374c54cce45816c (MD5) / CNPq / O presente trabalho realizado em amostras coletadas na Baía de Todos os Santos, Bahia, Brasil, propõe estratégias para determinação multielementar e análise de especiação de vanádio em amostras de moluscos bivalves e sedimentos, empregando ICP OES e ICP-MS. Na primeira parte da tese, foram avaliadas as potencialidades de dois espectrômetros de massa com plasma indutivamente acoplado para determinação de As, Cr, Ni, Se e V em amostras de moluscos bivalves. Para efeito de comparação, três materiais de referência certificados (CRM) foram analisados: NIST 1566b SRM Oyster tissue, NRCC TORT-2 Lobster Hepatopancreas e NIST 2977 SRM Mussel tissue. A condição recomendada, que consistiu na determinação no modo padrão com adição de padrão interno, foi aplicada na determinação dos analitos em amostras de Macoma constricta coletadas em diferentes regiões da BTS. Os resultados apontaram que a maioria dos níveis de As, Cr, Ni, Se e V presentes na espécie Macoma constricta apresentaram valores acima dos níveis estabelecidos pela legislação vigente brasileira que são: 1,0 (As); 0,1 (Cr); 5,0 (Ni); 0,3 (Se) e 0,77 (V) g g-1 (peso úmido), o que representa risco à população e ao ecossistema. Na segunda parte da tese, foram investigadas as condições para a análise de especiação de vanádio em amostras de sedimentos, empregando extração alcalina e determinação por ICP OES. A concentração de V(IV) foi determinada por diferença, considerando a determinação de vanádio total, também realizada por ICP OES, após digestão das amostras com ácido nítrico em forno de micro- ondas com cavidade, usando o procedimento EPA 3051A. O procedimento foi validado através de testes de adição e recuperação e de análise do seguinte material de referência certificado: NIST 2702 (Inorganics in marine sediment). Pelos resultados obtidos pode-se concluir que os compostos contendo V(IV) são mais dominantes nas amostras dos sedimentos analisados. / The present work conducted on samples collected in Todos os Santos Bay, Bahia, Brasil, proposes strategies for multielement determination and speciation analysis of vanadium in bivalve molluscs and sediments samples, using ICP OES and ICP-MS. In the first part of the thesis, the potential of two inductively coupled plasma mass spectrometers for determination of As, Cr, Ni, Se and V in bivalve molluscs samples were evaluated. For comparison, three certified reference materials (CRM) were analyzed: NIST 1566b SRM Oyster tissue, NRCC TORT-2 Lobster Hepatopancreas e NIST 2977 SRM Mussel tissue. The recommended condition, which consisted in determining the standard mode with addition of internal standard, was applied in the determination of analytes in samples of Macoma constricta collected in different regions of the BTS. The results showed that most levels of As, Cr, Ni, Se and V present in the species Macoma constricta have values above the levels established by the Brazilian legislation, that are: 1.0 (As); 0.1 (Cr); 5.0 (Ni); 0.3 (Se) and 0.77 (V) g g-1 (wet weight), which represents risk to the population and the ecosystem. In the second part of the thesis, the conditions for speciation analysis of vanadium in soil and sediment samples were investigated, using alkaline extraction and determining by ICP OES. The V (IV) concentration was determined by difference, considering the total vanadium determination, also performed by ICP OES, after the samples digestion with nitric acid in cavity microwave oven, using the procedure EPA 3051A. The procedure was validated through addition and recovery tests and analysis of the following certified reference material: NIST 2702 (Inorganics in marine sediment). From the obtained results it can be concluded that compounds containing V (IV) are majority in the sediment samples analyzed.

Made available in DSpace on 2014-06-11T19:26:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-10-05Bitstream added on 2014-06-13T19:54:17Z : No. of bitstreams: 1 neves_jp_me_rcla.pdf: 9688721 bytes, checksum: e459ad8bf7d442ee8695ccc1b4c3da9f (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Bivalves fósseis são comuns em rochas siliciclásticas e carbonáticas das formações Teresina e Rio do Rasto (Permiano Médio/Superior), Bacia do Paraná. Entretanto, a gênese das camadas de conchas carbonáticas ainda é pouco conhecida. Cinco camadas de calcários do centro-sul e norte do Estado do Paraná foram selecionados para estudo tafonômico: dois grainstones oolíticos a bivalves (Prudentópolis) e um grainstone a peloides e bivalves com intraclastos e oncoides (Rio Preto) da Formação Teresina; um packstone e um wackestone a bivalves e oncoides (Ribeirão Claro) da Formação Rio do Rasto. As camadas carbonáticas (≤ 45 cm de espessura), estão intercaladas entre rochas pelíticas, possuem contato basal erosivo, quantidade variável de intraclastos pelíticos, valvas caoticamente distribuídas com gradação vertical descontínua e empacotamento denso a disperso. As conchas encontram-se desarticuladas, comumente fragmentadas, às vezes incrustadas por cianobactérias, e correspondem a espécies alóctonas que viviam em substratos distintos. As assembléias de bivalves representam, portanto, tanatocenoses em tempestitos proximais amalgamados, gerados em água rasa com frequentes modificações por bioturbação e baixa taxa de sedimentação. A presente pesquisa corrobora outras evidências de que acumulações de conchas paleozóicas em mares epicontinentais resultaram de história tafonômica complexa com acentuada mistura temporal e espacial dos bioclastos sob influência de tempestades / Bivalve fossils are common in siliciclastic and carbonate rocks of the Teresina and Rio do Rasto formations (Middle to Late Permian), Paraná Basin. However, little is known about shell concentrations in carbonates. Five limestone beds in south-central and northern Paraná State were selected for a taphonomic study: two oolite-bivalve grainstones at Prudentópolis and one peloid-bivalve grainstone with intraclasts and oncoids at Rio Preto, from Teresina Formation; one packstone and one wackestone with bivalves and oncoids at Ribeirão Claro, from Rio do Rasto Formation. The limestone beds (≤ 45 cm thick) are intercalated with pelitic rocks; the basal contact is sharp and erosive, with variable amount of pelitic intraclasts. Shells are randomly oriented (many nested/stacked), showing dense to disperse packing, and discontinuous grading. The shells are disarticulated, commonly fragmented, sometimes encrusted by stromatolites, and correspond to allochthonous specimens winnowed from distinct life-substrates. The bivalve concentrations were generated in shallow water settings punctuated by storms, under very low sedimentation rates, with frequent intrastratal bioturbation. Hence, the concentrations are amalgamated proximal tempestites. This work corroborates previous evidences that Paleozoic shell beds from the epeiric seas have complex taphonomic histories and result from strong temporal/spatial mixing of bioclasts under storm influence

Dans le contexte des changements climatiques, les écosystèmes marins arctiques sont confrontés à des modifications environnementales accélérés, dont les conséquences sur les communautés biotiques sont encore débattues. La diminution du couvert de glace, l’augmentation de la turbidité et des apports d’eau douce vont affecter les producteurs primaires arctiques, avec des effets en cascade sur un processus-clé de ces écosystèmes : la relation trophique entre producteurs primaires et consommateurs benthiques (à laquelle se réfère généralement l’expression « couplage pélagos-benthos »). L’étude directe de ces interactions complexes n’est pas aisée dans ces milieux. Le modèle biologique des bivalves filtreurs peut permettre de contourner ces problèmes en remplissant une fonction d’ « intermédiaire » pour la compréhension de ces processus écologiques. Parmi les avantages de ce modèle d’étude, il y a tout d’abord le fait que ces organismes enregistrent au sein de leur coquille, dans les couches de biocarbonates, certaines dynamiques de leurs environnements. Les informations contenues dans ces « bioarchives » sont interprétées grâce aux méthodes de la sclérochronologie et de la sclérochimie et concernent une fenêtre temporelle correspondante à la vie des individus (de quelques années à plus que 500 ans). Un autre avantage de ce modèle biologique est que, s’agissant d’organismes qui sont des consommateurs primaires, l’étude de leur régime alimentaire peut apporter des éléments sur la relation trophique avec les producteurs primaires. Avec les méthodes de l’écologie trophique, en particulier les acides gras et les isotopes stables, l’étude des tissus permet d’obtenir des informations sur les sources assimilées à l’échelle de quelques semaines/mois. L’objectif de cette thèse est de tester le potentiel des bivalves Astarte moerchi (complexe borealis) comme modèle biologique pour l’étude des écosystèmes marins arctiques. Pour ce faire, une approche couplée est utilisée combinant l’analyse de la coquille par les méthodes de la sclérochronologie et de la sclérochimie (ratios élémentaires) et l’analyse des tissus par les méthodes de l’écologie trophique (acides gras, isotopes stable du carbone et de l’azote, isotopes du carbone sur acides gras individuels). Deux populations actuelles d’Astarte moerchi ont été étudiées dans deux fjords présentant des caractéristiques environnementales contrastées : le Young Sound au Nord-Est du Groenland (considéré comme fjord « arctique ») et le Kongsfjorden à l’Ouest de l’Archipel du Svalbard (considéré comme site « sub-arctique »). L’étude des tissus d’A. moerchi a permis de mettre en évidence la plasticité trophique de cette espèce, avec des différences dans les sources d’alimentation de deux populations liées aux dynamiques locales de production primaire. L’étude de la coquille d’A. moerchi a permis de : a) corroborer l’hypothèse de la formation annuelle des stries de croissance, confirmant la longévité de cette espèce pouvant atteindre 150 ans ; b) montrer l’intérêt potentiel de l’étude des ratios élémentaires dans les biocarbonates et en particulier du ratio Barium sur Calcium (Ba/Ca), qui pourrait être relié aux efflorescences phytoplanctoniques et c) montrer que les conditions environnementales contrastées du site arctique et sub-arctique se traduisent dans des patrons de croissance coquillère différents. Des perspectives pour l’utilisation ultérieure de ce modèle en écologie sont discutées. Pour conclure, une réflexion épistémologique est amorcée sur la spécificité du modèle biologique d’étude des bivalves filtreurs. En opposition à une notion plus classique d’ « organisme modèle » utilisée en biologie expérimentale, nous proposons que les bivalves filtreurs (à l’instar d’autres « bioarchives » comme les arbres, des coraux et des algues corallines) appartiennent à une catégorie de modèles biologiques qu’on pourrait qualifier « in situ » et qui semble être spécifique à la discipline écologique. / In the context of climate change, Arctic marine ecosystems are affected by rapid environmental modifications, whose effects on biotic communities are still debated. The sea-ice decline and the increase in freshwater inputs and turbidity are likely to impact Arctic primary producers, with cascade effects on a key-process in those ecosystems: the trophic relationship between primary producers and benthic consumers (generally referred as “pelagic-benthic coupling”). The direct study of such complex interaction is not straightforward in the Arctic. The biological model of filter-feeding bivalves offers the possibility to get around these problems, allowing to study those ecological processes indirectly. Among the advantages of this model, there is first of all the fact that these organisms record in their shell, in the carbonate layers, some dynamics of their environments. The information recorded in such “bioarchives” are interpreted through the methods of sclerochronology and sclerochemistry and relate to a time window corresponding to the organism lifespan (from some years to more than 500 years). Given that these organisms are primary consumers, another advantage of this biological model is that the study of their diet can provide information about the trophic relationship with primary producers. With the methods of trophic ecology, especially fatty acids and stable isotopes, the study of the tissues allows the investigation of sources assimilated at a timescale of weeks/months.The main objective of this thesis is to test the potential of bivalves Astarte moerchi (borealis complex) as a biological model for the study of marine Arctic ecosystems. A coupled approach is used to combine shell analysis by the methods of sclerochronology and sclerochemistry (elemental ratios) and tissue analysis by the methods of trophic ecology (fatty acids, carbon and nitrogen stable isotopes, compound-specific carbon stable isotopes on individual fatty acids). Two living A. moerchi populations have been studied in two fjords presenting contrasted environmental conditions: Young Sound in North-East Greenland (considered as “Arctic” site) and Kongsfjorden in the West coast of the Svalbard Archipelago (considered as a “sub-Arctic” site). The study of the tissues of A. moerchi allowed to show the trophic plasticity of this species, with differences in food sources of the two populations linked to local primary production dynamics. The analysis of the shell of A. moerchi allowed to: a) corroborate the hypothesis of annual growth lines formation, thus confirming the longevity of this species that can attain 150 years; b) show the potential interest of the analysis of elemental ratios and particularly the ratio between Barium and Calcium (Ba/Ca), which could be relied to phytoplanktonic blooms and c) show that contrasted environmental conditions in the Arctic and sub-Arctic sites result in different shell growth patterns. Some perspectives for the further use of this model study in ecology are discussed. To conclude, an epistemological reflection is sketched about the specificity of the biological model study of filter-feeding bivalves. In contrast to the classical notion of “model organism” used in experimental biology, we suggest that filter-feeding bivalves (as well as other “bioarchives” like trees, corals and coralline algae) belong to a category of biological models that could be named “in situ” and seems specific to the ecological discipline.

Os bivalves marinhos sÃo muitos apreciados como alimento e no aproveitamento de suas conchas para vÃrias utilidades especialmente como enfeites. A presente pesquisa teve como objetivo estudar o bivalve marinho venerÃdeo Anomalocardia brasiliana, quanto a densidade populacional e ciclo de reproduÃÃo, correlacionando este Ãltimo aos parÃmetros ambientais (salinidade, temperatura e precipitaÃÃo pluviomÃtrica). As amostragens foram realizadas mensalmente durante o perÃodo de maio/02 a abril/03, na praia do Canto da Barra, municÃpio de Fortim â CE, no estuÃrio do Rio Jaguaribe. Os indivÃduos Anomalocardia brasiliana apresentaram um padrÃo de distribuiÃÃo bem caracterÃstico, com os juvenis concentrados em Ãrea com sedimento fino e mais Ãmido e os adultos em banco arenosos com sedimento mais grosseiro. A diferenciaÃÃo sexual ocorreu provavelmente entre 12,6mm e 17,9mm de comprimento de conchas. A identificaÃÃo dos sexos foi realizada utilizando-se tÃcnicas histolÃgicas de rotina. Durante a gametogÃnese, foram reconhecidas em fÃmeas, ovogÃnias, ovÃcitos prÃ-vitelogÃnicos, ovÃcitos vitelogÃnicos e ovÃcitos maduros, e, em machos, espermatogÃnias, espermatÃcitos I, espematÃcitos II, espermÃtides e espermatozÃides. A reproduÃÃo da espÃcie à contÃnua, entretanto foram observados perÃodos de maior liberaÃÃo de gametas de julho a outubro/2002 e de fevereiro a abril/2003. O ciclo reprodutivo caracterizou-se pelas seguintes as fases do desenvolvimento gonadal: âem MaturaÃÃoâ, âMaduroâ, âem EliminaÃÃoâ, âEliminadoâ, para ambos os sexos. Dentre os fatores ambientais observados neste estudo, a precipitaÃÃo pluviomÃtrica e a salinidade foram as que apresentaram as maiores amplitudes de variaÃÃo. Entretanto, uma influÃncia significativa do efeito da diminuiÃÃo da salinidade foi observada somente em machos, provavelmente causada pela alta precipitaÃÃo pluviomÃtrica verificada em abril/03. Durante este estudo, observou-se a ocorrÃncia de parasitas do gÃnero Bucephalus sp. em 6,5% dos indivÃduos analisados. Apesar da baixa incidÃncia de organismos parasitados, a ocorrÃncia de parasitismo na populaÃÃo de Anomalocardia brasiliana, no estuÃrio do Rio Jaguaribe, alerta para necessidade de estudo sobre as condiÃÃes fisiolÃgicas desta espÃcie na regiÃo. / Marine bivalves are very appreciated as food and its shell is used for several purposes, especially for ornament. The present research aims to study the marine bivalve Anomalocardia brasiliana in relation to its population density and reproductive cycle, correlating them with environmental parameters (salinity, temperature and precipitation). Monthly samplings were realized from May 2002 to April 2003 at Canto da Barra Beach, in the Jaguaribe River Estuary, Fortim, CearÃ. The Anomalocardia brasiliana individuals presented a quite characteristic distribution pattern, with the juvenile ones concentrated in areas with fine and moist sediment (fine sand) and the adult ones concentrated in sandy beds with coarse sediment (medium sand). Sexual differentiation probably occurred when the shell length was between 12,6mm and 17,9mm. Sex determination was realized using custom histological techniques. During gametogenesis, oogonia, previtellogenic oocytes, vitellogenic oocytes and mature oocytes were recognized in females, and spermatogonia, spermatocytes I, spermatocytes II, spermatids and spermatozoa were recognized in males. The reproductive cycle is continuous. Nevertheless, periods of intense gamete liberation were observed from July to October 2002 and form February to April 2003. For both sexes the reproduction cycle was characterized by the following stages in the development: âin Ripeningâ, âRipeâ, âin Spawningâ, âSpawnedâ. Among the environmental agents observed in this study, precipitation and salinity were the ones that presented the largest fluctuation in amplitude. However, a major influence by the salinity reduction effect was only observed in males, probably caused by the high precipitation registered in April 2003. During this study, the occurrence of parasites from the genus Bucephalus sp. was observed in 6,5% of the analyzed individuals. Despite the low influence of organisms infected with parasites, the occurrence of parasitism in the Anomalocardia brasiliana population at the Jaguaribe River Estuary alerts the necessity for further studies on the physiological conditions of this species in that area.

Salmonella spp. foi isolada em 20% (18) amostras de água de regiões portuárias (Portos de Belém/PA, Fortaleza/CE, Itaguaí/RJ, Paranaguá/PR, Recife/PE, Rio Grande/RS e Santos/SP) e em 19% (04) amostras de moluscos bivalves. Dentre os 214 isolados de Salmonella spp. em amostras de água (206) e bivalves (8), o sorotipo S. Saintpaul O:4 [B] foi o mais freqüente (53/214). Na avaliação dos isolados quanto à suscetibilidade a 14 antimicrobianos, 204 (95,3%) apresentaram resistência até a dez agentes antimicrobianos. O método de PCR foi utilizado para caracterizar os sorogrupos de Salmonella spp. (A-D1, B, e C2-C3), sendo eficiente em 93,6% dos isolados. A presença de Salmonella spp., inclusive multiresistentes, em áreas portuárias reflete em risco para saúde pública. Programas de vigilância ambiental, epidemiológica e sanitária poderiam ser contempladas nas áreas portuárias, evitando o transporte de Salmonella spp., através da água de lastro de navios e que podem atingir áreas de balneabilidade ou aqüicultura. / Salmonella spp. was isolated in 20% (18) samples of port regions water (Ports of Belém/PA, Fortaleza/CE, Itaguaí/RJ, Paranaguá/PR, Recife/PE, Rio Grande/RS and Santos/SP) and in 19% (04) samples of mollusk bivalves. Amongst the 214 isolated of Salmonella spp. in water samples (206) and bivalves (8), serotype S. Saintpaul O: 4 [B] was most frequent (53/214). In the evaluation of these isolated to the susceptibility for the 14 antimicrobial agents profile, 204 (95.3%) had presented resistance until ten agents. The PCR method was used to characterize the serogroups of Salmonella spp. (A-D1, B, and C2-C3), being efficient in 96,3% of the isolated ones. The presence of Salmonella spp., also multiresistant, in port areas reflects at risk for public health. Programs of environment survey, epidemiological and sanitary could be contemplated in the port areas, preventing the transport of Salmonella spp. through at the ballast water of ships and that they can reaching recreational or aquaculture areas.

Global change poses new threats for life in the oceans forcing marine organisms to respond through molecular acclimatory and adaptive strategies. Although bivalve molluscs are particularly tolerant and resilient to environmental stress, they must now face the challenge of more frequent and severe Harmful Algal Blooms (HABs) episodes. These massive outbreaks of microalgae produce toxins that accumulate in the tissues of these filter-feeder organisms, causing changes in their gene expression profiles, which in turn modify their phenotype in order to maintain homeostasis. Such modifications in gene expression are modulated by epigenetic mechanisms elicited by specific environmental stimuli, laying the foundations for long-term adaptations. The present work aims to examine the links between environmental stress in bivalve molluscs (with especial emphasis on Harmful Algal Blooms) and specific epigenetic marks triggering responses through modifications in gene expression patterns. Overall, a better understanding of the molecular strategies underlying the conspicuous stress tolerance observed in bivalve molluscs will provide a framework for developing a new generation of biomonitoring strategies. In addition, this strategy will represent a valuable contribution to our knowledge in acclimatization, adaptation and survival. With that goal in mind, the present work has generated transcriptomic data using RNA-Seq and microarray technologies, facilitating the characterization and investigation of the epigenetic mechanisms used by the Mediterranean mussel Mytilus galloprovincialis during responses to HAB exposure. That information was made publicly available through a specialized online resource (the Chromevaloa Database, chromevaloa.com) assessing the response of chromatin-associated transcripts to Okadaic Acid. Specific epigenetic marks have been assessed under lab-controlled exposure experiments simulating the natural development of the HAB Florida Red Tide (FRT). Results demonstrate a role for the phosphorylation of histone H2A.X and DNA methylation in the response to FRT in the Eastern oyster Crassostrea virginica. Lastly, the study of co-expression networks based on RNA-Seq data series from the Pacific oyster Crassostrea gigas reveals dynamic transcriptomic patterns that vary with time, stressor and tissue. However, consistent functional profiles support the existence of a core response to general conditions of environmental stress. Such response involves metabolic and transport processes, response to oxidative stress and protein repair or disposal, as well as the activation of immune mechanisms supporting a tightly intertwined neuroendocrine-immune regulatory system in bivalves.

Les mitochondries et leur génome, l'ADN mitochondrial (ADNmt), sont généralement transmis uniquement par la mère aux fils et aux filles chez les métazoaires (transmission strictement maternelle, SMI). Une exception à la règle générale de la SMI se trouve dans environ 100 espèces de bivalves, qui se caractérise par une double transmission uniparentale (DUI) des mitochondries. Chez les espèces DUI, deux lignées d'ADNmt très divergentes et liées au sexe coexistent. Une lignée mitochondriale maternelle (type F), présente dans les ovocytes et les tissus somatiques des individus femelles et males, et une lignée paternelle (type M), présente dans les spermatozoïdes. Dans les tissus somatiques mâles, les deux lignées coexistent parfois, une condition appelée hétéroplasmie. En sachant que les variations génétiques dans l’ADNmt peuvent avoir un impact sur les fonctions mitochondriales, et en donnant l'association stricte des ADNmt de type M et F avec différents gamètes, il est imaginable que la forte divergence entre les deux lignées DUI puisse entraîner des adaptations bioénergétiques avec répercussion sur la reproduction. Le système DUI apporte également la nécessité pour les mitochondries paternelles de préserver leur propre intégrité génétique, ainsi que pour les cellules somatiques de faire face à l'hétéroplasmie. L'objectif de ma thèse était de lier le génotype mitochondrial des espèces bivalves DUI et SMI au phénotype. Plus précisément, j'ai exploré l'impact des variations de l'ADN mitochondrial spécifiques au sexe sur un large éventail de traits phénotypiques, allant de la bioénergétique mitochondriale et cellulaire à la performance des spermatozoïdes, en étudiant la valeur adaptative du système DUI à la lumière du fitness reproductif, de la sélection et de la transmission mitochondriales. Les résultats issus de ce projet de thèse ont révélé une nette divergence phénotypique entre les espèces DUI et SMI, reflétant peut-être les différentes pressions sélectives agissant sur les deux lignées mitochondriales. Contrairement aux espèces SMI, l'évolution sexo-spécifique des variants d'ADNmt DUI entraîne l'expression de différents phénotypes bioénergétiques mâles et femelles. Au niveau de la fonctionnalité mitochondriale, les mitochondries DUI de type M présentent une phosphorylation oxydative (OXPHOS) remodelée, caractérisée par un contrôle respiratoire inhabituel à l'extrémité de la chaîne respiratoire. La réorganisation générale de la bioénergétique des spermes DUI entraîne également une variation de l'équilibre entre les principales voies de production d'énergie, incluant la glycolyse, la glycolyse anaérobique, le métabolisme des acides gras, le cycle de l'acide tricarboxylique, l'OXPHOS, ainsi que la capacité antioxydante. Enfin, les spermatozoïdes DUI comptent entièrement sur l'énergie produite par OXPHOS pour maintenir une motilité inhabituelle caractérisée par une vitesse lente et une trajectoire plus curviligne, traits potentiellement associés à un plus grand succès de reproduction chez les organismes marins sessiles. Aussi, ils conservent la capacité de passer à une stratégie de production d'énergie mixte (aérobique et anaérobie) après la détection des ovocytes. Dans l'ensemble, ces résultats suggèrent que la variation de l'ADNmt dans les espèces DUI pourrait être adaptative, incluant adaptation bioénergétique sexo-spécifiques avec un effet en aval sur la performance des spermatozoïdes, la capacité de reproduction, la sélection et transmission des mitochondries paternelles. / Mitochondria and their genome, the mitochondrial DNA (mtDNA), are usually transmitted only by the mother to both sons and daughters in metazoan (i.e. strict maternal inheritance, SMI). An exception to the general rule of SMI is found in around 100 species of bivalves, which are characterized by a doubly uniparental inheritance (DUI) of mitochondria. In DUI species, two highly divergent and sex-linked mtDNA lineages coexist. One mitochondrial lineage is maternally inherited (F-type) and is present in oocytes and somatic tissues of both female and male individuals. The other lineage is paternally inherited (M-type) and is present in sperm. In male somatic tissues both lineages sometimes coexist, a condition named heteroplasmy. Knowing that variations in mitochondrially-encoded genes might impact mitochondrial functions, and giving the strict association of M and F-type mtDNAs with different gametes, it is conceivable that the variation between the two DUI lineages might result in sex-specific bioenergetic adaptations with repercussion on reproduction. Despite providing an unprecedented opportunity for the mtDNA to evolve for male functions, the DUI system also brings the need for sperm mitochondria to preserve their genetic integrity, as well as for somatic cells to deal with heteroplasmy. The objective of my PhD was to link the mitochondrial genotype of DUI and SMI bivalve species to the phenotype. I explored the impact of sex-specific mtDNA variations upon a wide set of phenotypic traits, ranging from mitochondrial and cellular bioenergetics to sperm performance, investigating the adaptive value of DUI system in the light of reproductive fitness, mitochondrial selection, preservation and transmission. The results stemming from this PhD project revealed a clear phenotypic divergence between DUI and SMI species, possibly reflecting the different selective pressures acting on their mitochondria as a result of their different mode of mitochondria transmission. Conversely to SMI species, the sex-specific evolution of DUI mtDNA variants results in the expression of different male and female bioenergetic phenotypes. At the level of mitochondrial functionality, M-type mitochondria exhibit a remodelled OXPHOS characterized by unusual respiratory control at the terminus of the respiratory chain. The general reorganization of DUI sperm bioenergetics also entails variation in the balance between the main energy producing pathways, including glycolysis, anaerobic glycolysis, fatty acid metabolism, tricarboxylic acid cycle, OXPHOS, as well as the antioxidant capacity. Finally, DUI sperm exhibit an unusual motility phenotype characterized by slow speed and high curvilinear trajectory, traits potentially associated with a higher reproductive success in sessile broadcast spawning marine organisms. They also completely rely on the energy produced by OXPHOS to sustain their performance, although maintaining the ability to switch to a more combined aerobic/anaerobic strategy of energy production after oocyte detection. Altogether, these results suggest that the mtDNA variation in DUI species might be adaptive, resulting in the expression of sex-specific bioenergetic adaptation with downstream effect on sperm performance, reproductive fitness, paternal mitochondria selection, preservation and transmission. The results also suggest that heteroplasmy has an impact onto the bioenergetics of male soma, and that a functional compensation between genomes might minimize any potential deleterious outcome.

Freshwater bivalves of the family Unionidae have filter-feeding traits that alter freshwater ecosystems on several trophic levels. Filtration of organic and inorganic particulates in the water column transform matter through the mussel’s egestion before it is sequestered in the sediment, making it available as food for benthic invertebrates. Rates of filtration and deposition vary depending on species-specific traits, however, the filtration rate of Anodonta anatina (duck mussel) and Unio tumidus (swollen river mussel) are equivalent. This experimental aquarium study was performed by feeding duck mussel and swollen river mussel, to assess the biodeposition rate for both species. I hypothesized (1) that the biodeposition rate decreases for both species over a period of 180 minutes after feeding, and (2) there is no difference in deposited mass between A. anatina and U. tumidus. Results confirmed a decrease of organic and inorganic deposition over 180 minutes after feeding, with no significant difference of deposition mass between species. This study will contribute to valuable feeding methodology and knowledge for future studies of biodeposition of the family Unionidae. / Sötvattensmusslor av familj Unionidae har filtrerande egenskaper som förändrar sötvattensekosystem på flera trofiska nivåer. Filtrering av organiska och icke-organiska partiklar i de fria vattenmassorna transformerar material genom musslans matsmältningssystem som sedan lagras i sedimenten, och möjliggör materialet för födoupptag av bentiska evertebrater. Filtrering- och depositionshastigheter varierar beroende på artspecifika egenskaper, men filtreringshastigheten för Anodonta anatina (allmän dammussla) och Unio tumidus (spetsig målarmussla) är jämförbara. Denna experimentella akvariumstudie utfördes genom matning av allmän dammussla och spetsig målarmussla, med syftet att bedöma biodepositionshastigheten för de båda arterna. Jag ställde hypoteserna (1) biodepositionshastigheten minskar för båda arter över 180 minuter efter matning, och (2) det finns ingen skillnad i deponerad massa mellan A. anatina och U. tumidus. Resultaten bekräftar en minskning av organiskt och icke-organiskt material över 180 minuter efter matning, med ingen signifikant skillnad av depositionsmängd mellan arter. Denna studie bidrar med viktig kunskap om matningsmetoder och kunskaper för framtida studier om biodeposition för familjen Unionidae.

[pt] Os hidrocarbonetos policíclicos aromáticos (HPA) são compostos orgânicos de origem petrogênica e pirogênica, ubíquos, persistentes, bioacumuláveis em tecidos adiposos e comprovadamente carcinogênicos. O Monumento Natural do Arquipélago das Ilhas Cagarras (MONA) é uma unidade de conservação de proteção integral localizada a 4,5 km da cidade do Rio de Janeiro, cujas potenciais fontes de HPA para o local incluem o Emissário Ipanema, a Baía de Guanabara, o trânsito de grandes e pequenas embarcações, e a deposição atmosférica proveniente do alto trânsito de veículos e emissões industriais. Considerando que moluscos bivalves são biomonitores efetivos da exposição aos HPA, em fev/22 (C1) e jul/22 (C2) coletou-se cerca de 240 mexilhões da espécie Perna perna em três ilhas do MONA para avaliar o aporte de HPA oriundos de atividades antrópicas. Amostras compostas (3-5 animais) foram extraídas em triplicata utilizando extração acelerada por solvente (ASE) com purificação na célula, e purificação adicional por cromatografia em coluna. A fração de HPA foi determinada por cromatógrafo a gás acoplado a um espectrômetro de massas (GC-MS). As concentrações de (Somatório)39HPA variaram de 236 a 1378 ng g-1 , com os maiores valores encontrados em C1, níveis semelhantes aos de estudos anteriores com P. perna na região da Baía de Guanabara e algumas das ilhas do MONA (68-6271 ng g-1), sendo caracterizado, no entanto, aumento no aporte ao longo dos últimos 10 anos. O perfil geral dos contaminantes demonstra uma mistura de fontes pirolíticas e petrogênicas na região do MONA, sendo fontes prováveis a queima de combustíveis fósseis na região metropolitana e o aporte crônico de óleo, incluindo vazamentos irregulares, pelo trânsito constante de embarcações e fundeios próximos ao MONA. As concentrações de benzo[a]pireno, o (Somatório)4HPA – benz[a]antraceno, benzo[a]pireno, criseno e benzo[b]fluoranteno – e o Quociente de Equivalência Tóxica de Benz[a]pireno (TEQ BaPy) ficaram abaixo dos limiares de 5 ng g-1, 30 ng g-1 e 18 ng g-1, respectivamente, porém mais altos na amostragem realizada no verão, sugerindo um risco potencial para a qualidade de frutos do mar (moluscos) e da população humana. O cenário fornecido por um conjunto de dados limitado (apenas duas amostragens) reforça a necessidade de um programa de biomonitoramento no MONA Cagarras que inclua HPA e outras classes de contaminantes orgânicos, como as bifenilas policloradas (PCB) e pesticidas. / [en] Polycyclic aromatic hydrocarbons (PAH) are organic compounds of petrogenic and pyrogenic origin that are ubiquitous, persistent, bioaccumulative in adipose tissue, and proven to be carcinogenic. The Cagarras Islands Natural Monument (MONA) is a protected area located 4,5 km from the city of Rio de Janeiro, whose potential sources of PAH to the site include the Ipanema Outfall, Guanabara Bay, the transit of large and small vessels, and atmospheric deposition from high traffic of vehicles and industrial emissions. Considering that bivalve mollusks are effective biomonitoring organisms of exposure to PAH, in Feb/22 (C1) and Jul/22 (C2), approximately 240 mussels of the species Perna perna were collected from three MONA islands to assess the input of PAHs from anthropogenic activities. Composite samples (3-5 animals) were extracted in triplicate using accelerated solvent extraction (ASE) with purification in the cell, and additional purification by column chromatography. The PAH fraction was determined by gas chromatography-mass spectrometry (GC-MS). The concentrations of (summation)39PAH ranged from 236 to 1378 ng g-1, with the highest values found in C1, similar to previous studies with P. perna in the region of Guanabara Bay and some of the MONA islands (68-6271 ng g-1), but characterized by an increase in input over the last 10 years. The general profile of contaminants demonstrates a mixture of pyrolytic and petrogenic sources in the MONA region, with probable sources being fossil fuel combustion in the metropolitan region and chronic oil input, including irregular leaks, from the constant traffic of vessels and anchoring near the MONA. The concentrations of Benzo[a]pyrene, the (summation)4PAH – benzo[a]anthracene, chrysene, benz[b]fluoranthene, and benzo[a]pyrene – and the Toxic Equivalent Quotient of Benzo[a]pyrene (TEQ BaPy) were below the thresholds of 5 ng g-1, 30 ng g-1, and 18 ng g-1, respectively, but higher in the sampling performed in summer, suggesting a potential risk to the quality of seafood (mollusks) and the human population. The scenario provided by a limited data set (only two samplings) reinforces the need for a biomonitoring program in the Cagarras Islands Natural Monument that includes PAHs and other classes of organic contaminants, such as polychlorinated biphenyls (PCBs) and pesticides.

Globaler Temperaturanstieg, Ozeanversauerung und Sauerstoffmangel (temperaturbedingte Stressoren), spielen eine wichtige Rolle während Klimawandel. Die kombinierten Auswirkungen wirken sich negativ auf marine Lebensgemeinschaften und Ökosystemen aus und verschärfen die Effekte anderer Stressoren. Es gibt für den aktuellen und vergangenen Erwärmungsphasen Belege für veränderte Artenverteilung, Lebensraumverlust, Artensterben und verminderte physiologische Leistungen von Organismen. Die spezifischen Mechanismen werden diskutiert. Umweltveränderungen in der Erdvergangenheit bieten die Gelegenheit, die Dynamik von Ökosystemkrisen vor, zu untersuchen. Diese Arbeit integriert verschiedenen Disziplinen um unser Verständnis der Rolle temperaturbedingter Stressoren für marine benthische Ökosysteme zu verbessern. Das Toarcian Event (Unterjura) wurde aufgrund seiner geologischen, geochemischen und paläontologischen Überlieferung ausgewählt. Hochaufgelöste quantitative taxonomische, paläoökologische und geochemische Daten von benthischen Makroinvertebraten wurden analysiert. Ziel ist es, die Mechanismen ökologischer Veränderungen unter Temperaturstress zu bewerten, mit Schwerpunkt auf Körpergrößenmuster sowie Veränderungen in der Zusammensetzung und Struktur von Lebensgemeinschaften, und die Rolle von Umweltstressoren als Ursachen der biotischen Krise abzuschätzen. Der Temperaturanstieg führte zu dem Verlust an biologischer Vielfalt und zu der Verringerung der Körpergröße der Organismen vor und während der biotischen Krise. Die Struktur der Gemeinschaften wurde in Bezug auf die taxonomische und ökologische Zusammensetzung neu organisiert. Die Reaktion auf Umweltstress war innerhalb und zwischen den taxonomischen Gruppen unterschiedlich, wobei Brachiopoden stark betroffen waren. Diese Ergebnisse verbessern unser Verständnis der biotischen Reaktionen auf temperaturbedingte Stressoren und können zur Vorhersage Veränderungen Ökosysteme bei den aktuellen Erwärmungstrends beitragen. / Global warming, oceanic acidification and deoxygenation (temperature-related stressors) play an important role during climate change. The combined impact of these stressors is known to negatively affect marine biota and ecosystems, often exacerbating the impact of other stressors. Under the current and past climate change, there is record of altered species distribution, habitat loss, extinctions and decreased physiological performance of organisms. Despite the increasing evidence, the specific mechanisms through which climate change influences ecological patterns are debated. Past environmental perturbations represent an opportunity to investigate the dynamics of ecosystem and biotic crises across Earth history. This work integrates different disciplines to increase our understanding on the role played by temperature-related stressors on marine benthic biota and ecosystems. The Toarcian Event (Early Jurassic) was chosen for its well-preserved geological, geochemical and palaeontological record. High-resolution quantitative faunal and geochemical data from benthic marine macroinvertebrates were investigated. The aims are to identify and evaluate the mechanisms of faunal and ecological shifts under a temperature-related stressor scenario, with focus on body size patterns and ecosystem change and recovery, and to estimate the role of environmental stressors as proximate causes of the biotic crisis. Temperature increase led to biodiversity loss and reduced body size of organisms during and possibly before the event. Moreover, the structure of marine communities was reorganized in terms of taxonomic and ecological composition. The response to environmental stress was variable within and between taxonomical groups, with brachiopods severely affected. These findings increase our understanding of biotic responses and ecosystem and biodiversity shifts under temperature stress, hopefully contributing to the prediction of ecosystem changes under the current warming scenario.