Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / A introdução de espécies invasoras marinhas tem causado danos econômicos e ecológicos consideráveis em todo o mundo. Algumas destas espécies incluindo corais escleractíneos possuem adaptações, tais como metabólitos secundários utilizados para evitar a predação e competição por espaço por outros organismos. Este arsenal químico e as interações entre espécies invasoras e nativas podem causar alterações na distribuição das espécies e na estrutura das comunidades de costões rochosos tropicais. Os objetivos deste estudo foram (1) caracterizar os metabólitos secundários produzidos pelos corais invasores Tubastraea tagusensis e T. coccinea na Baía da Ilha Grande, Brasil, (2) detectar os compostos químicos liberados pelos tecidos de Tubastraea tagusensis in situ utilizando um aparelho submersível; (3) testar no campo os extratos metanólicos produzidos por ambas as espécies de Tubastraea contra a predação por peixes generalistas e assentamento de outros organismos, (4) testar no campo se os compostos químicos produzidos por ambos os corais invasores variaram na concentração ou tipo quando os corais foram colocados próximos de competidores nativos e (5) determinar como as comunidades de costões rochosos da Baía da Ilha Grande foram afetadas pela expansão de Tubastraea coccinea e T. tagusensis em 8 locais estudados durante 2 anos. As principais classes de substâncias encontradas nos extratos metanólicos de Tubastraea foram identificados como esteróis, ácidos graxos, hidrocarbonetos, alcalóides, ésteres e alcoóis, entretanto, o aparelho submersível identificou somente hidrocarbonetos liberados por Tubastraea na água do mar. O extrato metanólico de T. tagusensis reduziu a predação por peixes generalistas e já os extratos de ambas as espécies mostraram efeitos espécie-específicos sobre organismos incrustantes no campo. No experimento de interação competitiva foi detectada a presença de necrose nos tecidos do coral endêmico Mussismilia hispida e isso provocou variação nas concentrações de esteróis, alcalóides e ácidos graxos nos tecidos de Tubastraea. Em contraste, a esponja Desmapsamma anchorata cresceu sobre os tecidos das colônias de ambos os corais invasores. A presença de Tubastraea nas comunidades bentônicas causou uma dissimilaridade média de 4,8% nas comunidades invadidas. Uma forte relação positiva foi encontrada entre a cobertura de Tubastraea e a mudança na estrutura da comunidade da Baía da Ilha Grande. Portanto, os efeitos negativos de ambos os corais invasores são suficientes para acarretar mudanças na estrutura das comunidades bentônicas tropicais. / Invasive marine species has caused economic and ecological damage around the world. Some scleractinian corals possess secondary metabolites used to avoid predation and to help them in competition for space with other organisms. The negative action of chemical substances in the interactions between invasive and native species may produce changes on community structure in tropical rocky shores. This study showed (1) the secondary metabolites produced by the invasive corals Tubastraea tagusensis and T. coccinea in Ilha Grande Bay, Brazil; (2) chemical compounds released by Tubastraea tagusensis tissues in situ using a submersible apparatus; (3) the action of MeOH extracts produced by both species of Tubastraea against predation by generalist fish and settlement of other organisms in the field; (4) variation on class and concentration of chemical compounds produced by these invasive corals when these species were placed in proximity to native competitors and (5) changes on rocky shore community structure due to expansion of Tubastraea coccinea and T. tagusensis after 2 years of study. The main classes of compounds detected by GC / MS in the MeOH extracts of Tubastraea were identified as sterols, fatty acids, hydrocarbons, alkaloids, esters and alcohols. The submersible apparatus identified only hydrocarbons released by Tubastraea in situ. The MeOH extract of T. tagusensis reduced generalist fish predation and the MeOH extracts of both Tubastraea species showed species-specific effects on fouling organisms in the field. The competition experiment showed necrosis in the tissues of the native coral Mussismilia hispida and variation in sterols, alkaloids and fatty acid concentrations in Tubastraea tissues. The opposite behavior was sawn, the sponge Desmapsamma anchorata overgrew Tubastraea colonies. The presence of Tubastraea in the benthic communities caused a mean dissimilarity of 4.8% in the invaded communities. A positive relationship between invader cover and change in Ilha Grande community structure was found and, therefore, Tubastraea is a threat to native benthic communities throughout the tropical Oceans.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / A introdução de espécies invasoras marinhas tem causado danos econômicos e ecológicos consideráveis em todo o mundo. Algumas destas espécies incluindo corais escleractíneos possuem adaptações, tais como metabólitos secundários utilizados para evitar a predação e competição por espaço por outros organismos. Este arsenal químico e as interações entre espécies invasoras e nativas podem causar alterações na distribuição das espécies e na estrutura das comunidades de costões rochosos tropicais. Os objetivos deste estudo foram (1) caracterizar os metabólitos secundários produzidos pelos corais invasores Tubastraea tagusensis e T. coccinea na Baía da Ilha Grande, Brasil, (2) detectar os compostos químicos liberados pelos tecidos de Tubastraea tagusensis in situ utilizando um aparelho submersível; (3) testar no campo os extratos metanólicos produzidos por ambas as espécies de Tubastraea contra a predação por peixes generalistas e assentamento de outros organismos, (4) testar no campo se os compostos químicos produzidos por ambos os corais invasores variaram na concentração ou tipo quando os corais foram colocados próximos de competidores nativos e (5) determinar como as comunidades de costões rochosos da Baía da Ilha Grande foram afetadas pela expansão de Tubastraea coccinea e T. tagusensis em 8 locais estudados durante 2 anos. As principais classes de substâncias encontradas nos extratos metanólicos de Tubastraea foram identificados como esteróis, ácidos graxos, hidrocarbonetos, alcalóides, ésteres e alcoóis, entretanto, o aparelho submersível identificou somente hidrocarbonetos liberados por Tubastraea na água do mar. O extrato metanólico de T. tagusensis reduziu a predação por peixes generalistas e já os extratos de ambas as espécies mostraram efeitos espécie-específicos sobre organismos incrustantes no campo. No experimento de interação competitiva foi detectada a presença de necrose nos tecidos do coral endêmico Mussismilia hispida e isso provocou variação nas concentrações de esteróis, alcalóides e ácidos graxos nos tecidos de Tubastraea. Em contraste, a esponja Desmapsamma anchorata cresceu sobre os tecidos das colônias de ambos os corais invasores. A presença de Tubastraea nas comunidades bentônicas causou uma dissimilaridade média de 4,8% nas comunidades invadidas. Uma forte relação positiva foi encontrada entre a cobertura de Tubastraea e a mudança na estrutura da comunidade da Baía da Ilha Grande. Portanto, os efeitos negativos de ambos os corais invasores são suficientes para acarretar mudanças na estrutura das comunidades bentônicas tropicais. / Invasive marine species has caused economic and ecological damage around the world. Some scleractinian corals possess secondary metabolites used to avoid predation and to help them in competition for space with other organisms. The negative action of chemical substances in the interactions between invasive and native species may produce changes on community structure in tropical rocky shores. This study showed (1) the secondary metabolites produced by the invasive corals Tubastraea tagusensis and T. coccinea in Ilha Grande Bay, Brazil; (2) chemical compounds released by Tubastraea tagusensis tissues in situ using a submersible apparatus; (3) the action of MeOH extracts produced by both species of Tubastraea against predation by generalist fish and settlement of other organisms in the field; (4) variation on class and concentration of chemical compounds produced by these invasive corals when these species were placed in proximity to native competitors and (5) changes on rocky shore community structure due to expansion of Tubastraea coccinea and T. tagusensis after 2 years of study. The main classes of compounds detected by GC / MS in the MeOH extracts of Tubastraea were identified as sterols, fatty acids, hydrocarbons, alkaloids, esters and alcohols. The submersible apparatus identified only hydrocarbons released by Tubastraea in situ. The MeOH extract of T. tagusensis reduced generalist fish predation and the MeOH extracts of both Tubastraea species showed species-specific effects on fouling organisms in the field. The competition experiment showed necrosis in the tissues of the native coral Mussismilia hispida and variation in sterols, alkaloids and fatty acid concentrations in Tubastraea tissues. The opposite behavior was sawn, the sponge Desmapsamma anchorata overgrew Tubastraea colonies. The presence of Tubastraea in the benthic communities caused a mean dissimilarity of 4.8% in the invaded communities. A positive relationship between invader cover and change in Ilha Grande community structure was found and, therefore, Tubastraea is a threat to native benthic communities throughout the tropical Oceans.

Cette étude a pour but d'identifier les principales maladies coralliennes, de quantifier leurs prévalences sur les récifs de la Réunion, d'Afrique du Sud et de Mayotte et de déterminer leurs variations spatiales et saisonnières. Les résultats ont révélé la présence de six grands types de maladies coralliennes. Les prévalences totales moyennes des maladies ont été plus élevées à la Réunion (7.5 ± 2.2%) qu'en Afrique du Sud (3.9 ± 0.8%) et qu'à Mayotte (2.7 ± 0.3%). Les genres coralliens Acropora et Porites ont été les plus vulnérables aux maladies dans les trois régions étudiées. Des variations spatiales et saisonnières ont été détectées pour la maladie de la bande noire (BBD) et les syndromes blancs (WS) à la fois à la Réunion et en Afrique du Sud avec des prévalences plus élevées sur les récifs peu profonds et pendant la saison chaude. Ce travail a également pour but de caractériser deux maladies non décrites à ce jour : le syndrome des patches blancs (PWPS) et le syndrome du patch noir (PBPS) chez Porites. PWPS, observé à fois à la Réunion, en Afrique du Sud et à Mayotte est caractérisé par d'importantes fragmentations tissulaires généralement associées à des corps ovoïdes basophiles ressemblant à des agrégats bactériens. L'analyse bactérienne a révélé une grande variabilité entre les tissus infectés et sains. L'identification des pathogènes responsables de PWPS a également été réalisée en laboratoire. Sur les 14 souches bactériennes sélectionnées et testées, seule la P180R, génétiquement proche de Vibrio tubiashii a généré des signes de PWPS. Pour finir, PBPS, une forme atypique de BBD a été observée sur les récifs de la Réunion. Les suivis réalisés ont montré des variations spatiales et saisonnières avec des prévalences plus élevées sur le platier et pendant l'été. L'histologie a mis en évidence la présence de cyanobactéries et de corps basophiles dans les tissus infectés du corail. L'analyse génétique a révélé la présence d'une plus grande diversité de taxons dans les tissus infectés par PBPS que dans les tissus sains représentés par le genre Vibrio. / During the past two decades, the emergence and spread of infectious diseases have caused substantial declines in the biodiversity and abundance of reef-building corals. Despite their increased global prevalence and virulence, little is known about coral diseases on Indian Ocean coral reefs. This study aims to fill this gap in knowledge by identifying the main coral diseases and quantify their prevalence at three localities Reunion, South Africa and Mayotte, determining their spatial distribution and seasonal variation. Principal findings of this study demonstrated the presence of six main coral diseases including black band disease (BBD), white syndromes (WS), pink line syndrome (PLS), growth anomalies (GA), skeleton eroding band (SEB) and Porites white patch syndrome (PWPS). The overall disease prevalence was higher in Reunion (7.5 ± 2.2%; mean ± SE) compared to South Africa (3.9 ± 0.8%; mean ± SE) and Mayotte (2.7 ± 0.3%; mean ± SE). Acropora and Porites were the genera most vulnerable to disease. Spatial variability was detected in both Reunion and South Africa with BBD and WS more prevalent on shallow than deep reefs. There was also evidence of seasonality in two diseases: BBD and WS, their prevalence being higher in summer than winter. Corals exhibiting signs of PWPS revealed extensive tissue fragmentation, generally associated with ovoid basophilic bodies resembling bacterial aggregates within the mesoglea of the body wall. Other organisms, including Cyanobacteria, Nematoda, Ciliata and endophytic algae, were also observed on diseased tissues and were generally associated with the dead epidermis and cell debris. Results of 16S rRNA sequence analysis revealed a high variability between bacterial communities associated with PWPS-infected and healthy tissues in Porites lutea. Several bacterial ribotypes affiliated to potential putative pathogens notably Shimia marina and Vibrio hepatarius were consistently found among the 16S rRNA sequences derived from the PWPS lesions, and absent and/or poorly represented in HT. Primary pathogens involved in the PWPS were also investigated in this study using traditional culturing techniques and laboratory infection trials. Of the 14 isolates selected for the inoculation trials, only the bacterial strain P180R mostly phylogenetically closely related8 to Vibrio tubiashii with its closest known sister taxon, V. hepatarius, was shown to cause signs resembling those of PWPS and satisfied the four Henle-Koch’s postulates. P180R displayed focalised and progressive tissue paling 12 h after inoculation and visible lesions of PWPS were observed 12 h thereafter. Signs of PWPS appeared on 90% of the exposed coral fragments (27 of 30) under controlled environmental conditions. Moreover, the virulence of this marine pathogen was tested and seemed to be strongly dependent on seawater temperature, resulting in significantly higher tissue loss at 30°C than 28°C and 26°C. Finally, a multidisciplinary approach involving field surveys, gross lesion monitoring, histopathology and 454-pyrosequencing was investigated to characterize an atypical form of BBD named PBPS. Histology revealed cyanobacterial penetration of the compromised tissue as well as the presence of basophilic bodies resembling bacterial aggregates in the living tissue, adjacent to the bacterial mat. Bacterial 16S rRNA sequences yielded a broader diversity of bacterial taxa in PBPS-infected tissues than in healthy tissue, represented by the genus Vibrio (24.9%), followed by sulfate-reducers or sulfide-oxidizers such as Desulfovibrio (20%), Clostridium (12.9%) and Arcobacter (9.9%). PBPS appears to be a multi-stage disease triggered by cyanobacterial invasion and resulting in secondary infections by environmental bacteria that grow in mucus-like decomposing tissue.

La formation d’un squelette de CaCO3 par les coraux Scléractiniaires est à la base de l’édification des récifs coralliens. Nombre de ces coraux constructeurs de récif vivent en symbiose avec des Dinoflagellés photosynthétiques. Ces deux processus reposent sur le transport et l’utilisation de carbone inorganique (Ci) provenant de l’eau de mer pour la photosynthèse, et du métabolisme animal pour la calcification. Cette thèse s’est intéressée à l’étude moléculaire et physiologique des mécanismes, permettant l’utilisation de ce carbone inorganique.Malgré l’importance des transports de HCO3-, aucun transporteur n’a été caractérisé à cejour et leur implication dans la physiologie des coraux n’est que suggérée par la pharmacologie. Durant cette thèse nous avons cloné un gène codant pour un transporteur deHCO3- chez le corail Acropora sp. La conversion de ce HCO3- en CO2 pour la photosynthèse est facilitée par l’acidification de l’environnement proche du Dinoflagellé dans la cellule animale. Cette acidification est causée par une H+-ATPase de type P que nous avons caractérisée. Ce gène est le premier à montrer une expression dépendante de la vie en symbiose chez le symbiote.Nous avons aussi cloné et localisé deux anhydrases carboniques (AC). L’une impliquée dans la calcification et l’autre dans la régulation du pH intracellulaire et l’équilibre entre leCO2 et HCO3-. Une étude pharmacologique de ces deux AC, a identifié des molécules inhibitrices et activatrices qui ont permis des expériences de physiologie in vivo. Celles-ci permettent une analyse plus discriminante du rôle des AC dans la calcification. / Coral reefs edification is based on the formation of a calcium carbonate skeleton byscleractinian corals. Many of these reef-building corals establish a symbiotic association with photosynthetic Dinoflagellates. Both processes involve the transport and utilization of inorganic carbon (Ci) coming from seawater for photosynthesis, and from animal metabolismfor calcification. This work focused on the molecular and physiological study of poorlyknown mechanisms that allow the utilization of Ci.Despite the importance of bicarbonate transport, no transporter has been characterized and their role in coral physiology is only suggested by pharmacological experiments. We have cloned a gene encoding a bicarbonate transporter in the coral Acropora sp. The conversion of this bicarbonate into CO2 for photosynthesis is mediated by the acidification of the are asurrounding the Dinoflagellate in the animal cell. This is performed by a P type H+-ATPasethat we characterized here. This is the first gene with a symbiosis-dependent expression in the symbiont.This work also allowed the cloning and the localization of two carbonic anhydrases (CA).The first one is involved in calcification, the second one plays a role in the intracellular pHregulation and the CO2 / HCO3- equilibrium. A pharmacological study of these two enzymes identified inhibitor and activator compounds that have been then used in physiology experiments. This last approach represents a more accurate study of the role of CAs incalcification.

Ce travail de thèse porte sur la connectivité des populations de coraux du genre Pocillopora dans le Sud-Ouest de l'océan Indien et l'océan Pacifique tropical. Ces coraux sont répartis sur toute la frange tropicale des océans Indien et Pacifique. Traditionnellement, les espèces étaient identifiées sur la base critères morphologies [17 espèces décrites dans Veron (2000)]. Différentes études utilisant des données génétiques ont révélé que la délimitation des espèces était parfois floue chez ces coraux. Ainsi, au cours de ce travail, l'utilisation de méthodes de délimitation d'espèces à partir d'ADN mitochondrial (ABGD, GMYC, PTP) et nucléaire (haplowebs) 16 hypothèses primaires d'espèces (PSH) ont été identifiées. Ces PSH ont ensuite été confrontées à des tests d'assignement à partir de marqueurs microsatellites, révélant un minimum de 18 hypothèses d'espèces secondaires (SSH). Une fois que les hypothèses d'espèces sont définies, il est possible de réaliser des études de connectivité. Au cours de ce travail, deux hypothèses d'espèces présentant des écologies différentes ont été choisies pour mener ces analyses. La première, Pocillopora damicornis type β (SSH05) a été échantillonnée dans les lagons et la seconde, Pocillopora eydouxi (SSH09) a, quant à elle, été échantillonnée sur la pente externe. L'estimation de la structure génétique des populations a permis d'estimer les modes de reproduction (sexuée ou asexuée) chez ces deux hypothèses d'espèces et les analyses de connectivité ont révélé des patterns de structuration complexes pour chacune des SSHs. / This work focuses on the connectivity among populations of the coral genus Pocillopora in the Southwestern Indian Ocean and the Southwestern Pacific Ocean. These corals are widely distributed throughout the tropical fringe of the Indian and Pacific oceans. Traditionally, species were identified on the basis of morphological criteria [17 species described in Veron (2000)]. Different studies using genetic data revealed that the delimitation of species was sometimes blurred in these corals. Thus, in this work, the use of species delineation methods from mitochondrial (ABGD, GMYC, PTP) and nuclear (haplowebs) DNA, 16 primary species hypotheses (PSH) were identified. These PSHs were then confronted to assignment tests from microsatellite loci, revealing a minimum of 18 secondary species hypotheses (SSH). Once the species hypotheses are defined, it is possible to conduct connectivity studies. In this work, two SSHs with different ecologies were chosen to carry out these analyses. The first, Pocillopora damicornis type β (SSH05) was sampled in the lagoons and the second, Pocillopora eydouxi (SSH09) was sampled on the outer slope. The estimation of the genetic structure of the populations made possible to estimate the reproductive modes (sexual or asexual) in these two SSHs and the connectivity analyzes revealed complex structuring patterns for each of the SSHs.