Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Cold seeps have recently been discovered in the Nile deep-sea fan (Eastern Mediterranean), and data regarding associated fauna are still scarce. In this study, two bivalve species associated with carbonate crusts and reduced sediment are identified based on sequence analysis of their 18S and 28S rRNAencoding genes, and associated bacterial symbioses are investigated using 16S rRNA gene sequencing and microscopy-based approaches. The specimens are closely related to Myrtea spinifera and Thyasira flexuosa, two species previously documented at various depths from other regions but not yet reported from the Eastern Mediterranean. Both species harbour abundant gammaproteobacterial endosymbionts in specialized gill epithelial cells. The Myrtea-associated bacterium is closely related to lucinid symbionts from both deep-sea and coastal species, whereas the Thyasira-associated bacterium is closely related to the symbiont of a T. flexuosa from coastal waters off the U.K. An epsilonproteobacterial sequence has also been identified in Thyasira which could correspond to a helicoid-shaped morphotype observed by electron microscopy, but this was not confirmed using fluorescent in situ hybridization. Virus-like particles were observed within some symbionts in Thyasira, mostly in bacteriocytes localized close to the ciliated zone of the gill filament. Overall, results indicate that very close relatives of shallow species M. spinifera and T. flexuosa occur at cold seeps in the Eastern Mediterranean and harbour chemoautotrophic symbioses similar to those found in their coastal relatives.
ISSN: 0173-9565

hal-00743765
https://hal.univ-antilles.fr/hal-00743765
DOI : 10.1111/j.1439-0485.2010.00413.x

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
This work relates to the characterisation of the predominant gill protein of the white clam, Codakia orbicularis (Linné, 1758), which harbours endosymbiotic sulphur-oxidising chemoautotrophic bacteria. Total RNA was extracted from the clam to perform 3'rapid amplification of cDNA ends (3'RACE) using degenerate oligonucleotides prepared from a partial sequence of a predominant protein of about 14kDa, termed codakine. The partial peptide sequence was obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and Edman degradation. Clones isolated from the cDNA library and containing the gene of interest were used in polymerase chain reaction (PCR). The PCR and RACE-PCR products were sequenced to determine the entire coding DNA sequence of codakine. BLAST analysis revealed about 23% to 29% sequence identity between codakine and various animal C-type lectins that are often involved in symbiosis and immune defences. Codakine also contains the motifs and domains of Ca(2+)-dependent C-type lectins, and in particular the tripeptide EPN motif frequently found in mannose-binding lectins (MBLs). Analysis of the protein by affinity chromatography on a mannose-agarose column is consistent with the findings that codakine is a dimeric Ca(2+)-dependent MBL of about 29kDa. Based on the present results, it is hypothesised that this novel C. orbicularis gill protein is involved in the recognition of symbiotic and pathogenic bacteria.
ISSN: 1050-4648

hal-00746015
https://hal.univ-antilles.fr/hal-00746015
DOI : 10.1016/j.fsi.2006.06.013
PUBMED : 17169576

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Background information. Marine nematodes belonging to the Stilbonematidae (Desmodoridae) family are described as living in obligatory association with sulfur-oxidizing chemoautotrophic ectosymbionts. The symbiotic bacteria carrying out this chemosynthesis should contain elemental sulfur in periplasmic granules as sulfur granules of chemoautotrophic endosymbionts described in various marine invertebrates. Results. Based on TEM analyses, extracellular bacteria surrounding Eubostrichus dianae possess these spherical periplasmic granules. Few investigative techniques can be used to identify elemental sulfur S8 such as EDXS (energy dispersive X-ray spectroscopy) and EELS (electron energy loss spectroscopy) associated with cryo-fixation of the sample to avoid sulfur loss. These techniques are time consuming, expensive and require technical skills. Raman microspectrometry applied to the analysis of E. dianae allowed us to detect elemental sulfur S8 and confirmed the location of these sulfur clusters in the bacterial coat. In the same way, Raman spectrometry was positively applied to the endosymbiotic bivalve Codakia orbicularis, suggesting that this technique can be used to characterize sulfur in ecto- as well as in endo-symbiotic sulfur-oxidizing bacteria. Conclusions. As Raman spectrometry can be used on living organisms (without preliminary fixation) without sample damage and preserving the molecular structure of the sulfur (denatured during chemical fixation) it represents a very well-adapted investigative tool for biologists. This technique therefore permits us to detect quickly and easily (in a few seconds and on entire living animals) the presence of sulfur compounds in the symbiotic nematode.
ISSN: 0248-4900

hal-00753017
https://hal.univ-antilles.fr/hal-00753017

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
In chemoautotrophic associations, sequence comparison of 16S rRNA has been the method of choice to study bacterial diversity in the context of host/symbiont coevolution. However, the relative low rate of evolution of 16S rRNA has been shown to result in a diminished capacity to discriminate between closely related bacterial strains or species. Within chemoautotrophic associations, as described in several studies, the use of other genetic markers may reveal previously unobserved strain diversity among gill endosymbionts. Herein, we conducted a survey of symbionts harbored by six species within the Lucinidae family using five genetic markers (dnaE, gyrB, aprA, cbbL genes and the 16S-23S internal transcribed spacer). Thus, within the already described SoLuc₁ bacterial species shared by six host species we observed an obvious bacterial strain diversity. This diversity is structured in function of the geographic location of the hosts and not in function of ecological parameters or host phylogeny. Interestingly at the local level (same seagrass bed environment), we observed an unexpected specificity in host species/bacterial strain association (i.e., all individuals of the same host species harbor the same symbiotic sequence type). This specificity of association implied that there was a control of symbiont strain acquisition by the host, which was confirmed by a cross-infection experiment of starved adults performed in our study. Based upon our results and other evidence from the literature, we hypothesize that this pattern may be due to a ``capture/escape'' type antagonist evolution of the two partners.
ISSN: 0025-3162

hal-01544753
https://hal.archives-ouvertes.fr/hal-01544753
DOI : 10.1007/s00227-016-2949-0

Éditeur(s) : HAL CCSD Nature Publishing Group
Résumé : International audience
The shallow water bivalve Codakia orbicularis lives in symbiotic association with a sulfur-oxidizing bacterium in its gills. The endosymbiont fixes CO2 and thus generates organic carbon compounds, which support the host's growth. To investigate the uncultured symbiont's metabolism and symbiont-host interactions in detail we conducted a proteogenomic analysis of purified bacteria. Unexpectedly, our results reveal a hitherto completely unrecognized feature of the C. orbicularis symbiont's physiology: the symbiont's genome encodes all proteins necessary for biological nitrogen fixation (diazotrophy). Expression of the respective genes under standard ambient conditions was confirmed by proteomics. Nitrogenase activity in the symbiont was also verified by enzyme activity assays. Phylogenetic analysis of the bacterial nitrogenase reductase NifH revealed the symbiont's close relationship to free-living nitrogen-fixing Proteobacteria from the seagrass sediment. The C. orbicularis symbiont, here tentatively named `Candidatus Thiodiazotropha endolucinida', may thus not only sustain the bivalve's carbon demands. C. orbicularis may also benefit from a steady supply of fixed nitrogen from its symbiont-a scenario that is unprecedented in comparable chemoautotrophic symbioses.
ISSN: 2058-5276

hal-01545904
https://hal.archives-ouvertes.fr/hal-01545904
DOI : 10.1038/nmicrobiol.2016.193

Éditeur(s) : HAL CCSD Wiley-Blackwell
Résumé : International audience
Associations between marine invertebrates and chemoautotrophic bacteria constitute a wide field for the study of symbiotic associations. In these interactions, symbiont transmission must represent the cornerstone allowing the persistence of the association throughout generations. Within Bivalvia, in families such as Solemyidae or Vesicomyidae, symbiont transmission is undoubtedly vertical. However, in Lucinidae, symbiont transmission is described in the literature as 'environmental', symbionts being acquired from the environment by the new host generations. Hence, if there is transmission, symbionts should be transmitted from adults to juveniles via the environment. Consequently, we should observe a release of the symbiont by adults. We attempted to detect such a release within two Lucinidae species of the genus Codakia. We sampled 10 Codakia orbicularis and 20 Codakia orbiculata distributed in 10 crystallizing dishes containing filtered seawater. During 1 month of investigation, we analyzed water of the dishes in order to detect any release of a symbiont using catalyzed report deposition-FISH techniques. For 140 observations realized during this period, we did not observe any release of symbionts. This suggests that the idea of host-to-host passage in Lucinidae is inaccurate. We could therefore consider that the transmission mode from generation to generation does not occur within Lucinidae, symbiosis appearing to be advantageous in this case only for the host, and constitutes an evolutionary dead-end for the bacteria.
ISSN: 0168-6496

hal-00751560
https://hal.univ-antilles.fr/hal-00751560
DOI : 10.1111/j.1574-6941.2008.00626.x

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
The shallow-water bivalve Codakia orbiculata which harbors gill-endosymbiotic sulfur-oxidizing ?-proteobacteria can lose and acquire its endosymbionts throughout its life. Long-term starvation and recolonization experiments led to changes in the organization of cells in the lateral zone of gill filaments. This plasticity is linked to the presence or absence of gill-endosymbionts. Herein, we propose that this reorganization can be explained by three hypotheses: (a) a variation in the number of bacteriocytes and granule cells due to proliferation or apoptosis processes, (b) a variation of the volume of these two cell types without modification in the number, and (c) a combination of both number and cell volume variation. To test these hypotheses, we analyzed cell reorganization in terms of proliferation and apoptosis in adults submitted to starvation and returned to the field using catalyzed reporter deposition fluorescence in situ hybridization, immunohistochemistry, and structural analyses. We observed that cell and tissue reorganization in gills filaments is due to a variation in cell relative abundance that maybe associated with a variation in cell apparent volume and depends on the environment. In fact, bacteriocytes mostly multiply in freshly collected and newly recolonized individuals, and excess bacteriocytes are eliminated in later recolonization stages. We highlight that host tissue regeneration in gill filaments of this symbiotic bivalve can occur by both replication of existing cells and division of undifferentiated cells localized in tissular bridges, which might be a tissue-specific multipotent stem cell zone.
ISSN: 1059-910X

hal-01545915
https://hal.archives-ouvertes.fr/hal-01545915
DOI : 10.1002/jemt.22041

Éditeur(s) : HAL CCSD Wiley-Blackwell
Résumé : International audience
Here, the first description of an epsilon sulfur-oxidizing bacterium from sulfide-rich sediments of marine mangrove in Caribbean is reported. According to TEM, this new strain contains intracytoplasmic large internal sulfur granules as confirmed by EDXS analyses performed using ESEM. The sulfur distribution obtained for this sulfur-oxidizing bacterial strain allows to conclude that elemental sulfur is formed as an intermediate oxidation product and stored intracellularly. According to conventional SEM observations, bacterial cells are ovoid, extremely motile by lophotrichous flagella. Phylogenetic analyses based on the analysis of partial sequence of 16S rDNA confirm that such bacterial strain belongs to the Thiovulum cluster and could be a representative of a new species in this poorly studied genus of sulfur-oxidizing free-living bacteria. Thus, reduced sediment of marine mangrove represents sulfide-rich environment sustaining development of both gamma and epsilon sulfur-oxidizing Proteobacteria
ISSN: 0378-1097

hal-01682113
https://hal.univ-antilles.fr/hal-01682113
DOI : 10.1093/femsle/fnx172

Éditeur(s) : HAL CCSD Wiley-Blackwell
Résumé : International audience
The shallow-water bivalve Codakia orbiculata which harbors gill-endosymbiotic sulfur-oxidizing g-proteobacteria can lose and acquire its endosymbionts throughout its life. Longterm starvation and recolonization experiments led to changes in the organization of cells in the lateral zone of gill filaments. This plasticity is linked to the presence or absence of gill-endosymbionts. Herein, we propose that this reorganization can be explained by three hypotheses: (a) a variation in the number of bacteriocytes and granule cells due to proliferation or apoptosis processes, (b) a variation of the volume of these two cell types without modification in the number, and (c) a combination of both number and cell volume variation. To test these hypotheses, we analyzed cell reorganization in terms of proliferation and apoptosis in adults submitted to starvation and returned to the field using catalyzed reporter deposition fluorescence in situ hybridization, immunohistochemistry, and structural analyses. We observed that cell and tissue reorganization in gills filaments is due to a variation in cell relative abundance that maybe associated with a variation in cell apparent volume and depends on the environment. In fact, bacteriocytes mostly multiply in freshly collected and newly recolonized individuals, and excess bacteriocytes are eliminated in later recolonization stages. We highlight that host tissue regeneration in gill filaments of this symbiotic bivalve can occur by both replication of existing cells and division of undifferentiated cells localized in tissular bridges, which might be a tissue-specific multipotent stem cell zone.
Microscopy Research and Technique

hal-00743036
https://hal-uag.archives-ouvertes.fr/hal-00743036

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
The Caribbean bivalves Codakia orbicularis (Linnée, 1758) and C. orbiculata (Montagu, 1808) live in seagrass beds of Thalassia testudinum and harbor intracellular sulfuroxidizing gamma-proteobacteria. These bacterial symbionts fix CO2 via the Calvin Benson cycle and provide organic compounds to the bivalve. During experimentally induced starvation, no reduced sulfur compounds and no organic particle food are available; the symbionts could be considered as the sole nutrient source of the host bivalve. A previous study has shown that the intracellular bacterial population decreased considerably during starvation and that bacterial endosymbionts were not released by the bivalves. In this study, the activity of two lysosomal marker enzymes (acid phosphatase and arylsulfatase) was detected using cytochemical experiments coupled with energy-dispersive X-ray transmission electron microscopy during sulfide and organic particle starvation. The degradation of bacterial endosymbionts began after 2 weeks of starvation in C. orbiculata and after 3 weeks in C. orbicularis. Degradation processes seem to be continuous over several months and could be responsible for the disappearance of the bacterial endosymbionts within the gills during starvation. These data suggest that the host use symbionts as a nutrient source to survive a hunger crisis. The carbon transfer from the symbionts to the host could be flexible and could consist in transfer of organic matter, “milking,” under normal feeding conditions and digestion of the symbionts under starved conditions.
ISSN: 1059-910X

Droits : info:eu-repo/semantics/OpenAccess
hal-01206367
https://hal.univ-antilles.fr/hal-01206367
https://hal.univ-antilles.fr/hal-01206367/document
https://hal.univ-antilles.fr/hal-01206367/file/revised%20version%20to%20be%20submitted%20to%20the%20editor3oct.pdf
DOI : 10.1002/jemt.22458

Résumé : La zone latérale des filaments branchiaux des bivalves Codakia orbiculata et Lucina pensylvanica est le lieu d'une symbiose chimioautotrophe avec des bactéries sulfo-oxydantes hébergées dans des cellules spécialisées, appelées bactériocytes. Dans cette étude, nous nous sommes proposés de déterminer les mécanismes qui sous-tendent la plasticité cellulaire et tissulaire observée au cours des processus de décolonisation et de recolonisation bactérienne. Pour ce faire, des individus collectés dans leur milieu naturel, ont été maintenus au laboratoire dans des bacs d'eau de mer filtrée en absence de nourriture et de soufre réduit, afin de provoquer la décolonisation bactérienne des filaments branchiaux. Lorsque la branchie apparaissait purgée de ses symbiotes, les individus ont été remis dans leur habitat naturel afin de provoquer sa recolonisation. L'analyse des branchies au cours de ces processus a fait appel à des techniques variées (histologie, immunohistochimie, hybridation in situ, cytométrie en flux, dosage des protéines et spectrométrie de fluorescence X). Les résultats obtenus ont permis de montrer que l'acquisition environnementale mise en évidence chez les juvéniles des Codakia se poursuivait tout au long de la vie des adultes. Cette étude a également permis une meilleure compréhension des mécanismes tissulaires sous-jacents à la plasticité du filament branchial en mettant en évidence les processus d'apoptose et de prolifération cellulaire qui ont lieu au cours des processus de décolonisation et de recolonisation. Ces processus s'accompagnent d'une variation de la teneur en soufre élémentaire, ainsi que de la taille relative et du contenu génomique des symbiotes
The lateral zone of gills filaments of coastal bivalves Codakia orbiculata and Lucina pensylvanica is the site of chemoautotrophic symbiosis with sulfur-oxidizing bacteria, housed in specialized cells called bacteriocytes. The objective of this thesis is to determine the mechanisms underlying cel1 plasticity and tissue plasticity observed in the lateral zone of gills filaments during the processes of bacterial decolonization and recolonization. In order to do this, the individuals collected in their natural habitat were maintained at the laboratory in seawater filtered tanks, without food and reduced sulfur, to cause bacterial decolonization. When the gills seemed to be purged, the individuals were returned to their natural habitat in order to cause the bacterial recolonization of gills filaments. The analysis of the gills during these processes involves several techniques (histology, immunohistochemistry, molecular hybridization, flow cytometry, total protein assays, protein sulfur assays, X-ray fluorescence spectrometry).This study shows that symbiont acquisition can occur during the entire life of Codakia bivalves. It also allows a better understanding of gills filaments plasticity by highlighting apoptosis and cell proliferation during decolonization and recolonization processes.Theses processes are accompanied with of elemental sufur, relative size and genomic content of symbiontes.

http://www.theses.fr/2011AGUY0461/document

Éditeur(s) : Elsevier
Résumé : The discovery of deep-sea hydrothermal vent fauna, kilometres deep in the oceans, is a great achievement of 20th-century marine biology. The deep-sea hydrothermal food web does not directly depend on the sun's energy. Vent communities rely primarily on trophic associations between chemoautotrophic bacteria and consumers A small. number of endemic taxa are adapted to the inhospitable vent environments that are distributed along ridge crests. Where these vent communities originated and how they dispersed are among the important questions ecologists must answer. Here, by statistical analysis of the most comprehensive database ever assembled about deep-sea hydrothermal fauna, we delineate six major hydrothermal provinces in the world ocean and identify seven possible dispersal pathways between adjacent provinces. Our model suggests that the East-Pacific Rise may have played a pivotal role as a centre of dispersal for the hydrothermal fauna. Our data-driven conclusion will have to be tested by phylogenetic studies and completed by surveys of less-explored fields.
Deep Sea Research Part I: Oceanographic Research Papers (0967-0637) (Elsevier), 2009-08 , Vol. 56 , N. 8 , P. 1371-1378

Droits : 2009 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/2009/publication-6936.pdf
http://archimer.ifremer.fr/doc/2009/sup-6936.xls
DOI:10.1016/j.dsr.2009.01.009
http://archimer.ifremer.fr/doc/00000/6936/