Sylvie M. Gaudron

Colonization of organic substrates deployed in deep-sea reducing habitats by symbiotic species and associated fauna.

In this study, our goal was to test whether typical vent/seep organisms harbouring symbionts or not, would be able to settle on organic substrates deployed in the vicinity of chemosynthetic ecosystems. Since 2006, a series of novel standardized colonization devices (CHEMECOLI: CHEMosynthetic Ecosystem COlonization by Larval Invertebrates) filled with three types of substrates (wood, alfalfa and carbonate) have been deployed in different types of reducing habitats including cold seeps in the eastern Mediterranean, a mud volcano in the Norwegian Sea, and hydrothermal vents on the Mid-Atlantic Ridge for durations of 2 weeks to 1 year. For all deployments, highest species diversities were recovered from CHEMECOLIs filled with organic substrates. Larvae from species associated with thiotrophic symbionts such as thyasirid, vesicomyid and mytilid bivalves, were recovered in the eastern Mediterranean and at the Mid-Atlantic Ridge. At the Haakon Mosby Mud Volcano, larvae of symbiotic siboglinids settled on both organic and carbonate substrates. Overall, novel colonization devices (CHEMECOLI) filled with organic substrates attracted both fauna relying on chemosynthesis-derived carbon as well as fauna relying on heterotrophy the latter being opportunistic and tolerant to sulphide.

Estimating dispersal distance in the deep sea: challenges and applications to marine reserves

Population connectivity refers to the exchange of individuals among populations: it affects gene flow, regulates population size and function, and mitigates recovery from natural or anthropogenic disturbances. Many populations in the deep sea are spatially fragmented, and will become more so with increasing resource exploitation. Understanding population connectivity is critical for spatial management. For most benthic species, connectivity is achieved by the planktonic larval stage, and larval dispersal is, in turn, regulated by complex interactions between biological and oceanographic processes. Coupled biophysical models, incorporating ocean circulation and biological traits, such as planktonic larval duration (PLD), have been used to estimate population connectivity and generate spatial management plans in coastal and shallow waters. In the deep sea, knowledge gaps in both the physical and biological components are delaying the effective use of this approach. Here, we review the current efforts in conservation in the deep sea and evaluate (1) the relevance of using larval dispersal in the design of marine protected areas and (2) the application of biophysical models in the study of population connectivity. Within biophysical models, PLD can be used to estimate dispersal distance. We propose that a PLD that guarantees a minimum dispersal distance for a wide range of species should be used in the planning of marine protected areas in the deep sea. Based on a review of data on species found at depths > 200 m, a PLD of 35 and 69 days ensures a minimum distance for 50% and 75%, respectively, of eurybathic and deep-sea species. We note that more data are required to enhance accuracy and address the high variability in PLD between and within taxonomic groups, limiting generalizations that are often appealing to decision-makers. Given the imminent expansion of resource exploitation in the deep sea, data relevant to spatial management are needed urgently.

Desulfovibrio piezophilus sp. nov., a piezophilic, sulfate-reducing bacterium isolated from wood falls in the Mediterranean Sea

A novel sulfate-reducing bacterium, designated C1TLV30(T), was isolated from wood falls at a depth of 1693 m in the Mediterranean Sea. Cells were motile vibrios (2-4 × 0.5 µm). Strain C1TLV30(T) grew at temperatures between 15 and 45 °C (optimum 30 °C) and at pH 5.4-8.6 (optimum 7.3). It required NaCl for growth (optimum at 25 g NaCl l(-1)) and tolerated up to 80 g NaCl l(-1). Strain C1TLV30(T) used as energy sources: lactate, fumarate, formate, malate, pyruvate and ethanol. The end products from lactate oxidation were acetate, H(2)S and CO(2) in the presence of sulfate as terminal electron acceptor. Besides sulfate, thiosulfate and sulfite were also used as terminal electron acceptors, but not elemental sulfur, fumarate, nitrate or nitrite. Strain C1TLV30(T) possessed desulfoviridin and was piezophilic, growing optimally at 10 MPa (range 0-30 MPa). The membrane lipid composition of this strain was examined to reveal an increase in fatty acid chain lengths at high hydrostatic pressures. The G+C content of the genomic DNA was 49.6 % and the genome size was estimated at 3.5 ± 0.5 Mb. Phylogenetic analysis of the SSU rRNA gene sequence indicated that strain C1TLV30(T) was affiliated to the genus Desulfovibrio with Desulfovibrio profundus being its closest phylogenetic relative (similarity of 96.4 %). On the basis of SSU rRNA gene sequence comparisons and physiological characteristics, strain C1TLV30(T) ( = DSM 21447(T) = JCM 1548(T)) is proposed to be assigned to a novel species of the genus Desulfovibrio, Desulfovibrio piezophilus sp. nov.

The role of chemical signals in the spawning induction of polychaete worms and other marine invertebrates

Extracts of gonad, body fluid and spawning water (into which gametes have been released by the process of spawning) were tested for the presence of spawning-inducing activity (SIA) on both sexes of the polychaetes Nereis succinea (Frey and Leuckhart) and Platynereis dumerilii (Audouin and Milne-Edwards). Gonadal and body fluid extracts from Asterias rubens (L.) and Echinus esculentus (L.) (echinoderms), Arenicola marina (L.) (polychaete), Clupea harengus (L.) (teleost) and body fluid and spawning water of Nereis virens (Sars) (polychaete) all exhibit SIA on male N. succinea. C. harengus extract from male gonads also had SIA on male P. dumerilii. Phytoplankton monoculture extracts were also tested. The cryptomonad (Rhodomonas baltica) (Karsten) had SIA on male P. dumerilii, and the diatom Phaeodactylum tricornutum (Bohlin) had some activity on Mytilus edulis (L.). The active fraction of R. baltica is chromatographically similar to uric acid (the natural sperm-release pheromone produced by females of P. dumerilii) [Nature 382 (1996) 214], but has a different retention time and UV absorbance spectra on reverse phase HPLC. Conspecific extracts of spawning water and gonadal material did not have SIA on the same and opposite sex in A. marina and N. virens. The role of chemical signals in marine invertebrates is discussed in relation to their mode of spawning.

Bacterial communities associated with the wood-feeding gastropod Pectinodonta sp. (Patellogastropoda, Mollusca)

Even though their occurrence was reported a long time ago, sunken wood ecosystems at the deep-sea floor have only recently received specific attention. Accumulations of wood fragments in the deep sea create niches for a diverse fauna, but the significance of the wood itself as a food source remains to be evaluated. Pectinodonta sp. is a patellogastropod that exclusively occurs on woody substrates, where individuals excavate deep depressions, and is thus a potential candidate for a wood-eating lifestyle. Several approaches were used on Pectinodonta sampled close to Tongoa island (Vanuatu) to investigate its dietary habits. Host carbon is most likely derived from the wood material based on stable isotopes analyses, and high cellulase activity was measured in the digestive mass. Electron microscopy and FISH revealed the occurrence of two distinct and dense bacterial communities, in the digestive gland and on the gill. Gland-associated 16S rRNA gene bacterial phylotypes, confirmed by in situ hybridization, included members of three divisions (Alpha- and Gammaproteobacteria, Bacteroidetes), and were moderately related (90-96% sequence identity) to polymer-degrading and denitrifying bacteria. Gill-associated phylotypes included representatives of the Delta- and Epsilonproteobacteria. The possible involvement of these two bacterial communities in wood utilization by Pectinodonta sp. is discussed.

Reproduction of the sea cucumber Holothuria leucospilota in the Western Indian Ocean: biological and ecological aspects

Summary Reproduction of Holothuria leucospilota was investigated for the first time in the Western Indian Ocean in 2005-2006 in the reef of La Réunion. Two spawning events occurred revealed by gonad index (GI) and histology. Gametogenesis of H. leucospilota was divided into five stages of maturity: resting, immature, growing, maturation and post-spawning. It was synchronous within the population and initiated in July when solar radiation increased. From July to December, the growing stage was dominant, followed by a majority of specimen in maturation in January. The first spawning event occurred in February whereas the second one occurred in May. Breeding season of H. leucospilota takes place during Austral summer when temperature and tropical rainfall reach their maximal. Between March and June, only 22% of specimens were found with a completely resorbed gonad in resting stage. ‘Atresia’ appeared more frequently in females in an advanced stage of maturity. In male gonad ‘atresia’ was only observed in the post-spawning stage. Sexual dimorphism was significantly measured in body weight, gonad weight, tubule diameter being all greater in females. An unbalanced sex ratio was skewed (17:191) towards females. Weight at the first maturity was defined for a total weight above 180 g. All of these biological aspects are relevant for the conservation of H. leucospilota species in the Western Indian Ocean that is under fishing pressure.

Reproductive Traits of the Cold-Seep Symbiotic Mussel Idas modiolaeformis: Gametogenesis and Larval Biology

We describe the first reproductive features of a chemosynthetic mussel collected at cold seeps from the eastern Mediterranean Sea. Idas modiolaeformis (Bivalvia, Mytilidae) is a hermaphroditic species in which production of male and female gametes likely alternates, a feature regarded as an adaptation to patchy and ephemeral habitats. By using fluorescent in situ hybridization, we demonstrate that bacterial symbionts, while present within the gills, are absent within acini that enclose female gametes and male gametes. This supports the hypothesis of environmental acquisition of symbionts in chemosynthetic mytilids. Prodissoconch I (PI) is relatively small compared to prodissoconch II (PII), suggesting a planktotrophic larval stage. Diameters of the two larval shells are in the range of sizes reported for mytilids, with a PII size between that of the shallow Mytilus edulis and that of the cold-seep mussel “Bathymodiolus” childressi.

Relative abundances of methane- and sulphur-oxidising symbionts in the gills of a cold seep mussel and link to their potential energy sources

Bathymodiolus mussels are key species in many deep-sea chemosynthetic ecosystems. They often harbour two types of endosymbiotic bacteria in their gills, sulphur- and methane oxidisers. These bacteria take up sulphide and methane from the environment and provide energy to their hosts, supporting some of the most prolific ecosystems in the sea. In this study, we tested whether symbiont relative abundances in Bathymodiolus gills reflect variations in the highly spatially dynamic chemical environment of cold seep mussels. Samples of Bathymodiolus aff. boomerang were obtained from two cold seeps of the deep Gulf of Guinea, REGAB (5°47.86S, 9°42.69E, 3170 m depth) and DIAPIR (6°41.58S, 10°20.94E, 2700 m depth). Relative abundances of both symbiont types were measured by means of 3D fluorescence in situ hybridisation and image analysis and compared considering the local sulphide and methane concentrations and fluxes assessed via benthic chamber incubations. Specimens inhabiting areas with highest methane content displayed higher relative abundances of methane oxidisers. The bacterial abundances correlated also with carbon stable isotope signatures in the mussel tissue, suggesting a higher contribution of methane-derived carbon to the biomass of mussels harbouring higher densities of methane-oxidising symbionts. A dynamic adaptation of abundances of methanotrophs and thiotrophs in the gill could be a key factor optimising the energy yield for the symbiotic system and could explain the success of dual symbiotic mussels at many cold seeps and hydrothermal vents of the Atlantic and Gulf of Mexico.

A tale of two chitons: is habitat specialisation linked to distinct associated bacterial communities?

Although most chitons (Mollusca: Polyplacophora) are shallow-water molluscs, diverse species also occur in deep-sea habitats. We investigated the feeding strategies of two species, Leptochiton boucheti and Nierstraszella lineata, recovered on sunken wood sampled in the western Pacific, close to the Vanuatu Islands. The two species display distinctly different associations with bacterial partners. Leptochiton boucheti harbours Mollicutes in regions of its gut epithelium and has no abundant bacterium associated with its gill. Nierstraszella lineata displays no dense gut-associated bacteria, but harbours bacterial filaments attached to its gill epithelium, related to the Deltaproteobacteria symbionts found in gills of the wood-eating limpet Pectinodonta sp. Stable carbon and nitrogen isotope signatures and an absence of cellulolytic activity give evidence against a direct wood-feeding diet; both species are secondary consumers within the wood food web. We suggest that the distinct associations with bacterial partners are linked to niche specialisations of the two species. Nierstraszella lineata is in a taxonomic family restricted to sunken wood and is possibly adapted to more anoxic conditions thanks to its gill-associated bacteria. Leptochiton boucheti is phylogenetically more proximate to an ancestral form not specialised on wood and may itself be more of a generalist; this observation is congruent with its association with Mollicutes, a bacterial clade comprising gut-associated bacteria occurring in several metazoan phyla.

Diversity of symbioses between chemosynthetic bacteria and metazoans at the Guiness cold seep site (Gulf of Guinea, West Africa)

Fauna from deep‐sea cold seeps worldwide is dominated by chemosymbiotic metazoans. Recently, investigation of new sites in the Gulf of Guinea yielded numerous new species for which symbiosis was strongly suspected. In this study, symbioses are characterized in five seep‐specialist metazoans recently collected from the Guiness site located at ∼600 m depth. Four bivalve and one annelid species belonging to families previously documented to harbor chemosynthetic bacteria were investigated using bacterial marker gene sequencing, fluorescence in situ hybridization, and stable isotope analyses. Results support that all five species display chemosynthetic, sulfur‐oxidizing γ‐proteobacteria. Bacteria are abundant in the gills of bivalves, and in the trophosome of the siboglinid annelid. As observed for their relatives occurring at deeper sites, chemoautotrophy is a major source of carbon for animal nutrition. Although symbionts found in each host species are related to symbionts found in other metazoans from the same families, several incongruencies are observed among phylogenetic trees obtained from the different bacterial genes, suggesting a certain level of heterogeneity in symbiont strains present. Results provide new insights into the diversity, biogeography, and role of symbiotic bacteria in metazoans from the Gulf of Guinea, at a site located at an intermediate depth between the continental shelf and the deep sea.