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The current report focuses on shrimps from deep hydrothermal vents of the Mid-Atlantic Ridge that live in an environment characterized by high hydrostatic pressure, lack of sunlight, and with hot and potentially toxic emissions of black smoker vents. Malacostracan crustaceans display a large diversity of lifestyles and life histories and a rich repertoire of complex behavioral patterns including sophisticated social interactions. These aspects promote this taxon as an interesting group of organisms for those neurobiologists interested in evolutionary transformation of brain structures and evolutionary diversification of neuronal circuits. Here, we explore how analyzing the nervous system of crustacean species from extreme habitats can provide deeper insights into the functional adaptations that drive the diversification of crustacean brain structure., Innerhalb der höheren Krebstiere (Malacostraca) finden sich Vertreter mit einer großen morphologischen und ökologischen Diversität, die sich außerdem durch eine Vielfalt an unterschiedlichen Lebenszyklen und komplexen Verhaltensweisen auszeichnen. Diese Diversität bietet sehr gute Voraussetzungen, um evolutive Transformationen von Gehirnstrukturen und Sinnessystemen im Spannungsfeld zwischen funktionalen und phylogenetischen Zwängen zu analysieren. Insbesondere die Neurobiologie von Tieren aus extremen Lebensräumen kann uns neue Einblicke in funktionale Anpassungen ermöglichen, die die evolutive Diversifizierung von Gehirnstrukturen antreiben. In diesem Beitrag stellen wir die Sinnessysteme von Garnelen der Tiefsee vor, die eng assoziiert mit „black smoker“ hydrothermalen Quellen des Mittelatlantischen Rückens leben, einem lichtlosen und auf den ersten Blick lebensfeindlichem Habitat, dass durch hohen hydrostatischen Druck und die toxischen Emissionen der heißen Quellen geprägt ist.

Deep-sea species endemic to hydrothermal vents face the critical challenge of detecting active sites in a vast environment devoid of sunlight. This certainly requires specific sensory abilities, among which olfaction could be a relevant sensory modality, since chemical compounds in hydrothermal fluids or food odors could potentially serve as orientation cues. The temperature of the vent fluid might also be used for locating vent sites. The objective of this study is to observe the following key behaviors of olfaction in hydrothermal shrimp, which could provide an insight into their olfactory capacities: (1) grooming behavior, (2) attraction to environmental cues (food odors and fluid markers). We designed experiments at both deep-sea and atmospheric pressure to assess the behavior of the vent shrimp Rimicaris exoculata and Mirocaris fortunata, as well as of the coastal species Palaemon elegans and Palaemon serratus for comparison. Here, we show that hydrothermal shrimp groom their sensory appendages similarly to other crustaceans, but this does not clean the dense bacterial biofilm that covers the olfactory structures. These shrimp have previously been shown to possess functional sensory structures, and to detect the environmental olfactory signals tested, but we do not observe significant attraction behavior here. Only temperature, as a signature of vent fluids, clearly attracts vent shrimp and thus is confirmed to be a relevant signal for orientation in their environment.

International audience; The detection of chemical signals is involved in a variety of crustacean behaviors, such as social interactions, search and evaluation of food and navigation in the environment. At hydrothermal vents, endemic shrimp may use the chemical signature of vent fluids to locate active edifices, however little is known on their sensory perception in these remote deep-sea habitats. Here, we present the first comparative description of the sensilla on the antennules and antennae of 4 hydrothermal vent shrimp (Rimicaris exoculata, Mirocaris fortunata, Chorocaris chacei, and Alvinocaris markensis) and of a closely related coastal shrimp (Palaemon elegans). These observations revealed no specific adaptation regarding the size or number of aesthetascs (specialized unimodal olfactory sensilla) between hydrothermal and coastal species. We also identified partial sequences of the ionotropic receptor IR25a, a co-receptor putatively involved in olfaction, in 3 coastal and 4 hydrothermal shrimp species, and showed that it is mainly expressed in the lateral flagella of the antennules that bear the unimodal chemosensilla aesthetascs.

The marine species sensitivity to climate change will depend on the ways by which these species can adapt to thermal increase and heterogeneity. Here, we present evidence that the intertidal shrimp Palaemon elegans acclimates its thermal tolerance, in response to environmental water temperature, through a significant shift of its upper thermal limit with no concomittant acclimation of the heat shock response ( hsp70 stress gene expression threshold). This species is less thermotolerant than its congener Palaemonetes varians, and would therefore potentially be more sensitive to an increase in environmental temperature, such as imposed by global warming. In P. elegans life cycle, physiological adjustments like the shift of the thermal limit and the acquisition of a significant HSR, occurred during the metamorphosis from larvae to post-larvae. This suggests that this step is a genetically-programmed milestone in the process of thermal tolerance acquisition.

The present study questions the ability of marine fauna to modulate the molecular composition of their membrane lipids, as a function of environmental hydrostatic pressure conditions. Specific variations in the cell lipidome composition are presented for the deep-sea hydrothermal vent shrimp Mirocaris fortunata, acclimated at the laboratory for several months at both natural (18 MPa) and atmospheric pressures, but fed with the same diet. These animals display physiological capabilities of acclimation to pressure variations, possibly driven by a particular composition of their lipid species. In particular, it is shown that shrimps exposed to atmospheric pressure exhibit a significantly lower level of the mono-unsaturated fatty acid, vaccenic acid (C18:1n-7), compared to those maintained at natural pressure. The observed fatty acid variations are consistent with a homeoviscous response, i.e. a modulation of the lipidome composition, in response to physical constraints, in order to balance the effects of pressure on membrane order. Additionally, supervised multivariate data analysis was performed for complex lipids and revealed an increase in levels of sterols, sphingomyelin, and ether-phosphatidylethanolamine lipids for animals exposed to atmospheric pressure, with respect to natural pressure. These observations strongly suggest that lipidic domains are involved in the shrimp's response to experimental conditions, and their consistency with a homeoviscous response to pressure is discussed.

International audience; Chemoreception might play an important role for endemic shrimp that inhabit deep and dark hydrothermal vents to find food sources and to locate active edifices that release specific chemicals. We compared the chemosensory abilities of the hydrothermal shrimp Mirocaris fortunata and the coastal related species, Palaemon elegans. The detection of diverse ecologically relevant chemical stimuli by the antennal appendages was measured with electroantennography. The 2 species can detect food-related odor and sulfide, a short-distance stimulus, via both their antennae and antennules. Neither iron nor manganese, considered as long-distance stimuli, was detected by the antennal appendages. Investigation of the ultrastructure of aesthetasc sensilla revealed no specific features of the hydrothermal species regarding innervation by olfactory sensory neurons. Pore-like structures occurring in the aesthetasc cuticle and dense bacterial covering seem to be unique to hydrothermal species, but their potential link to chemoreception remains elusive.

In light of deep-sea mining industry development, particularly interested in massive-sulphide deposits enriched in metals with high commercial value, efforts are increasing to better understand potential environmental impacts to local fauna. The aim of this study was to assess the natural background levels of biomarkers in the hydrothermal vent shrimp Rimicaris exoculata and their responses to copper exposure at in situ pressure (30MPa) as well as the effects of depressurization and pressurization of the high-pressure aquarium IPOCAMP. R. exoculata were collected from the chimney walls of the hydrothermal vent site TAG (Mid Atlantic Ridge) at 3630m depth during the BICOSE cruise in 2014. Tissue metal accumulation was quantified in different tissues (gills, hepatopancreas and muscle) and a battery of biomarkers was measured: metal exposure (metallothioneins), oxidative stress (catalase, superoxide dismutase, glutathione-S-transferase and glutathione peroxidase) and oxidative damage (lipid peroxidation). Data show a higher concentration of Cu in the hepatopancreas and a slight increase in the gills after incubations (for both exposed groups). Significant induction of metallothioneins was observed in the gills of shrimps exposed to 4μM of Cu compared to the control group. Moreover, activities of enzymes were detected for the in situ group, showing a background protection against metal toxicity. Results suggest that the proposed method, including a physiologically critical step of pressurizing and depressurizing the test chamber to enable the seawater exchange during exposure to contaminants, is not affecting metal accumulation and biomarkers response and may prove a useful method to assess toxicity of contaminants in deep-sea species. info:eu-repo/semantics/publishedVersion

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.

In the context of climate change, there is a sustained interest in understanding better the functional mechanisms by which marine ectotherms maintain their physiological scope and define their ability to cope with thermal changes in their environment. Here, we present evidence that the variable shrimp Palaemonetes varians shows genuine acclimation capacities of both the thermal limit (CT(max)) and the heat shock response (hsp70 induction temperature). During cold acclimation to 10 °C, the time lag to adjust the stress gene expression to the current environmental temperature proved to exceed 1 week, thereby highlighting the importance of long-term experiments in evaluating the species' acclimation capacities. Cold and warm-acclimated specimens of P. varians can mobilise the heat shock response (HSR) at temperatures above those experienced in nature, which suggests that the species is potentially capable of expanding its upper thermal range. The shrimp also survived acute heat shock well above its thermal limit without subsequent induction of the HSR, which is discussed with regard to thermal adaptations required for life in highly variable environments.

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.

A new genus of a deep-sea ascomycete with one new species, Alisea longicolla, is described based on analyses of 18S and 28S rDNA sequences and morphological characters. A. longicolla was found together with Oceanitis scuticella, on small twigs and sugar cane debris trawled from the bottom of the Pacific Ocean off Vanuatu Islands. Molecular and morphological characters indicate that both fungi are members of Halosphaeriaceae. Within this family, O. scuticella is phylogenetically related to Ascosalsum and shares similar ascospore morphology and appendage ontogeny. The genus Ascosalsum is considered congeneric with Oceanitis and Ascosalsum cincinnatulum, Ascosalsum unicaudatum and Ascosalsum viscidulum are transferred to Oceanitis, an earlier generic name.

Wood falls in the deep sea have recently become the focus of studies showing their importance as nutrients on the deep-sea floor. In such environments, Crustaceans constitute numerically the second-largest group after Mollusks. Many questions have arisen regarding their trophic role therein. A careful examination of the feeding appendages, gut contents, and gut lining of Munidopsis andamanica caught with wood falls revealed this species as a truly original detritivorous species using wood and the biofilm covering it as two main food sources. Comparing individuals from other geographic areas from substrates not reported highlights the galatheid crab as specialist of refractory substrates, especially vegetal remains. M. andamanica also exhibits a resident gut microflora consisting of bacteria and fungi possibly involved in the digestion of wood fragments. The results suggest that Crustaceans could be full-fledged actors in the food chains of sunken-wood ecosystems and that feeding habits of some squat lobsters could be different than scavenging.

Symposium on Marine Benthic Ecology and Biodiversity - A Compilation of Recent Advances held in Honor of J Frederick Grassle, Rutgers Univ, Inst Marine Coastal Sci, New Brunswick, NJ, NOV 20-21, 2008; International audience; Several species of the polychaete family Alvinellidae may be considered as `ecosystem engineer' because, by building their tubes, they modify the architecture of the hydrothermal fluid-seawater interface on the walls of vent chimneys. This affects the thermal and chemical gradients, and creates a mosaic of micro-niches, which could enable colonization by a variety of less-tolerant species. On high temperature vents of the Juan de Fuca Ridge, Alvinellid-dominated communities colonizing first mineral surfaces are followed by a succession of communities with different species composition. On the East Pacific Rise (EPR), tubes of Alvinella spp, may seal the mineral surface on which they grow and decrease fluid seepage, or tubes may become encrusted in mineral precipitations. An alvinellid colony may therefore persist for only a restricted time period at a given place. Here we investigated the development of alvinellid colonies on the EPR vent sites in order to detect whether a succession of new species less tolerant would follow colonization by Alvinella spp. or if different assemblages are forming depending on local conditions. Using a specially designed device called TRAC (titanium ring for alvinellid colonization), we described the evolution of newly formed colonies. Fifteen experiments were conducted on several chimneys of the 9 degrees N and 13 degrees N vent fields of the EPR, over durations ranging from 5 days up to 5 months. Through video analysis, different types of colonies were identified, characterized by increasing thickness of the Alvinella coverage, decreasing fluid flow bathing the colony, and decreasing surface temperatures. We showed that the assemblage formed by minerals, tubes, and organisms is produced at a very high rate. While animals may colonize the new surface in less than a week, and tubes are also quickly produced (Alvinella species may grow their tube up to 1 cm day(-1) during the early stages of colonization), mineral precipitation progressively increases the mineral content of the assemblage. Active colonies do not seem to persist longer than 5 months, highlighting the rapid turnover of this habitat. Colonies collected had different species compositions, but community structures exhibited no significant variations with vent field (9 degrees N/13 degrees N), deployment duration (< or > 1 month), or type of colony. However, multivariate analysis revealed that mineral content would best explain community structure variations. Biodiversity indexes increased quickly within the first weeks of TRAC deployment time, as well as with the amount of Alvinella tube. The development of a complex architecture may thus promote the installation of species lacking adaptations to temperature. However, our results do not support a succession pattern but rather the development of communities with slight differences in species compositions that may reflect local environmental conditions. (C) 2009 Elsevier Ltd. All rights reserved.

Specimens of Lamellibrachia (Annelida: Siboglinidae) were recently discovered at cold seeps in the eastern Mediterranean. In this study, we have investigated the phylogeny and function of intracellular bacterial symbionts inhabiting the trophosome of specimens of Lamellibrachia sp. from the Amon mud volcano, as well as the bacterial assemblages associated with their tube. The dominant intracellular symbiont of Lamellibrachia sp. is a gammaproteobacterium closely related to other sulfide-oxidizing tubeworm symbionts. In vivo uptake experiments show that the tubeworm relies on sulfide for its metabolism, and does not utilize methane. Bacterial communities associated with the tube form biofilms and occur from the anterior to the posterior end of the tube. The diversity of 16S rRNA gene phylotypes includes representatives from the same divisions previously identified from the tube of the vent species Riftia pachyptila, and others commonly found at seeps and vents.

The Rimicaris exoculata shrimp is considered as a primary consumer that dominates the fauna of most Mid-Atlantic Ridge (MAR) hydrothermal ecosystems. These shrimps harbour in their gill chambers an important ectosymbiotic community of chemoautotrophic bacteria associated with iron oxide deposits. The structure and elemental composition of the mineral concretions associated with these bacteria have been investigated by using LM, ESEM, TEM STEM and EDX microanalyses. The nature of the iron oxides in shrimps obtained from the Rainbow vent field has also been determined by Mössbauer spectroscopy. This multidisciplinary approach has revealed that the three layers of mineral crust in the Rimicaris exoculata shrimps consist of large concretions formed by aggregated nanoparticles of two-line ferrihydrite and include other minor elements as Si, Ca, Mg, S and P, probably present as silicates cations, sulphates or phosphates respectively that may contribute to stabilise the ferrihydrite form of iron oxides. TEM-observations on the bacteria have revealed their close interactions with these minerals. Abiotic and biotic precipitation could occur within the gill chamber of Rimicaris exoculata, suggesting the biologically-mediated formation of the iron oxide deposits. The difference of the bacterial density in the three-mineral crust layers could be correlated to the importance of the iron oxide concretions and suggest that the first mineral particles precipitates on the lower layer which could be considered as the most likely location of iron-oxidizing bacteria.

Antennular chemoreception in aquatic decapods is well-studied via the recording of single chemoreceptor neuron activity in the antennule, but global responses of the antennule (or antennae in insects) by electroantennography (EAG) was so far mainly restricted to aerial conditions. We present here a well-established underwater EAG method to record the global antennule activity in the marine shrimp Palaemon elegans in natural (aqueous) conditions. EAG responses to food extracts, recorded as net positive deviations of the baseline, are reproducible, dose-dependent and exhibit sensory adaptation. This new EAG method opens a large field of possibilities for studying in vivo antennular chemoreception in aquatic decapods, in a global approach to supplement current more specific techniques.

Two trawl samples of natural sunken wood collected near Vanuatu were identified based on histological studies. Detailed descriptions were made and microphotographs of the cell types were taken, using the three classical sections (cross, tangential and radial). The botanical characters were compared first to the native flora of Vanuatu, then also to the introduced species. The possibility of transportation by ocean currents with a mainly southern and eastern direction is discussed. The first sample lacks apparent colonization marks. Its main histologic character is the presence of `paedomorphic type I rays' which relates it to the shrubby genera Fitchia (Asteraceae) and Fuchsia (Onagraceae): both are known in Polynesia but they seem not to be recorded from Vanuatu. The second wood sample is densely colonized by molluscs and other fauna. It comes from a tree close to the Fabaceae-Mimosoideae, possibly belonging to the genera Leucaena or Serianthes, both known from Vanuatu. Our work shows for the first time that, even after prolonged submergence in water at substantial depth (> 500 m), the wood structure may be well preserved. Therefore, the botanical identification and the determination of the origin of the sunken wood were possible. The two selected samples show completely different colonization patterns, which could be related to differences in chemical composition or to time elapsed since sinking.

Identifying the factors driving community dynamics in hydrothermal vent communities, and in particular biological interactions, is challenged by our ability to make direct observations and the difficulty to conduct experiments in those remote ecosystems. As a result, we have very limited knowledge on species׳ behaviour and interactions in these communities and how they in turn influence community dynamics. Interactions such as competition or predation significantly affect community structure in vent communities, and video time-series have successfully been used to gain insights in biological interactions and species behaviour, including responses to short-term changes in temperature or feeding strategies. In this study, we combined in situ and ex situ approaches to characterise the behaviour and interactions among two key species encountered along the Mid-Atlantic Ridge (MAR): the shrimp Mirocaris fortunata and the crab Segonzacia mesatlantica. In situ, species small-scale distribution, interactions and behaviour were studied using the TEMPO observatory module deployed on the seafloor at the base of the active Eiffel Tower edifice in the Lucky Strike vent field as part of the EMSO-Acores MoMAR observatory. TEMPO sampled 2 min of video four times a day from July 2011 to April 2012. One week of observations per month was used for ‘long-term’ variations, and a full video data set was analysed for January 2012. In addition, observations of crab and shrimp individuals maintained for the first time under controlled conditions in atmospheric pressure (classic tank) and pressurised (AbyssBox) aquaria allowed better characterisation and description of the different types of behaviour and interactions observed in nature. While the identified in situ spatial distribution pattern was stable over the nine months, both species displayed a significant preference for mussel bed and anhydrite substrata, and preferentially occupied the area located directly in the fluid flow axis. The aggregation behaviour of M. fortunata resulted in the occurrence of numerous intraspecific interactions mainly involving the use of two pairs of sensory organs (antenna/antennule) and fleeing behaviours when in contact or close to individuals of S. mesatlantica. The higher level of passiveness observed in the ex situ artificial environment compared to the in situ environment was attributed to the lack of stimulation related to low densities of congeners and/or of sympatric species compared to the natural environment and the absence of continuous food supply, as both species displayed a significant higher level of activity during feeding time. This result emphasises the role of food supply as a driver of species distribution and behaviour. Direct in situ observations using cameras deployed on deep-sea observatories, combined with experimental set-up in pressurised aquaria, will help investigators understand the factors influencing community dynamics and species biology at vents as well as their underlying mechanisms.

International audience; The AbyssBox project aims to provide the first permanent public exhibition of live deep-sea hydrothermal fauna maintained at in situ pressure. AbyssBox is a pressurized aquarium designed to function permanently. Here we present details of the project after the public exhibition functioned for more than three years at Océanopolis aquarium in Brest, France. We also describe the AbyssBox pressure aquarium, and provide data and observations on vent shrimp (Mirocaris fortunata) and crabs (Segonzacia mesatlantica) that were sampled from 1700 m depth at the Lucky Strike vent field (Mid-Atlantic Ridge) during different cruises. While mortalities exceeded 50% during the first days following sampling, the remaining animals appeared to acclimate fairly well. Some crabs have now been kept for more than 2 years, and some shrimp have spent more than 3 years in captivity. Primarily designed for a public exhibition, the AbyssBox is already used for scientific purposes, since it provides one of the most effective tools for long-term rearing of deep-sea fauna. AbyssBox is a first step towards maintaining a variety of deep-sea fauna year-round at in situ pressure, which will serve both scientific and public interests.

Animal communities on the walls of deep-sea hydrothermal chimneys are distributed in mosaics of patches that may evolve as local environmental conditions change and biological inter- actions develop. Alvinella pompejana Desbruyeres et Laubier, 1980 is one of the first metazoan colonisers of new surfaces created by mineral precipitation, and therefore may be particularly impor- tant in community establishment in active parts of smokers. Here our goal was to investigate the colonisation mechanisms of A. pompejana in new patches and determine whether these mechanisms may influence population structure and reproductive patterns in this species. We deployed a series of TRAC (Titanium Ring for Alvinellid Colonisation) devices at East Pacific Rise (EPR) vent sites to com- pare the size and stage (i.e. reproductive maturity) distribution of A. pompejana individuals between recently colonised patches (TRACs) and established patches (grabbed by submersible). TRACs deployed for short time periods (11 d to 1 mo) were generally colonised by smaller individuals than those found in background populations or in TRACs deployed for longer time periods (>1 mo). Colonists into new patches were a mix of juveniles and individuals that were sexually mature but non-reproductive (i.e. not producing gametes), whereas background population and older patches harboured a mixture of individuals at different stages including reproductive females. Although some individuals may have recruited on TRACs as larvae, the major colonisation process involved was probably immigration of post-larval stages. In long-term TRAC, reproductive females were not reproductively synchronised. In this dynamic environment, reproduction would be triggered by the disturbance/migration processes, explaining the heterogeneity observed in reproductive patterns.

In this study, we have investigated the mineralogy present on the scaphognatite of the Rimicaris exoculata shrimp collected at a Mid-Atlantic Ridge hydrothermal vent. The structure, morphology and impurities of the minerals were analyzed by transmission electron microscopy (TEM) and electron energy loss spectroscopy (EELS). The main minerals observed were clusters of iron oxides found attached to the bacterial cells present on the setae of the scaphognatite. Structural investigations based on high-resolution electron microscopy and electron diffraction affirm that these clusters are composed of two-line ferrihydrite. The EELS measurements demonstrate that the ferrihydrite is bearing a mixed valence iron distribution and P, Si, Ca, N and Ni ions at impurity levels. The stability of this uncommon phase is then discussed based on work previously reported in literature for analogous poorly ordered iron oxides.

Hydrothermal vent environments, particularly those associated with the vestimentiferan Riftia pachyptila, are believed to be among the highest chitin-producing systems. In order to elucidate the chitin cycle in these environments, we estimate the in situ chitin degradation rates of tube-worm exoskeletons. Our in situ experiments show that the tubes of Riftia are highly stable structures. Comparative measurements of the degradation rates of Riftia tubes and crab shells immersed at deep-sea vents show that the tubes would be degraded within 2.5 years, whereas the time for the total degradation of the vent crab (Bythograea thermydron) carapaces would not exceed 36 days. The importance of the microbial participation in this degradation was estimated for Riftia tubes. Based on previous work, we calculated chitin production by a population of Riftia tubes of about 750 g m-2 year-1 (763). From our in situ experiments, we estimated a microbial chitinolysis rate of about 500 g m-2 year-1 (496) (65% of the chitin produced).Exoskeletons containing β-chitin appear more stable in natural environments than those containing α-chitin and would thus be less available as carbon and nitrogen sources. In contrast, isolated β-chitin was hydrolysed faster than α-chitin during in vitro degradation experiments; for instance, Riftia β-chitin was degraded about 3- to 4-fold faster than Bythograea α-chitin. A stabilization process by disulfide bonds of the proteins-chitin link, rather than the crystalline form of the chitin (α/β), accounts for the resistance of Riftia tubes to enzymatic attacks.

A novel thermophilic, chemolithoautotrophic bacterium, designated as NE1206(T), was isolated from a Juan de Fuca Ridge hydrothermal vent sample (tubes of the annelid polychaete Paralvinella sulfincola attached to small pieces of hydrothermal chimney). The cells were rod-shaped (1.2-3.5 x 0.4-0.7 microm), occurring as single motile rods or forming macroscopic aggregates visible as pinkish to brownish streamers. The new isolate was anaerobic. It grew between 50 and 70 degrees C (optimum 60-65 degrees C; doubling time approximately 1 h 15 min at 60 degrees C), between pH 5.0 and 7.5 (optimum pH around 6.0-6.5) and at sea salts concentrations between 20 and 40 g l(-1 )(optimum 30 g l(-1)). Cells grew chemolithoautotrophically in an H(2)/CO(2) atmosphere (80/20, v/v; 200 kPa). Molecular hydrogen was the sole electron donor used by the strain. Nitrate and elemental sulfur served as electron acceptors, yielding ammonia and hydrogen sulfide, respectively (nitrate reduction supported higher growth rates than sulfur reduction). The G+C content of the genomic DNA was 36.7+/-0.8 mol%. Phylogenetic analyses of the 16S rRNA gene located the strain within the genus Desulfurobacterium. However, the novel isolate possesses physiological and biochemical characteristics that differ from the previously described species of this genus. We propose that the isolate represents a novel species, Desulfurobacterium crinifex sp. nov. The type strain is NE1206(T) (DSM 15218(T), CIP 107649(T)). An amendment of the genus Desulfurobacterium description is proposed, based on the phenotypic characteristics of the novel species.

International audience; During the past decade, a large number of multi-gene analyses aimed at resolving the phylogenetic relationships within Decapoda. However relationships among families, and even among sub-families, remain poorly defined. Most analyses used an incomplete and opportunistic sampling of species, but also an incomplete and opportunistic gene selection among those available for Decapoda. Here we test in the Caridea if improving the taxonomic coverage following the hierarchical scheme of the classification, as it is currently accepted, provides a better phylogenetic resolution for the inter-families relationships. The rich collections of the Muséum National d’Histoire Naturelle de Paris are used for sampling as far as possible at least two species of two different genera for each family or subfamily. All potential markers are tested over this sampling. For some coding genes the amplification success varies greatly among taxa and the phylogenetic signal is highly saturated. This result probably explains the taxon-heterogeneity among previously published studies. The analysis is thus restricted to the genes homogeneously amplified over the whole sampling. Thanks to the taxonomic sampling scheme the monophyly of most families is confirmed. However the genes commonly used in Decapoda appear non-adapted for clarifying inter-families relationships, which remain poorly resolved. Genome-wide analyses, like transcriptome-based exon capture facilitated by the new generation sequencing methods might provide a sounder approach to resolve deep and rapid radiations like the Caridea.

A novel thermophilic, anaerobic, strictly chemoorganoheterotrophic bacterium, designated as AM1114(T), was isolated from a deep-sea hydrothermal vent sample from the East-Pacific Rise (EPR 13degrees N). The cells were long (3-10 mum) rods, motile with peritrichous flagella, and exhibited a Gram-negative cell wall ultrastructure. In the late stationary phase of growth, cells formed an ovoid, refractile, terminal endospore. They grew at 45-65 degreesC inclusive (optimum 55-60 degreesC; doubling time approx. 45 min), at pH 4.5-8.0 inclusive (optimum pH 7.5-8.0) and at sea salt concentrations of 20-60 g l(-1) inclusive (optimum 25-30 g l(-1)). Strain AM1114(T) was an obligately heterotrophic bacterium able to ferment a mixture of 20 amino acids, complex proteinaceous substrates (such as yeast extract, brain-heart infusion or peptone), and carbohydrates such as glucose, galactose or maltose. The main fermentation products on glucose/yeast extract/peptone/sulfur medium were hydrogen, carbon dioxide, butyrate, ethanol, acetate, formate and L-alanine. The G+C content of the genomic DNA (determined by thermal denaturation) was 24.2+/-1 mol %. Phylogenetic analyses of the 16S rRNA gene located the strain within cluster XI of the lineage encompassing the genus Clostridium and related genera (sensu Collins et al., 1994), in the bacterial domain. On the basis of 16S rDNA sequence comparisons and physiological and biochemical characteristics, it is proposed that the isolate should be described as a novel genus, namely Caminicella gen. nov., of which Caminicella sporogenes sp. nov. is the type species. The type strain is AM114(T) (= DSM 14501(T) = CIP 107141(T)).

Nanocrystalline zinc-iron sulphide minerals were found within tubes of Alvinella pompejana , collected at 9°N on the East Pacific Rise, in a preliminary study carried out by conventional analytical transmission electron microscopy. Their structure is consistent with either the sphalerite or wurtzite polymorphs of ZnS. In contrast with sulphides usually observed in deep-sea hydrothermal environments, the unique sulphide observed within the exoskeleton of the pompeii worms has a (Zn 0.88 Fe 0.12 )S composition. The zinc-iron sulphide nanocrystals are grouped in submicrometer-sized clusters which form layers, concentrically to the proteinaceous tube axis. This is the first characterization of minerals precipitated within a biological matrix from hydrothermal vents organisms. These minerals represent a new example of zinc-iron biologically induced mineralization. Such layers of nanocrystalline zinc-iron sulphide minerals could be used as valuable markers of annelid tubes, in the growing field of studies focusing on fossilized paleo-hydrothermal vent systems.

The shrimp Rimicaris exoculata dominates several hydrothermal vent ecosystems of the Mid-Atlantic Ridge and is thought to be a primary consumer harbouring a chemoautotrophic bacterial community in its gill chamber. The aim of the present study was to test current hypotheses concerning the epibiont’s chemoautotrophy, and the mutualistic character of this association. In-vivo experiments were carried out in a pressurised aquarium with isotope-labelled inorganic carbon (NaH13CO3 and NaH14CO3) in the presence of two different electron donors (Na2S2O3 and Fe2+) and with radiolabelled organic compounds (14C-acetate and 3H-lysine) chosen as potential bacterial substrates and/or metabolic by-products in experiments mimicking transfer of small biomolecules from epibionts to host. The bacterial epibionts were found to assimilate inorganic carbon by chemoautotrophy, but many of them (thick filaments of epsilonproteobacteria) appeared versatile and able to switch between electron donors, including organic compounds (heterotrophic acetate and lysine uptake). At least some of them (thin filamentous gammaproteobacteria) also seem capable of internal energy storage that could supply chemosynthetic metabolism for hours under conditions of electron donor deprivation. As direct nutritional transfer from bacteria to host was detected, the association appears as true mutualism. Import of soluble bacterial products occurs by permeation across the gill chamber integument, rather than via the digestive tract. This first demonstration of such capabilities in a decapod crustacean supports the previously discarded hypothesis of transtegumental absorption of dissolved organic matter or carbon as a common nutritional pathway.

Marine waterlogged woods on the ocean floor provide the foundation for an ecosystem resulting in high biomass and potentially high macrofaunal diversity, similarly to other large organic falls. However, the microorganisms forming the base of wood fall ecosystems remain poorly known. To study the microbial diversity and community structure of sunken woods, we analyzed over 2800 cloned archaeal and bacterial 16S rRNA gene sequences from samples with different geographic locations, depths, and immersion times. The microbial communities from different wood falls were diverse, suggesting that sunken woods provide wide-ranging niches for microorganisms. Microorganisms dwelling at sunken woods change with time of immersion most likely due to a change in chemistry of the wood. We demonstrate, for the first time in sunken woods, the co-occurrence of free-living sulfate-reducing bacteria and methanogens and the presence of sulfide oxidizers. These microorganisms were similar to those of other anaerobic chemoautotrophic environments suggesting that large organic falls can provide similar reduced habitats. Furthermore, quantification of phylogenetic patterns of microbial community assembly indicated that environmental forces (habitat filtering) determined sunken wood microbial community structure at all degradation phases of marine woodfalls. We also include a detailed discussion on novel archaeal and bacterial phylotypes in this newly explored biohabitat.

The caridean shrimp Rimicaris exoculata dominates the fauna at several Mid-Atlantic Ridge hydrothermal vent sites. This shrimp has an enlarged gill chamber, harboring a dense ectosymbiotic community of chemoautotrophic bacteria associated with mineral oxide deposits. Until now, their acquisition is not fully understood. At three hydrothermal vent sites, we analyzed the epibionts diversity at different moult stages and also in the first stages of the shrimp life (eggs, hatched eggs (with larvae) and juveniles). Hatched eggs associated with young larvae were collected for the first time directly from gravid females at the Logachev vent site during the Serpentine cruise. An approach using 16S rRNA clone libraries, scanning and transmission electron microscopy, and fluorescent in situ hybridization was used. Molecular results and microscope observations indicated a switch in the composition of the bacterial community between early R. exoculata life cycle stage (egg libraries dominated by the Gammaproteobacteria) and later stages (juvenile/adult libraries dominated by the Epsilonproteobacteria). We hypothesized that the epibiotic phylotype composition could vary according to the life stage of the shrimp. Our results confirmed the occurrence of a symbiosis with Gammaproteobacteria and Epsilonproteobacteria, but more complex than previously assumed. We revealed the presence of active type-I methanotrophic bacteria colonizing the cephalothorax of shrimps from the Rainbow site. They were also present on the eggs from the Logachev site. This could be the first 'epibiotic' association between methanotrophic bacteria and hydrothermal vent crustacean. We discuss possible transmission pathways for epibionts linked to the shrimp life cycle. The ISME Journal (2012) 6, 597-609; doi: 10.1038/ismej.2011.133; published online 13 October 2011

International audience; Squat lobsters of the deep-sea genus Munidopsis are among the most regularly reported crustaceans associated with deep-sea wood falls. They are often thought to indirectly use these substrates for preying or scavenging wood-associated molluscs or annelids, albeit the species M. andamanica has been recently highlighted as a xylophagous specialist. In this work, we examined the feeding appendages, gut contents and gut lining of M. nitida, M. bispinoculata and M. pilosa specimens from natural sunken woods and compared them with specimens of the same species having survived and grown on different hard-to-digest substrates (i.e. woods, turtle shells and whale bones) experimentally submerged in the deep South Pacific. In both cases, all three species directly ingest large wood fragments deeply degraded by microorganisms, but M. nitida also feeds on experimentally submerged whale bone and turtle shell fragments. Munidopsis nitida is also the only species to host a resident gut microflora, but the bacterial morphotypes vary according to the ingested substrate. The results suggest that the three species are most probably opportunistic, bacterivorous detritivores and that M. nitida could be at the beginning of an evolutionary process towards xylophagy within the genus Munidopsis.

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.

En libre-accès sur Archimer : http://archimer.ifremer.fr/doc/00000/11142/7919.pdf; International audience; Rimicaris exoculata dominates the megafauna of several Mid-Atlantic Ridge hydrothermal sites. Its gut is full of sulphides and iron-oxide particles and harbours microbial communities. Although a trophic symbiosis has been suggested, their role remains unclear. In vivo starvation experiments in pressurized vessels were performed on shrimps from Rainbow and Trans-Atlantic Geotraverse sites in order to expel the transient gut contents. Microbial communities associated with the gut of starved and reference shrimps were compared using 16S rRNA gene libraries and microscopic observations (light, transmission and scanning electron microscopy and FISH analyses). We show that the gut microbiota of shrimps from both sites included mainly Deferribacteres, Mollicutes, Epsilon- and Gammaproteobacteria. For the first time, we have observed filamentous bacteria, inserted between microvilli of gut epithelial cells. They remained after starvation periods in empty guts, suggesting the occurrence of a resident microbial community. The bacterial community composition was the same regardless of the site, except for Gammaproteobacteria retrieved only in Rainbow specimens. We observed a shift in the composition of the microbiota of long-starved specimens, from the dominance of Deferribacteres to the dominance of Gammaproteobacteria. These results reinforce the hypothesis of a symbiotic relationship between R. exoculata and its gut epibionts.

The definitive version is available at ww3.interscience.wiley.com; International audience; The shrimp Rimicaris exoculata from hydrothermal vents on the Mid-Atlantic Ridge (MAR) harbours bacterial epibionts on specialized appendages and the inner surfaces of its gill chamber. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization (FISH), we examined the R. exoculata epibiosis from four vents sites along the known distribution range of the shrimp on the MAR. Our results show that R. exoculata lives in symbiosis with two types of filamentous epibionts. One belongs to the Epsilonproteobacteria, and was previously identified as the dominant symbiont of R. exoculata. The second is a novel gammaproteobacterial symbiont that belongs to a clade consisting exclusively of sequences from epibiotic bacteria of hydrothermal vent animals, with the filamentous sulfur oxidizer Leucothrix mucor as the closest free-living relative. Both the epsilon- and the gammaproteobacterial symbionts dominated the R. exoculata epibiosis at all four MAR vent sites despite striking differences between vent fluid chemistry and distances between sites of up to 8500 km, indicating that the symbiosis is highly stable and specific. Phylogenetic analyses of two mitochondrial host genes showed little to no differences between hosts from the four vent sites. In contrast, there was significant spatial structuring of both the gamma- and the epsilonproteobacterial symbiont populations based on their 16S rRNA gene sequences that was correlated with geographic distance along the MAR. We hypothesize that biogeography and host-symbiont selectivity play a role in structuring the epibiosis of R. exoculata.

International audience; With an increased appreciation of the frequency of their occurrence, large organic falls such as sunken wood and whale carcasses have become important to consider in the ecology of the oceans. Organic-rich deep-sea falls may play a major role in the dispersal and evolution of chemoautotrophic communities at the ocean floor, and chemosynthetic symbiotic, free-living, and attached microorganisms may drive the primary production at these communities. However, little is known about the microbiota thriving in and around organic falls. Our aim was to investigate and compare free-living and attached communities of bacteria and archaea from artificially immersed and naturally sunken wood logs with varying characteristics at several sites in the deep sea and in shallow water to address basic questions on the microbial ecology of sunken wood. Multivariate indirect ordination analyses of capillary electrophoresis single-stranded conformation polymorphisms (CE-SSCP) fingerprinting profiles demonstrated high similarity of bacterial and archaeal assemblages present in timbers and logs situated at geographically distant sites and at different depths of immersion. This similarity implies that wood falls harbor a specialized microbiota as observed in other ecosystems when the same environmental conditions reoccur. Scanning and transmission electron microscopy observations combined with multivariate direct gradient analysis of Bacteria CE-SSCP profiles demonstrate that type of wood (hard vs. softwood), and time of immersion are important in structuring sunken wood bacterial communities. Archaeal populations were present only in samples with substantial signs of decay, which were also more similar in their bacterial assemblages, providing indirect evidence of temporal succession in the microbial communities that develop in and around wood falls.

The phylogenetic diversity of the gut microbial population of the hydrothermal shrimp Rimicaris exoculata was determined. The presence of micro-organisms in the hindgut of the shrimp was determined, and their morphology illustrated for the first time by transmission electron microscopy. DNA was extracted from the fore-, mid- and hindgut of shrimps collected at the Rainbow site, at the Mid-Atlantic Ridge, and the sequences of the small-subunit rRNA (16S rDNA) were analyzed. Seven different bacterial phylotypes were identified from the 90 clones screened. The sequences were affiliated to three major groups: epsilon-Proteobacteria (48,8%), Entomoplasmatales (23.3%) and representatives of the new phylum Deferribacteres (26.6%). These results show that the diversity in the shrimp gut is very low compared to that of the surrounding medium. Furthermore, the presence of groups that are not found in the external medium (Spiroplasma sp. and Geovibrio sp.) suggests the existence of a local microflora. The potential roles of these bacteria are discussed, involving the shrimp diet and metal bioremediation.

The shrimp Rimicaris exoculata is considered a primary consumer that dominates the fauna of most Mid-Atlantic Ridge hydrothermal ecosystems. The shrimps harbour in their gill chamber an important ectosymbiotic community of chemoautotrophic bacteria associated with iron oxide deposits. The settlement and development of this ectosymbiosis was investigated using microscopy techniques (light microscopy, LM; and scanning, transmission and environmental scanning electron microscopy: SEM and ESEM, respectively) for shrimps from 2 different vent fields (Rainbow, 36 degrees 14.0' N and TAG, 26 degrees 08.0' N). The results revealed a bacterial re-colonisation after each exuviation and a development of the bacterial community in 5 steps in relation to the moult stages, which were used as a reference time scale. In 287 shrimps from both vent fields, pre-ecdysial stages prevailed in the population, suggesting a short anecdysis and high moulting rate, probably to renew the ectosymbiosis. Comparisons with moult cycles of littoral shrimps suggest that the interval between successive exuviations in R. exoculata may be as short as 10 d. The colours of R. exoculata result from accumulation of iron oxide, which forms a bacteria-associated mineral crust in the gill chambers. The close correspondence between moult stages, the development of the ectosymbiont community and shrimp colours indicate that colour could be used to rapidly determine shrimp moult stages.

En libre-accès sur Archimer : http://archimer.ifremer.fr/doc/2008/publication-4563.pdf; International audience; The Rimicaris exoculata dominates the megafauna of some of the Mid Atlantic ridge hydrothermal vent sites. This species harbors a rich community of bacterial epibionts inside its gill chamber. Literature data indicate that a single 16S rRNA phylotype dominates this epibiotic community, and is assumed to be a sulfide-oxidizing bacteria. However attempts of cultivation were not successful and did not allow to confirm it. The aim of our study was to test the hypothesis of sulfide oxidation in the gill chamber, by a multidisciplinary approach, using in vivo experiments at in situ pressure in the presence of sulfide, microscopic observations and a molecular survey. Morphology of microorganisms, before and after treatment, was analyzed to test the effect of sulfide depletion and re-exposure. Our observations, as well as molecular data indicate a wider diversity than previously described for this shrimp's epibiotic community. We observed occurrence of bacterial intracellular sulfur- and iron-enriched granules and some methanotrophic-like bacteria cells for the first time. Genes that are characteristic of methane- oxidizing (pmoA) and sulfide-oxidizing (APS) bacteria were identified. These results suggest that three metabolic types (iron, sulfide and methane oxidation) may co-occur within the epibiont community associated with Rimicaris exoculata. As this shrimp colonizes chemically contrasted environments, the relative abundance of each metabolic type could vary according to the local availability of reduced compounds.

Sunken woods are abundant in deep oceanic environments, housing a huge faunal diversity. Studies on that substrate firstly focused on the associated organisms, but since a few years, identification of wood is a further aim. The purpose is to appreciate its degradation state, diversity, geographical origin and to identify specific associations between colonizing organisms and substrates. The first determinations were made on sunken woods from Taiwan/Philippines, the Vanuatu Archipelago, and the Mediterranean Sea. Samples’ identification was based on histological studies. Different preparation techniques were used, depending on their degradation state. Detailed anatomy descriptions were made and compared to the native flora and the introduced species. Wood samples were well preserved. Diversified species were encountered, seemingly originating from local floras. In situ settlements of known wood species will enhance the knowledge of degradation and colonization degrees.

International audience; Rimicaris exoculata is a deep-sea hydrothermal vent shrimp whose enlarged gill chamber houses a complex trophic epibiotic community. Its gut harbours an autochthonous and distinct microbial community. This species dominates hydrothermal ecosystem megafauna along the Mid-Atlantic Ridge, regardless of contrasting geochemical conditions prevailing in them. Here, the resident gut epibiont community at four contrasted hydrothermal vent sites (Rainbow, TAG, Logatchev and Ashadze) was analysed and compiled with previous data to evaluate the possible influence of site location, using 16S rRNA surveys and microscopic observations (transmission electron microscopy, scanning electron microscopy and fluorescence in situ hybridization analyses). Filamentous epibionts inserted between the epithelial cell microvilli were observed on all examined samples. Results confirmed resident gut community affiliation to Deferribacteres, Mollicutes, Epsilonproteobacteria and to a lesser extent Gammaproteobacteria lineages. Still a single Deferribacteres phylotype was retrieved at all sites. Four Mollicutes-related operational taxonomic units were distinguished, one being only identified on Rainbow specimens. The topology of ribotype median-joining networks illustrated a community diversification possibly following demographic expansions, suggesting a more ancient evolutionary history and/or a larger effective population size at Rainbow. Finally, the gill chamber community distribution was also analysed through ribotype networks based on sequences from R. exoculata collected at the Rainbow, Snake Pit, TAG, Logatchev and Ashadze sites. Results allow the refining of hypotheses on the epibiont role and transmission pathways.

Alvinella pompejana is a tube-dwelling polychaete colonizing hydrothermal smokers of the East Pacific Rise. Extreme temperature, low pH and millimolar sulfide levels have been reported in its immediate surroundings. The conditions experienced by this organism and its associated microbes are, however, poorly known and the processes controlling the physico-chemical gradients in this environment remain to be elucidated. Using miniature in situ sensors coupled with close-up video imagery, we have characterized fine-scale pH and temperature profiles in the biogeoassemblage constituting A. pompejana colonies. Steep discontinuities at both the individual and the colony scale were highlighted, indicating a partitioning of the vent fluid–seawater interface into chemically and thermally distinct micro-environments. The comparison of geochemical models with these data furthermore reveals that temperature is not a relevant tracer of the fluid dilution at these scales. The inner-tube micro-environment is expected to be supplied from the seawater-dominated medium overlying tube openings and to undergo subsequent conductive heating through the tube walls. Its neutral pH is likely to be associated with moderately oxidative conditions. Such a model provides an explanation of the atypical thermal and chemical patterns that were previously reported for this medium from discrete samples and in situ measurements. Conversely, the medium surrounding the tubes is shown to be dominated by the fluid venting from the chimney wall. This hot fluid appears to be gradually cooled (120–30 °C) as it passes through the thickness of the worm colony, as a result of a thermal exchange mechanism induced by the tube assemblage. Its pH, however, remains very low (pH∼4), and reducing conditions can be expected in this medium. Such a thermal and chemical buffering mechanism is consistent with the mineralogical anomalies previously highlighted and provides a first explanation of the exceptional ability of these animals to colonize this hostile biotope. It furthermore suggests that A. pompejana , in providing various buffered micro-niches, would act as a primary player of microbial and related biogeochemical processes in this environment.

The shrimp Rimicaris exoculata dominates the megafauna of some Mid-Atlantic Ridge hydrothermal vent fields. This species harbours a rich bacterial epibiosis inside its gill chamber. At the 'Rainbow' vent site (36° 14.0' N), the epibionts are associated with iron oxide deposits. Investiga- tion of both bacteria and minerals by scanning electron microscopy (SEM) and X-ray microanalysis (EDX) revealed 3 distinct compartments in the gill chamber: (1) the lower pre-branchial chamber, housing bacteria but devoid of minerals; (2) the 'true' branchial chamber, containing the gills and devoid of both bacteria and minerals; and (3) the upper pre-branchial chamber, housing the main ectosymbiotic bacterial community and associated mineral deposits. Our chemical and temperature data indicated that abiotic iron oxidation appears to be kinetically inhibited in the environment of the shrimps, which would explain the lack of iron oxide deposits in the first 2 compartments. We propose that iron oxidation is microbially promoted in the third area. The discrepancy between the spatial dis- tribution of bacteria and minerals suggests that different bacterial metabolisms are involved in the first and third compartments. A possible explanation lies in the modification of physico-chemical con- ditions downstream of the gills that would reduce the oxygen content and favours the development of bacterial iron-oxidizers in this Fe II -rich environment. A potential role of such iron-oxidizing symbionts in the shrimp diet is suggested. This would be unusual for hydrothermal ecosystems, in which most previously described symbioses rely on sulphide or methane as an energy source.

Alvinella pompejana and Alvinella caudata live in organic tubes on active sulphide chimney walls at deep-sea hydrothermal vents. These polychaete annelids are exposed to extreme thermal and chemical gradients and to intense mineral precipitation. This work points out that mineral particles associated with Pompeii worm (A. pompejana and A. caudata) tubes constitute useful markers for evaluating the chemical characteristics of their micro-environment. The minerals associated with these worm tubes were analysed on samples recovered from an experimental alvinellid colony, at different locations in the vent fluid-seawater interface. Inhabited tubes from the most upper and lower parts of the colony were analysed by light and electron microscopies, X-ray microanalysis and X-ray diffraction. A change was observed from a Fe-Zn-S mineral assemblage to a Zn-S assemblage at the millimeter scale from the outer to the inner face of a tube. A similar gradient in proportions of minerals was observed at a decimeter scale from the lower to the upper part of the colony. The marcasitc/pyrite ratio of iron disulphides also displays a steep decrease along the few millimeters adjacent to the external tube surface. The occurrence of these gradients indicates that the microenvironment within the tube differs from that outside the tube, and suggests that the tube wall acts as an efficient barrier to the external environment.