Bathymodiolus Puteoserpentis Research Articles

Genome Study of α-, β-, and γ-Carbonic Anhydrases from the Thermophilic Microbiome of Marine Hydrothermal Vent Ecosystems.

Carbonic anhydrases (CAs) are metalloenzymes that can help organisms survive in hydrothermal vents by hydrating carbon dioxide (CO2). In this study, we focus on alpha (α), beta (β), and gamma (γ) CAs, which are present in the thermophilic microbiome of marine hydrothermal vents. The coding genes of these enzymes can be transferred between hydrothermal-vent organisms via horizontal gene transfer (HGT), which is an important tool in natural biodiversity. We performed big data mining and bioinformatics studies on α-, β-, and γ-CA coding genes from the thermophilic microbiome of marine hydrothermal vents. The results showed a reasonable association between thermostable α-, β-, and γ-CAs in the microbial population of the hydrothermal vents. This relationship could be due to HGT. We found evidence of HGT of α- and β-CAs between Cycloclasticus sp., a symbiont of Bathymodiolus heckerae, and an endosymbiont of Riftia pachyptila via Integrons. Conversely, HGT of β-CA genes from the endosymbiont Tevnia jerichonana to the endosymbiont Riftia pachyptila was detected. In addition, Hydrogenovibrio crunogenus SP-41 contains a β-CA gene on genomic islands (GIs). This gene can be transferred by HGT to Hydrogenovibrio sp. MA2-6, a methanotrophic endosymbiont of Bathymodiolus azoricus, and a methanotrophic endosymbiont of Bathymodiolus puteoserpentis. The endosymbiont of R. pachyptila has a γ-CA gene in the genome. If α- and β-CA coding genes have been derived from other microorganisms, such as endosymbionts of T. jerichonana and Cycloclasticus sp. as the endosymbiont of B. heckerae, through HGT, the theory of the necessity of thermostable CA enzymes for survival in the extreme ecosystem of hydrothermal vents is suggested and helps the conservation of microbiome natural diversity in hydrothermal vents. These harsh ecosystems, with their integral players, such as HGT and endosymbionts, significantly impact the enrichment of life on Earth and the carbon cycle in the ocean.

Mitochondrial genomic analyses provide new insights into the “missing” atp8 and adaptive evolution of Mytilidae

BackgroundMytilidae, also known as marine mussels, are widely distributed in the oceans worldwide. Members of Mytilidae show a tremendous range of ecological adaptions, from the species distributed in freshwater to those that inhabit in deep-sea. Mitochondria play an important role in energy metabolism, which might contribute to the adaptation of Mytilidae to different environments. In addition, some bivalve species are thought to lack the mitochondrial protein-coding gene ATP synthase F0 subunit 8. Increasing studies indicated that the absence of atp8 may be caused by annotation difficulties for atp8 gene is characterized by highly divergent, variable length.ResultsIn this study, the complete mitochondrial genomes of three marine mussels (Xenostrobus securis, Bathymodiolus puteoserpentis, Gigantidas vrijenhoeki) were newly assembled, with the lengths of 14,972 bp, 20,482, and 17,786 bp, respectively. We annotated atp8 in the sequences that we assembled and the sequences lacking atp8. The newly annotated atp8 sequences all have one predicted transmembrane domain, a similar hydropathy profile, as well as the C-terminal region with positively charged amino acids. Furthermore, we reconstructed the phylogenetic trees and performed positive selection analysis. The results showed that the deep-sea bathymodiolines experienced more relaxed evolutionary constraints. And signatures of positive selection were detected in nad4 of Limnoperna fortunei, which may contribute to the survival and/or thriving of this species in freshwater.ConclusionsOur analysis supported that atp8 may not be missing in the Mytilidae. And our results provided evidence that the mitochondrial genes may contribute to the adaptation of Mytilidae to different environments.

First data on benthic and fish communities from the Mid-Atlantic Ridge, 16°40′− 17°14′N

The first ecological survey in the tropical part of the northern Mid-Atlantic Ridge (MAR) was conducted between 17°14′N and 16°40′N in February–March 2015 during the 37th cruise of the RV Professor Logachev. Significant effort of the survey was aimed at studies of benthic non-vent soft sediment fauna at depths from 2270m to 3900m. Samples of this fauna at seven stations revealed ~780 specimens and at least 136 species. The most common species was the hermit crab Parapagurus cf. nudus. Also common were the ophiuroids Ophiotypa simplex and Ophiomusium cf. lymani. Most benthic organisms were <3cm in size. Collection of fish taken in the depth range from 0 to 3850m included 1128 specimens represented by 48 species from 16 families. The highest number of species was in the families Myctophidae (18), Gonostomatidae (9) and Sternoptychidae (5). In the course of the cruise, two new fields of massive sulphide deposits were discovered on the south-western slope of the seamount on the eastern flank of the rift valley, one at 17°08.7′N and second at 17°07.45′N. During video profiling in this area indications of modern hydrothermal activity were recorded. Extensive fields of shells of Bathymodiolus puteoserpentis and Thyasira sp. were discovered and samples of bivalves were taken using the TV-grab and geological square corer. Surprisingly high concentration of drowned pelagic algae Sargassum sp. (Phaeophyceae, Fucales) in different state of degradation was observed at TV transects at the seafloor both on soft and hard substrates. Thalli of Sargassum fluitans appeared in trawl catches. Massive aggregations of Sargassum also were abundant at the sea surface in the area from December 2014 to April 2015.

Estimating Symbiont Abundances and Gill Surface Areas in Specimens of the Hydrothermal Vent Mussel Bathymodiolus puteoserpentis Maintained in Pressure Vessels

The hydrothermal vent mussel Bathymodiolus puteoserpentis hosts gill-associated sulfur- and methane-oxidizing bacteria which sustain host nutrition and allow it to reach high densities at various sites along the northern Mid-Atlantic Ridge. Previous studies have demonstrated that in similar dual symbioses, relative abundances of each bacterial type could change following variations in symbiont substrate availabilities. In this study, pressurized recovery and incubations in pressure vessels were used to test whether B. puteoserpentis symbionts displayed similar behavior in the presence of symbiont substrates. The relative abundances of both types of symbionts were analyzed using fluorescence in situ hybridization (FISH) and group-specific gene copy numbers were assessed using qPCR. Specimens sampled using isobaric and non-isobaric recovery contained similar relative proportions (in surface coverage) of sulfur- and methane-oxidizing bacteria indicating that recovery type did not have impact on measured relative areas. Similarly, pressurized incubations with different substrates did lead to significant differences in the relative surface coverage of the two types of bacteria, although slight variations were measured with qPCR, suggesting changes in relative abundances of gene copy numbers but not in relative areas covered. Total gill surface areas and total bacterial numbers in specimens were estimated for the first time. Symbiont bearing-mussels display exchange surfaces about 20-fold higher than those found in similar-sized coastal mussels, and mean bacterial numbers of 2.5*1012 per specimen were estimated. This emphasizes that symbiotic mussels are a major reservoir of bacteria in vent ecosystems.

In situ measurements of hydrogen sulfide, oxygen, and temperature in diffuse fluids of an ultramafic-hosted hydrothermal vent field (Logatchev, 14°45′N, Mid-Atlantic Ridge): Implications for chemosymbiotic bathymodiolin mussels

The Logatchev hydrothermal vent field (14°45′N, Mid-Atlantic Ridge) is located in a ridge segment characterized by mantle-derived ultramafic outcrops. Compared to basalt-hosted vents, Logatchev high-temperature fluids are relatively low in sulfide indicating that the diffuse, low-temperature fluids of this vent field may not contain sufficient sulfide concentrations to support a chemosymbiotic invertebrate community. However, the high abundances of bathymodiolin mussels with bacterial symbionts related to free-living sulfur-oxidizing bacteria suggested that bioavailable sulfide is present at Logatchev. To clarify, if diffuse fluids above mussel beds of Bathymodiolus puteoserpentis provide the reductants and oxidants needed by their symbionts for aerobic sulfide oxidation, in situ microsensor measurements of dissolved hydrogen sulfide and oxygen were combined with simultaneous temperature measurements. High temporal fluctuations of all three parameters were measured above the mussel beds. H2S and O2 coexisted with mean concentrations between 9 and 31 μM (H2S) and 216 and 228 μM (O2). Temperature maxima (≤7.4°C) were generally concurrent with H2S maxima (≤156 μM) and O2 minima (≥142 μM). Long-term measurements for 250 days using temperature as a proxy for oxygen and sulfide concentrations indicated that the mussels were neither oxygen limited nor sulfide limited. Our in situ measurements at Logatchev indicate that sulfide may also be bioavailable in diffuse fluids from other ultramafic-hosted vents along slow and ultraslow spreading ridges.

Expression patterns of mRNAs for methanotrophy and thiotrophy in symbionts of the hydrothermal vent mussel Bathymodiolus puteoserpentis.

The hydrothermal vent mussel Bathymodiolus puteoserpentis (Mytilidae) from the Mid-Atlantic Ridge hosts symbiotic sulfur- and methane-oxidizing bacteria in its gills. In this study, we investigated the activity and distribution of these two symbionts in juvenile mussels from the Logatchev hydrothermal vent field (14°45'N Mid-Atlantic Ridge). Expression patterns of two key genes for chemosynthesis were examined: pmoA (encoding subunit A of the particulate methane monooxygenase) as an indicator for methanotrophy, and aprA (encoding the subunit A of the dissimilatory adenosine-5'-phosphosulfate reductase) as an indicator for thiotrophy. Using simultaneous fluorescence in situ hybridization (FISH) of rRNA and mRNA we observed highest mRNA FISH signals toward the ciliated epithelium where seawater enters the gills. The levels of mRNA expression differed between individual specimens collected in a single grab from the same sampling site, whereas no obvious differences in symbiont abundance or distribution were observed. We propose that the symbionts respond to the steep temporal and spatial gradients in methane, reduced sulfur compounds and oxygen by modifying gene transcription, whereas changes in symbiont abundance and distribution take much longer than regulation of mRNA expression and may only occur in response to long-term changes in vent fluid geochemistry.

Widespread occurrence of an intranuclear bacterial parasite in vent and seep bathymodiolin mussels

Many parasitic bacteria live in the cytoplasm of multicellular animals, but only a few are known to regularly invade their nuclei. In this study, we describe the novel bacterial parasite "Candidatus Endonucleobacter bathymodioli" that invades the nuclei of deep-sea bathymodiolin mussels from hydrothermal vents and cold seeps. Bathymodiolin mussels are well known for their symbiotic associations with sulfur- and methane-oxidizing bacteria. In contrast, the parasitic bacteria of vent and seep animals have received little attention despite their potential importance for deep-sea ecosystems. We first discovered the intranuclear parasite "Ca. E. bathymodioli" in Bathymodiolus puteoserpentis from the Logatchev hydrothermal vent field on the Mid-Atlantic Ridge. Using primers and probes specific to "Ca. E. bathymodioli" we found this intranuclear parasite in at least six other bathymodiolin species from vents and seeps around the world. Fluorescence in situ hybridization and transmission electron microscopy analyses of the developmental cycle of "Ca. E. bathymodioli" showed that the infection of a nucleus begins with a single rod-shaped bacterium which grows to an unseptated filament of up to 20 microm length and then divides repeatedly until the nucleus is filled with up to 80,000 bacteria. The greatly swollen nucleus destroys its host cell and the bacteria are released after the nuclear membrane bursts. Intriguingly, the only nuclei that were never infected by "Ca. E. bathymodioli" were those of the gill bacteriocytes. These cells contain the symbiotic sulfur- and methane-oxidizing bacteria, suggesting that the mussel symbionts can protect their host nuclei against the parasite. Phylogenetic analyses showed that the "Ca. E. bathymodioli" belongs to a monophyletic clade of Gammaproteobacteria associated with marine metazoans as diverse as sponges, corals, bivalves, gastropods, echinoderms, ascidians and fish. We hypothesize that many of the sequences from this clade originated from intranuclear bacteria, and that these are widespread in marine invertebrates.

Life‐history traits of the symbiotic scale‐worm Branchipolynoe seepensis and its relationships with host mussels of the genus Bathymodiolus from hydrothermal vents

AbstractAssociations between scale‐worms and giant mussels are common constituents of hydrothermal vent and cold seep ecosystems, but very little is known about their nature and ecology. Here, we analyze the ecological characteristics of the associations between Branchipolynoe seepensis, an obligate symbiotic polychaete, and their host mytilid mussels Bathymodiolus puteoserpentis and B. azoricus inhabiting hydrothermal vent fields on the Mid‐Atlantic Ridge. Infested mussels generally harbored a single symbiont (&lt;4% had two to six worms). Infestation rate varied from 7.2% to 76.5%, increasing with mussel size, and was significantly lower for B. puteoserpentis. Symbiont density at Lucky Strike ranged between 1071 and 1191 individuals m−2. Female symbiont size was always positively correlated with host size, while only males and juveniles from small mussels showed the same trend. This suggested a relatively long‐lasting host/symbiont association for females and short‐lasting association with successive reproductive migrations for adult males. The sex ratio of symbionts was always biased in favor of females. Males were smaller and more slender than females and had one mode in their size distributions, whereas females typically had three or more modes, suggestive of a longer life span in females. Between 59.1% and 72.2% of mussels had damaged soft tissues with substantially higher incidence of trauma in infested ones, suggesting that symbionts may cause trauma. The symbionts also induce tunnel‐like structures among the ctenidia, indicating fidelity to a particular location inside the host. Based on our data, together with the fact that infested mussels became relatively wider than non‐infested ones, this association is considered parasitic (likely kleptoparasitic). Our data, together with those from previous studies, allowed us to define the main life‐history traits of B. seepensis: (i) the relationship with their host is parasitic, (ii) the association begins at the smallest mytilid size classes, (iii) there is sexual dimorphism in body size, (iv) sex ratio deviates from 1:1 in favor of females, (v) fertilization occurs through temporal pairing and pseudocopulation, (vi) sperm are stored by females, (vii) eggs are large (likely lecithotrophic or with direct development), (viii) females have a longer life span than males, (ix) adult males may be semalparous, undertaking reproductive migrations followed by a short period of pairing and then death, and (x) females have a semi‐continuous iteroparous reproductive cycle.

Hydrothermal vent meiobenthos associated with mytilid mussel aggregations from the Mid-Atlantic Ridge and the East Pacific Rise

Deep-sea hydrothermal vents occur along the mid-ocean ridges and back-arc basins around the globe. There are very few community analyses of vent meiobenthos. The central objectives of this study were to identify and quantify for the first time the entire metazoan meiobenthic community associated with mussel aggregations of Bathymodiolus thermophilus Kenk and Wilson, 1985 from the EPR, 11°N and of Bathymodiolus puteoserpentis Cosel et al., 1994 from the Mid-Atlantic Ridge (MAR), 23°N. Using a quantitative sampling method, abundance, biomass, sex ratio, species richness, diversity, evenness, and trophic structure were studied based on three samples from each site. Meiobenthic abundance in each sample was unexpectedly low, but similar between sites. The community was composed of nematodes, copepods, ostracods, and mites, with a total of 24 species at EPR vents, and 15 species at MAR vents. While most copepod species were vent endemics within the family Dirivultidae, nematodes and harpacticoid copepods belonged to generalist genera, which occur at a variety of habitats and are not restricted to hydrothermal vents or the deep sea. The meiobenthos of hydrothermal-vent mussel beds constitutes a unique community unlike those of other sulfidic habitats, including the thiobios of shallow-water sediments and the meiobenthos of deep-sea, cold-seep sediments. The trophic structure was dominated by primary consumers, mainly deposit feeders, followed by parasites. Predatory meiofaunal species were absent.

A dual symbiosis shared by two mussel species, Bathymodiolus azoricus and Bathymodiolus puteoserpentis (Bivalvia: Mytilidae), from hydrothermal vents along the northern Mid‐Atlantic Ridge

Bathymodiolus azoricus and Bathymodiolus puteoserpentis are symbiont-bearing mussels that dominate hydrothermal vent sites along the northern Mid-Atlantic Ridge (MAR). Both species live in symbiosis with two physiologically and phylogenetically distinct Gammaproteobacteria: a sulfur-oxidizing chemoautotroph and a methane-oxidizer. A detailed analysis of mussels collected from four MAR vent sites (Menez Gwen, Lucky Strike, Rainbow, and Logatchev) using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization (FISH) showed that the two mussel species share highly similar to identical symbiont phylotypes. FISH observations of symbiont distribution and relative abundances showed no obvious differences between the two host species. In contrast, distinct differences in relative symbiont abundances were observed between mussels from different sites, indicating that vent chemistry may influence the relative abundance of thiotrophs and methanotrophs in these dual symbioses.

Development and cross‐amplification of nine polymorphic microsatellite markers in the deep‐sea hydrothermal vent polychaete Branchipolynoe seepensis

AbstractThe hydrothermal vent polychaete Branchipolynoe seepensis lives commensally inside the mantel cavity of the bivalves Bathymodiolus azoricus and Bathymodiolus puteoserpentis that form dense mussel beds around hydrothermal vents on the Mid‐Atlantic Ridge. In order to study its dispersal capabilities and the way individuals develop and reproduce, nine microsatellite polymorphic markers were developed. Polymorphism was tested from three well‐separated populations ranging from 14°N to 35°N along the ridge and showed significant levels of genetic differentiation. Cross‐amplification tests in other polynoid species revealed that most loci could also be useful to study closely related species from both Atlantic and Pacific sites.

Community structure in mussel beds at Logatchev hydrothermal vents and a comparison of macrofaunal species richness on slow‐ and fast‐spreading mid‐ocean ridges

AbstractSpecies lists for vent fields on the Mid‐Atlantic Ridge (MAR) from 14°N to 38°N suggest that there is a northern (&gt;27°N), shallow (&lt;2000 m) fauna and a southern (&lt;27°N), deeper (&gt;3000 m) endemic vent fauna, but little is known about how community structure varies along the ridge axis. In this study, quantitative samples of macrofaunal invertebrates associated with mussels (Bathymodiolus puteoserpentis) were collected at Logatchev (14°45′N), the southern‐most explored vent field on the MAR. Community structure (including species composition, species richness, diversity, and relative species abundances) in mussel beds at Logatchev was compared with that of Snake Pit (23°22′N) and Lucky Strike (37°17′N) mussel beds. The most striking feature of the Logatchev mussel‐bed macrofaunal invertebrate community was the tremendous abundance (up to 2390 individuals per liter of mussel‐volume sampled) and biomass of the ophiuroid, Ophioctenella acies. Logatchev and Snake Pit mussel beds share &gt;50% of their associated macrofaunal species; these two sites share only 20–25% of their macrofaunal species with Lucky Strike. Species–effort curves and univariate measures of diversity (H′, J′) do not support the claim that diversity of vent organisms on the MAR is highest at Logatchev, at least when one assesses this within a habitat type. Multivariate analysis readily differentiates the species‐abundance characteristics of Logatchev, Snake Pit, and Lucky Strike mussel‐bed macrofaunas. The relationship between sea‐floor spreading rate and diversity was explored through comparison of species richness in mussel‐bed habitats on slow‐spreading (MAR), fast‐spreading [northern East Pacific Rise (EPR)], and ultra‐fast‐spreading (southern EPR) mid‐ocean ridges. Species richness was greater in samples from the faster‐spreading ridge axes, where vents are more closely spaced but shorter lived, than on slow‐spreading centers, where vents are further apart but longer lived.

Environmental acquisition of thiotrophic endosymbionts by deep-sea mussels of the genus bathymodiolus.

Deep-sea Bathymodiolus mussels, depending on species and location, have the capacity to host sulfur-oxidizing (thiotrophic) and methanotrophic eubacteria in gill bacteriocytes, although little is known about the mussels' mode of symbiont acquisition. Previous studies of Bathymodiolus host and symbiont relationships have been based on collections of nonoverlapping species across wide-ranging geographic settings, creating an apparent model for vertical transmission. We present genetic and cytological evidence for the environmental acquisition of thiotrophic endosymbionts by vent mussels from the Mid-Atlantic Ridge. Open pit structures in cell membranes of the gill surface revealed likely sites for endocytosis of free-living bacteria. A population genetic analysis of the thiotrophic symbionts exploited a hybrid zone where two Bathymodiolus species intergrade. Northern Bathymodiolus azoricus and southern Bathymodiolus puteoserpentis possess species-specific DNA sequences that identify both their symbiont strains (internal transcribed spacer regions) and their mitochondria (ND4). However, the northern and southern symbiont-mitochondrial pairs were decoupled in the hybrid zone. Such decoupling of symbiont-mitochondrial pairs would not occur if the two elements were transmitted strictly vertically through the germ line. Taken together, these findings are consistent with an environmental source of thiotrophic symbionts in Bathymodiolus mussels, although an environmentally "leaky" system of vertical transmission could not be excluded.

Utilization of methane and carbon dioxide by symbiotrophic bacteria in gills of Mytilidae (Bathymodiolus) from the Rainbow and Logachev hydrothermal fields on the Mid-Atlantic Ridge

Bivalve mollusks Bathymodiolus asoricus and Bathymodiolus puteoserpentis collected from the Rainbow and Logachev hydrothermal fields during dives of Mir 1 and Mir 2 deep-sea manned submersibles were studied. Rates of methane oxidation and carbon dioxide assimilation in mussel gill tissue were determined by radiolabel analysis. During oxidation of 14Ch4, radiocarbon was detected in significant quantities not only in carbon dioxide but also in dissolved organic matter, most notably 14C-formate and 14C-acetate, occurring in a 2:1 ratio. Activities of hexulose-phosphate synthase, phosphoribulokinase, and ribulose 1,5-bisphosphate carboxylase were shown in the soluble fraction of gill tissue cells. At the same time, no activity of hydroxypyruvate reductase--the key enzyme of the serine pathway of C1-assimilation--was detected. The results of PCR amplification using genetic probes for membrane-bound methane monooxygenase (pmoA) and methanol dehydrogenase (mxaF) attest to the presence of the genes of these enzymes in the total DNA extracted from gill samples. However, no appropriate PCR responses were obtained with the mmoX primer system, which is a marker for soluble methane monooxygenase. All samples studied showed amplification with primers for the genera Methylobacter and Methylosphaera. At the same time, no genes specific to the genera Methylomonas, Methylococcus, Methylomicrobium, or Methylosinus and Methylocystis were detected. Electron microscopic examinations revealed the presence of two groups of endosymbiotic bacteria in the mussel gill tissue. The first group was represented by large cells possessing a complex system of cytoplasmic membranes, typical of methanotrophs of morphotype I. The other type of endosymbionts, having much smaller cells and lacking intracellular membrane structures, is likely to be constituted by sulfur bacteria.

Different energy sources for three symbiont-dependent bivalve molluscs at the Logatchev hydrothermal site (Mid-Atlantic Ridge)

The vent mussel Bathymodiolus puteoserpentis, a large vesicomyid clam and a smaller thyasirid were collected from an area of sediment subject to diffuse hydrothermal flow. The mussels live on the surface, the vesicomyids are partly buried and the thyasirids burrow in the sediment. The fine structure of the gills differs in the three bivalves. Bathymodiolus puteoserpentis hosts two types of bacterial symbiont, one methanotrophic, and another probably thiotrophic. The other two bivalves have single types of symbiont of different shapes. Stable isotope ratios of carbon and nitrogen indicate thiotrophy in the vesicomyid and thyasirid, but a predominance of methanotrophy in the mussel. This is the first time that such an assemblage has been found at a hydrothermal site on the Mid-Atlantic Ridge (MAR), with the different faunistic elements exploiting different energy resources.

Molecular and morphological identification of settlement‐stage vent mussel larvae, Bathymodiolus azoricus (Bivalvia: Mytilidae), preserved in situ at active vent fields on the Mid‐Atlantic Ridge

This paper describes the first successful attempt to trap and identify the larvae of a deep‐sea vent organism using a combination of sediment traps and molecular analysis. During the European Union‐funded MARVEL cruise in August and September 1997, sediment traps containing a high‐salt DNA preservative were deployed around active black smoker chimneys on the newly discovered Rainbow vent field in an attempt to collect larval stages of the dominant vent bivalve Bathymodiolus azoricus. A total of 2,055 shelled mussel larvae was collected within a 2‐week period, all of which were of identical size and shell morphology that indicated that they were at the settlement, prodissoconch II stage. Scanning electron microscopy of the shell hinge indicated that they belonged to the family Mytilidae, but it required molecular analysis to confirm their species identity. Polymerase chain reaction (PCR) amplification of the ITS2 region was performed on a subset of the larvae, and the resulting PCR products were cut with diagnostic restriction endonucleases to allow comparison with a DNA database based on adult specimens. The DNA restriction patterns typifying the Rainbow larvae were identical to those of adult B. azoricus from the Menez Gwen, Lucky Strike, and Rainbow vent sites and were significantly different from those that typified Bathymodiolus thermophilus (Pacific) and Bathymodiolus puteoserpentis (Snake Pit vent field on the Mid‐Atlantic Ridge), which clearly points to the Rainbow larvae having their origin on that part of the ridge close to the Azores. These findings point to the value of sediment traps as a way to study the temporal and spatial aspects of larval settlement in deep‐sea hydrothermal vent environments.

Genetic and morphometric characterization of mussels (Bivalvia: Mytilidae) from mid-atlantic hydrothermal vents.

Mussels were collected from deep-sea hydrothermal vents along the Mid-Atlantic Ridge. Specimens from the Snake Pit site were previously identified genetically and anatomically as Bathymodiolus puteoserpentis, but the relationships of mussels from other sites (Logatchev and Lucky Strike) were unclear. Molecular genetic and morphological techniques were used to assess differences among these mussel populations. The results indicate that the range for B. puteoserpentis extends from Snake Pit to Logatchev, and that an unnamed second species, B. n. sp., occurs at Lucky Strike. Analysis of mitochondrial NADH dehydrogenase subunit 4 (ND4) revealed 13% sequence divergence between the two species. Nei's genetic distance (D) based on 14 allozyme loci was 0.112. A multivariate morphometric analysis yielded a canonical discriminant function that correctly identified individuals from these sites to species 95% of the time.

Physiological and immunological evidence for two distinct C 1 -utilizing pathways in Bathymodiolus puteoserpentis (Bivalvia: Mytilidae), a dual endosymbiotic mussel from the Mid-Atlantic Ridge

The existence of endosymbiotic sulfur-oxidizing chemoautotrophic and methanotrophic bacteria associating with marine mytilid mussels has previously been inferred by 16S rDNA analysis in Bathymodiolus puteoserpentis Von Cosel et al. 1994, a hydrothermal vent mussel from a site on the Mid-Atlantic Ridge. In mussels collected in June 1993, we found evidence of enzymes diagnostic of two distinct C1 assimilation pathways in this symbiosis. Assays for the utilization of radiolabelled methane and for immunodetection of methanol dehydrogenase were positive, indicating that oxidation and incorporation of this substrate are occurring in this symbiosis. Sulfide or thiosulfate had no detectable stimulatory effect on CO2 incorporation, and assays for the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), an enzyme diagnostic for the Calvin–Benson cycle, were negative. RubisCO was detected in all samples examined by immunoblot analysis, indicating this enzyme is expressed in the B. puteoserpentis symbiosis. Stable isotope data showed that carbon isotope values were in agreement with previously reported values, and nitrogen isotope values were among the most depleted ever reported for bivalve symbioses. The carbon isotope values do not preclude the utilization of vent-derived methane. These data could be explained by the presence of two metabolically distinct bacterial symbionts or a Type X methanotrophic symbiont.

Arsenic speciation in shrimp and mussel from the Mid-Atlantic hydrothermal vents

Specimens of shrimp (Rimicaris exoculata) and mussel (Bathymodiolus puteoserpentis) were collected 3500 m below the ocean surface at the hydrothermal vents of the mid-Atlantic Ridge (TAG and Snake Pit sites, respectively). Arsenic, a potentially toxic element, is among the substances emitted by the hydrothermal vents. The hydrothermal vent shrimp, which are known to be a primary consumer of the primary producing chemolithoautotrophic bacteria, contained arsenic at 13 μg g−1 almost exclusively as arsenobetaine (AsB). Arsenic was present in the soft tissues of the mussel at 40 μg g−1 and the major part of the extractable arsenic species in the adductor muscle/mantle tissues and in the gill were present as dimethylarsinylriboside-derivatives (arsenosugars), while AsB was present at 16 and 3.6%, respectively, in these tissues. In spite of the absence of biosynthetically active algae, the pattern of arsenic species found in the shrimp and mussel species in the deep-sea is similar to that found in their counterparts from the ocean surface. It is concluded that the autotrophic bacteria of the hydrothermal vent ecosystem and the symbiotic bacteria harboured in the mussel species are responsible for the biosynthesis of the organoarsenicals detected in the two hydrothermal vent animal species. It is not known if the formation of the same organoarsenicals in the surface and deep sea ecosystems is primarily a detoxification process of inorganic arsenic, or whether these compounds play a physiological role.