Bathymodiolus Thermophilus Research Articles

Bioaccumulation des métaux chez le mytilidae hydrothermal Bathymodiolus sp. de la ride médio-atlantique

The hydrothermal environment is characterised, among other things, by a high metal concentration, related to the convective sea-water circulation inside the oceanic crust and its interaction with basaltic rocks. The biological communities associated with the hydrothermal system can survive in this toxic environment owing to their ability to regulate their intracellular metal levels by excretion or accumulation of metal ions in non-toxic forms. Various detoxification processes have been previously studied within an hydrothermal organism: immobilisation and precipitation of the metal in lysosomal systems, or binding to specific and soluble ligands such as metallothioneins. The later are heat stable and characterised by a high content of cysteine. The quantification of Fe, Zn, Cu, Mn, Cd, Ag, Ba and Sr in the tissues of the vent mussel Bathymodiolus sp. indicates a high accumulation of these metals in two target organs, the gill and the digestive gland. This accumulation is in relation with high concentrations of metallothioneins, but the subcellular distribution of metals indicates a higher contribution of the insoluble compartment for detoxification processes. These results are compared with the literature data about metal bioaccumlation in Bathymodiolus thermophilus collected at the Galapagos Rift.

Temporal and spatial patterns of biological community development at nascent deep-sea hydrothermal vents (9°50′N, East Pacific Rise)

The April 1991 discovery of newly formed hydrothermal vents in areas of recent volcanic eruption between 9°45′N and 9°52′N on the East Pacific Rise provided a unique opportunity to follow temporal changes in biological community structure from the “birth” of numerous deep-sea hydrothermal vents. In March l992, DSV Alvin was used to deploy an on-bottom observatory, the Biologic–Geologic Transect, to monitor faunal succession along a 1.37 km segment of the axial summit caldera between 9°49.61′N and 9°50.36′N (depth ∼2520 m). Photo- and videographic documentation of megafaunal colonization and chemical analyses of diffuse hydrothermal fluids associated with many of these developing communities within the Transect were performed in March 1992, December 1993, October 1994, and November 1995. Photographic and chemical time-series analyses revealed the following sequence of events in low-temperature venting areas. (1) Immediately following the 1991 eruption, hydrogen sulfide and iron concentrations in diffuse fluids were extremely high (>1 mmol kg -1) and microbially derived material blanketed active areas of venting in the form of thick microbial mats. (2) Mobile vent fauna (e.g. amphipods, copepods, octopods, and galatheid and brachyuran crabs) and non-vent fauna (e.g. nematocarcinid shrimp) proliferated in response to this increased biological production. (3) Within 1 yr of the eruption, areal coverage of microbial mat was reduced by ∼60% and individuals of the vestimentiferan tube worm Tevnia jerichonana settled gregariously in areas where diffuse flow was most intense. (4) Two years after the eruption, maximum levels of H 2S decreased by almost half (from 1.90 to 0.97 mmol kg -1) and dense thickets of the vestimentiferan Riftia pachyptila dominated vent openings previously inhabited by Tevnia jerichonana. (5) Three years after the eruption, maximum hydrogen sulfide levels declined further to 0.88 mmol kg -1 and mussels ( Bathymodiolus thermophilus) were observed on basaltic substrates. (6) Four years after the eruption, galatheid crabs and serpulid polychaetes increased in abundance and were observed close to active vent openings as maximum hydrogen levels decreased to 0.72 mmol kg -1. Also by this time mussels had colonized on to tubes of Riftia pachyptila. (7) Between 3 and 5 yr after the eruption, there was a 2- to 3-fold increase in the number of species in the faunal assemblages. In the absence of additional volcanic/tectonic disturbance, we predict that mytilid and vesicomyid bivalves will gradually replace vestimentiferans as the dominant megafauna 5–10 yr following the eruption. We also anticipate that the abundance of suspension feeders will decline during this period while the abundance of carnivores will increase. We hypothesize that the above series of events (1–7) represents a general sequence of biological successional changes that will occur at newly formed low-temperature deep-sea hydrothermal vents along the northern East Pacific Rise and contiguous ridge axes. Megafaunal colonization at deep-sea hydrothermal vents is considered to be the consequence of an intimate interaction of the life-history strategies of individual species, physical oceanographic processes, and the dynamic hydrothermal environment. Our observations indicate that the successful sequential colonization of dominant megafaunal vent species, from Tevnia jerichonana to Riftia pachyptila to Bathymodiolus thermophilus, also may be strongly influenced by temporal changes in geochemical conditions. Additional evidence demonstrating the close link between diffuse vent flux, fluid geochemistry, and faunal succession included the rapid death of several newly formed biological assemblages coincident with abrupt changes in the geochemical composition of the venting fluid and the local refocusing or cessation of vent flow. These correlations suggest that future models of faunal succession at hydrothermal vents along intermediate to fast-spreading mid-ocean ridges should consider not only the interplay of species-specific life-history strategies, community productivity, and physical oceanographic processes, but also the influence of changing geochemical conditions on the sequential colonization of megafaunal species.

Ultrastructural characteristics of spermatogenesis in three species of deep-sea hydrothermal vent mytilids

To enhance our understanding of the reproductive biology of deep-sea hydrothermal vent mytilids, the histology of the male gonad and the ultrastructure of its gametes were studied in Bathymodiolus thermophilus, B. puteoserpentis, and B. elongatus. Specimens of B. thermophilus were collected at the 13°N site on the East Pacific ridge, while B. puteoserpentis were sampled from the Snake Pit site of the mid-Atlantic ridge and B. elongatus were obtained from the North Fiji Basin. Gonad histology conformed to the typical bivalve profile; the differences in the proportions of acinal and interacinal tissue, as well as differences in acinal fullness in B. puteoserpentis, indicate that gametogenesis is discontinuous in these deep-sea mytilids. Evidence of protandric hermaphroditism was observed in B. elongatus, which exhibited acini containing both maturing and residual male gametes and immature oocytes. The ultrastructural characteristics of the male gametes conform to those described for littoral bivalve species, and the spermatozoon is of the primitive type. No species-specific differences in spermatozoon ultrastructure were discerned. No evidence of bacterial inclusions was found in either the gametes or the associated gonad cells in any of the species examined. The male gametes are thus probably not vectors for the endosymbiotic bacteria that characterize the nutritional biology of the adults in this genus.

Extensive gene flow among mytilid (Bathymodiolus thermophilus) populations from hydrothermal vents of the eastern Pacific

Prior studies of the hydrothermal vent mussel Bathymodiolus thermophilus (Bivalvia: Mytilidae), provided conflicting predictions about the dispersal ability and population structure of this highly specialized species. Analyses of morphological features associated with its larval shells revealed a feeding larval stage that might facilitate dispersal between ephemeral vent habitats. In contrast, an allozyme study revealed substantial genetic differentiation between samples taken from populations 2370 km apart on Galapagos Rift (Latitude 0°N) and the East Pacific Rise (13°N). To resolve the discrepancy between these studies, we examined allozyme and mitochondrial (mt) DNA variation in new samples from the same localities plus more recently discovered sites (9° and 11°N) along the East Pacific Rise. Although analysis of 26 enzyme-determining loci revealed relatively low levels of genetic variation within the five populations, no evidence existed for significant barriers to dispersal among populations. We estimated an average effective rate of gege flow (Nm) of ≃ 8 migrants per population per generation. Two common mtDNA variants predominated at relatively even frequencies in each population, and similarly provided no evidence for barriers to gene flow or isolation-by-distance across this species' known range. Larvae of this species appear to be capable of dispersing hundreds of kilometers along a continuous ridge system and across gaps separating non-contiguous spreading centers.

Phylogenetic diversity of the bacterial community from a microbial mat at an active, hydrothermal vent system, Loihi Seamount, Hawaii.

The phylogenetic diversity of small-subunit rRNA genes associated with the domain Bacteria was examined (by using previously defined operational taxonomic units [C. L. Moyer, F.C. Dobbs, and D. M. Karl, Appl. Environ. Microbiol. 60:871-879, 1994]; those for Pele's Vents Bacteria are hereafter abbreviated PVB OTUs) with samples from a microbial mat at an active, deep-sea hydrothermal vent system. A cluster of phylogenetically related PVB OTUs (OTUs 2, 3, 6, and 8) was closely affiliated with Thiovulum sp. contained within the epsilon subclass of the class Proteobacteria and accounted for 60.5% of the small-subunit rRNA bacterial clone library from Pele's Vents. A second, smaller cluster of PVB OTUs (OTUs 1 and 11) was closely affiliated with Xanthomonas sp., contained within the gamma subclass of the Proteobacteria and accounted for a total of 27.1% of the bacterial clone library. The remaining five PVB OTUs each accounted for 2.1% of the clones recovered and were affiliated with the following phylogenetic groups: PVB OTU 5 was a member of the Alteromonas group; PVB OTU 12 was a member of the Colwellia assemblage; PVB OTU 4 was loosely determined to be a member of the Thiothrix group, with the endosymbiotic bacteria from Bathymodiolus thermophilus and Calyptogena magnifica as the nearest relatives; PVB OTU 10B was a member of the Myxobacterium group; and PVB OTU 9A was a member of the Paraphyletic assemblage, with the Octopus Spring microbial mat type K clone as the closest known relative.(ABSTRACT TRUNCATED AT 250 WORDS)

Digestive enzymes in marine invertebrates from hydrothermal vents and other reducing environments

The present study demonstrates the potential hydrolytic activities in the symbiont-containing tissues of the vent invertebrates Riftia pachyptila, Bathymodiolus thermophilus (collected in 1991 at the East Pacific Rise) and the shallow-water bivalve Lucinoma aequizonata (collected in 1991 from the Santa Barbara Basin). Activities of phosphatases, esterases, β-glucuronidase and leucineaminopeptidase were comparable to those of digestive tract tissues of other marine invertebrates. A lack in most glycosidases as well as in trypsin and chymotrypsin was observed. Activities of lysozyme and chitobiase were rather high. In all vent invertebrates with symbionts and in L. aequizonata, the symbiont-containing tissues and the symbiont-free tissues had similar levels of enzymatic activities, indicating that polymeric nutrients could be hydrolysed after release from the symbionts and cellular uptake. The high activities of α-fucosidase in all vent invertebrates as well as in the shallow-water bivalve L. aequizonata could point to the existence of a yet undescribed substrate available to hydrolysation. The ectosymbionts-carrying polychaete Alvinella pompejana (collected in 1991 at the East Pacific Rise, EPR) shows high lysozyme activities in its gut, consistent with the proposed food source of bacteria. For the vent crab Bythogrea thermydron (also collected in 1991 at the EPR) hydrolytic activities were highest in the gut, dominated by esterase and peptidase activities which support their proposed carnivorous food source. A snail and a limpet collected from R. pachyptila tubes showed high levels of chitobiase suggesting a food source of grazed bacteria or ingested R. pachyptila tube.

Lipid characteristics of hydrothermal vent organisms from 9°N, East Pacific Rise

Abstract Lipid compositions are reported for three distinctive deep-sea hydrothermal vent invertebrate species collected around 9°N East Pacific Rise: Riftia pachyptila Jones, a vestimentiferan tubeworm; Bathymodiolus thermophilus Kenk and Wilson, a mussel; and Halice hesmonectes Martin et al. , an amphipod crustacean. The lipid compositions of all these organisms were dominated by components characteristic of diets based on bacteria, with only very minor contributions from carbon derived from the oceanic photic zone. In all the organisms studied, large abundances of n-7 fatty acids, polyunsaturated fatty acids with unsaturations separated by more than one methylene bond, and sterol distributions dominated by cholesterol were observed. Branched fatty acids were generally of low abundance, whereas polyunsaturated fatty acids separated by single methylene groups were either absent, as in the case of R. pachyptila , or in very low abundance, as in the case of B. thermophilus and H. hesmoncetes . Monounsaturated fatty acids were the most abundant component of R. pachyptila lipids, whereas non-methylene interrupted fatty acids were particularly abundant in the lipids of B. thermophilus (up to 450f total fatty acids). The lipids of H. hesmonectes were dominated by storage lipids (e.g. wax esters). Stable carbon isotope analyses of individual sterols from the organisms examined allow specific sources to be proposed for these biochemicals. The δ 13 C values of sterols from R. pachyptila were consistent with de novo biosynthesis, whereas that of cholesterol from B. thermophilus corresponded to that from marine phytoplankton. The δ 13 C values of sterols from H. hesmonectes fell into two different groups and suggest that at least two distinct sources of sterols are available to these crustacea in the vent ecosystem, one of which derives from phytoplankton. Overall, the combination of the interpretation of lipid structure and distribution with compound specific isotope analyses can lead to valuable insights into trophic relationships within the deep-sea hydrothermal ecosystem.

The gill symbiont of the hydrothermal vent mussel Bathymodiolus thermophilus is a psychrophilic, chemoautotrophic, sulfur bacterium

Certain hydrothermal vent invertebrates, e.g. Riftia pachyptila and Calyptogena magnifica, are clearly established as harboring dense populations of chemoautotrophic sulfur bacteria in specialized tissues. By contrast, the physiological characteristics of the abundant intracellular gill symbiont of the vent mussel Bathymodiolus thermophilus have been questioned. The low activities of enzymes diagnostic for CO2 fixation (Calvin cycle) and for sulfur-driven energy generation, as measured by other investigators, have been attributed to bacterial contamination of the gill surface. Based on research at the Galapagos Rift hydrothermal vents in 1988 and subsequent laboratory experiments, the current study confirms that the B. thermophilus symbiont is a psychrophile for which thiosulfate and sulfide stimulate CO2 fixation. It strongly indicates that the symbiont is a chemoautotroph by establishing the following: (1) Sulfide and thiosulfate can stimulate CO2 fixation by partially purified symbionts by up to 43-fold and 120-fold, respectively; (2) the ribulose-1,5-bisphosphate carboxylase/oxygenase activity of the symbiont is sufficient to account for its sulfide- or thiosulfate-stimulated CO2 incorporation; (3) the symbiont's molar growth yield on thiosulfate, as judged by CO2 incorporation, is indistinguishable from that of free-living chemoautotrophs. Due to the high protein-degrading activity of B. thermophilus gill lysate, it is also suggested that host lysis of symbionts plays a more important role in the nutrition of the vent mussel than in R. pachyptila or C. magnifica, for which no comparable protein-degrading activity was found.

Genetic differentiation across the Western Pacific populations of the hydrothermal vent bivalve Bathymodiolus spp. and the Eastern Pacific (13°N) population of Bathymodiolus thermophilus

Deep-sea mussels, closely related to the Bathymodiolus genus and individuals of Bathymodiolus thermophilus, were collected on hydrothermal vents of the Lau and North-Fiji back-arc basins (Western Pacific) and at 13°N on the East Pacific Rise, respectively. Separate tissue homogenates were submitted to starch gel electrophoresis in order to study the genetic structure of the populations at 11 enzyme loci. Significant departures from the Hardy-Weinberg equilibrium were found at Aat-1, Gpi and Pgm loci without any location specificity. All the observed departures corresponded to heterozygote deficiencies that are common in mollusc bivalves. The genetic variability of these mussels was low in all samples, especially for the North-Fiji population. The genetic distances measured between populations demonstrated that significant genetic differentiation occurs between the Western Pacific mussel populations and the Eastern Pacific B. thermophilus, whereas gene flow appeared to be maintained between the two back-arc basins. The presence of both shared heterozygotes and genetic distances found between the western and the eastern populations do not allow us to separate these allopatric forms as distinct sibling species.

Calcium carbonate dissolution rates in hydrothermal vent fields of the Guaymas Basin

Analysis of bivalve shell fragments that were embedded in epoxy blocks, mounted on titanium stakes, and deployed by DSRV Alvin at 5 sites in the Southern Trough of the Guaymas Basin (27”00’N, lll”24.55’W; depth 2012 m) indicates significant variation of calcium carbonate dissolution in in situ exposures of more than 900 days. Arrays of shell fragments of six bivalve species (i.e., Bathymodiolus thermophilus, Calyptogena magnijka, Calyptogena sp., Corbicula Jluminea, Crassostrea virginica and Mytilus edulis) were positioned -17 cm, -7 cm and -2.5 cm below the sediment-water interface and 2.5 cm, 7 cm and 17 cm above the sediment-water interface in hydrothermal vent fields of the basin. Maximum dissolution rates for both calcite (mean = 86 pm/yr) and aragonite (mean = 312 urnr clam area) to 227 p,m/yr (site 3; clam area), and those of calcite dissolution ranged from 13 km/yr (site 3; clam area) to 94 um/yr (site 4; bacterial mat area). Dissolution rates were consistently highest in the bacterial mat area (site 4; mean = 94 um/yr for calcite and 223 p,m/yr for aragonite). Rates of calcium carbonate dissolution reported here for hydrothermal vent fields of the Guaymas Basin compare favorably with those of Rose Garden (Galapagos Rift) and 21N (East Pacific Rise) hydrothermal vent sites. These results have important implications for assessing biological rate processes in deep-sea hydrothermal vent environments.

Biogeochemistry of hydrothermal vent mussel communities: the deep-sea analogue to the intertidal zone

Continuous measurements of sulfide, silicate and temperature were made in situ from the submersible Alvin in the Rose Garden and New Vent hydrothermal fields of the Galapagos Spreading Center. Continuous measurements of temperature also were made for an 18 day period in the Rose Garden field. The results demonstrate several adaptations that appear to have contributed to the success of the vent mussel Bathymodiolus thermophilus in the Rose Garden. Dense clusters of B. thermophilus can disperse the hydrothermal fluids laterally for distances of several meters. This results in a large increase in the area of the redox transition zone, where both dissolved oxygen and hydrogen sulfide are available. As a result, the animal communities can grow to occupy areas that would not otherwise provide adequate reduced substrates. Measurements of the temperature demonstrate a distinct tidal periodicity. This periodicity will result in a large range of environmental conditions within the vent community. The mussel can tolerate these wide ranges in condition because of its ability to accept long periods of anoxia and to filter feed.

Enzymologie du tractus digestif de la modiole hydrothermale Bathymodiolus thermophilus (Mollusque Bivalve)

Although the structure and ultrastructure of the digestive tract of the hydrothermal vent mytilid Bathymodiolus thermophilus conform to those of other bivalves, enzymological data are lacking. To address this question, digestive enzyme assays and histoenzymological tests were performed on different regions of the digestive tract: labial palps, oesophagus, stomach, digestive gland, intestine, and rectum. Carbohydrases, mainly present in the digestive gland and the stomach, were the most active of the 33 enzymes studied. These enzymes would allow substantial digestion of particles from the immediate environment as well as those descending from the photic zone. Acid phosphatases present in all the compartments of the digestive tract indicate intracellular digestion, whereas alkaline phosphatase activity, mainly in the digestive gland and the stomach, demonstrates absorption phenomena. We conclude that, in addition to the nutrient supply furnished by chemoautotrophic bacteria in the gill bacteriocytes, the digestive tract is functional and provides at least some of the nutritive requirements of this species.

Microscopical observations on the excretory organs of the hydrothermal mussel Bathymodiolus thermophilus (Mollusca: Bivalvia)

The pericardial gland of the deep-sea hydrothermal mussel Bathymodiolus thermophilus (Mollusca: Bivalvia: Mytilidae) differs from other mytilids by (1) a narrow lumen and (2) conspicuous polyhedral inclusion bodies in large cytoplasmic vacuoles of the epithelial cells. These inclusions do not contain metals as revealed by microanalysis but are possibly made of proteins. The cells of this gland exhibit a fenestrated basal border which indicates a site of haemolymph ultrafiltration. The kidney is histologically comparable to that of the coastal mussel Mytilus edulis.

Experimental evidence for filter-feeding by the hydrothermal vent mussel, Bathymodiolus thermophilus

We provide experimental evidence, using a high-pressure recirculating aquarium and radiolabeled bacteria, that the hydrothermal vent mussel. Bathymodiolus thermophilus, can clear and assimilate particulate organic matter. Our results support previous evidence that this mussel can filter-feed on particulate organic matter to supplement nutrients provided by endosymbiotic chemoautotrophic bacteria.

Temperature Effects on Mitochondria from Hydrothermal Vent Invertebrates: Evidence for Adaptation to Elevated and Variable Habitat Temperatures

The effects of elevated temperatures on mitochondrial respiration and the activities of selected mitochondrial enzymes were measured for several invertebrates endemic to the deep-sea hydrothermal vents to determine whether the differences in distribution patterns noted among these species are related to differences in temperature adaptation. Parallel studies were done with related (congeneric or confamilial) species from shallow marine habitats. Mitochondrial respiration of hydrothermal vent species living in zones with high water flux (high, variable temperatures), including the large tube worm Riftia pachyptila, the Pompeii worm Alvinella pompejana, and the brachyuran crab Bythograea thermydron, was more resistant to high temperatures than was mitochondrial respiration of hydrothermal vent species or shallow-living species in cooler waters, for example, the vent clam Calyptogena magnifica and mussel Bathymodiolus thermophilus. The temperatures at which Arrhenius plots of respiration rate exhibited sharp breaks were correlated with the apparent maximal habitat temperature of the species. The apparent Arrhenius activation energies of respiration also correlated with maximal habitat temperature, although not as closely as the Arrhenius break temperatures. Temperature inactivation of mitochondrial enzymes generally adhered to the pattern noted in the interspecific comparisons of mitochondrial respiration. These findings suggest that diferences in thermal adaptation exist among endemic vent species and that these differences are correlated with the microhabitat conditions the diferent species encounter at the vent sites.

Elemental concentrations of hydrothermal vent organisms from the Gal�pagos Rift

Concentrations of 40 elements were determined in tissues and whole bodies of mussels (Bathymodiolus thermophilus) and/or whole bodies of limpets (Neomphalus fretterae) collected from the submarine thermal springs on the Galapagos Rift in 1977. Concentrations of Ag, As, Fe, Mn, Mo, Se, Sr, and Zn were comparable or elevated relative to those of intertidal mollusks, which have been utilized as indicators of anthropogenic trace-metal contamination. Both species contained concentrations of some elements (e.g. Ag, As, Fe, Mn, Mo, Se, Zn) which corresponded with the elevated levels of those elements discharged from hydrothermal vents. Many elements were most enriched in the gills and digestive glands of the mussels. Iron:manganese ratios of those tissues were similar to ratios of pelagic and transitional sediments, indicating that metal-rich suspended particulates had been absorbed onto mucous membranes. Comparisons with other benthopelagic organisms were precluded, since the present data represent essentially the only trace-element analyses of deep-sea macroinvertebrates.

Variation in the hydrothermal vent clam, Calyptogen magnifica, at the Rose Garden vent on the Galapagos spreading center

Abstract Calyptogena magnifica occupy a relatively restricted habitat at the Rose Garden hydrothermal vent site on the Galapagos Rift. These clams are found in areas with very low flow of vent water and gain exposure to hydrogen sulfide by inserting their well-vascularized foot into cracks that contain this flow. Vent water is undetectable around the siphons of many of the individuals, and they therefore probably take up sulfide through their foot, and oxygen and inorganic carbon through their gills. Age estimates indicate that the bulk of the recruitment of C. magnifica occured between 1971 and 1976. Isotopic evidence indicates that symbionts are the main source of both nutritional carbon and nitrogen for the clams, and that the symbionts assimilate both of these substrates from inorganic sources. Carbohydrate and protein in the clam soft tissues, as well as the elemental sulfur content of their gills, decrease with increasing clam size. There is only slight variation in most of the parameters measured, and none of the parameters show nearly the variation seen in the other hydrothermal vent bivalve, Bathymodiolus thermophilus . However, several parameters, such as δ 13 C, condition index, and some bacterial enzyme activities, vary significantly with habitat.

Microhabitat variation in the hydrothermal vent mussel, Bathymodiolus thermophilus, at the Rose Garden vent on the Galapagos Rift

Clumps of Bathymodiolus thermophilus were collected from three discrete areas at the ‘Rose Garden’ site on the Galapagos Rift using the deep submersible Alvin. Two mussel collections were made from the central Riftia mass, an area associated with very active venting, and three other collections were of two different peripheral mussel clumps. Before collection the clumps were extensively photographed and the water at two of the ‘microhabitats’ was analysed in situ for oxygen silica, sulfide and temperature. Sulfide levels of up to 300 μM were recorded at the central collection site, while the highest sulfide level recorded at the peripheral site assayed was 35 μM. Levels of RuBP carboxylase activity in the gills were significantly higher in mussels collected from the central ‘Riftia site’ than in either peripheral site. ATP sulfurylase was significantly higher in the gills of mussels from the central clump than in one of the peripheral clump collections. The chemical composition (% water, protein, carbohydrate, lipid and ash) and stable carbon isotope ratios (δ13C) of the mussels showed the same trends, with highest lipid and carbohydrate and the lowest water content and δ13C in the central site mussels. Similarly, the mussels from the central site were significantly depleted in stable nitrogen (δ15N) when compared with the peripheral site mussels. Variations between sites and tissues of the same animal may be indicative of differential utilization of inorganic or dissolved molecular nitrogen sources. The condition index (CI = soft tissue dry mass / internal shell volume) was similar for all animals collected at Rose Garden. The presence of a commensal polychaete, Branchipolynoe symmytilida, in the mantle cavity of the mussels was also correlated with the collection site, with the highest incidence of occurrence in the central clump.

Sulfur-oxidizing bacterial endosymbionts: analysis of phylogeny and specificity by 16S rRNA sequences.

The 16S rRNAs from the bacterial endosymbionts of six marine invertebrates from diverse environments were isolated and partially sequenced. These symbionts included the trophosome symbiont of Riftia pachyptila, the gill symbionts of Calyptogena magnifica and Bathymodiolus thermophilus (from deep-sea hydrothermal vents), and the gill symbionts of Lucinoma annulata, Lucinoma aequizonata, and Codakia orbicularis (from relatively shallow coastal environments). Only one type of bacterial 16S rRNA was detected in each symbiosis. Using nucleotide sequence comparisons, we showed that each of the bacterial symbionts is distinct from the others and that all fall within a limited domain of the gamma subdivision of the purple bacteria (one of the major eubacterial divisions previously defined by 16S rRNA analysis [C. R. Woese, Microbiol. Rev. 51: 221-271, 1987]). Two host specimens were analyzed in five of the symbioses; in each case, identical bacterial rRNA sequences were obtained from conspecific host specimens. These data indicate that the symbioses examined are species specific and that the symbiont species are unique to and invariant within their respective host species.

The importance of methane and thiosulfate in the metabolism of the bacterial symbionts of two deep-sea mussels

Undescribed hydrocarbon-seep mussels were collected from the Louisiana Slope, Gulf of Mexico, during March 1986, and the ultrastructure of their gills was examined and compared to Bathymodiolus thermophilus, a mussel collected from the deep-sea hydrothermal vents on the Galapagos Rift in March 1985. These closely related mytilids both contain abundant symbiotic bacteria in their gills. However, the bacteria from the two species are distinctly different in both morphology and biochemistry, and are housed differently within the gills of the two mussels. The symbionts from the seep mussel are larger than the symbionts from B. thermophilus and, unlike the latter, contain stacked intracytoplasmic membranes. In the seep mussel three or fewer symbionts appear to be contained in each host-cell vacuole, while in B. thermophilus there are often more than twenty bacteria visible in a single section through a vacuole. The methanotrophic nature of the seep-mussel symbionts was confirmed in 14C-methane uptake experiments by the appearance of label in both CO2 and acid-stable, non-volatile, organic compounds after a 3 h incubation of isolated gill tissue. Furthermore, methane consumption was correlated with methanol dehydrogenase activity in isolated gill tissue. Activity of ribulose-1,5-biphosphate (RuBP) carboxylase and 14CO2 assimilation studies indicate the presence of either a second type of symbiont or contaminating bacteria on the gills of freshly captured seep mussels. A reevaluation of the nutrition of the symbionts in B. thermophilus indicates that while the major symbiont is not a methanotroph, its status as a sulfur-oxidizing chemoautotroph, as has been suggested previously, is far from proven.