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 Galápagos 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.     

Role of filter-feeding in the nutritional biology of a deep-sea mussel with methanotrophic symbionts

The ability of an undescribed deep-sea hydrocarbon-seep mussel which contains endosymbiotic methanotrophic bacteria to clear, ingest, and assimilate radiolabeled bacteria (Vibrio pelagicus andEscherichia coli) and algae (Dunaliella tertiolecta) was compared with that of the bay musselMytilus edulis. The seep mussel, collected in August 1987 from the Louisana Slope in the Gulf of Mexico, was slower to clear bacteria and algae thanM. edulis. The ingestion and assimilation of filtered bacteria and algae was established from the presence of radiolabel in mussel tissues and feces. The seep mussel was somewhat less efficient in assimilating radiolabeled components from bacteria and algae thanM. edulis. The dietary carbon maintenance-requirement of the seep mussel could potentially be met at environmental concentrations of greater than 10⁶ bacteria ml^–1. At lower concentrations of particulate organic matter, filter-feeding could be an important source of nitrogen and essential nutrients not supplied by the endosymbionts.     

Physiological ecology of a mussel with methanotrophic endosymbionts at three hydrocarbon seep sites in the Gulf of Mexico

In situ growth rates were determined, using two, 1-yr mark/recapture experiments, conducted between September 1991 and July 1993, for an undescribed mytilid, Seep Mytilid Ia, at three hydrocarbon seep sites in the Gulf of Mexico. The sites are located at depths of 540 to 730m, approximately 27°45N; 91°30W, and are separated by distances of 6 to 18 miles. These seep mytilids harbor methanotrophic endosymbionts and use methane as both a carbon and energy source. The mussel habitats were chemically characterized by analysis of water samples taken from precisely located microenvironments over, among and below the mussels, using small-volume, interstitial water samplers and the Johnson Sea Link submersible. Substantial differences were found in habital conditions, growth rates, and population structure for the mussels at the three sites examined. The growth rate of these seep mytilids reflects the methane concentration in their immediate habitat. Mussels at sites with abundant methane had growth rates that were comparable to shallow water mytilids at similar temperatures (5 to 8°C) with increases in shell length up to 17 mm yr^–1 documented for smaller mussels (<40 mm shell length). In conjunction with measurements of growth rates, three condition indices (glycogen content, tissue water content, and the ratio of ash-free dry weight to shell volume) were used to determine the relationship between the condition of the mussels, their growth rates, and their habitat chemistry. The three condition indices were correlated with growth rate and were often significantly different between mussels in different samples.     

Physiological and immunological evidence for two distinct C1-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. Received: 3 October 1997 / Accepted: 23 July 1998     

Community structure in Florida Escarpment seep and Snake Pit (Mid-Atlantic Ridge) vent mussel beds

Comparisons between invertebrate communities hosted by similar foundation species under different environmental conditions permit identification of patterns of species distributions that might be characteristic of the different ecosystems. Similarities and differences in community structure between two major types of chemosynthetic ecosystems were assessed by analyzing samples of invertebrates associated with Bathymodiolus heckerae Gustafson et al. mussel beds at the Florida Escarpment seep (Gulf of Mexico, 26°01.8N; 84°54.9W; October 2000) and B. puteoserpentis von Cosel et al. mussel beds at the Snake Pit vent (Mid-Atlantic Ridge, 23°22.1N; 44°56.9W; July 2001). Macrofaunal species richness was nearly twice as high in the seep mussel bed compared to the vent mussel bed, and only a single morphospecies, the ophiuroid Ophioctenella acies Tyler et al., was shared between the sites. Similarities between the two faunas at higher taxonomic levels (genus and family) were evident for only a small percentage of the total number of taxa, suggesting that evolutionary histories of many of these seep and vent macrofaunal taxa are not shared. The taxonomic distinctiveness of the seep and vent mussel-bed macrofaunal communities supports the hypothesis that environmental and oceanographic barriers prevent most taxa from occupying both types of habitats. Macrofaunal community heterogeneity among samples was similar in seep and vent mussel beds, indicating that spatial scales of processes regulating community variability may be similar in the two types of ecosystems. Suspension feeders were not represented in the macrofauna of seep or vent mussel beds. Primary consumers (deposit feeders and grazers) contributed more to the total abundance of macrofauna of seep mussel beds than vent mussel beds; secondary consumers (polychaetes and shrimp) were more abundant in the vent mussel beds.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00227-004-1304-z.Communicated by J.P. Grassle, New Brunswick     

Molecular phylogeny of vestimentiferans collected around Japan, revealed by the nucleotide sequences of mitochondrial DNA

The phylogenetic relationships among local populations of undescribed vestimentiferan species that belong to the genera Lamellibrachia and Escarpia and had been collected at six sites around Japan were analyzed on the basis of the partial (1023 bp) nucleotide sequences of the mitochondrial gene for cytochrome oxidase I (COI), using a pogonophoran and a polychaete as outgroups. The identical amino acid sequence was deduced from the nucleotide sequence obtained from each of the vestimentiferans analyzed. The strong similarity among deduced amino acid sequences of COI suggested a close relationship between vestimentiferans and pogonophorans. On the basis of the phylogenetic relationships, the analyzed vestimentiferan populations were classified tentatively as five species. The genetic differentiation of vestimentiferans was suggested to occur bathymetrically as well as being a consequence of horizontal segregation. Two of these tentatively identified species inhabit both a hydrothermal area and a cold seep area, as is the case for some species of bivalves that belong to the genera Bathymodiolus and Calyptogena. Received: 28 August 1996 / Accepted: 2 October 1996     

Heterogeneous energetic pathways and carbon sources on deep eastern Mediterranean cold seep communities

Cold seep communities in the Mediterranean Sea have only been discovered two decades ago, and their trophic ecology has been the subject of very few studies. We investigated the benthic food web of two deep chemosynthesis-based ecosystems on the Napoli and Amsterdam mud volcanoes (MVs) in the eastern Mediterranean Sea (~2,000 m depth). Seeping methane has been detected at the surface of both MVs during pioneering cruises and has been hypothesised to be assimilated by benthic fauna as observed in other oceans’ margins. Given the extreme oligotrophic character of the eastern Mediterranean Sea, we a priori expected that chemosynthetic food sources, especially methane-derived carbon (MDC), played a major trophic role in these deep seep communities relative to what has been observed in other seep systems worldwide. We aimed at unravelling the trophic relationships on Napoli and Amsterdam MVs through the analysis of carbon, nitrogen and sulphur isotopes both in the dominant benthic invertebrates including the small endofauna (300 μm < size < 1 cm) and in the sedimented organic matter. In particular, we assessed the fraction of MDC in the tissue of several heterotrophic and symbiotic species. Low mean δ³⁴S and δ¹³C values (0.4 ± 4.8‰ and −31.6 ± 5.7‰, respectively) obtained for mega- and macrofauna suggested that the investigated benthic food webs are virtually exclusively fuelled by carbon of chemosynthetic origin. A few grazer invertebrates (δ³⁴S up to 11‰) depart from this trend and could complement their diet with sedimented and decayed phytoplanktonic organic matter. Faunal δ¹³C values indicated that the oxidation of sulphur is likely the predominant energetic pathway for biosynthesis on both MVs. Nevertheless, mytilid bivalves and small capitellid, ampharetid and spionid polychaetes were ¹³C-depleted (δ¹³C < −37‰) in a way indicating they assimilated a significant portion of MDC. For these later heterotrophic species, MDC ranged between 21 and 31% (lower estimates) and 97 and 100% (upper estimates). However, our results highlighted that the origin of assimilated carbon may be complex for some symbiotic species. The vestimentiferan tubeworm Lamellibrachia sp., which exclusively depends on its sulphur-oxidising endosymbionts, showed a ~20‰ inter-individual δ¹³C variability on a very small spatial scale (<1 m) at the summit of Napoli MV. This mostly reflects the variable isotopic composition of pore-water-dissolved inorganic carbon (DIC) and evidenced that tubeworms (and subsequently their endosymbionts) uptake DIC derived from multiple methane oxidation processes in varying proportions. The lower and upper MDC estimates for the vestimentum of Napoli’s individuals were 11–38 and 21–73%, respectively. Finally, data on trophic ecology of Napoli and Amsterdam MVs clearly corroborate previous geophysical results evidencing the spatial heterogeneity of Mediterranean MV environmental conditions.     

A new bathymodioline mussel symbiosis at the Juan de Fuca hydrothermal vents

Until recently, the only major hydrothermal vent biogeographic province not known to include bathymodioline mussels was the spreading centers of the northeast Pacific, but deep-sea dives using DSV Alvin on the Endeavor segment of the Juan de Fuca Ridge (47°56N 129°06W; ∼2,200 m depth) in August 1999 yielded the only recorded bathymodioline mytilids from these northeastern Pacific vents. One specimen in good condition was evaluated for its relatedness to other deep-sea bathymodioline mussels and for the occurrence of chemoautotrophic and/or methanotrophic symbionts in the gills. Phylogenetic analyses of the host cytochrome oxidase I gene show this mussel shares evolutionary alliances with hydrothermal vent and cold seep mussels from the genus Bathymodiolus, and is distinct from other known species of deep-sea bathymodiolines, suggesting this mussel is a newly discovered species. Ultrastructural analyses of gill tissue revealed the presence of coccoid bacteria that lacked the intracellular membranes observed in methanotrophic symbionts. The bacteria may be extracellular but poor condition of the fixed tissue complicated conclusions regarding symbiont location. A single gamma-proteobacterial 16S rRNA sequence was amplified from gill tissue and directly sequenced from gill tissue. This sequence clusters with other mussel chemoautotrophic symbiont 16S rRNA sequences, which suggests a chemoautotrophic, rather than methanotrophic, symbiosis in this mussel. Stable carbon (δ¹³C = −26.6%) and nitrogen (δ¹⁵N = +5.19%) isotope ratios were also consistent with those reported for other chemoautotroph-mussel symbioses. Despite the apparent rarity of these mussels at the Juan de Fuca vent sites, this finding extends the range of the bathymodioline mussels to all hydrothermal vent biogeographic provinces studied to date.     

Stable isotopic compositions of hydrothermal vent organisms

Stable isotopic analyses were used to study trophic relationships in two communities of deep-sea hydrothermal vent organism in the Pacific Ocean. The community at Hanging Gardens on the East Pacific Rise (21°N), sampled in 1985, is dominated by two species of vestimentiferan tubeworms; communities at Alice Springs and Snail Pits on the Marianas Back Arc Spreading Center (western Pacific), sampled in 1987, are dominated by gastropod mollusks, barnacles, and anemones. In both locations, carbon and nitrogen isotopic values of vent invertebrates are significantly different from those of non-vent invertebrates collected at 11°N on the East Pacific Rise and elsewhere in the deep-sea. These distinct isotopic compositions reflect local sources of organic carbon and nitrogen used by vent consumers. Many vent invertebrates lacking chemoautotrophic endosymbionts have ¹³C-enriched values of-11 to-16%. compared to values of-17 to-22%. normally observed in deep-sea fauna. This suggests that a ¹³C-enriched food source is trophically important in both vent communities. Free-living bacteria colonizing surfaces and suspended in the water column may constitute this food resource. Nitrogen isotopic analyses show that the food web of the East Pacific Rise community has more trophic levels than the Marianas vent community.     

The infaunal benthos of a natural oil seep in the Santa Barbara channel

We are studying a diverse infaunal benthic community that exists in the fine sand sediments of a shallow (16 m) natural oil seep near Santa Barbara, California, USA. The study area and sampling methods are described in this introductory paper. Data presented indicate the adequacy of sampling in revealing horizontal patchiness and vertical faunal distributions. The infauna of the seep and of a nearby comparison area, without seepage but of similar depth and sediment type, are compared. The data indicate a consistently larger but fluctuating density of organisms at the seep station. However, Shannon-Weaver diversity (H=1.6 to 1.7), Peilou's evenness (J=0.80 to 0.81), and measures of dominance-diversity with estimates of graphical skewness (0.66 to 0.68) and kurtosis (1.2 to 1.4) are all similar for the two stations. The rank correlation of common species at the stations is significant according to Spearman's rho. Species common to both stations account for 85 to 95% of the individuals, further indicating the high degree of similarity between stations. Denser populations of oligochaetes and the maldanid polychaete Praxillella affinis pacifica suggest some advantages for deposit feeders at the seep station. Mats of the bacterium Beggiatoa sp. are associated with localized intense oil seepage. Hypotheses suggesting trophic enrichment and biochemical adaptation at the seep are presented.     

Evolutionary relationships among deep-sea mytilids (Bivalvia: Mytilidae) from hydrothermal vents and cold-water methane/sulfide seeps

A protein electrophoretic survey of mytilids inhabiting deep-sea hydrothermal vents and cold-water methane/sulfide seeps revealed electromorph patterns diagnostic of 10 distinct species. From hydrothermal vents located at sites on the Galápagos Rift, the Mid-Atlantic Ridge, and the Mariana Back Arc Basin, we detected four species of mytilids. Six additional species were detected from three cold-water seep sides in the Gulf of Mexico. The patchy distribution and temporal stability of seeps may provide a greater opportunity for mytilid diversification and persistence than vent sites Nei's genetic distances (D) between species were relatively large (range: 0.528 to ) both within and among habitat types. This pronounced degree of genetic differentiation suggests a relatively ancient common ancestor for the group. Phylogenetic trees were generated using distance Wagner and parsimony analyses of allozyme and morphological characters. The tree topologies obtained from both methods support: (1) the hypothesis that a seep ancestor gave rise to the deep-sea hydrothermal vent mytilids, (2) a historical progression from shallow-water to deep-water habitats, and (3) a co-evolutionary progression from external to internal localization of bacterial symbionts. Whether the seep mytilid taxa constitute paraphyletic or polyphyletic groups remains unresolved. Our phylogenetic hypotheses also provide a benchmark for the phylogeny of mytilid bacterial symbionts.     

Comparison of proton-specific ATPase activities in plume and root tissues of two co-occurring hydrocarbon seep tubeworm species Lamellibrachia luymesi and Seepiophila jonesi

Lamellibrachia luymesi and Seepiophila jonesi are co-occurring species of vestimentiferan tubeworms found at hydrocarbon seepage sites on the upper Louisiana slope of the Gulf of Mexico. Like all vestimentiferans, they rely on internal sulfide-oxidizing symbiotic bacteria for nutrition. These symbionts produce hydrogen ions as a byproduct of sulfide oxidation, which the host tubeworm needs to eliminate to prevent acidosis. The hydrothermal vent tubeworm Riftia pachyptila uses a high activity of P- and V-type H⁺-ATPases located in its plume epithelium to excrete protons. Unlike R. pachyptila, the seep species grow a posterior root, which they can use in addition to their plumes as a nutrient exchange surface. In this study we measured the ATPase activities of plume and root tissues collected from L. luymesi and S. jonesi, and used a combination of inhibitors to determine the relative activities of P- and V-type H⁺-ATPases. We found that the total H⁺-ATPase activity of their plumes was approximately 14 μmol h⁻¹ g⁻¹ wet weight, and that of their roots was between 5 and 7 μmol h⁻¹ g⁻¹ wet weight. These activities were more than ten times lower than those measured in R. pachyptila. We suggest that seep tubeworms might use passive channels to eliminate protons across their roots, in addition to ATP-dependant proton pumps located in their plumes and roots. In addition, we found strong differences between the types of ATPase activities in the plumes of L. luymesi and S. jonesi. While the H⁺-ATPase activity of L. luymesi plumes is dominated by P-type ATPases, S. jonesi has an unusually high activity of V-type H⁺-ATPases. We suggest that S. jonesi relies on its high V-type H⁺-ATPase activity to drive carbon dioxide uptake across its plume surface. L. luymesi, on the other hand, might rely partially on bicarbonate uptake across its root.     

Ecology of the bivalve-inhabiting hydroid Eugymnanthea inquilina in the coastal sounds of Taranto (Ionian Sea,SE Italy)

The life history of the bivalve-inhabiting hydroid Eugymnanthea inquilina was investigated in two different hosts, the mussel Mytilus galloprovincialis (collected from 1989–1991 from the Ionian Sea, Italy) and the clam Ruditapes decussatus (collected from 1991–1992, Ionian Sea, Italy). Hydroids living in mussels revealed a size selection for hosts longer than 40 mm, being almost completely absent in mussels below this size. This might be controlled by signals linked to the sexual maturity of the bivalve. The proportion of molluscs inhabited showed a seasonal trend in mussels only, with a sharp decline at the onset of medusoid liberation. Production of medusoids was high in mussels, whereas medusoids were rare and often abortive in clams. This indicates a higher degree of fitness of E. inquilina in mussels than in clams. The hydroid life cycle was not completed in clams, which therefore were presumably supplied by planulae from medusoids produced by mussel-inhabiting hydroids. Mussels played a key role in the stability and persistence of E. inquilina populations in the studied area. Selective ingestion of trematode sporocysts by E. inquilina hydroids indicated a protective role of the hydroid against mussel parasites, leading to reconsideration of this symbiotic association as a possible mutualism rather than a simple inquilinism.     

Composition of free amino acids and related compounds in invertebrates with symbiotic bacteria at hydrocarbon seeps in the Gulf of Mexico

Five species of bivalves and two species of vestimentiferan tubeworms were collected from hydrocarbon seeps in the Gulf of Mexico, and the composition of their free amino acid and related compounds analysed. Like other marine molluscs, taurine, glycine, glutamic acid, and alanine were abundant in the seep bivalves, but, unlike other molluscs, hypotaurine and thiotaurine were also abundant in the seep species. The relative levels of the most abundant amino compounds indicate that glycine is likely to be an important osmoregulatory compound in the bivalves, but not in the vestimentiferans. A consistent pattern of decreasing taurine:glycine ratio with increasing depth was evident in both vent and seep bivalves, and attributed to differences in the relative availability of taurine and glycine in their diet. Additionally, the generally high glutamate levels and higher levels in the symbiont-containing gills are interpreted as consistent with the proposed role of glutamate as a nutritive transfer molecule in these symbioses. The distribution of hypotaurine and thiotaurine in the seep species is discussed in relation to previously proposed hypotheses on the function of these compounds: hypotaurine as an antioxidant, and thiotaurine as a binding and transport molecule for reduced-sulphur species. Received: 22 October 1998 / Accepted: 4 November 1999     

Ingestion of bivalve larvae by<Emphasis Type="Italic"> Mytilus edulis</Emphasis>: experimental and field demonstrations of larviphagy in farmed blue mussels

Ingestion of bivalve larvae by Mytilus edulis was investigated. Laboratory experiments revealed that ~ 90% of bivalve larvae offered to mussels was ingested and apparently fully digested. The shell of the bivalve larvae offered no protection against digestive processes, resulting in high larval mortality once inside the stomach. Stomach content analysis (September 2001–January 2003) showed that bivalve larvae were ingested by farmed mussels year-round, with the exception of March 2002. Numbers of ingested larvae were highest in October 2001 and May 2002, which coincides with known spawning times of farmed mussels in Ireland. Mussels ingested a large size-range of bivalve larvae, suggesting that all stages of the bivalve life cycle are vulnerable to predation. It is suggested that adult bivalves routinely filter larvae from the surrounding water and that, given the high biomass of mussels present in mussel farms, filtration by adult bivalves significantly reduces numbers of bivalve larvae in nearby waters.Communicated by J.P. Thorpe, Port Erin     

Feeding biology of the shrimp Rimicaris exoculata at hydrothermal vents on the Mid-Atlantic Ridge

A newly described species of shrimp, Rimicaris exoculata Williams and Rona, 1986, dominates the megafaunal community at two hydrothermal vent sites on the Mid-Atlantic Ridge. Behavioral observations and gut-content analyses indicate, that these shrimp ingest large amounts of sulfide particles from black smoker chimneys. We found no evidence for chemoautotrophic endosymbionts in R. exoculata, based on analyses of morphology, stable isotopes, lipopolysaccharides, and ribulose-1,5-bisphosphate carboxylase (RuBPCase) activity. Instead we suggest that the shrimp, are normal heterotrophs, grazing on free-living microorganisms associated with black smoker chimneys. We infer that high bacterial productivity is required to sustain populations of R. exoculata at these vent sites.     

Trophic relationships among invertebrates at the Kairei hydrothermal vent field (Central Indian Ridge)

Exploration of hydrothermal vent systems in locations remote from well-studied sites allows ecologists to determine the degree of site-specific variation in trophic relationships among communities. A preliminary outline of the trophic structure of the Kairei hydrothermal vent community on the Central Indian Ridge (25°19.23′S; 70°02.42′E) is provided here, based on analysis of collections from an April 2001 expedition. Invertebrate biomass at Kairei is dominated by organic carbon with a δ¹³C isotopic value of about –13‰, due to the abundance of primary consumers (shrimp: Rimicaris aff. exoculata) and secondary consumers (anemones: Marianactis n. sp.) with this isotopic composition. Filamentous thiotrophic episymbionts on shrimp have been interpreted to be the major diet items of the shrimp and hence are the dominant primary producers within the community. Free-living autotrophic microorganisms are implicated as the dietary base for other invertebrate species. Four trophic groups are identified within the Kairei invertebrates based on carbon- and nitrogen-isotope ratios, but these groups do not always define discrete trophic levels. Ontogenetic shifts in diet are documented for R. aff. exoculata and brachyuran crabs (Austinograea n. sp.). Diets of symbiont-bearing mussels (Bathymodiolus aff. brevior) and two species of gastropods are isotopically constant throughout the range of sizes analyzed. There is a consistent but unexplained pattern of increasing nitrogen isotopic composition with increasing carbon isotopic composition in vent communities from geographically disjunct oceanic regions. Given the assumptions associated with interpretations of isotopic data, there remains a missing pool of carbon (presumably unsampled bacterial biomass) that contributes to the maintenance of the ¹³C- and ¹⁵N-enriched primary consumers in these ecosystems.     

Bioaccumulation of heavy metals by bivalves from Lim Fjord (North Adriatic Sea)

Trace metal concentrations (Zn, Cd, Pb and Cu) were studied in two different bivalve species of the same age, the mussel Mytilus galloprovincialis, Lmk., and the oyster Ostrea edulis, Linnaeus, which had been grown in the water of Lim Fjord, North West Yugoslavia (Peninsula Istria), i.e. under the same physicochemical conditions. The study offers a realistic view on the metal accumulation ability of oysters and mussels. The distribution of trace metals over the different organs and the edible parts of the mussels and oysters, collected in June 1979, was determined and is discussed in detail. The results are supported by the determination of the trace metal levels in the dissolved state and in the suspended material in the ambient sea water of the bivalves. Concentration factors for zinc, cadmium, lead and copper in the mussels of: 95 000, 9 100, 1 500 and 4 000; and in the oysters of: 95 500, 30 400, 3 400 and 64 500 were found, respectively. The values were evaluated comparing the metal concentration in the bivalve soft part and the dissolved trace metal levels in the adjacent water.     

Molecular taxonomy of vestimentiferans of the western Pacific,and their phylogenetic relationship to species of the eastern Pacific III. Alaysia-like vestimentiferans and relationships among families

The phylogenetic status of Alaysia-like vestimentiferans, which were collected at eight sites in the western Pacific, was analyzed on the basis of the nucleotide sequence of part of a mitochondrial gene for cytochrome oxidase I. The 123 individuals analyzed were tentatively classified into four species, which inhabit, respectively, seep areas off the central part of the Japanese mainland, hydrothermal vent fields in the Mid-Okinawa Trough, hydrothermal vent fields in the southern Okinawa Trough, and both seep and vent sites in the Bismarck Sea. From six additional lamellibrachiid specimens from two sites in the Bismarck Sea, two new tentative species were recognized, namely, one from a seep area off Papua New Guinea and another from the hydrothermal vent field at the DESMOS site in the Manus Basin. Phylogenetic analysis using all available sequences of six vestimentiferan families and Alaysia-like vestimentiferans revealed that vestimentiferans could be divided into three groups, namely, lamellibrachiids, escarpiids, and others, even though the monophyly of the third group was supported by only low bootstrap probabilities. Within the first group, the earliest divergence of a tentative lamellibrachiid species from the DESMOS site was apparent. All Alaysia-like vestimentiferans formed a monophyletic group with Arcovestia ivanovi from the Manus Basin, and it is suggested that this group might be derived from vestimentiferans that are endemic to the hydrothermal sites in the Eastern Pacific Rise.