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.     

Evolution of habitat use by deep-sea mussels

Previous phylogenetic studies proposed that symbiont-bearing mussels of the subfamily Bathymodiolinae (Bivalvia: Mytilidae) invaded progressively deeper marine environments and evolved from lineages that decomposed wood and bone to specialized lineages that invaded cold-water hydrocarbon seeps and finally deep-sea hydrothermal vents. To assess the validity of the hypotheses, we examined two nuclear (18S and 28S rRNA) and two mitochondrial genes (COI and ND4) from a broad array of bathymodiolin species that included several recently discovered species from shallow hydrothermal seamounts. Bayesian phylogenetic analysis and maximum-likelihood estimates of ancestral character states revealed that vent species evolved multiple times, and that reversals in vent and seep habitat use occurred within the sampled taxa. Previous hypotheses regarding evolution from wood/bone-to-seeps/vents are supported in that mid-ocean hydrothermal vent species may represent a monophyletic group with one noticeable reversal. Earlier hypotheses about progressive evolution from shallow-to-deep habitats appear to hold with a few instances of habitat reversals.     

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.     

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.     

Distribution of the Lamellibrachia spp. (Siboglinidae,Annelida) and their trophosome endosymbiont phylotypes in the Mediterranean Sea

During the 2010/2011 Exploration vessel Nautilus expedition to the Mediterranean Sea, samples of Lamellibrachia (Siboglinidae, Annelida) were imaged in situ and collected from hydrothermal vent and methane “cold seeps.” An analysis of these Lamellibrachia and their endosymbiotic thioautotrophic gammaproteobacteria reveals two distinct endosymbiont phylotypes. Phylotype 1 was present in Lamellibrachia specimens from 947 m at the Eratosthenes seamount seep (a seep off Cyprus in the Eastern Mediterranean), and Phylotype 2 was found in siboglinids from 618 m at a hydrothermal vent within Palinuro volcanic complex in Tyrrhenian Sea. Both phylotypes coexist in siboglinids at 1,036 m from the Palmachim disturbance, a cold seep in the Eastern Mediterranean’s Levantine basin. Our results, combined with existing knowledge of siboglinid host and endosymbiotic bacteria biogeography, reveal that two major groups of endosymbionts coexist within lamellibranchids and escarpids. The phylogenetic clustering of these bacteria is primarily influenced by geographic location, rather than selection by the siboglinid host.     

The effect of cadmium on antioxidant responses and the susceptibility to oxidative stress in the hydrothermal vent mussel Bathymodiolus azoricus

Hydrothermal vents are a unique environment of extreme physical–chemical characteristics and biological species composition. Cd is a toxic non-essential metal present in high concentrations in the hydrothermal vent environment, contrary to those found in marine coastal areas. Cd toxicity has been related, among other things, with reactive oxygen species production, even though this is a non-redox metal. Bathymodiolus azoricus is a deep-sea Mytilid bivalve very common in the Mid Atlantic Ridge (MAR) hydrothermal vent fields and very little is known about the antioxidant defence system in this specie. Because lethal Cd concentration in B. azoricus is unknown, the aim of this study was to assess the effects of a Cd concentration higher than that found in the hydrothermal vents on oxidative stress biomarkers, such as antioxidant enzymes. Mussels were exposed to 100 μg l⁻¹ Cd during 24, 48 and 144 h, respectively, in a pressurized aquarium (IPOCAMP). Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx), total oxyradical scavenging capacity (TOSC), metallothionein (MT) and lipid peroxidation (LPO) were measured in the gills and mantle of B. azoricus. The results indicate that gills are first affected by Cd toxicity. This may be due to different physiological functions of the tissues and by the presence of thio and methanotrophic symbiotic bacteria in the gills. The SOD and CAT are inhibited during the first day of exposure in the gills, although TOSC and MT concentrations were the same in control and exposed mussels. In the mantle, enzymatic activation only occurred after 6 days, and no significant differences in MT concentrations were found in the control and exposed mussels during the first day, as observed in the gills.     

Persistence,morphology, and nutritional state of a gastropod hosted bacterial symbiosis in different levels of hydrothermal vent flux

The limpet, Lepetodrilus fucensis McLean, is found in prominent stacks around hydrothermal vents on the Juan de Fuca Ridge. L. fucensis hosts a filamentous episymbiont on its gill lamellae that may be ingested directly by the gill epithelium. To assess the persistence of this symbiosis I used microscopy to examine the gills of L. fucensis from sites representing its geographic range and different habitats. The symbiosis is present on all the specimens examined in this study, including both sexes and a range of juvenile and adult sizes. Next, I aimed to determine if patterns in bacterial abundance, host condition, and gill morphology support the hypotheses that the bacteria are chemoautotrophic and provide limpets with a food resource. To do so, I compared specimens from high and low flux locations at multiple vents. My results support the above hypotheses: (1) gill bacteria are significantly less abundant in low flux where the concentrations of reduced chemicals (for chemoautotrophy) are negligible, (2) low flux specimens have remarkably poor tissue condition, and (3) the lamellae of high flux limpets have greater surface area: the blood space and bacteria-hosting epithelium are deeper and have more folds than low flux lamellae, modifications that support higher symbiont abundances. I next asked if the morphology of the lamellae could change. To test this, I moved high flux limpets away from a vent and after 1 year the lamellar depth and shape of the transplanted specimens resembled low flux gills. Last, I was interested in whether bacterial digestion by the gill epithelium is a significant feeding mechanism. As bacteria-like cells are rarely apparent in lysosomes of the gill epithelium, I predicted that lysosome number would be unrelated to bacterial abundance. My data support this prediction, suggesting that digestion of bacteria by the gill epithelium probably contributes only minimally to the limpet’s nutrition. Overall, the persistence and morphology of the L. fucensis gill symbiosis relates to the intensity of vent flux and indicates that specimens from a variety of habitats may be necessary to characterize the morphological variability of gill-hosted symbioses in other molluscs.     

Na+-K+-adenosine triphosphatase activities in gills of marine teleost fishes: Changes with depth,size and locomotory activity level

Activities of the primary enzyme responsible for monovalent ion regulation, Na⁺-K⁺-adenosine triphosphatase (Na⁺-K⁺-ATPase), were measured in gills of marine teleost fishes with different depths of occurrence (0 to 4800 m), body weights (a range of five orders of magnitude), and locomotory capacities. Specimens were collected off the coasts of California and Oregon in 1983–1989, and at the Galápagos Spreading Center and 13°N East Pacific Rise hydrothermal vent sites in 1987 and 1988, respectively. Except for two hydrothermal vent fishes, deep-sea species had much lower Na⁺-K⁺-ATPase activities g^–1 gill filament than shallow-living species, indicating that osmoregulatory costs, like total metabolic rate, are greatly reduced in most deep-living fishes. Within a species, the total branchial Na⁺-K⁺-ATPase activity per individual was dependent on size; the average allometric scaling exponent was 0.83. Using published values for oxygen consumption rates, and the total branchial Na⁺-K⁺-ATPase activities as an index of osmoregulatory costs, we estimated the maximal cost (as percent of ATP turnover) for osmoregulation in ten teleosts. Osmoregulatory costs averaged about 10% of total ATP turnover among these species, and maximal costs were no greater than about 20%. The percent costs of osmoregulation did not differ between shallow- and deep-living fishes. The reduced total ATP expenditure for osmoregulation in deep-living fishes is proposed to result from the sluggish locomotory habits of these fishes, not from selection for reduced osmotic coastper se. Thus, the reduced swimming abilities of these fishes lead to lower rates of water flow over the gills and less blood flow through the gills due to reduced demands for oxygen. Consequently, passive flux of water and ions through the gills is much lower than in more active fishes, and osmotic costs are thereby minimized. The relatively high activities of Na⁺-K⁺-ATPase in gills of the two hydrothermal vent fishes suggest that these fishes may be more active and have higher metabolic rates than other deep-sea fishes.     

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     

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.     

Gill structure in Lucina pectinata (Bivalvia: Lucinidae) with reference to hemoglobin in bivalves with symbiotic sulphur-oxidizing bacteria

Lucina pectinata is a large tropical Lucinidae which is characterized by abundant tissue hemoglobin in its deep-red gills. In the present paper, hemoglobin is described as being located in cytoplasmic dark patches of the bacteriocytes together with a cystine-rich protein. Large microbodies contain a non-hemoglobin heme-compound which is identified with a previously described non-protein-bound hematin; however, it has not been established whether this heme is involved in a sulphur-oxidizing system or represents a catabolic by-product of hemoglobin. Electron-lucent vesicles are associated with the basal microbodies but their function is, so far, unknown. In addition, the bacteriocytes have been observed to have direct contact with sea water, modulated by large intercalary cells which overlap the bacteriocytes on their margin. Such relationships between bacteriocytes and intercalary cells, as well as their cytological features, are different from those observed in lucinid species inhabiting sea-grass beds, but very similar to those observed in Calyptogena magnifica. From the congruence between the shallow-water Lucinidae L. pectinata, inhabiting mangrove swamps, and the deep-sea Vesicomyidae C. magnifica, found at hydrothermal vents, we conclude that such features are likely to be adaptative to high-sulphide environments, notwithstanding the phylogenetic distance.     

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 Galápagos 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.     

Ecotoxicological responses to chalcopyrite exposure in a proxy for deep-sea hydrothermal vent shrimp: implications for seafloor massive sulphide mining

Deep-sea mineral prospecting has raised concerns regarding potential ecotoxicological impacts of deep-sea mineral extraction. Although metal mineral phases are predicted to exhibit low bioavailability, few data explore the relative toxicity of mineral phases and dissolved constituent metals. Acute 96?h chalcopyrite (CuFeS₂) (<250?µm grain size) exposures using the shallow-water ecophysiological model organism Palaemon varians as an ecotoxicological proxy for deep-sea hydrothermal vent shrimp revealed no effect in both lethal and sublethal assays up to 2.888?g?L^?1, suggesting that chalcopyrite is not bioavailable. Deep-sea species, therefore, appear at greater ecotoxicological risk from dissolved metals during seafloor massive sulphide (SMS) mining. Consequently, an approach combining modelling the release, and spatial and temporal dilution of dissolved metals during SMS mining, with data on sublethal effects of dissolved metals on shallow-water proxies, may best constrain the potential ecotoxicological impacts of SMS mining, and deliver ecotoxicological threshold concentrations for active SMS extraction.     

Chromosomal and nuclear characteristics of deep-sea hydrothermal-vent organisms: correlates of increased growth rate

A range of tissue and cell types from adult and juvenile stages of vent- and non-vent-dwelling deep-sea and shallow-water organisms were compared for signs of cell division, in preparation for a cytogenetic study of the different groups. Virtually all the Mid-Atlantic Ridge (MAR) vent species (bresiliid shrimp, bathymodiolid mussel, branchipolynoid polychaete, and a range of small gastropods) showed an abundance of metaphase chromosome spreads, indicating a generally high intrinsic rate of cell division, irrespective of their phylogenetic group. In comparison, similar tissues and life-history stages (i.e. gonad, developing eggs and gill) from two deep-sea bathypelagic crustaceans, Hymenodora and Gnathophausia, failed to yield any convincing evidence of cell division. This indicated that rates of growth and reproduction in the rest of the deep sea are extremely low. While much has been written about the contrasting growth rates of vent versus non-vent organisms, this is the first direct evidence of differences in cell-division rate between vent and non-vent species, which undoubtedly reflects contrasting conditions relating to food availability and temporal stability in these two deep-sea environments. The cells of shallow-water marine invertebrates (Mytilus, Nucella, Littorina, Pomatoceros and Crangon), based on the results of previous cytogenetic investigations, occupy an intermediate position with respect to their cell-division kinetics. When chromosome numbers, DNA content and nuclear diameters were compared between vent and non-vent species, this showed that no change had occurred in these characters since the time of vent colonisation. Representative chromosome spreads from hydrothermal-vent organisms are shown here for the first time.     

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.     

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     

Effect of ambient oxygen concentration on activities of enzymatic antioxidant defences and aerobic metabolism in the hydrothermal vent worm, Paralvinella grasslei

The alvinellid Paralvinella grasslei is a common endemic polychaete from the deep-sea hydrothermal vent communities located on the East Pacific Rise (EPR). These organisms colonise a large range of microhabitats around active sites where physico-chemical conditions are thought to generate reactive oxygen species (ROS). Furthermore, in this aerobic organism, ROS could also be generated by the activity of the mitochondrial respiratory chain. In this paper, we investigated the effect of ambient oxygen concentration on the activities of three essential antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX) and their relationships with the activity of enzymes involved in aerobic metabolism (cytochrome c oxidase, COX; citrate synthase, CS). Results of incubation of P. grasslei in a high-pressure vessel with circulating seawater at different oxygen partial pressures indicate that this worm regulates COX and CS activities differently in gills and body wall. CAT and GPX activities increase in these tissues when animals are maintained in filtered surface seawater. Moreover, levels of malondialdehyde increase in gills, testifying that oxidative damage occurs under these conditions. CAT and GPX activities are positively related to COX and CS activities, but no correlation was detected between SOD and the metabolic enzyme activities. In comparison with littoral annelids, SOD activities are very high whereas CAT activities are very low or absent in P. grasslei. The possible reasons for the occurrence of such differences are discussed.     

Trophic ecology of Rimicaris exoculata : a combined lipid abundance/stable isotope approach

The alvinocaridid shrimp Rimicaris exoculata is an abundant component of the biota of Mid-Atlantic Ridge hydrothermal vents. To determine the nutritional strategy of this organism, we analysed the molecular abundance and carbon isotopic composition of its phospholipid fatty acids. High abundances of n-7 fatty acids (>40% total fatty acids) were observed in R. exoculata muscle tissues, in bacterial epibionts scraped from its gill bailers, and from the bacterially infested metal sulphides that the shrimp ingest. The phospholipid fatty acid abundance data indicates that the bacteria in the sulphides are closely related to the bacterial epibiota inhabiting the shrimp gill bailers, carapace and other body parts. Compound specific δ¹³C analyses of the phospholipid fatty acids gave average values of −12‰ for the epibiont bacteria and −21‰ for the sulphide bacteria. This difference may be largely due to the expression of different forms of RuBisCO (Forms I and II) which fractionate against ¹³C to different extents. Carbon limitation within the shrimp epibiont population may be an additional factor. The δ¹³C values (mean = −13‰) of the saturated and monounsaturated fatty acids isolated from the muscle tissues of R. exoculata were very close to those of the epibionts, indicating that the predominant source of dietary carbon for the shrimp is their epibionts, with a lesser contribution from free-living bacteria. The δ¹³C values (−26‰) of shrimp cholesterol were much more negative than those of the fatty acids, and this cholesterol is likely to have derived from the oceanic photic zone. Received: 26 June 1997 / Accepted: 6 November 1998     

Spatial and interannual variation in the faunal distribution at Broken Spur vent field (29°N, Mid-Atlantic Ridge)

Initial ecological observations at Broken Spur in 1993 suggested a low biomass relative to other deep-water vent communities known along the Mid-Atlantic Ridge. The persistence of a low shrimp biomass over 15 mo at Broken Spur vents appears to refute the hypothesis that the community is expanding through reproduction and immigration to occupy habitat vacated during a recent period of hydrothermal quiescence or catastrophe. Although the absence of “swarms” of shrimp similar to those found at the hydrothermal mounds of TAG (26°N) and Snake Pit (23°N) is a visually striking feature of the majority of venting structures known at Broken Spur, the biomass of fauna other than shrimp may not be significantly less than that of other Mid-Atlantic sites. The discovery of “swarms” of shrimp at Bogdanov Site, visited for the first time in 1994, suggests that availability of substratum exposed to the flow of hydrothermal fluids, which is a function of the topography of venting structures, may be a prerequisite for the development of these dense aggregations. Two testable predictions arise from this hypothesis. Firstly, dense aggregations of Rimicaris exoculata should occur at any other structures with a morphology similar to Bogdanov Site that may be discovered in the Broken Spur vent field, and should not occur at other isolated chimney structures that may be found. Secondly, “swarms” of shrimp should appear at any Broken Spur chimneys that develop into structures with a morphology more similar to that of Bogdanov Site in future. Received: 2 July 1997 / Accepted: 9 July 1997     

Scientists as Stakeholders in Conservation of Hydrothermal Vents

Abstract: Hydrothermal vents are deep‐sea ecosystems that are almost exclusively known and explored by scientists rather than the general public. Continuing scientific discoveries arising from study of hydrothermal vents are concommitant with the increased number of scientific cruises visiting and sampling vent ecosystems. Through a bibliometric analysis, we assessed the scientific value of hydrothermal vents relative to two of the most well‐studied marine ecosystems, coral reefs and seagrass beds. Scientific literature on hydrothermal vents is abundant, of high impact, international, and interdisciplinary and is comparable in these regards with literature on coral reefs and seagrass beds. Scientists may affect hydrothermal vents because their activities are intense and spatially and temporally concentrated in these small systems. The potential for undesirable effects from scientific enterprise motivated the creation of a code of conduct for environmentally and scientifically benign use of hydrothermal vents for research. We surveyed scientists worldwide engaged in deep‐sea research and found that scientists were aware of the code of conduct and thought it was relevant to conservation, but they did not feel informed or confident about the respect other researchers have for the code. Although this code may serve as a reminder of scientists’ environmental responsibilities, conservation of particular vents (e.g., closures to human activity, specific human management) may effectively ensure sustainable use of vent ecosystems for all stakeholders.