Deep-sea Hydrothermal Vent Sites Research Articles

The soundscape of two deep-sea hydrothermal vent sites

Hydrothermal vents are sea floor structures where geothermally heated seawater is discharged. The high-temperature, chemically rich environment is host to uniquely adapted marine life. Vent soundscapes may contain important bioacoustic cues as well as signals enabling passive acoustic monitoring of hydrothermal vent dynamics. Proposals for deep-sea mining of seafloor massive sulfides near hydrothermal vents have elicited concern over potential environmental impacts due to disturbance from industrial activity, including changes to the soundscape. This study assesses the baseline soundscape at two sites, the Main Endeavour Field on the Juan de Fuca Ridge and the Lucky Strike vent field on the Mid-Atlantic Ridge over 12 months and 3 months respectively. To facilitate comparison with future studies at other sites, the most recently proposed standard soundscape analysis methodologies are employed, including terminology in alignment with ISO 18405:2017. In accordance with the latest soundscape standard literature, metrics quantifying the amplitude, impulsiveness, periodicity, and uniformity are reported. Spectral probability densities, percentiles, and long-term spectrograms are computed in hybrid millidecade frequency bands. Finally, a qualitative analysis is included to describe the source types contributing to the hydrothermal vent sound field.

Deep-sea hydrothermal vent metagenome-assembled genomes provide insight into the phylum Nanoarchaeota.

Ectosymbiotic Nanoarchaeota live on the surface of diverse archaeal hosts. Despite being broadly distributed in global geothermal systems, only three Nanoarchaeota have been successfully co-cultivated with their hosts, and until now no nanoarchaeotal cultures or genomes have been described from deep-sea hydrothermal vents. We recovered three nanoarchaeotal metagenome-assembled genomes (MAGs) from deep-sea hydrothermal vent sites at the Eastern Lau Spreading Center (M10-121), Guaymas Basin (Gua-46) and the Mid-Cayman Rise (MC-1). Based on average amino acid identity analysis, M10-121 is a novel species in the candidate genus Nanoclepta, while the other two MAGs represent novel genera in the Nanoarchaeota. Like previously sequenced Nanoarchaeota, each MAG encodes at least one split protein-coding gene. The MAGs also contain a mosaic of key nanoarchaeotal features, including CRISPR repeat regions and marker genes for gluconeogenesis and archaeal flagella. MC-1 also encodes the pentose bisphosphate pathway, which may allow the nanoarchaeote to bypass several steps in glycolysis and produce ATP.

Fungi associated with chimney and sulfide samples from a South Mid-Atlantic Ridge hydrothermal site: Distribution, diversity and abundance

Deep-sea hydrothermal vent ecosystems are known to support remarkably diverse microbial communities, ranging from chemoautotrophic prokaryotes to heterotrophic prokaryotes and microeukaryotes. While fungi have generally been identified as an important component of various microbial communities in the environment, little is known about the species richness and abundance of such microorganisms in deep-sea hydrothermal vent ecosystems. In this study, a combined culture-dependent and culture-independent sequence-based approach was used to investigate fungal distribution and diversity at a deep-sea hydrothermal vent site located at the Mid-Atlantic Ridge of the South Atlantic Ocean. Sequence analyses showed that the fungal community was dominated by members of the Ascomycota and the Basidiomycota. Several new phylotypes (28 of 65 total fungal OTUs and 2 of 19 culturable fungal phylotypes) were identified, contributing to the literally unknown diversity of fungi in this understudied habitat. The fungal community structures in the chimney samples were distinct from those in three sulfide samples. The qPCR results revealed that fungal LSU rRNA gene copy numbers ranged from 5.88×105 to 6.77×106 copies/gram rock (wet weight), and the Ascomycota was significantly more abundant 2–3 orders) than the Basidiomycota. Our findings provide new insights into the diversity and abundance of fungi in deep-sea hydrothermal vent ecosystems, which increases our knowledge and understanding of the fungal diversity in deep-sea environments.

Copepod colonization of organic and inorganic substrata at a deep-sea hydrothermal vent site on the Mid-Atlantic Ridge

The few existing studies on deep-sea hydrothermal vent copepods indicate low connectivity with surrounding environments and reveal high endemism among vents. However, the finding of non-endemic copepod species in association with engineer species at different reduced ecosystems poses questions about the dispersal of copepods and the colonization of hydrothermal vents as well as their ecological connectivity.The objective of this study is to understand copepod colonization patterns at a hydrothermal vent site in response to environmental factors such as temperature and fluid flow as well as the presence of different types of substrata. To address this objective, an in situ experiment was deployed using both organic (woods, pig bones) and inorganic (slates) substrata along a gradient of hydrothermal activity at the Lucky Strike vent field (Eiffel Tower, Mid-Atlantic Ridge). The substrata were deployed in 2011 during the MoMARSAT cruise and were recovered after two years in 2013.Overall, copepod density showed significant differences between substrata types, but was similar among different hydrothermal activity regimes. Highest densities were observed on woods at sites with moderate or low fluid input, whereas bones were the most densely colonized substrata at the 2 sites with higher hydrothermal influence. Although differences in copepod diversity were not significant, the observed trends revealed overall increasing diversity with decreasing temperature and fluid input. Slates showed highest diversity compared to the organic substrata. Temperature and fluid input had a significant influence on copepod community composition, resulting in higher similarity among stations with relatively high and low fluid inputs, respectively. While vent-specialists such as dirivultids and the tegastid Smacigastes micheli dominated substrata at high vent activity, the experiment demonstrated increasing abundance and dominance of non-vent taxa with decreasing temperature and fluid input. Effects of the substratum type on community composition were not significant, although at sites with moderate or low fluid input, woods exhibited distinctive communities with high densities and relative abundance of the taxon Nitocrella sp. In conclusion, copepod colonization and species composition were mainly influenced by hydrothermal fluid input and temperature rather than the type of substratum. The outcome of this study provides fundamental knowledge to better understand copepod colonization at hydrothermal vents.

Identification and characterization of carboxyl esterases of gill chamber-associated microbiota in the deep-sea shrimp Rimicaris exoculata by using functional metagenomics.

The shrimp Rimicaris exoculata dominates the fauna in deep-sea hydrothermal vent sites along the Mid-Atlantic Ridge (depth, 2,320 m). Here, we identified and biochemically characterized three carboxyl esterases from microbial communities inhabiting the R. exoculata gill that were isolated by naive screens of a gill chamber metagenomic library. These proteins exhibit low to moderate identity to known esterase sequences (≤52%) and to each other (11.9 to 63.7%) and appear to have originated from unknown species or from genera of Proteobacteria related to Thiothrix/Leucothrix (MGS-RG1/RG2) and to the Rhodobacteraceae group (MGS-RG3). A library of 131 esters and 31 additional esterase/lipase preparations was used to evaluate the activity profiles of these enzymes. All 3 of these enzymes had greater esterase than lipase activity and exhibited specific activities with ester substrates (≤356 U mg(-1)) in the range of similar enzymes. MGS-RG3 was inhibited by salts and pressure and had a low optimal temperature (30°C), and its substrate profile clustered within a group of low-activity and substrate-restricted marine enzymes. In contrast, MGS-RG1 and MGS-RG2 were most active at 45 to 50°C and were salt activated and barotolerant. They also exhibited wider substrate profiles that were close to those of highly active promiscuous enzymes from a marine hydrothermal vent (MGS-RG2) and from a cold brackish lake (MGS-RG1). The data presented are discussed in the context of promoting the examination of enzyme activities of taxa found in habitats that have been neglected for enzyme prospecting; the enzymes found in these taxa may reflect distinct habitat-specific adaptations and may constitute new sources of rare reaction specificities.

Water column imaging with multibeam echo-sounding in the mid-Okinawa Trough: Implications for distribution of deep-sea hydrothermal vent sites and the cause of acoustic water column anomaly

Water column imaging with multibeam echo-sounding in the mid-Okinawa Trough: Implications for distribution of deep-sea hydrothermal vent sites and the cause of acoustic water column anomaly

Bacteria dominate the ammonia-oxidizing community in a hydrothermal vent site at the Mid-Atlantic Ridge of the South Atlantic Ocean.

Ammonia oxidation is the first and rate-limiting step of nitrification, which is carried out by two groups of microorganisms: ammonia-oxidizing bacteria (AOB) and the recently discovered ammonia-oxidizing archaea (AOA). In this study, diversity and abundance of AOB and AOA were investigated in five rock samples from a deep-sea hydrothermal vent site at the Mid-Atlantic Ridge (MAR) of the South Atlantic Ocean. Both bacterial and archaeal ammonia monooxygenase subunit A (amoA) gene sequences obtained in this study were closely related to the sequences retrieved from deep-sea environments, indicating that AOB and AOA in this hydrothermal vent site showed typical deep ocean features. AOA were more diverse but less abundant than AOB. The ratios of AOA/AOB amoA gene abundance ranged from 1/3893 to 1/242 in all investigate samples, indicating that bacteria may be the major members responding to the aerobic ammonia oxidation in this hydrothermal vent site. Furthermore, diversity and abundance of AOA and AOB were significantly correlated with the contents of total nitrogen and total sulfur in investigated samples, suggesting that these two environmental factors exert strong influences on distribution of ammonia oxidizers in deep-sea hydrothermal vent environment.

Deep transcriptome-sequencing and proteome analysis of the hydrothermal vent annelid Alvinella pompejana identifies the CvP-bias as a robust measure of eukaryotic thermostability

BackgroundAlvinella pompejana is an annelid worm that inhabits deep-sea hydrothermal vent sites in the Pacific Ocean. Living at a depth of approximately 2500 meters, these worms experience extreme environmental conditions, including high temperature and pressure as well as high levels of sulfide and heavy metals. A. pompejana is one of the most thermotolerant metazoans, making this animal a subject of great interest for studies of eukaryotic thermoadaptation.ResultsIn order to complement existing EST resources we performed deep sequencing of the A. pompejana transcriptome. We identified several thousand novel protein-coding transcripts, nearly doubling the sequence data for this annelid. We then performed an extensive survey of previously established prokaryotic thermoadaptation measures to search for global signals of thermoadaptation in A. pompejana in comparison with mesophilic eukaryotes. In an orthologous set of 457 proteins, we found that the best indicator of thermoadaptation was the difference in frequency of charged versus polar residues (CvP-bias), which was highest in A. pompejana. CvP-bias robustly distinguished prokaryotic thermophiles from prokaryotic mesophiles, as well as the thermophilic fungus Chaetomium thermophilum from mesophilic eukaryotes. Experimental values for thermophilic proteins supported higher CvP-bias as a measure of thermal stability when compared to their mesophilic orthologs. Proteome-wide mean CvP-bias also correlated with the body temperatures of homeothermic birds and mammals.ConclusionsOur work extends the transcriptome resources for A. pompejana and identifies the CvP-bias as a robust and widely applicable measure of eukaryotic thermoadaptation.ReviewerThis article was reviewed by Sándor Pongor, L. Aravind and Anthony M. Poole.

Complete Genome Sequence of the Thermophilic, Piezophilic, Heterotrophic Bacterium Marinitoga piezophila KA3

Marinitoga piezophila KA3 is a thermophilic, anaerobic, chemoorganotrophic, sulfur-reducing bacterium isolated from the Grandbonum deep-sea hydrothermal vent site at the East Pacific Rise (13°N, 2,630-m depth). The genome of M. piezophila KA3 comprises a 2,231,407-bp circular chromosome and a 13,386-bp circular plasmid. This genome was sequenced within Department of Energy Joint Genome Institute CSP 2010.

Hippea jasoniae sp. nov. and Hippea alviniae sp. nov., thermoacidophilic members of the class Deltaproteobacteria isolated from deep-sea hydrothermal vent deposits

Thirteen novel, obligately anaerobic, thermoacidophilic bacteria were isolated from deep-sea hydrothermal vent sites. Four of the strains, designated EP5-r(T), KM1, Mar08-272r(T) and Mar08-368r, were selected for metabolic and physiological characterization. With the exception of strain EP5-r(T), all strains were short rods that grew between 40 and 72 °C, with optimal growth at 60-65 °C. Strain EP5-r(T) was more ovoid in shape and grew between 45 and 75 °C, with optimum growth at 60 °C. The pH range for growth of all the isolates was between pH 3.5 and 5.5 (optimum pH 4.5 to 5.0). Strain Mar08-272r(T) could only grow up to pH 5.0. Elemental sulfur was required for heterotrophic growth on acetate, succinate, Casamino acids and yeast extract. Strains EP5-r(T), Mar08-272r(T) and Mar08-368r could also use fumarate, while strains EP5-r(T), KM1 and Mar08-272r(T) could also use propionate. All isolates were able to grow chemolithotrophically on H(2), CO(2), sulfur and vitamins. Phylogenetic analysis of 16S rRNA gene sequences placed all isolates within the family Desulfurellaceae of the class Deltaproteobacteria, with the closest cultured relative being Hippea maritima MH(2)(T) (~95-98 % gene sequence similarity). Phylogenetic analysis also identified several isolates with at least one intervening sequence within the 16S rRNA gene. The genomic DNA G+C contents of strains EP5-r(T), KM1, Mar08-272r(T) and Mar08-368r were 37.1, 42.0, 35.6 and 37.9 mol%, respectively. The new isolates differed most significantly from H. maritima MH(2)(T) in their phylogenetic placement and in that they were obligate thermoacidophiles. Based on these phylogenetic and phenotypic properties, the following two novel species are proposed: Hippea jasoniae sp. nov. (type strain Mar08-272r(T) = DSM 24585(T) = OCM 985(T)) and Hippea alviniae sp. nov. (type strain EP5-r(T) = DSM 24586(T) = OCM 986(T)).

Fossil evidence for serpentinization fluids fueling chemosynthetic assemblages

Among the deep-sea hydrothermal vent sites discovered in the past 30 years, Lost City on the Mid-Atlantic Ridge (MAR) is remarkable both for its alkaline fluids derived from mantle rock serpentinization and the spectacular seafloor carbonate chimneys precipitated from these fluids. Despite high concentrations of reduced chemicals in the fluids, this unique example of a serpentinite-hosted hydrothermal system currently lacks chemosynthetic assemblages dominated by large animals typical of high-temperature vent sites. Here we report abundant specimens of chemosymbiotic mussels, associated with gastropods and chemosymbiotic clams, in approximately 100 kyr old Lost City-like carbonates from the MAR close to the Rainbow site (36 °N). Our finding shows that serpentinization-related fluids, unaffected by high-temperature hydrothermal circulation, can occur on-axis and are able to sustain high-biomass communities. The widespread occurrence of seafloor ultramafic rocks linked to likely long-range dispersion of vent species therefore offers considerably more ecospace for chemosynthetic fauna in the oceans than previously supposed.

Pathways of Carbon and Energy Metabolism of the Epibiotic Community Associated with the Deep-Sea Hydrothermal Vent Shrimp Rimicaris exoculata

BackgroundThe shrimp Rimicaris exoculata dominates the faunal biomass at many deep-sea hydrothermal vent sites at the Mid-Atlantic Ridge. In its enlarged gill chamber it harbors a specialized epibiotic bacterial community for which a nutritional role has been proposed.Methodology/Principal FindingsWe analyzed specimens from the Snake Pit hydrothermal vent field on the Mid-Atlantic Ridge by complementing a 16S rRNA gene survey with the analysis of genes involved in carbon, sulfur and hydrogen metabolism. In addition to Epsilon- and Gammaproteobacteria, the epibiotic community unexpectedly also consists of Deltaproteobacteria of a single phylotype, closely related to the genus Desulfocapsa. The association of these phylogenetic groups with the shrimp was confirmed by fluorescence in situ hybridization. Based on functional gene analyses, we hypothesize that the Gamma- and Epsilonproteobacteria are capable of autotrophic growth by oxidizing reduced sulfur compounds, and that the Deltaproteobacteria are also involved in sulfur metabolism. In addition, the detection of proteobacterial hydrogenases indicates the potential for hydrogen oxidation in these communities. Interestingly, the frequency of these phylotypes in 16S rRNA gene clone libraries from the mouthparts differ from that of the inner lining of the gill chamber, indicating potential functional compartmentalization.ConclusionsOur data show the specific association of autotrophic bacteria with Rimicaris exoculata from the Snake Pit hydrothermal vent field, and suggest that autotrophic carbon fixation is contributing to the productivity of the epibiotic community with the reductive tricarboxylic acid cycle as one important carbon fixation pathway. This has not been considered in previous studies of carbon fixation and stable carbon isotope composition of the shrimp and its epibionts. Furthermore, the co-occurrence of sulfur-oxidizing and sulfur-reducing epibionts raises the possibility that both may be involved in the syntrophic exchange of sulfur compounds, which could increase the overall efficiency of this epibiotic community.

Culturability and Secondary Metabolite Diversity of Extreme Microbes: Expanding Contribution of Deep Sea and Deep-Sea Vent Microbes to Natural Product Discovery

Microbes from extreme environments do not necessarily require extreme culture conditions. Perhaps the most extreme environments known, deep-sea hydrothermal vent sites, support an incredible array of archaea, bacteria, and fungi, many of which have now been cultured. Microbes cultured from extreme environments have not disappointed in the natural products arena; diverse bioactive secondary metabolites have been isolated from cultured extreme-tolerant microbes, extremophiles, and deep-sea microbes. The contribution of vent microbes to our arsenal of natural products will likely grow, given the culturability of vent microbes; their metabolic, physiologic, and phylogenetic diversity; numerous reports of bioactive natural products from microbes inhabiting high acid, high temperature, or high pressure environments; and the recent isolation of new chroman derivatives and siderophores from deep-sea hydrothermal vent bacteria.

Novel uncultured Epsilonproteobacteria dominate a filamentous sulphur mat from the 13°N hydrothermal vent field, East Pacific Rise

Rapid growth of microbial sulphur mats have repeatedly been observed during oceanographic cruises to various deep-sea hydrothermal vent sites. The microorganisms involved in the mat formation have not been phylogenetically characterized, although the production of morphologically similar sulphur filaments by a Arcobacter strain coastal marine has been documented. An in situ collector deployed for 5 days at the 13 degrees N deep-sea hydrothermal vent site on the East Pacific Rise (EPR) was rapidly colonized by a filamentous microbial mat. Microscopic and chemical analyses revealed that the mat consisted of a network of microorganisms embedded in a mucous sulphur-rich matrix. Molecular surveys based on 16S rRNA gene and aclB genes placed all the environmental clone sequences within the Epsilonproteobacteria. Although few 16S rRNA gene sequences were affiliated with that of cultured organisms, the majority was related to uncultured representatives of the Arcobacter group (< or = 95% sequence similarity). A probe designed to target all of the identified lineages hybridized with more than 95% of the mat community. Simultaneous hybridizations with the latter probe and a probe specific to Arcobacter spp. confirmed the numerical dominance of Arcobacter-like bacteria. This study provides the first example of the prevalence and ecological significance of free-living Arcobacter at deep-sea hydrothermal vents.

Mineralization of major lateral teeth in the radula of a deep-sea hydrothermal vent limpet (Gastropoda:Neolepetopsidae)

Radular teeth of a neolepetopsid patellogastropod limpet, Paralepetopsis ferrugivora, from a deep-sea hydrothermal vent site have similar elemental composition (O, Si, P, S, Cl, K and Fe) to radular teeth of the shallow-waters patellogastropod limpets. However, in contrast to shallow waters limpets, the fully mature teeth of P. ferrugivora do not show any crystalline phases. Amorphous silica was found in the cusp of the teeth and amorphous iron oxide in the junction zone and base of the teeth. Ferritin-rich vesicles were observed in cells adjacent to the junction zone of the early mature teeth, suggesting that these vesicles can mediate the delivery of iron to the tooth matrix. The similarity in the elemental composition between the hydrothermal limpet P. ferrugivora and shallow-waters patellogastropod limpets shows that the extreme hydrothermal environment did not alter the elemental composition of the radular teeth in the deep-sea species.

Caminibacter profundus sp. nov., a novel thermophile of Nautiliales ord. nov. within the class 'Epsilonproteobacteria', isolated from a deep-sea hydrothermal vent.

A novel moderately thermophilic, microaerobic to anaerobic, chemolithoautotrophic bacterium, designated strain CRT, was isolated from a deep-sea hydrothermal vent site at 36 degrees N on the Mid-Atlantic Ridge. Cells were Gram-negative, non-motile rods. The organism grew at 45-65 degrees C and pH 6.5-7.4, with optimum growth at 55 degrees C and pH 6.9-7.1. The NaCl range for growth was 5-50 g l(-1) (optimum 30 g l(-1)). Strain CRT was an obligate chemolithoautotroph, growing with H2 as energy source, sulfur, nitrate or oxygen as electron acceptors and CO2 as carbon source. Hydrogen sulfide and ammonium were the respective products of sulfur and nitrate reduction. The G+C content of the genomic DNA was 32.1 mol%. Based on 16S rRNA gene sequence analysis, this organism was most closely related to Caminibacter hydrogeniphilus (94.9% similarity). On the basis of phenotypic and phylogenetic data, it is proposed that the isolate represents a novel species, Caminibacter profundus sp. nov. The type strain is CRT (=DSM 15016T=JCM 11957T). The phylogenetic data also correlate well with the significant phenotypic differences between the lineage encompassing the genera Nautilia and Caminibacter and other members of the class 'Epsilonproteobacteria'. The lineage encompassing the genera Nautilia and Caminibacter is therefore proposed as a new order, Nautiliales ord. nov., represented by a single family, Nautiliaceae fam. nov.

Oceanithermus profundus gen. nov., sp. nov., a thermophilic, microaerophilic, facultatively chemolithoheterotrophic bacterium from a deep-sea hydrothermal vent.

A novel moderately thermophilic, organotrophic, microaerophilic, facultatively chemolithotrophic bacterium, designated strain 506(T), was isolated from a deep-sea hydrothermal vent site at 13 degrees N in the East Pacific Rise. Cells were Gram-negative, non-motile rods. The organism grew in the temperature range 40-68 degrees C, with an optimum at 60 degrees C, and in the pH range 5.5-8.4, with an optimum around pH 7.5. The NaCl concentration for growth was in the range 10-50 g l(-1), with an optimum at 30 g l(-1). Strain 506(T) grew chemoorganoheterotrophically with carbohydrates, proteinaceous substrates, organic acids and alcohols using oxygen or nitrate as electron acceptor. Alternatively, strain 506(T) was able to grow lithoheterotrophically with molecular hydrogen as the energy source. The G +C content of the genomic DNA was 62.9 mol%. Phylogenetic analysis of the 16S rDNA sequence placed strain 506(T) in the family Thermaceae. On the basis of phenotypic and phylogenetic data, strain 506(T) (= DSM 14977(T) = VKM B-2274(T)) is proposed as the type strain of a novel species in a new genus, Oceanithermus profundus gen. nov., sp. nov.

Persephonella marina gen. nov., sp. nov. and Persephonella guaymasensis sp. nov., two novel, thermophilic, hydrogen-oxidizing microaerophiles from deep-sea hydrothermal vents.

Two thermophilic, strictly chemolithoautotrophic, microaerophilic, hydrogen-oxidizing members of the Bacteria designated strain EX-H1T and strain EX-H2T were isolated from two separate deep-sea hydrothermal vent sites at 9 degrees N 104 degrees W in the Pacific Ocean and Guaymas Basin. The motile 2-4-microm-long rods were Gram-negative and non-sporulating. The temperature range for growth was between 55 and 80 degrees C for EX- H1T (optimum at 73 degrees C) and 55-75 C for EX-H2T (optimum at 70 C). Both strains grew fastest at 2.5% (w/v) NaCl and at pH 6, although growth was observed from pH 4.7 to pH 7.5. EX-H1T and EX-H2T were able to use elemental sulfur, thiosulfate or hydrogen as an electron donor, and oxygen (2-3%, v/v) or nitrate as an electron acceptor. EX-H1T was also able to use elemental sulfur as an electron acceptor. EX-H1T and EX-H2T further differed in their genomic G+C content (38.5 and 37.4 mol%, respectively) and 16S rRNA sequences (4% difference). Maximum-likelihood analysis of the 16S rRNA phylogeny placed both isolates within the Aquificales as a distinct lineage and showed them to be only about 85% similar to Aquifex pyrophilus. On the basis of phenotypic and phylogenetic characteristics, it is proposed that EX-H1T and EX-H2T belong to a new genus within the Aquificales, namely Persephonella gen. nov. It is further proposed that EX-H1T be named Persephonella marina sp. nov., the type species of the genus, and that EX-H2T be named Persephonella guaymasensis sp. nov., a second species in this genus.

Characterization of a Novel Spirochete Associated with the Hydrothermal Vent Polychaete Annelid, Alvinella pompejana

A highly integrated, morphologically diverse bacterial community is associated with the dorsal surface of Alvinella pompejana, a polychaetous annelid that inhabits active high-temperature deep-sea hydrothermal vent sites along the East Pacific Rise (EPR). Analysis of a previously prepared bacterial 16S ribosomal DNA (rDNA) library identified a spirochete most closely related to an endosymbiont of the oligochete Olavius loisae. This spirochete phylotype (spirochete A) comprised only 2.2% of the 16S rDNA clone library but appeared to be much more dominant when the same sample was analyzed by denaturing gradient gel electrophoresis (DGGE) and the terminal restriction fragment length polymorphism procedure (12 to 18%). PCR amplification of the community with spirochete-specific primers used in conjunction with DGGE analysis identified two spirochete phylotypes. The first spirochete was identical to spirochete A but was present in only one A. pompejana specimen. The second spirochete (spirochete B) was 84.5% similar to spirochete A and, more interestingly, was present in the epibiont communities of all of the A. pompejana specimens sampled throughout the geographic range of the worm (13 degrees N to 32 degrees S along the EPR). The sequence variation of the spirochete B phylotype was less than 3% for the range of A. pompejana specimens tested, suggesting that a single spirochete species was present in the A. pompejana epibiotic community. Additional analysis of the environments surrounding the worm revealed that spirochetes are a ubiquitous component of high-temperature vents and may play an important role in this unique ecosystem.

New Erebonasteridae (Copepoda) from Vilkitzky Strait in the Arctic and from a Pacific Hydrothermal Vent Site (Northern Fiji Basin)

A new genus of the poecilostomatoid family Erebonasteridae is described from Vilkitzky Strait in the Arctic, the channel connecting the Kara Sea and the Laptev Sea. Within the Erebonasteridae, Nansennaster, new genus, belongs to the Centobnaster-group. The new genus is characterized by the presence of only 2 setae on the mandibular palp, the fusion of the second and third antennal segments, the presence of a single copulatory pore, and the relatively short furca lacking seta I. A previously overlooked sexual dimorphism in Centobnaster is described, the males of this genus having a modified outer spine on the third endopodal segment of leg 3. Amphicrossus tuerkayi, new species, is described from a deep-sea hydrothermal vent site in the northern Fiji basin. The new species has retained some remarkable plesiomorphic characters, such as an incompletely fused genital double-somite in the female, and 2 setae representing the endopod of the maxilla. An identification key to all known genera and species of the Erebonasteridae is provided.