Deep-sea Cold Seeps Research Articles

High-resolution isotopic records (δ 18O and δ 13C) and cathodoluminescence study of lucinid shells from methane seeps of the Eastern Mediterranean

We present high-resolution isotopic records and cathodoluminescence studies of recently dead and live bivalve specimens from cold seeps, in an attempt to reconstruct environmental conditions during organism growth, and thereby the possible variability of fluid-venting activity at the seafloor. Shells of the burrowing lucinid Myrtea aff. amorpha were collected at three localities near actively venting methane seeps in the Eastern Mediterranean deep sea, using the Nautile submersible during two French oceanographic cruises: from the Kazan mud volcano, in the vicinity of the Anaximander mounts (MEDINAUT cruise, 1998), and from the central pockmark province and the Amon mud volcano of the Nile deep-sea fan (NAUTINIL cruise, 2003). The oxygen and carbon isotope compositions of 18 shells from the various localities, and also from different sites at the same locality show a rather strong scatter (1.8 < δ18O‰ < 3.4; −10.2 < δ13C‰ < 2.2), and values lower than those expected for carbonate precipitated at equilibrium with present-day bottom waters. This means that warm methane-rich fluids were mixed with bottom seawater during precipitation of shell carbonates. We have tried to determine ontogenetic age of two shells by using cathodoluminescence as a sclerochronological proxy, because the direct counting of carbonate increments was not possible in these specimens. There is a relatively good correspondence between cathodoluminescence trends and oxygen isotope profiles that might support the link between manganese incorporation during growth and temperature. Eight specimens of lucinid shells were selected for high-resolution isotopic profiling. A few shells exhibit decreasing δ18O and δ13C values from the umbo to the actively growing ventral shell margin, which can be attributed to the commonly observed physiologically controlled deceleration of growth with increasing organism age, this metabolic effect corresponding to the increase of incorporation of respiratory CO2. A few shells exhibit high-frequency δ18O variations with an amplitude of about 1.5‰ that might be related to temperature variations controlled by fluid-venting activity. One shell from the pockmark province of the Nile deep-sea fan records a strong, sharp δ13C decrease of about 9‰, and extending over a 5-mm interval in the shell that can be related to a major methane release event. Another shell from the Kazan mud volcano exhibits a progressive increase of δ13C values from −10‰ to 0‰ with age, which might indicate decreasing methane flow throughout the organism’s life. This study has demonstrated that bivalve shells from deep-sea cold seeps represent good indicators of variability in seepage activity of methane-rich fluids, at various scales in both space and time. Although the precise chronology of the observed events was not established, because shell growth rate is not known in this case, this remains a priority for future studies in such environments.

Molecular phylogenetic and chemical analyses of the microbial mats in deep-sea cold seep sediments at the northeastern Japan Sea

Microbial communities inhabiting deep-sea cold seep sediments at the northeastern Japan Sea were characterized by molecular phylogenetic and chemical analyses. White patchy microbial mats were observed along the fault offshore the Hokkaido Island and sediment samples were collected from two stations at the southern foot of the Shiribeshi seamount (M1 site at a depth of 2,961 m on the active fault) and off the Motta Cape site (M2 site at a depth of 3,064 m off the active fault). The phylogenetic and terminal-restriction fragment polymorphism analyses of PCR-amplified 16S rRNA genes revealed that microbial community structures were different between two sampling stations. The members of ANME-2 archaea and diverse bacterial components including sulfate reducers within Deltaproteobacteria were detected from M1 site, indicating the occurrence of biologically mediated anaerobic oxidation of methane, while microbial community at M2 site was predominantly composed of members of Marine Crenarchaeota group I, sulfate reducers of Deltaproteobacteria, and sulfur oxidizers of Epsilonproteobacteria. Chemical analyses of seawater above microbial mats suggested that concentrations of sulfate and methane at M1 site were largely decreased relative to those at M2 site and carbon isotopic composition of methane at M1 site shifted heavier ((13)C-enriched), the results of which are consistent with molecular analyses. These results suggest that the mat microbial communities in deep-sea cold seep sediments at the northeastern Japan Sea are significantly responsible for sulfur and carbon circulations and the geological activity associated with plate movements serves unique microbial habitats in deep-sea environments.

Sulfurovum lithotrophicum gen. nov., sp. nov., a novel sulfur-oxidizing chemolithoautotroph within the epsilon-Proteobacteria isolated from Okinawa Trough hydrothermal sediments.

A novel mesophilic sulfur- and thiosulfate-oxidizing bacterium, strain 42BKTT, was isolated from the gas-bubbling sediment at the Iheya North hydrothermal system in the mid-Okinawa Trough, Japan. The isolate was a Gram-negative, non-motile and coccoid to oval-shaped bacterium. Growth was observed at 10-40 degrees C (optimum 28-30 degrees C) and in the pH range 5.0-9.0 (optimum 6.5-7.0). Strain 42BKTT grew chemolithoautotrophically with elemental sulfur or thiosulfate as a sole electron donor and oxygen (optimum 5 % in gas phase) or nitrate as an electron acceptor. The G + C content of the genomic DNA was 48.0 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the isolate belonged to the previously uncultivated Group F within the epsilon-Proteobacteria, which includes phylotypes of vent epibiont and environmental sequences from global deep-sea cold seep and hydrothermal vent fields. On the basis of the physiological and molecular characteristics of this isolate, the type species of a novel genus, Sulfurovum lithotrophicum gen. nov., sp. nov., is proposed. The type strain is 42BKTT (= ATCC BAA-797T = JCM 12117T).

Evolutionary relationships of deep-sea vent and cold seep clams (Mollusca: Vesicomyidae) of the "pacifica/lepta" species complex

Vesicomyid clams are among the dominant invertebrates of chemosynthesis-based communities found at deep-sea cold seeps and hydrothermal vents, yet there is considerable taxonomic confusion within the family Vesicomyidae. The present study examined phylogenetic relationships among vesicomyid clams be- longing to a cryptic-species complex that includes Ve- sicomya (previously Calyptogena) pacifica (Dall, 1891) and Vesicomya lepta (Dall, 1896). Mitochondrial (mt) COI sequences from clams collected between 1991 and 2001 along the western margin of North America from the San Clemente Basin (32� N latitude) to the Juan de Fuca Ridge (48� N latitude) revealed five discrete evolutionary lineages: V. pacifica (mt-type I), V. lepta (mt-type IV), and three undescribed species defined by mttypes II, III, and V. Separation of the three lineages defined by mt-types I, II, and III was also revealed by an independent nuclear gene (ribosomal ITS-1), and by DNA sequences from vertically transmitted bacterial endosymbionts found in the gills of these clams. Lin- eages I, II, and III occur in Monterey Bay, California, and the data suggest that bathymetric segregation may have played a role in their isolation and divergence.

Molecular phylogenetic analyses of reverse-transcribed bacterial rRNA obtained from deep-sea cold seep sediments.

A depth profile of naturally occurring bacterial community structures associated with the deep-sea cold seep push-core sediment in the Japan Trench at a depth of 5343 m were evaluated using molecular phylogenetic analyses of RNA reverse transcription-PCR (RT-PCR) amplified 16S crDNA fragments. A total of 137 clones of bacterial crDNA (complimentary rDNA) phylotypes (phylogenetic types) obtained at three different depths (2-4, 8-10 and 14-16 cm) were identified in partial crDNA sequencings. crDNA phylotypes from the cold seep sediment were dominantly composed of delta- and epsilon-Proteobacteria (36% and 42% respectively). Phylotype analysis of crDNA clone libraries and terminal restriction fragment length polymorphism (T-RFLP) analysis revealed that the majority of bacterial components shifted from delta- Proteobacteria to epsilon-Proteobacteria with increasing depth. Among the delta-proteobacterial crDNA clones, the sequences related to the genus Desulfosarcina were dominant. Almost all sequences of crDNA belonging to epsilon-Proteobacteria were affiliated with the same cluster (epsilon-CSG: epsilon-proteobacterial cold seep group), and were closely related with rDNA sequences from deep-sea hydrothermal vent environments.

Monterey Bay cold seep biota: Euglenozoa with chemoautotrophic bacterial epibionts

Summary Deep-sea cold seeps in Monterey Bay, California, are prominent sites of chemosynthetic production, mediated through bacterial oxidation of sulfur. Bacterial mats and vesicomyid clams rely upon hydrogen-sulfide oxidation. Euglenozoa comprise a large and diverse component of the protistan assemblage associated with the dense chemoautotrophic bacterial mats found in the top 1 cm of sediment at some of these seeps. A high percentage of these euglenozoans have conspicuous coats of rod-shaped bacteria that cover virtually their entire surface. In at least one of the three morphotypes identified that possess these epibionts, the epibiotic bacteria contain membrane bound translucent sulfur vesicles. Total sulfide concentration in the top 1 cm of the sediment ranged from undetectable at the sediment-water interface to a maximum of 350 uM. The presence of other hydrogen sulfide oxidizing bacteria (bacterial mats and endosymbiotic bacteria of the clams), the measurable levels of hydrogen sulfide and the distinct membrane surrounding the sulfur inclusion suggests that the epibiotic bacteria on at least some of the euglenozoans are sulfur oxidizers. This is the first report of flagellates possessing sulfur oxidizing epibiotic bacteria. No evidence has been found that the euglenozoans ingest their epibiotic bacteria, however, and the degree and mode of any nutritional contribution the bacteria might make to the host is unknown.

Characterization of the endosymbiont of a deep-sea bivalve, Calyptogena soyoae

We have purified DNA from gill tissue of a marine bivalve, Calyptogena soyoae, collected from the deep-sea cold seep communities in Sagami Bay, Japan. An rRNA gene was amplified, cloned, and sequenced. In situ hybridization revealed that the sequence is that of a bacterial endosymbiont within the gill of C. soyoae.

A new poecilostomatoid copepod (Erebonasteridae) from deep-sea cold seeps at the West Florida Escarpment

A new species of poecilostomatoid copepod, Erebonaster spinulosus (Erebonasteridae), is described on the basis of a single female from a depth of 3 266 m at cold seeps near the base of the West Florida Escarpment in the eastern Gulf of Mexico. Among its distinctive features are: the large body highly ornamented with setules and spinules, the rostrum with small median teeth and lateral setules, the claw of the second maxilla with a prominent knob on the concave margin, the maxilliped ornamented with setules and its claw bearing a digitiform process near the seta, the third segment of the exopod of leg 4 with the formula II,I,4, and the highly ornamented leg 5 having on the free segment a very long seta 9 times the length of the segment. The genus Erebonaster now contains two species, the previously described Erebonaster protentipes found in 2000 m at the Guaymas Basin, Gulf of California, and the new Erebonaster spinulosus.