Astronomical and atmospheric impacts on deep-sea hydrothermal vent invertebrates.

Ocean tides and winter surface storms are among the main factors driving the dynamics and spatial structure of marine coastal species, but the understanding of their impact on deep-sea and hydrothermal vent communities is still limited. Multidisciplinary deep-sea observatories offer an essential tool to study behavioural rhythms and interactions between hydrothermal community dynamics and environmental fluctuations. Here, we investigated whether species associated with a Ridgeia piscesae tubeworm vent assemblage respond to local ocean dynamics. By tracking variations in vent macrofaunal abundance at different temporal scales, we provide the first evidence that tides and winter surface storms influence the distribution patterns of mobile and non-symbiotic hydrothermal species (i.e. pycnogonids Sericosura sp. and Polynoidae polychaetes) at more than 2 km depth. Local ocean dynamics affected the mixing between hydrothermal fluid inputs and surrounding seawater, modifying the environmental conditions in vent habitats. We suggest that hydrothermal species respond to these habitat modifications by adjusting their behaviour to ensure optimal living conditions. This behaviour may reflect a specific adaptation of vent species to their highly variable habitat.

Fluid chemistry alters faunal trophodynamics but not composition on the deep-sea Capelinhos hydrothermal edifice (Lucky Strike vent field, Mid-Atlantic Ridge).

The recently discovered deep-sea Capelinhos hydrothermal edifice, ~ 1.5 km of the main Lucky Strike (LS) vent field (northern Mid-Atlantic Ridge), contrasts with the other LS edifices in having poorly-altered end-member hydrothermal fluids with low pH and chlorine, and high metal concentrations. Capelinhos unique chemistry and location offer the opportunity to test the effects of local abiotic filters on faunal community structure while avoiding the often-correlated influence of dispersal limitation and depth. In this paper, we characterize for the first time the distribution patterns of the Capelinhos faunal communities, and analyze the benthic invertebrates (> 250 µm) inhabiting diffusive-flow areas and their trophic structures (δ13C, δ15N and δ34S). We hypothesized that faunal communities would differ from those of the nearest LS vent edifices, showing an impoverished species subset due to the potential toxicity of the chemical environment. Conversely, our results show that: (1) community distribution resembles that of other LS edifices, with assemblages visually dominated by shrimps (close to high-temperature focused-fluid areas) and mussels (at low-temperature diffuse flow areas); (2) most species from diffuse flow areas are well-known LS inhabitants, including the bed-forming and chemosymbiotic mussel Bathymodiolus azoricus and (3) communities are as diverse as those of the most diverse LS edifices. On the contrary, stable isotopes suggest different trophodynamics at Capelinhos. The high δ15N and, especially, δ13C and δ34S values suggest an important role of methane oxidation (i.e., methanotrophy), rather than the sulfide oxidation (i.e., thiotrophy) that predominates at most LS edifices. Our results indicate that Capelinhos shows unique environmental conditions, trophic structure and trophodynamics, yet similar fauna, compared to other LS edifices, which suggest a great environmental and trophic plasticity of the vent faunal communities at the LS.

Biological rhythms in the deep-sea hydrothermal mussel Bathymodiolus azoricus.

Biological rhythms are a fundamental property of life. The deep ocean covers 66% of our planet surface and is one of the largest biomes. The deep sea has long been considered as an arrhythmic environment because sunlight is totally absent below 1,000 m depth. In the present study, we have sequenced the temporal transcriptomes of a deep-sea species, the ecosystem-structuring vent mussel Bathymodiolus azoricus. We reveal that tidal cycles predominate in the transcriptome and physiology of mussels fixed directly at hydrothermal vents at 1,688 m depth at the Mid-Atlantic Ridge, whereas daily cycles prevail in mussels sampled after laboratory acclimation. We identify B. azoricus canonical circadian clock genes, and show that oscillations observed in deep-sea mussels could be either a direct response to environmental stimulus, or be driven endogenously by one or more biological clocks. This work generates in situ insights into temporal organisation in a deep-sea organism.

Speciation of dissolved copper within an active hydrothermal edifice on the Lucky Strike vent field (MAR, 37 degrees N).

The objective of this study was to determine the concentrations of different fractions of dissolved copper (after filtration at 0.45 microm) along the cold part of the hydrothermal fluid-seawater mixing zone on the Tour Eiffel edifice (MAR). Dissolved copper was analyzed by stripping chronopotentiometry (SCP) after chromatographic C(18) extraction. Levels of total dissolved copper (0.03 to 5.15 microM) are much higher than those reported for deep-sea oceanic waters but in accordance with data previously obtained in this area. Speciation measurements show that the hydrophobic organic fraction (C(18)Cu) is very low (2+/-1%). Dissolved copper is present mainly as inorganic and hydrophilic organic complexes (nonC(18)Cu). The distribution of copper along the pH gradient shows the same pattern for each fraction. Copper concentrations increase from pH 5.6 to 6.5 and then remain relatively constant at pH>6.5. Concentrations of oxygen and total sulphides demonstrate that the copper anomaly corresponds to the transition between suboxic and oxic waters. The increase of dissolved copper should correspond to the oxidative redissolution of copper sulphide particles formed in the vicinity of the fluid exit. The presence of such a secondary dissolved copper source, associated with the accumulation of metal sulphide particles, could play a significant role in the distribution of fauna in the different habitats available at vents.

Changes of gill and hemocyte-related bio-indicators during long term maintenance of the vent mussel Bathymodiolus azoricus held in aquaria at atmospheric pressure.

The deep-sea hydrothermal vent mussel Bathymodiolus azoricus has been the subject of several studies aimed at understanding the physiological adaptations that vent animals have developed in order to cope with the particular physical and chemical conditions of hydrothermal environments. In spite of reports describing successful procedures to maintain vent mussels under laboratory conditions at atmospheric pressure, few studies have described the mussel's physiological state after a long period in aquaria. In the present study, we investigate changes in mucocytes and hemocytes in B. azoricus over the course of several months after deep-sea retrieval. The visualization of granules of mucopolysaccharide or glycoprotein was made possible through their inherent auto-fluorescent property and the Alcian blue-Periodic Acid Schiff staining method. The density and distribution of droplets of mucus-like granules was observed at the ventral end of lamellae during acclimatization period. The mucus-like granules were greatly reduced after 3 months and nearly absent after 6 months of aquarium conditions. Additionally, we examined the depletion of endosymbiont bacteria from gill tissues, which typically occurs within a few weeks in sea water under laboratory conditions. The physiological state of B. azoricus after 6 months of acclimatization was also examined by means of phagocytosis assays using hemocytes. Hemocytes from mussels held in aquaria up to 6 months were still capable of phagocytosis but to a lesser extent when compared to the number of ingested yeast particles per phagocytic hemocytes from freshly collected vent mussels. We suggest that the changes in gill mucopolysaccharides and hemocyte glycoproteins, the endosymbiont abundance in gill tissues and phagocytosis are useful health criteria to assess long term maintenance of B. azoricus in aquaria. Furthermore, the laboratory set up to which vent mussels were acclimatized is an applicable system to study physiological reactions such as hemocyte immunocompetence even in the absence of the high hydrostatic pressure found at deep-sea vent sites.

Food-Web Complexity in Guaymas Basin Hydrothermal Vents and Cold Seeps.

In the Guaymas Basin, the presence of cold seeps and hydrothermal vents in close proximity, similar sedimentary settings and comparable depths offers a unique opportunity to assess and compare the functioning of these deep-sea chemosynthetic ecosystems. The food webs of five seep and four vent assemblages were studied using stable carbon and nitrogen isotope analyses. Although the two ecosystems shared similar potential basal sources, their food webs differed: seeps relied predominantly on methanotrophy and thiotrophy via the Calvin-Benson-Bassham (CBB) cycle and vents on petroleum-derived organic matter and thiotrophy via the CBB and reductive tricarboxylic acid (rTCA) cycles. In contrast to symbiotic species, the heterotrophic fauna exhibited high trophic flexibility among assemblages, suggesting weak trophic links to the metabolic diversity of chemosynthetic primary producers. At both ecosystems, food webs did not appear to be organised through predator-prey links but rather through weak trophic relationships among co-occurring species. Examples of trophic or spatial niche differentiation highlighted the importance of species-sorting processes within chemosynthetic ecosystems. Variability in food web structure, addressed through Bayesian metrics, revealed consistent trends across ecosystems. Food-web complexity significantly decreased with increasing methane concentrations, a common proxy for the intensity of seep and vent fluid fluxes. Although high fluid-fluxes have the potential to enhance primary productivity, they generate environmental constraints that may limit microbial diversity, colonisation of consumers and the structuring role of competitive interactions, leading to an overall reduction of food-web complexity and an increase in trophic redundancy. Heterogeneity provided by foundation species was identified as an additional structuring factor. According to their biological activities, foundation species may have the potential to partly release the competitive pressure within communities of low fluid-flux habitats. Finally, ecosystem functioning in vents and seeps was highly similar despite environmental differences (e.g. physico-chemistry, dominant basal sources) suggesting that ecological niches are not specifically linked to the nature of fluids. This comparison of seep and vent functioning in the Guaymas basin thus provides further supports to the hypothesis of continuity among deep-sea chemosynthetic ecosystems.

Rhythms and community dynamics of a hydrothermal tubeworm assemblage at main endeavour field - a multidisciplinary deep-sea observatory approach.

The NEPTUNE cabled observatory network hosts an ecological module called TEMPO-mini that focuses on hydrothermal vent ecology and time series, granting us real-time access to data originating from the deep sea. In 2011-2012, during TEMPO-mini's first deployment on the NEPTUNE network, the module recorded high-resolution imagery, temperature, iron (Fe) and oxygen on a hydrothermal assemblage at 2186 m depth at Main Endeavour Field (North East Pacific). 23 days of continuous imagery were analysed with an hourly frequency. Community dynamics were analysed in detail for Ridgeia piscesae tubeworms, Polynoidae, Pycnogonida and Buccinidae, documenting faunal variations, natural change and biotic interactions in the filmed tubeworm assemblage as well as links with the local environment. Semi-diurnal and diurnal periods were identified both in fauna and environment, revealing the influence of tidal cycles. Species interactions were described and distribution patterns were indicative of possible microhabitat preference. The importance of high-resolution frequencies (<1 h) to fully comprehend rhythms in fauna and environment was emphasised, as well as the need for the development of automated or semi-automated imagery analysis tools.

Biological data extraction from imagery - How far can we go? A case study from the Mid-Atlantic Ridge.

In the past few decades, hydrothermal vent research has progressed immensely, resulting in higher-quality samples and long-term studies. With time, scientists are becoming more aware of the impacts of sampling on the faunal communities and are looking for less invasive ways to investigate the vent ecosystems. In this perspective, imagery analysis plays a very important role. With this study, we test which factors can be quantitatively and accurately assessed based on imagery, through comparison with faunal sampling. Twelve instrumented chains were deployed on the Atlantic Eiffel Tower hydrothermal edifice and the corresponding study sites were subsequently sampled. Discrete, quantitative samples were compared to the imagery recorded during the experiment. An observer-effect was tested, by comparing imagery data gathered by different scientists. Most factors based on image analyses concerning Bathymodiolus azoricus mussels were shown to be valid representations of the corresponding samples. Additional ecological assets, based exclusively on imagery, were included.

Diversity and function in microbial mats from the Lucky Strike hydrothermal vent field.

Diversity and function in microbial mats from the Lucky Strike hydrothermal vent field (Mid-Atlantic Ridge) were investigated using molecular approaches. DNA and RNA were extracted from mat samples overlaying hydrothermal deposits and Bathymodiolus azoricus mussel assemblages. We constructed and analyzed libraries of 16S rRNA gene sequences and sequences of functional genes involved in autotrophic carbon fixation [forms I and II RuBisCO (cbbL/M), ATP-citrate lyase B (aclB)]; methane oxidation [particulate methane monooxygenase (pmoA)] and sulfur oxidation [adenosine-5'-phosphosulfate reductase (aprA) and soxB]. To gain new insights into the relationships between mats and mussels, we also used new domain-specific 16S rRNA gene primers targeting Bathymodiolus sp. symbionts. All identified archaeal sequences were affiliated with a single group: the marine group 1 Thaumarchaeota. In contrast, analyses of bacterial sequences revealed much higher diversity, although two phyla Proteobacteria and Bacteroidetes were largely dominant. The 16S rRNA gene sequence library revealed that species affiliated to Beggiatoa Gammaproteobacteria were the dominant active population. Analyses of DNA and RNA functional gene libraries revealed a diverse and active chemolithoautotrophic population. Most of these sequences were affiliated with Gammaproteobacteria, including hydrothermal fauna symbionts, Thiotrichales and Methylococcales. PCR and reverse transcription-PCR using 16S rRNA gene primers targeted to Bathymodiolus sp. symbionts revealed sequences affiliated with both methanotrophic and thiotrophic endosymbionts.