Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • Data Processing Description
  • BCO-DMO Processing Description
  • Problem Description
• Related Publications
• Related Datasets
• Parameters
• Instruments
• Deployments
• Project Information
• Funding

Location: East Pacific Rise 9 N 104 W depth 2500m

Collection at the seafloor:

Sampling was conducted near 9°47’N on the East Pacific Rise (EPR) on dives with the Human Occupied Vehicle (HOV) Alvin during R/V Atlantis cruise AT50-06 (cruise DOI: 10.7284/909880); this dataset includes collections from HOV Alvin Dives 5134 and 5135 at Lucky’s Mound on 18 and 19 December 2022 and from Dive 5142 at Sentry Spire on 27 December 2022. This dataset also includes one sampling event from the Remotely Operated Vehicle (ROV) Jason II Dive 1311 at Sentry Spire on 9 April 2021 during R/V Revelle cruise RR2102 (cruise DOI: 10.7284/909120). Sampling events are specified in the dataset as “rock grab” or “slurp.” For rock grabs - the manipulators were used to break off a rock then place it into an individual compartment of a closable, vehicle-mounted container (biobox). For slurps - the substratum (but not the rock targeted for sampling) was vacuumed with a five-chambered hydraulic slurp sampler. The sampling event log was associated by time to renavigated vehicle data including latitude, longitude, and depth (for AT50-06 used renavigated vehicle data version June 2023). We thank the captain, crew, Alvin and Jason teams, and scientists on-board both cruises for enabling sample collection.

Shipboard sample processing:

After recovery on deck, animals were either rinsed or gently scraped off the surfaces of sulfides or siphoned out of the biobox compartment for the collected rock. Siphoned biobox water and slurps were poured over a sieve (60 micrometer on AT50-06). The animal samples were placed into 4°C filtered seawater, sorted in petri dishes to high rank taxonomic groups (e.g., gastropods), and placed into 2.5-mL cryotubes. On cruise AT50-06 gastropods were either preserved in 80% ethanol and stored at -20°C or fixed in 10% buffered formalin; the formalin material was transferred to 80% ethanol after 1-2 days, refreshed with 80% ethanol after 1 day, and again after 1 day, then accidentally placed in a -80°C freezer on-board the research vessel. On cruise RR2102 gastropods were preserved in 95% ethanol and stored at room temperature.

Laboratory sorting and morphological identification:

Cryotubes containing gastropod specimens from a total of 8 rock grabs and 2 slurps from cruise AT50-06 were thawed and poured into a dish for sorting by morphotype. A subset of sorted specimens was photographed and placed into new 1-mL tubes [recorded as “samples and images” metadata tables for the sorted tubes (1 for ethanol, 1 for formalin)]. The new tubes were shipped at room temperature from WHOI to JAMSTEC.

Specimens were identified morphologically, recorded into ID tables for cruise AT50-06 (1 for ethanol, 1 for formalin); specimens from 1 rock grab also were identified for cruise RR2102. Gastropod specimens were gently cleaned using small brushes, and observed under a stereo dissecting microscope (Olympus SZX9). To obtain the operculum and radula for scanning electron microscopy, dissections were done under the microscope using fine forceps and tungsten needles. As listed in the compiled data table column otherCatalogNumbers, a subset of specimens was deposited in Senckenberg Natural History Museum, Frankfurt (SMF), National Museum of Nature and Science, Tsukuba (NSMT), or Muséum National d’Histoire Naturelle, Paris (MNHN). For more information about gastropod morphological identification, please see Chen et al. (2024).

Genetic evidence for identification:

DNA was extracted from one whole specimen of each of the three Melanodrymia species using the DNeasy Blood & Tissue kit (Qiagen) following the manufacturer’s protocols. The same specimen's DNA extraction was used for both mitochondrial COI and mitogenome sequencing, with sequences deposited in National Center for Biotechnology Information (NCBI) GenBank under accession numbers OR828527-OR828529 (Chen et al. 2024) and under BioProject number PRJNA1048888 (Zhang et al. 2024), respectively.

Dataset compilation:

The compiled data table is a single table formatted as a Darwin Core Occurrence table. The table was created using a script to join the three types of input data files for cruise AT50-06 [sampling event log, sorted tube tables, and ID tables), standardize taxa to the World Register of Marine Species (WoRMS), and add a few Darwin Core terms required by the Ocean Biodiversity Information System (OBIS). The script AT50-06_snails_compilation.R is attached to this dataset as a supplemental file (the author contributed this file from https://github.com/sbeaulieu/EPR-inactive/blob/main/AT50-06_snails_compilation.R (access date 2024-10-28, commit:af07a9e). One row in the data table was manually added for the one sampling event included from cruise RR2102. A Darwin Core Event table can be created from the right-most columns of the compiled data table. 

* Data from source file gastropods_inactive_AT50-06_RR2102_BCO-DMO_20241022.csv were imported into the BCO-DMO data system as the primary data table for the dataset.

* character "é" in dataset values replaced with "e" to comply with BCO-DMO conventions.
* The data submitter indicated "The script is available online at https://github.com/sbeaulieu/EPR-inactive/blob/main/AT50-06_snails_compi... (access date 2024-10-28). " and that they would prefer it be attached to the BCO-DMO dataset as a supplemental file.
The corresponding commit SHA:af07a9e for AT50-06_snails_compilation.R at that time (access date 2024-10-28) was downloaded from the github repository and attached a supplemental file.

NSF Award Abstract:
Over four decades of research have shown that tiny free-swimming offspring of the unique inhabitants of hydrothermal vents can disperse effectively between their specialized habitats. Yet, we know almost nothing about how these larval animals complete the journey by locating and settling down in suitable locations. This question remains one of the key unresolved puzzles in the ecology of the deep sea and is becoming increasingly important to solve as hydrothermal vents are becoming threatened by human impacts. The investigators suggest that the films of bacteria that first form at vents are good signposts for settlement of larvae because they indicate that the hydrothermal vents are suitable for life. This project uses a combined program of field experiments, cutting-edge molecular biology techniques, and shipboard experiments with hydrothermal-vent larvae and cultured bacterial films. The project also connects undergraduate research interns at a primarily undergraduate institution (Western Washington University) with undergraduate research interns at two research institutions (Rutgers and Woods Hole Oceanographic Institution) while working on the project at sea together. Finally, the team is producing a science-in-action documentary filled with ocean science and exploration intended for television distribution and museum screenings. The investigators are using footage of the deep-sea vents, shipboard and diving operations, and laboratory work to create a documentary that highlights the foundation of scientific research—hypothesis-driven research, the application of the scientific method, and the importance of critical thinking—all in the framework of the study of an exciting, but threatened, ecosystem.

Hydrothermal vents are particularly tractable systems in which to study questions about the roles of biofilms in larval settlement because biofilms at vents are relatively low-complexity; vent animals are strictly dependent on vent microbes, often through symbiotic partnerships acquired after settlement; and environmental variations are present within the range of a common larval pool. Moreover, decades of research on settlement in model organisms give us good insight into biofilm cues; there is solid foundational understanding about colonization patterns at vents; we now have excellent tools to collect, identify, and culture vent larvae and microbes; and modern environmental "-omics" techniques are a good tool to characterize biological cues produced by biofilms. The project provides an unprecedented, quantitative look into the role of microbial biofilms in structuring larval settlement at hydrothermal vents, achieved only through the close collaboration of microbial and larval ecologists. The combined field program of short-term settlement experiments, microbial "-omics" work, and subsequent shipboard settlement experiments allows the investigative team to use field experiments to statistically model the factors that best predict larval settlement in the field, then test those predictions with shipboard experiments that decouple covarying conditions. This extensive characterization of putative larval settlement cues and their relationship to colonization success in heterogeneous vent habitat niches will contribute to a broader understanding of colonization success across diverse marine ecosystems. Understanding the role that the initial settlement of larvae plays in the recovery and resilience of hydrothermal-vent ecosystems is critical to developing informed management plans for deep-sea mining.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Award Abstract:
This project is investigating a newly discovered community of animals and microbes near deep-sea hydrothermal vents that appears to inhabit only cool, inactive sulfide features. The main objectives are to determine what species live on these features, whether they are new to science, and how they function in the community. The discovery of this novel community, which may be fueled by production of resident microbes, is likely to change the way we think about inactive vents and their contribution to deep-sea biodiversity and productivity. This project has broad impact in four different areas: 1) Informing policy for sustainable use (mining) of inactive sulfides; 2) Contributing to global data systems and the NSF-funded repository at BCO-DMO to make our data available for research use at other temporal, spatial, and taxonomic scales; 3) Increasing public scientific literacy by enhancing K-12 education in the sciences at Memorial Junior High in Eagle Pass TX with about 98% Hispanic and 2% Native American students and a high number of English Language Learners and migrants; and 4) Developing a diverse workforce by engaging students from under-represented and marginalized groups into undergraduate intern programs.

Hydrothermal venting of heated, reduced fluids from the seafloor occurs globally at plate tectonic boundaries and mid-plate hotspots and has been the subject of vigorous geological, chemical and biological research. However, this venting is ultimately transient, leaving behind only the sulfide mineral-rich deposits after the fluid flow stops. This project investigates the organisms living on these lesser studied inactive sulfide features in order to understand their ecology and associations with the mineral substratum. Recent discoveries indicate that some microbial and animal species inhabiting inactive sulfides are not found elsewhere in the marine environment, suggesting the sulfides serve as a unique habitat, distinct from other seafloor topographic features. The main project objectives are to characterize the species and functional diversity of the inactive sulfide ecosystem across all three domains of life (eukaryotic, bacterial, and archaeal), determine which animal species are endemic or predominantly associated with inactive sulfides, and explore the biological and geological characteristics governing those associations. The investigators are conducting field studies between 9-10 degrees N on the East Pacific Rise at sites within the axial summit trough as well as at recently discovered off-axis sites away from modern day venting features. The discovery of this novel community of organisms inhabiting inactive sulfide features at hydrothermal vent fields, fueled by resident chemolithotrophic microorganisms, is likely to change the way we think about the role of these ecosystems in deep-sea biodiversity and productivity.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.