Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • Data Processing Description
• Data Files
• Related Publications
• Related Datasets
• Parameters
• Instruments
• Deployments
• Project Information
• Funding

This dataset provides the sampling locations and identifications for larvae collected near three deep-sea hydrothermal vent fields and used in a microbiome study by Carrier et al. (2021). This dataset provides metadata for larval and microbiome genetic sequence data in another repository (Carrier et al., 2021, doi.org/10.5061/dryad.sqv9s4n18). The data table is structured as a Darwin Core occurrence table so that it can be harvested by the Ocean Biodiversity Information System (OBIS) and Global Biodiversity Information Facility (GBIF). In addition to the data table, photographs are provided for some of the specimens. A .zip file of images is attached containing the .JPG files referenced by column "associatedMedia".

Larvae were collected near three different hydrothermal vent fields with locality, Location, and waterBody (N. Pacific) specified in the data: East Pacific Rise, 9 50 N vent field, Tica vent site at N EPR; Mariana Back-Arc, Snail and Archaean vent fields in Mariana Trough; and Pescadero Basin, Auka vent field in Gulf of California.

Larval collection:
These larvae were collected near hydrothermal vent fields on the East Pacific Rise (EPR 9°50'N vent field), on the Mariana Back-Arc Spreading Center (near Snail or Archaean vent fields), and in Pescadero Basin (Auka vent field) in the Gulf of California. The EPR specimens were from McLane WTS-LV50 plankton pumps deployed on-axis near Tica vent site on LADDER 3 cruise AT15-26 in 2007. The pump sampled for 24 hr over a 63 micron mesh at a rate of 30 L/min at a position about 3 meters above bottom. The Mariana specimens were from the same plankton pumps with same configuration deployed on-site or off-site but within 300 m of active hydrothermal vents on R/V Yokosuka cruise YK10-11 in 2010. The Pescadero specimens were from the suction (slurp) sampler on ROV Hercules on E/V Nautilus cruise NA091 in 2017. The slurp sampled for 10 min over a 63 micron mesh at a rate of ~100 L/min at a position about 1 meter above bottom, while the ROV was parked on-site at an active vent ("intensive sampling station", or ISS). ISS 2 was during ROV dive H1657, and ISS 4 was during ROV dive H1658.

We acknowledge the sample collection permits CONAPESCA PPFE/DGOPA-010/17 and INEGI: Autorización EG0072017 associated to the Diplomatic Note number SRE 17-1087 (CTC/06727/17).

Shipboard sample processing:
Pump and slurp samples were processed within an hour upon recovery on deck, with many specimens alive. The sample was washed off the filter using 95% non-denatured ethanol into a 250 mL jar (brand new or rinsed with ethanol, but was not autoclaved).

Laboratory sorting and morphological identification:
Portions of the sample were extracted with a wide-mouthed pipette into a petri dish for sorting under a dissecting scope at magnifications up to 50X (although most tools were rinsed with ethanol, none were autoclaved). Specimens were sorted into vials by major taxa (i.e., all the gastropod larvae together), in some cases with separate vials for those retained on 300um or 63um sieve. Vials were brand new or rinsed with ethanol, but were not autoclaved. Individuals were identified to morphotypes at lowest taxonomic level. Some morphotypes contain morphologically indistinguishable larvae from multiple species (e.g., polychaete nectochaetes).

Specimens selected for DNA extraction:
We selected specimens in several major taxonomic groups (gastropods, polychaetes, crustaceans, and a bivalve) for DNA extraction. The amount of time the larval specimens had been stored in ethanol at the time of DNA extraction was ~10, ~7, and ~1 yrs, respectively, for EPR, Mariana, and Pescadero specimens. Only the Mariana specimens had been stored at 4 degrees C, while the others were at room temperature. Working in a laminar flow hood, we used sterile techniques to place 25 individuals from EPR and 25 individuals from Mariana into separate 1.5 mL vials with 95% ethanol. We thank Diana Franks for helping with sterile techniques. Pescadero specimens were provided in 2 vials each with 5 individuals of the same morphotype using sterile tools but not in a laminar flow hood.

DNA extraction and amplification are described by Carrier et al. (2021), and 28S rRNA larval genetic sequences and 16S rRNA bacterial genetic sequence data are in the Dryad repository (Carrier et al., 2021, doi.org/10.5061/dryad.sqv9s4n18). The 28S larval sequences were compared to those in the NCBI GenBank database using a BLAST search to assist with matching morphotypes to lowest taxonomic level. Results from the BLAST search were insufficient to provide additional taxonomic resolution; therefore, the 28S genetic results were primarily used as support for the morphological identification. We'd like to thank Bethany Fleming for conducting the BLAST searches.

Additional genetic evidence to assist with larval identification:
We'd like to thank Dr. Hiroka Hidaka, Dr. Shigeaki Kojima, and Dr. Hiromi Watanabe for providing histone 3 and Dr. Florence Pradillon for providing cytochrome oxidase subunit 1 (CO1) sequences for other Mariana larval specimens from the same collections. The Canadian Centre for DNA Barcoding provided CO1 sequences for other Pescadero specimens from the same collections; these results may be viewed in the Barcode of Life Data System (BOLD) Public Data Portal using the project search PESPL.

Data Processing:
The raw data table was created in a Google spreadsheet which was imported directly into a script that removed extraneous columns, added an asterisk to 6 of the TubeIDs, constrained the latitude and longitude to 4 decimal places, and exported the table to a comma-separated values file: https://github.com/sbeaulieu/EPR-traits/blob/master/subset_Google_Sheet_BCODMO_Beaulieu_Carrier.R. The data table is structured as a Darwin Core occurrence table so that it can be harvested by the Ocean Biodiversity Information System (OBIS) and Global Biodiversity Information Facility (GBIF). Note for the column linking to the microbiome genetic sequence data in the Dryad repository, we considered using the Darwin Core term associatedOrganisms but found other studies in which the link to microbiome data was made using an Occurrence Core with an extension for ResourceRelationships.

NSF Award Abstract:
Summary: Since the discovery of deep-sea hydrothermal vents over thirty years ago, scientists have been perplexed by the question: How are these vent sites colonized and, more specifically, How are the faunal populations established and maintained at these very discrete and often ephemeral habitats. For animals that are sessile or have limited mobility as adults, dispersal to these habitats occurs early in the life cycle, as planktonic larvae in the water column. Due to the difficulties in sampling deep-sea larvae, including low abundances (dilute concentrations), we have very few quantitative estimates of larval dispersal between or larval supply to hydrothermal vents. We also have little to no knowledge of the behavior of vent larvae. The PIs will use large-volume plankton pumps to collect larvae near vents in the southern Mariana Trough in a collaborative effort to quantify larval abundance, behavior, and dispersal in this little-studied region. The collaboration combines the PI's strengths in the collection and morphological identification of larvae and quantifying and modeling dispersal between deep-sea vents, and those of Japanese partners in rearing larvae of hydrothermal vent fauna, molecular genetic identification of larvae, and population genetics of vent fauna.

Intellectual merit: The southern Mariana Trough is a very interesting region in which to study dispersal of vent-endemic fauna, due to the proximity of vents in the back-arc spreading center to vents along the Mariana Arc. These two tectonic settings create different habitat conditions and support vent communities with different species composition. Vent sites the PIs will visit, in the axis and just off-axis of the back-arc spreading center are as close as 25 km to vents on the arc, yet 600 km south of the other known vents in the back-arc. In addition to the new information on larval abundance, diversity, behavior, and dispersal that will be gained for this little-studied region of the world's ridge system, this project has direct relevance to the integration and synthesis goals of the U.S. Ridge 2000 Program. The PI's lab group has conducted previous work at the Ridge 2000 East Pacific Rise (EPR) Integrated Studies Site (ISS). They will be making a direct comparison of the larval abundance and diversity at the EPR ISS to this very different setting along the global 'baseball seam' of oceanic spreading centers. No other such comparison has been possible due to the lack of sampling effort for larvae with large-volume pumps. Also, they are proposing the first experiments with live vent larvae (to the best of our knowledge - with the exception of brachyuran megalopae at 1 atm) to estimate swimming and sinking rates that are important for adding behavioral information to models of larval dispersal.

Broader Impacts: The project involves reciprocal training and cultural exchange - the PIs will learn field and laboratory research techniques from the Japanese PIs, and they will learn from the U.S. PIs. The project will also benefit the career development of a junior researcher (Beaulieu). The proposed activity broadens the participation of both U.S. and Japanese women scientists in sea-going, oceanographic research. The PIs will broadcast the cruise activities in a web log posted by the international InterRidge Program Office, and they anticipate at least three scientific publications will emerge. New species will be added to the online photographic identification guide for vent larvae and included in the second edition of the printed guide.

Additional cruise data and information are available from MGDS: http://www.marine-geo.org/tools/search/entry.php?id=YK10-11

NSF Award Abstract:
Hydrothermal vents support oases of life in the deep sea and are inhabited by unusual organisms that use chemical energy instead of photosynthesis as the basis of their food web. However, because the vents occur in geologically active areas of the seafloor, entire communities can be eradicated by catastrophic natural disturbances such as eruptions. The main objectives of this project are to quantify how quickly these communities recover from catastrophic disturbance and to determine what processes influence their resilience. The project focuses on both the structure (species diversity) and function (trait diversity) of the communities. The investigators will examine vents on an active segment of the East Pacific Rise where eruptive disturbance occurs on decadal time scales. These activities will create an unprecedented long-term (>14-year) quantitative time-series of colonist species composition and function. The application of trait-based analysis to the question of biological succession at vents has the potential to change the way we think about resilience in other patchy, transient and regionally-connected ecosystems. By considering how traits change over time, the researchers can untangle which species-level characteristics most influence abundance and distribution. The project objectives have broad significance with the growing potential for human-caused disturbances at deep-sea vents through deep-sea mining. Additional impacts include strengthening participation of under-represented minorities in marine science and contributing to international database development for functional traits of deep-sea vent species.

The unique, chemosynthesis-fueled fauna inhabiting deep-sea hydrothermal vents are subject to tectonic and eruptive disturbance that can eradicate entire communities. The main objectives of this project are to quantify how quickly these communities recover from catastrophic disturbance and to determine what processes influence their resilience. The focus is on vents on an active segment of the East Pacific Rise where eruptive disturbance occurs on decadal time scales. Field data on colonization and larval supply are used to characterize not only species succession but also the trajectory of functional diversity after a recent (2006) eruption. A new, promising approach to the colonization studies comes from incorporating trait-based analysis of functional diversity. Functional trait analysis is increasingly recognized in terrestrial and freshwater systems as a tool to holistically answer ecological questions, but trait analysis has not been often applied to marine systems. By considering how traits of incoming colonists change over time, the investigators can untangle which species-level factors most influence abundance and distribution. This project will create an unprecedented long-term (>14-year) quantitative time-series of colonist species composition and function. It includes multiple vent sites to encompass the full diversity of habitat conditions, and assesses both local processes and regional connectivity through larval supply. Field observations at individual sites contribute to broader questions when placed in the context of metacommunity theory. In this theoretical framework, field data such as this can be used to answer such questions as how the eradication of the vent community at a particular site affects the persistence of the metacommunity overall, and which vent sites contribute most to regional biodiversity.

NSF Award Abstract:
In this project, the researchers will develop new mathematical models to study the population dynamics of organisms that live at deep-sea hydrothermal vents, areas of the seafloor where volcanic activity causes hot, chemical-rich fluids to exit. The discovery of these vents in 1977 revealed unexpectedly novel and diverse organisms, challenging the prevailing view of the deep sea as a sparsely populated desert. Recent international efforts to mine hydrothermal vent deposits rich in copper, gold, silver and zinc are intensely debated, as deep-sea mineral mining can destroy diverse vent communities and alter the surrounding seafloor habitat. The investigators will extend their models to analyze the potential effects of mining activities. Results from new models will be synthesized to meet the needs of potential stakeholders, including organizations that advise, manage, and conduct activities related to seafloor mining and Marine Protected Areas. Research products will be disseminated as reports and in stakeholder meetings such as those organized by the International Seabed Authority and the Deep Ocean Stewardship Initiative. The research team will work with graphic artists, video producers, and educators to develop new educational presentations for the NOAA Science on a Sphere® (SOS) system. This new content will be distributed via open access to the entire SOS Users Network and incorporated into SOS programs at over 100 science centers across the U.S. and in 20 other countries. Undergraduate students will participate in the project through the Woods Hole Oceanographic Institution's Summer Student Fellowship Program and the Woods Hole Partnership Education Program. These two programs provide students with authentic research experiences; the PEP program attracts underrepresented minority students, and offers them a short course in marine science and a research internship along with a 6-wk research project.

Metacommunity theory offers important advantages over alternative approaches in modeling vent ecosystems, and the proposed work will advance both our understanding of these communities and strategies for developing models of metacommunities more generally. The proposed work substantially expands earlier metacommunity models for vent systems developed by the researchers in innovative ways. The analyses will remove many initial constraints and add important considerations including site-dependent transition probabilities and clustering of nearby sites with shared characteristics. The options for modeling dispersal properties with two alternative dispersal kernels will also advance understanding. The researchers will examine recognized successional patterns not considered in the previous work using patch occupancy models, and they will carry out sensitivity analyses to evaluate the role of parameters for which uncertainty is high, such as larval duration and dispersal distance, patch disturbance rates and recovery times. This element of the work plan will serve to prioritize future field research, emphasizing the role that models can play in guiding research programs.