Table of Contents

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

Pushcore, rocks, biotubes, and slurp samples were collected with HOV Alvin using the manipulator. Rocks were placed into individual compartments in a biobox on the Alvin basket. Before preserving, samples were kept cold and animals were handpicked to sample tissue for stable isotope analyses. The tissue (aprox. 0.5-2.5 mg) was placed in pre-weighted tin boats or combusted vials (500C overnight) and frozen at -70C.

In addition to natural abundance analyses, some animals were placed into mylar bags with addition of labeled C and N and protected from oxygen and light for incubation experiments. The bags were kept sealed in the ship's cold room for 3-8 days, and animals were then processed following the same method as for natural abundance samples.

 Surface and bottom seawater was collected using a CTD/Rosette system and filtered through microglass fiber filters. Filters were frozen for further processing in the laboratory.

In the laboratory, tissue samples were oven dried (60C) overnight, weighed and acidified with 12.5-25 ul 1N H3PO4 to remove inorganic C. Stable isotopes measurements (d13C and d15N) were carried out using an isotope ratio mass spectrometer at the Stable Isotope Facility at University of California, Davis. Microglass fiber filters were oven dried (60C) overnight, and the material on the filters was scrapped off and place into pre-weighed tin boats. Samples were then processed following the same method as for tissue samples.

- Imported "AT50-24_Isotope data_BCO-DMO.xlsx" into the BCO-DMO system without formatting
- Applied the rounding formats that existed in the Excel document
- Converted the "Date (2024)" field to YYYY-MM-DD ISO 8601 format and renamed the field to "Date"
- Removed text "open all dive" and "not logged" from the navigation fields and put them into a new "Navigation_Comment" field
- Removed "not an animal" value from the from the dataset
- Corrected "-118.0000333" with "-148.00003" in the "Longitude" parameter, upon request of submitter
- Renamed fields to remove spaces and special characters to conform to BCO-DMO naming conventions
- Exported file as "984699_v1_at50-24_isotope_data.csv"

Aphia IDs were checked and provided by submitter upon submission (2025-07-01).

NSF Award Abstract:
This research examines the role of deep-sea organisms in determining the fate and footprint of methane, a potent greenhouse gas, on Pacific continental margins. The investigators are evaluating the deep ocean methanosphere defined by the microbial communities that consume methane and the animals that directly feed on or form symbioses with methane-consuming microbes. They are also investigating animal communities that gain energy indirectly from methane, as well as those that take advantage of carbonate rocks, the physical manifestation of methane consumption in seafloor sediments. The study of methane seeps in the deep waters of both Alaska (4400-5500 meters) and Southern California (450-1040 meters) is enabling comparisons of the methanosphere under different food-limitation and oxygen regimes. By applying diverse chemical, isotopic, microscopy, and genetic-based analyses to seep microbes and fauna, this study is advancing understanding of the contribution of methane to deep-sea biodiversity and ecosystem function, information that can inform management and conservation actions in US waters. In addition to training for graduate and undergraduate students at their home institutions, the investigators are collaborating with the Alaska Native Science and Engineering Program (ANSEP). They are recruiting Alaskan undergraduates to participate in the research, contributing to ANSEP's online resources that promote interaction between scientists and middle and high school students, and participating in ANSEP's annual residential Career Exploration in Marine Science programs to engage middle school students in learning about deep-sea ecosystems and the variety of career pathways available in marine related fields.

Microbial production and consumption of methane is dynamic and widespread along continental margins, and some animals within deep-sea methane seeps rely on the oxidation and sequestration of methane for nutrition. At the same time, understanding of methane-dependent processes and symbioses in the deep-sea environment is still rudimentary. The goals of this study are to 1) examine the diversity of animals involved in methane-based symbioses and heterotrophic consumption of methane-oxidizing microbes and how these symbioses extend the periphery of seeps, contributing to non-seep, continental slope food webs; and 2) determine whether carbonates on the seep periphery sustain active methanotrophic microbial assemblages, providing a localized food source or chemical fuel for thiotrophic symbioses, via anaerobic oxidation of methane, or free-living, sulfide-oxidizing bacteria consumed by animals. The investigators are addressing these goals by surveying, sampling, and characterizing microbes, water, sediments, carbonates and animals at a deep seep site on the Aleutian Margin and a shallow site off Southern California. Shipboard experiments and laboratory analyses are using molecular, isotopic, geochemical, and radiotracer tools to understand transfer of methane-sourced carbon from aerobic methanotrophs under multiple oxygen levels, pressures, and photosynthetic food inputs. This approach offers a wide lens by which to examine the methane seep footprint, allow reinterpretation of past observations, and identify new scientific areas for future study. Improved characterization of the deep continental margin methanosphere informs climate science, biodiversity conservation, and resource management.