Table of Contents

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

Community samples were collected over a period of four days board the R/V Atlantic Explorer, during cruises AE2412 and AE2427 in April and November 2024. 

Replicate 250 mL polycarbonate bottles were filled with 200 mL of surface seawater collected in the Sargasso Sea. Each bottle received either 100 ul of 1x PBS buffer (control), 0.01% glucose (final w/v), or purified extracellular vesicles (approximately 1e9 EVs/mL final concentration). Replicates were placed in a deckboard flow-through incubator and sampled after 0, 24, 48, and 72 hours. The microbial community was sampled by filtering water through a 47mm 0.2 µm Supor filter and frozen at -80 C.  DNA was extracted using a phenol-chloroform method utilizing AMPureXP beads (Biller et al 2018). 16S amplicon generation (V4-V5 primers) and sequencing on an Illumina MiSeq was conduced at the Integrated Microbiome Resource, Dalhousie University. 

- Loaded TSV file "SRA_metadata_012628.tsv" using filename as resource name; treated empty strings and "nd" as missing values; headers from row 1
- Removed quotation marks from the title field using find/replace
- Reordered fields to: study, bioproject_accession, biosample_accession, accession, sample_name, library_ID, library_strategy, library_source, library_selection, library_layout, title, platform, instrument_model
- Output written to "997617_v1_sra_microbial_community_composition.csv"

NSF Award Abstract:
Microbial production and consumption of organic carbon play critical roles in the marine food web and global carbon cycling. Bacteria release organic matter in a variety of chemical forms and in diverse contexts, ranging from individual molecules to small aggregates and larger biological particles. In recent years we have come to understand that most, if not all, marine microbes release nanoscale structures called extracellular vesicles from their surfaces. These discrete particles, which are abundant in the oceans, are capable of transporting multiple classes of organic molecules between organisms and can serve as a potential nutrient source for other microbes. Extracellular vesicles thus represent a potentially important component of marine microbial food webs, but the magnitude and dynamics of this contribution are unknown. Further, the packaging of material within vesicles may influence the accessibility of this organic material as compared with truly ‘dissolved’ substances to different groups of marine organisms, potentially biasing nutrient exchanges. Broader impacts of this work is providing hands-on research experiences for female undergraduate students - including those from groups historically underrepresented in STEM fields - and training in data analysis tools.

The goal of this project is to advance the understanding of the role that extracellular vesicles play in marine dissolved organic carbon pools and microbial food webs. To determine the contribution of vesicles to organic matter release by marine microbes, the investigators are quantifying the fraction of excreted carbon and nitrogen associated with vesicles released by multiple marine cyanobacteria and heterotrophs. The project is examining how vesicles are ‘consumed’ by heterotrophs to calculate a mass balance of vesicle utilization and produce detailed gene expression data to explore how cells respond to the presence of vesicles. Finally, experiments with coastal and oligotrophic marine communities are providing insights into which organisms utilize vesicles in the field, and whether they are broadly accessible to all microbes or are instead preferentially consumed by a subset of microbes. Collectively, these experiments are opening up a new area of research into the mechanisms underlying the microbial loop and provide foundational insights into the roles of extracellular vesicles in ocean ecosystems.

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.