Table of Contents

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

Seawater was collected on the research vessel Fay Slover using 5-liter (L) Niskin bottles along a gradient from the open ocean near the Chesapeake light tower into the Chesapeake Bay. Conductivity, temperature, and depth were measured using a Seabird SBE 32 CTD equipped with a Wetlabs fluorometer and a Wetlabs transmissometer (650 nanoeters (nm)).

Field collection of ATP
Using biological oxygen demand (BOD) bottles, samples of 150 - 300 milliliters (mL) (depending on station) were taken from the Niskin bottles at five stations and six depths along the transect. Water was filtered through 25-millimeter (mm) GF/F filters using large capacity filter funnels at a vacuum of 200 millibars (mbar). Once filtered, and without keeping the filters running dry for more than a few seconds, the filters were quickly transferred into 1.5 mL cryovials filled with 1 mL of a solution of benzalkonium chloride (1% w/w fin. conc.) with phosphoric acid (5% w/w fin. conc.) in a 25 millimolar (mM) Tricine buffer (PBAC, Bochdansky et al. 2021). The filters were extracted in PBAC for 20 to 30 minutes before the cryovials (including the filters) were frozen and kept at -80 degrees Celsius (°C) before analysis. The location and methods were similar to those published in Bochdansky et al. 2024.

Laboratory analysis of ATP
Ten microliters (μL) of each sample (in triplicates) were transferred to 6 mL pony scintillation vials (Research Products International), and received 3 mL of ultrapure water, and 50 μL of CellTiter-Glo 2.0 (Promega Corporation). Internal standards were used by spiking a fourth vial with 50 μL of samples with 50 μL of 0.0164 micromolar (μM) ATP standard. Using internal standards instead of separate calibration curves corrects for matrix effects that change the luminescence signal caused by the presence of ions, acids, and organic material (Bochdansky et al., 2021). Luminescence was analyzed in a PerkinElmer liquid scintillation counter model Tricarb 3110 TR with a single photon counting protocol of 1 minute each. ATP was calculated using equation 2 in Bochdansky et al. (2021).

Particulate organic carbon and nitrogen
Between 250 - 500 mL of seawater (less in the inshore and more in the offshore stations) was filtered onto pre-combusted (450°C, 4 hours) GF/F filters. The filters were stored frozen (-20°C) and later dried at 50°C for approximately 24 hours, then acidified under concentrated HCl fumes in a desiccator, redried for another 24 hours at 50°C and then capped and stored in a desiccator. Filters were rolled in a tin wrap and pressed into pellets to be analyzed in a CHN analyzer (ODU Department of Chemistry Water Quality Laboratory).

ATP samples were analyzed in technical triplicates (three values for each sample) and then averaged. POC and PON values reflect single samples.

- Imported original file "CTD & Niskin bottle data COMBINED NaN 1Apr2024.csv" into the BCO-DMO system.
- Marked "NaN" as a missing data identifier (missing data are empty/blank in the final CSV file).
- Renamed fields to comply with BCO-DMO naming conventions.
- Created date-time fields (local and UTC) in ISO 8601 format.
- Saved the final file as "994268_v1_april_2024_slover_ctd_niskin.csv".

NSF Award Abstract:
In the ocean, most living organisms are microbes that are too small to be seen by the naked eye. Despite their small size, microbes play an important role in processes that govern marine ecosystems and food webs. For example, microbes affect the concentrations of nutrients and gases in the water and the atmosphere, thereby exerting a significant impact on the climate globally. Consequently, it is important to know how many microbes there are in any given environment because there is a direct causal connection between living mass and overall biological activity. Determining how “alive” any volume of water is, however, is a difficult task. The gold standard is to count microbial cells under the microscope. This method is extremely time consuming when done well and needs to be performed separately on many different types of microbial cells. In addition, standard microscopic techniques do not reveal whether the cells were alive when they were collected. In contrast, a chemical method based on the amount of adenosine triphosphate (ATP) offers distinct advantages. Notably, ATP is relatively easy to measure, and the method can be widely used because all living cells contain ATP in similar concentrations. This study tests and applies an improved method of ATP analysis to generate data at very high resolution in space and time. One PhD student and six undergraduate students will receive research training and the project fosters international research collaborations with European scientists. This research provides deeper insights into the distribution of live matter in different regions and depths of the world’s oceans.

Decades ago, adenosine triphosphate (ATP) was proposed as a universal biomass indicator. However, its application in the field of oceanography has been limited due to misconceptions regarding cellular ATP concentration. Recent evidence suggests that ATP functions as a hydrotrope requiring homeostatically controlled ATP levels much higher than those solely needed for energy metabolism. ATP occurs in surprisingly stable concentrations in cytoplasm across a wide range of microbes thus representing live cytoplasm volume. This project examines in detail the usefulness of particulate ATP (PATP) as a biomass marker over a large section of the North Atlantic Ocean with special emphasis on mesopelagic and deep-sea environments where chlorophyll is a poor indicator of biomass or associated biological processes. The project uses field collections of marine snow and ambient water in combination with particle cameras to examine the microscale heterogeny of biomass in the water column. Laboratory studies determine factors that may influence the recovery of PATP through filtration and extraction protocols and determine to what extent ATP concentrations potentially deviate from the typical cytoplasm concentration during phosphorus limitation. The improved PATP-biomass method offers numerous operational advantages, especially the fact that it can be employed at high spatial and temporal resolution. Once validated, the PATP biomass method could be widely adopted as a key variable for biomass in routine oceanographic surveys. This project supports graduate and undergraduate students from diverse backgrounds to contribute to laboratory and field research. Public outreach efforts include tours and presentations for middle and high-school students, as well as the general public.

This project is funded by the Chemical Oceanography and Biological Oceanography Programs in the Division of Ocean Sciences.

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.