Dataset: Adenosine triphosphate and microbial biomass measurements from the Chesapeake Bay, sampled aboard RV Fay Slover on April 11, 2018

ValidatedFinal no updates expectedDOI: 10.26008/1912/bco-dmo.939970.1Version 1 (2024-12-30)Dataset Type:Other Field Results

Principal Investigator, Contact: Alexander B. Bochdansky (Old Dominion University)

BCO-DMO Data Manager: Karen Soenen (Woods Hole Oceanographic Institution)


Project: Adenosine triphosphate as a master variable for biomass in the oceanographic context (ATP biomass indicator)


Abstract

This dataset contains values from instruments and Niskin bottle samples from a research expedition on the RV Slover on April 11, 2018. Six stations were sampled with five bottle samples each from the coastal ocean to tributaries of the Chesapeake Bay. The variables are latitude, longitude, station number, depth, sigma-t, beam attenuation, relative chlorophyll fluorescence, salinity, temperature, total ATP (dissolved and particulate), carbon of prokaryotic microbes (microscope estimates), and car...

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Field measurements and bottle collections: Seawater was collected on the research vessel Fay Slover using 5 L Niskin bottles along a gradient from the open ocean near the Chesapeake light tower into the Chesapeake Bay and the James River, in April 2018 and June 2019. Conductivity, temperature, and depth were measured using a SBE 32 CTD equipped with a Wetlabs fluorometer and a Wetlabs transmissometer (650 nm).

Field collection of ATP:  ATP samples were processed using the hot-water extraction method described by Bochdansky et al. (2021). Three types of ATP samples were collected: 1) pATP in 5 ml seawater filtered through 0.2 mm pore size polycarbonate filters (Isopore GTTP, 25 mm diam.), 2) dissolved ATP (ATP in the filtrate of 0.2 mm filtration), and 3) 0.5 ml ATP in whole water without filtration. Polycarbonate filters were chosen over GF/F filters because of their higher retention efficiency (Taguchi and Laws, 1988; Lee et al., 1995) and because the volumes filtered were small, to ensure filtration times were kept to a minimum even with the use of the smaller pore size. Five milliliters of each depth were immediately and simultaneously filtered using a filtration manifold (25 mm-diameter filters, stainless steel screen supports) preloaded with the polycarbonate filters. The filters were placed in 15 ml polypropylene centrifuge tubes (FalconTM) within seconds after the water passed. The dissolved ATP was captured in 15 ml polypropylene centrifuge vials placed underneath the filtration manifold and inside the vacuum flasks. The dissolved ATP samples (i.e., the 0.2 mm filtrates) were immersed in a boiling water bath for ~15 minutes immediately after filtration to sterilize the water and inactivate ATPases and then frozen at -80 °C until analysis.

In April 2018, approximately 4.5 ml of boiling ultrapure water was quickly added to each centrifuge tube. The tubes were stoppered and transferred to a beaker with boiling water for ~15 minutes for extraction of intracellular ATP and inactivation of ATPases. After extraction, the tubes were cooled to room temperature and then kept frozen at -20 °C for transport to the laboratory. Samples were subsequently kept at -80 °C until analysis.

Hot-water extraction was then performed on the still frozen samples (i.e., without prior thawing) in the laboratory (see Bochdansky et al. 2021 for details). The shock freezing-boiling treatment breaks up cells more efficiently than boiling alone, which results in higher extraction efficiencies (Bochdansky et al. 2021).

In 2018, 500 microliters of water from each bottle was collected and extracted in a boiling hot-water bath before the samples were cooled to room temperature, and then frozen at -20 °C. In 2019, 500 microliters of water from each bottle was placed in 15 ml centrifuge tubes and shock-frozen in liquid nitrogen. These unfiltered samples thus contained both particulate and dissolved ATP and were labeled total ATP (tATP). All samples were brought to the laboratory in a -20 °C freezer and subsequently stored at -80 °C in the laboratory.

For analysis of the 500 microliters shock-frozen samples, boiling hot water was added to the samples and extracted for ~15 minutes in a boiling-water bath.

Particulate and total ATP samples were topped up to 5 ml with ultrapure water using the gradations on the centrifuge tubes and mixed with a vortex mixer. The 500 microliters whole-water samples were also diluted to 5 ml with ultrapure water to reduce salt effects that strongly decrease the luminescence yield. It should be noted that the whole-water extraction method used here (for total ATP) requires sufficiently high ATP levels to produce a signal. This was possible because samples were taken in the mesotrophic coastal ocean and in a eutrophic estuary. Such a small amount of water (500 ml) would be insufficient in oligotrophic or deep-sea environments. Hot-water extraction is only one of two methods proposed by Bochdansky et al. (2021). Many of our subsequent collections were based on chemical extraction using phosphorus benzalkonium chloride (P-BAC) instead. Both methods give highly consistent results, with the values from the chemical extraction method exceeding that of the hot water extraction by 20% (Bochdansky et al. 2021). The hot-water method used here has the advantage that measurements of dissolved ATP can be added easily to the protocol as hot water for the inactivation of ATPases is already at hand.


Related Datasets

IsRelatedTo

Dataset: June 2019 Slover cruise data
Relationship Description: ATP data from 2019 cruise.
Bochdansky, A. B. (2025) Adenosine triphosphate and microbial biomass measurements from the Chesapeake Bay, sampled aboard RV Fay Slover on June 18, 2019. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2024-12-30 doi:10.26008/1912/bco-dmo.939509.1

Related Publications

Results

Bochdansky, A. B., Beecher, A. A., Calderon, J. R., Stouffer, A. N., & Washington, N. N. (2024). A comparison of adenosine triphosphate with other metrics of microbial biomass in a gradient from the North Atlantic to the Chesapeake Bay. Frontiers in Marine Science, 11. https://doi.org/10.3389/fmars.2024.1288812
Results

Bochdansky, A. B., Stouffer, A. N., & Washington, N. N. (2021). Adenosine triphosphate (ATP) as a metric of microbial biomass in aquatic systems: new simplified protocols, laboratory validation, and a reflection on data from the literature. Limnology and Oceanography: Methods, 19(2), 115–131. doi:10.1002/lom3.10409
Methods

Bochdansky, A. B., Clouse, M. A., & Hansell, D. A. (2017). Mesoscale and high-frequency variability of macroscopic particles (> 100 μm) in the Ross Sea and its relevance for late-season particulate carbon export. Journal of Marine Systems, 166, 120–131. doi:10.1016/j.jmarsys.2016.08.010
Methods

Ducklow, H. (2000). Bacterial production and biomass in the oceans. In: Kirchman, D.L. (Ed.), Microbial ecology of the oceans. Wiley-Liss, Inc, New York, 1, 85-120.
Methods

Fukuda, R., Ogawa, H., Nagata, T., & Koike, I. (1998). Direct Determination of Carbon and Nitrogen Contents of Natural Bacterial Assemblages in Marine Environments. Applied and Environmental Microbiology, 64(9), 3352–3358. https://doi.org/10.1128/aem.64.9.3352-3358.1998