Kujawinski Elizabeth B.

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Kujawinski

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Elizabeth B.

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0000-0001-8261-971X

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Identification of possible source markers in marine dissolved organic matter using ultrahigh resolution mass spectrometry

( 2009-04-30) Kujawinski, Elizabeth B. ; Longnecker, Krista ; Blough, Neil V. ; Del Vecchio, Rossana ; Finlay, Liam ; Kitner, Joshua B. ; Giovannoni, Stephen J.

Marine dissolved organic matter (DOM) is one of the most heterogeneous and largest pools of reactive carbon on earth, rivaling in mass the carbon in atmospheric carbon dioxide. Nevertheless, the molecular-level composition of marine DOM has eluded detailed description, impeding inquiry into the specific mechanisms that add or remove compounds from the DOM pool. Here we describe the molecular-level composition of C18-extracted DOM along an east-west transect of the North Atlantic Ocean. We examine the changes in DOM composition along this transect with ultrahigh resolution mass spectrometry and multivariate statistics. We use indicator species analysis (ISA) to identify possible source markers for photochemical degradation and heterotrophic bacterial metabolism. The inclusion of ISA in statistical evaluation of DOM mass spectral data allows investigators to determine the m/z values associated with significant changes in DOM composition. With this technique, we observe indicator m/z values in estuarine water that may represent components of terrestrially-derived chromophoric DOM subject to photo-chemical degradation. We also observe a unique set of m/z values in surface seawater and show that many of these are present in pure cultures of the marine α-proteobacterium Candidatus Pelagibacter ubique when grown in natural seawater. These findings indicate that a complex balance of abiotic and biotic processes controls the molecular composition of marine DOM to produce signatures that are characteristic of different environments.
The impact of microbial metabolism on marine dissolved organic matter

( 2010-05) Kujawinski, Elizabeth B.

Microbes mediate global biogeochemical cycles through their metabolism, and all metabolic processes begin with the interaction between the microbial cell wall or membrane and the external environment. For all heterotrophs and many autotrophs, critical growth substrates and factors are present within the dilute and heterogeneous mixture of compounds that constitutes dissolved organic matter (DOM). In short, the microbe--molecule interaction is one of the fundamental reactions within the global carbon cycle. Here, I summarize recent findings from studies that examine DOM--microbe interactions from either the DOM perspective (organic geochemistry) or the microbe perspective (microbial ecology). Gaps in our knowledge are highlighted and future integrative research directions are proposed.
Fate of dispersants associated with the Deepwater Horizon oil spill

( 2011-01-05) Kujawinski, Elizabeth B. ; Kido Soule, Melissa C. ; Valentine, David L. ; Boysen, Angela K. ; Longnecker, Krista ; Redmond, Molly C.

Response actions to the Deepwater Horizon oil spill included the injection of ~771,000 gallons (2,900,000 L) of chemical dispersant into the flow of oil near the seafloor. Prior to this incident, no deepwater applications of dispersant had been conducted and thus no data exists on the environmental fate of dispersants in deepwater. We used ultrahigh resolution mass spectrometry and liquid chromatography with tandem mass spectrometry (LC/MS/MS) to identify and quantify one key ingredient of the dispersant, the anionic surfactant DOSS (dioctyl sodium sulfosuccinate), in the Gulf of Mexico deepwater during active flow and again after flow had ceased. Here we show that DOSS was sequestered in deepwater hydrocarbon plumes at 1000-1200m water depth and did not intermingle with surface dispersant applications. Further, its concentration distribution was consistent with conservative transport and dilution at depth and it persisted up to 300 km from the well, 64 days after deepwater dispersant applications ceased. We conclude that DOSS was selectively associated with the oil and gas phases in the deepwater plume, yet underwent negligible, or slow, rates of biodegradation in the affected waters. These results provide important constraints on accurate modeling of the deepwater plume and critical geochemical contexts for future toxicological studies.
Analytical and computational advances, opportunities, and challenges in marine organic biogeochemistry in an era of "Omics"

(Frontiers Media, 2020-09-02) Steen, Andrew D. ; Kusch, Stephanie ; Abdulla, Hussain A. ; Cakić, Nevenka ; Coffinet, Sarah ; Dittmar, Thorsten ; Fulton, James M. ; Galy, Valier ; Hinrichs, Kai-Uwe ; Ingalls, Anitra ; Koch, Boris P. ; Kujawinski, Elizabeth B. ; Liu, Zhanfei ; Osterholz, Helena ; Rush, Darci ; Seidel, Michael ; Sepulveda, Julio ; Wakeham, Stuart G.

Advances in sampling tools, analytical methods, and data handling capabilities have been fundamental to the growth of marine organic biogeochemistry over the past four decades. There has always been a strong feedback between analytical advances and scientific advances. However, whereas advances in analytical technology were often the driving force that made possible progress in elucidating the sources and fate of organic matter in the ocean in the first decades of marine organic biogeochemistry, today process-based scientific questions should drive analytical developments. Several paradigm shifts and challenges for the future are related to the intersection between analytical progress and scientific evolution. Untargeted “molecular headhunting” for its own sake is now being subsumed into process-driven targeted investigations that ask new questions and thus require new analytical capabilities. However, there are still major gaps in characterizing the chemical composition and biochemical behavior of macromolecules, as well as in generating reference standards for relevant types of organic matter. Field-based measurements are now routinely complemented by controlled laboratory experiments and in situ rate measurements of key biogeochemical processes. And finally, the multidisciplinary investigations that are becoming more common generate large and diverse datasets, requiring innovative computational tools to integrate often disparate data sets, including better global coverage and mapping. Here, we compile examples of developments in analytical methods that have enabled transformative scientific advances since 2004, and we project some challenges and opportunities in the near future. We believe that addressing these challenges and capitalizing on these opportunities will ensure continued progress in understanding the cycling of organic carbon in the ocean.
Dissolved organic matter produced by Thalassiosira pseudonana

( 2014-10) Longnecker, Krista ; Kido Soule, Melissa C. ; Kujawinski, Elizabeth B.

Phytoplankton are significant producers of dissolved organic matter (DOM) in marine ecosystems but the identity and dynamics of this DOM remain poorly constrained. Knowledge on the identity and dynamics of DOM are crucial for understanding the molecular-level reactions at the base of the global carbon cycle. Here we apply emerging analytical and computational tools from metabolomics to investigate the composition of DOM produced by the centric diatom Thalassiosira pseudonana. We assessed both intracellular metabolites within T. pseudonana (the endo-metabolome) and extracellular metabolites released by T. pseudonana (the exo-metabolome). The intracellular metabolites had a more variable composition than the extracellular metabolites. We putatively identified novel compounds not previously associated with T. pseudonana as well as compounds that have previously been identified within T. pseudonana’s metabolic capacity (e.g. dimethylsulfoniopropionate and degradation products of chitin). The resulting information will provide the basis for future experiments to assess the impact of T. pseudonana on the composition of dissolved organic matter in marine environments.
Using stable isotope probing to characterize differences between free-living and sediment-associated microorganisms in the subsurface

( 2012-04-24) Longnecker, Krista ; Kujawinski, Elizabeth B.

Aquifers are subterranean reservoirs of freshwater with heterotrophic bacterial communities attached to the sediments and free-living in the groundwater. In the present study, mesocosms were used to assess factors controlling the diversity and activity of the subsurface bacterial community. The assimilation of 13C, derived from 13C-acetate, was monitored to determine whether the sediment-associated and free-living bacterial community would respond similarly to the presence of protozoan grazers. We observed a dynamic response in the sediment-associated bacterial community and none in the free-living community. The disparity in these observations highlights the importance of the sediment-associated bacterial community in the subsurface carbon cycle.
Molecular characterization of dissolved organic matter associated with the Greenland ice sheet

( 2010-03-16) Bhatia, Maya P. ; Das, Sarah B. ; Longnecker, Krista ; Charette, Matthew A. ; Kujawinski, Elizabeth B.

Subsurface microbial oxidation of overridden soils and vegetation beneath glaciers and ice sheets may affect global carbon budgets on glacial-interglacial timescales. The likelihood and magnitude of this process depends on the chemical nature and reactivity of the subglacial organic carbon stores. We examined the composition of carbon pools associated with different regions of the Greenland ice sheet (subglacial, supraglacial, proglacial) in order to elucidate the type of dissolved organic matter (DOM) present in the subglacial discharge over a melt season. Electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry coupled to multivariate statistics permitted unprecedented molecular level characterization of this material and revealed that carbon pools associated with discrete glacial regions are comprised of different compound classes. Specifically, a larger proportion of protein-like compounds were observed in the supraglacial samples and in the early melt season (spring) subglacial discharge. In contrast, the late melt season (summer) subglacial discharge contained a greater fraction of lignin-like and other material presumably derived from underlying vegetation and soil. These results suggest (1) that the majority of supraglacial DOM originates from autochthonous microbial processes on the ice sheet surface, (2) that the subglacial DOM contains allochthonous carbon derived from overridden soils and vegetation as well as autochthonous carbon derived from in situ microbial metabolism, and (3) that the relative contribution of allochthonous and autochthonous material in subglacial discharge varies during the melt season. These conclusions are consistent with the hypothesis that, given sufficient time (e.g., overwinter storage), resident subglacial microbial communities may oxidize terrestrial material beneath the Greenland ice sheet.
Protist community grazing on prokaryotic prey in deep ocean water masses

(Public Library of Science, 2015-04-20) Rocke, Emma ; Pachiadaki, Maria G. ; Cobban, Alec ; Kujawinski, Elizabeth B. ; Edgcomb, Virginia P.

Oceanic protist grazing at mesopelagic and bathypelagic depths, and their subsequent effects on trophic links between eukaryotes and prokaryotes, are not well constrained. Recent studies show evidence of higher than expected grazing activity by protists down to mesopelagic depths. This study provides the first exploration of protist grazing in the bathypelagic North Atlantic Deep Water (NADW). Grazing was measured throughout the water column at three stations in the South Atlantic using fluorescently-labeled prey analogues. Grazing in the deep Antarctic Intermediate water (AAIW) and NADW at all three stations removed 3.79% ± 1.72% to 31.14% ± 8.24% of the standing prokaryote stock. These results imply that protist grazing may be a significant source of labile organic carbon at certain meso- and bathypelagic depths.
Molecular signature of organic nitrogen in septic-impacted groundwater

( 2014-08) Arnold, William A. ; Longnecker, Krista ; Kroeger, Kevin D. ; Kujawinski, Elizabeth B.

Dissolved inorganic and organic nitrogen levels are elevated in aquatic systems due to anthropogenic activities. Dissolved organic nitrogen (DON) arises from various sources, and its impact could be more clearly constrained if specific sources were identified and if the molecular level composition of DON were better understood. In this work, the pharmaceutical carbamazepine was used to identify septic-impacted groundwater in a coastal watershed. Using ultrahigh resolution mass spectrometry data, the nitrogen-containing features of the dissolved organic matter in septic-impacted and non-impacted samples were compared. The septic impacted groundwater samples have a larger abundance of nitrogen-containing formulas. Impacted samples have additional DON features in the regions ascribed as ‘protein-like’ and ‘lipid-like’ in van Krevelen space and have more intense nitrogen-containing features in a specific region of a carbon versus mass plot. These features are potential indicators of dissolved organic nitrogen arising from septic effluents, and this work suggests that ultrahigh resolution mass spectrometry is a valuable tool to identify and characterize sources of DON.
Incubation experiments were conducted in St. John, US Virgin Islands to investigate the macronutrient drawdown response of reef seawater microbial communities to exudates released from the coral species Porites astreoides and Gorgonia ventalina.

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2022-11-10) Weber, Laura ; Apprill, Amy ; Kujawinski, Elizabeth

Incubation experiments were conducted in St. John, US Virgin Islands to investigate the composition of exudates released from different species of benthic organisms, and the response of reef seawater microbial communities to mixed exudates released from different species and to specific metabolites. Exudates were collected from the stony coral Porites astreoides, and the octocoral Gorgonia ventalina after an 8 hour incubation. Reef seawater microbial communities were incubated separately in the presence of exudates from P. astreoides and G. ventalina for 48 hours and samples were collected to monitor changes in macronutrient concentrations. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/865193
Scientific sampling event log from R/V Knorr cruise KN210-04 in the Western Atlantic Ocean between Uruguay and Barbados in 2013 (Deep Atlantic DOM project)

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2019-11-12) Kujawinski, Elizabeth

Scientific sampling event log from R/V Knorr cruise KN210-04 in the Western Atlantic Ocean between Uruguay and Barbados in 2013 (Deep Atlantic DOM project). For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/4008
Incubation experiments were conducted in St. John, US Virgin Islands to investigate the response of reef seawater microbial communities to the mixed exudates released from the coral species Porites astreoides and Gorgonia ventalina.

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2022-11-10) Weber, Laura ; Apprill, Amy ; Kujawinski, Elizabeth

Incubation experiments were conducted in St. John, US Virgin Islands to investigate the composition of exudates released from different species of benthic organisms, and the response of reef seawater microbial communities to mixed exudates released from different species and to specific metabolites. Exudates were collected from the stony coral Porites astreoides, and the octocoral Gorgonia ventalina after an 8 hour incubation. Reef seawater microbial communities were incubated separately in the presence of exudates from P. astreoides and G. ventalina for 48 hours and samples were collected to monitor changes in microbial abundance via flow cytometry and microbial community composition via 16S rRNA gene sequencing. Complementary Targeted and Untargeted metabolomic data from these incubation experiments is available on the MetaboLights database under accession number MTBLS2855. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/865739
Total organic sulfur (TOS) collected from Niskin bottle samples on R/V Knorr cruise KN210-04 in the Western Atlantic Ocean between Uruguay and Barbados from March to May 2013 (Deep Atlantic DOM project)

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2019-03-15) Kujawinski, Elizabeth ; Longnecker, Krista

This dataset contains the concentration of total organic sulfur (TOS) in seawater from samples collected during the R/V Knorr cruise KN210-04 between 29 Mar 2013 and 06 May 2013 along the eastern coast of South America. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/745536
Physico-chemical data from samples collected along the coast of Louisiana, USA during 2018.

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2019-06-11) Thrash, J. Cameron ; Kujawinski, Elizabeth

Physico-chemical data from samples collected along the coast of Louisiana, USA during 2018. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/745449
Benthic exometabolites and their ecological significance on threatened Caribbean coral reefs

(Springer, 2022-10-17) Weber, Laura ; Soule, Melissa Kido ; Longnecker, Krista ; Becker, Cynthia C. ; Huntley, Naomi ; Kujawinski, Elizabeth B. ; Apprill, Amy

Benthic organisms are the architectural framework supporting coral reef ecosystems, but their community composition has recently shifted on many reefs. Little is known about the metabolites released from these benthic organisms and how compositional shifts may influence other reef life, including prolific microorganisms. To investigate the metabolite composition of benthic exudates and their ecological significance for reef microbial communities, we harvested exudates from six species of Caribbean benthic organisms including stony corals, octocorals, and an invasive encrusting alga, and subjected these exudates to untargeted and targeted metabolomics approaches using liquid chromatography-mass spectrometry. Incubations with reef seawater microorganisms were conducted to monitor changes in microbial abundances and community composition using 16 S rRNA gene sequencing in relation to exudate source and three specific metabolites. Exudates were enriched in amino acids, nucleosides, vitamins, and indole-based metabolites, showing that benthic organisms contribute labile organic matter to reefs. Furthermore, exudate compositions were species-specific, and riboflavin and pantothenic acid emerged as significant coral-produced metabolites, while caffeine emerged as a significant invasive algal-produced metabolite. Microbial abundances and individual microbial taxa responded differently to exudates from stony corals and octocorals, demonstrating that exudate mixtures released from different coral species select for specific bacteria. In contrast, microbial communities did not respond to individual additions of riboflavin, pantothenic acid, or caffeine. This work indicates that recent shifts in benthic organisms alter exudate composition and likely impact microbial communities on coral reefs.
Incubation experiments were conducted in St. John, US Virgin Islands to investigate the response of reef seawater microbial communities to the specific metabolites riboflavin, pantothenic acid, and caffeine.

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2022-11-10) Weber, Laura ; Apprill, Amy ; Kujawinski, Elizabeth

Pre-filtered reef seawater microbial communities collected from Lameshur Bay, U.S. Virgin Islands were incubated separately in the presence of the individual metabolites riboflavin, pantothenic acid, and caffeine for 24 hours and samples were collected to monitor changes in microbial community composition using 16S rRNA gene sequencing and microbial abundances using flow cytometry. Targeted metabolomic data from these incubations is available on the MetaboLights database under accession number MTBLS3286. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/865159
Inorganic and organic nutrient data from Niskin bottles from R/V Knorr cruise KN210-04 in the Western Atlantic Ocean between Uruguay and Barbados in 2013 (Deep Atlantic DOM project)

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2019-11-20) Kujawinski, Elizabeth ; Longnecker, Krista

Inorganic and organic nutrient data from Niskin bottles from the KN210-04 cruise. Data include concentration of: phosphate, nitrate+nitrite, silicate, nitrite, ammonium, non-purgeable organic carbon, and total nitrogen. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/473296
Processed CTD data from all sensors mounted on the rosette from R/V Knorr cruise KN210-04 in the Western Atlantic Ocean between Uruguay and Barbados in 2013 (Deep Atlantic DOM project)

(Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu, 2019-11-21) Kujawinski, Elizabeth ; Longnecker, Krista

Processed CTD data from all sensors mounted on the rosette from the KN210-04 cruise. Data include temperature, salinity, dissolved oxygen, fluorometry, turbidity, PAR, and SPAR. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/481164
Characterization of dissolved organic matter in Lake Superior and its watershed using ultrahigh resolution mass spectrometry

( 2011-11-14) Minor, Elizabeth C. ; Steinbring, Carla J. ; Longnecker, Krista ; Kujawinski, Elizabeth B.

With the advent of ultrahigh resolution mass spectrometry, recent studies have begun to resolve molecular-level relationships between terrestrial and aquatic dissolved organic matter (DOM) in rivers, estuaries, mangrove swamps and their receiving oceans and lakes. Here, we extend ultrahigh resolution mass spectrometry techniques to Lake Superior, the largest freshwater lake in the world by area. Solid-phase extracted samples from the western arm of the lake and its watershed, including swamp, creek, river, lake-river confluence and offshore lake sites were compared using electrospray ionization (ESI) Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Results were analyzed using cluster analysis and van Krevelen diagrams. Chemical similarity appears related to hydrological proximity, terrestrial impact and flow conditions. For example, higher and lower flow samples from the same stream differ from one another. Toivola Swamp, Lake Superior, and the south shore river have diverse arrays of unique molecular formulae.relative to the north shore river and stream sampled in this data set. Lake Superior’s unique elemental formulae, relative to its watershed samples, are primarily in the lignin-like and reduced hydrocarbon regions of van Krevelen diagrams. ESI-amenable Lake Superior DOM also has a higher proportion of formulae containing nitrogen or sulfur relative to the other samples. The degree of overlap among formulae within our data set is consistent with previous ESI FT-ICR-MS characterization of terrestrial, estuarine and marine OM. There appears to be a conserved portion of formulae across natural OM samples, perhaps because these compounds are intrinsically refractory or because they are commonly generated as products of natural reworking processes.
Importance of passive diffusion in the uptake of polychlorinated biphenyls by phagotrophic protozoa

(American Society for Microbiology, 2000-05) Kujawinski, Elizabeth B. ; Farrington, John W. ; Moffett, James W.

Unicellular protozoan grazers represent a size class of organisms where a transition in the mechanism of chlorobiphenyl (CB) introduction, from diffusion through surface membranes to ingestion of contaminated prey, could occur. This study compares the relative importance of these two processes in the overall uptake of polychlorinated biphenyls by protists. Uptake rates and steady-state concentrations were compared in laboratory cultures of grazing and nongrazing protozoa. These experiments were conducted with a 10-µm marine scuticociliate (Uronema sp.), bacterial prey (Halomonas halodurans), and a suite of 21 CB congeners spanning a range of aqueous solubilities. The dominant pathway of CB uptake by both grazing and nongrazing protozoa was diffusion. Organic-carbon-normalized CB concentrations (in the protozoan cell) were equivalent in grazing and nongrazing protozoa for all congeners studied. Rate constants for uptake into and loss from the protozoan cell were independently determined by using [3,3',4,4'-14C]tetrachlorobiphenyl (IUPAC no. 77), 0.38 ± 0.03 min-1 and (1.1 ± 0.1) × 10-5 (g of organic carbon)-1 min-1, respectively. Magnitudes of the uptake and loss processes were calculated and compared by using a numerical model. The model result was consistent with data from the bioaccumulation experiment and supported the hypothesis that diffusive uptake is faster than ingestive uptake in phagotrophic unicellular protozoa.

Kujawinski Elizabeth B.

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