Table of Contents

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

These experiments were conducted with a strain of (strain NWFSC 731) was isolated from Long Beach, Washington State, USA on November 3, 2020. The temperature and salinity were 14°C and 27 ppt, respectively at the time of collection. The data was collected in laboratory experiments at the University of Southern California. The experiments began in September 2021 and finished in July of 2022.

These experiments were a cluster of combined temperature, nutrient concentration, and CO2 in order to reflect upwelling, heatwaves, and extreme heatwaves in the natural environment. LTCN was a treatment to test the interactive effects.
Upwelling: 13°C, high CO2, high nutrients
Heatwave: 19°C, low CO2, low nutrients
Extreme heatwave: 21°C, low CO2, low nutrients
LTCN (low temperature, CO2, and nitrogen):  13°C, low CO2, low nutrients

The following section provides a methodology summary for this dataset and references related datasets collected as part of the same experiment (see "Related Datasets" section for data access). A full methodology was published in "Simulated upwelling and marine heatwave events promote similar growth rates but differential domoic acid toxicity in Pseudo-nitzschia australis" in Harmful Algae (Kelly et al., 2023).

Pseudo-nitzschia australis was grown under upwelling heatwave, and extreme heatwave conditions (e.g., combined temperature, nutrient, and carbon dioxide levels specific to each condition) and in single-factor response curves for carbon dioxide, temperature, and varying nitrogen:phosphorus (N:P) ratios/total nutrient concentrations.

Samples for chlorophyll a (used to calculate growth rates) were filtered on GF/F filters, extracted in 6 mL of 90 % acetone at -20°C for 24 h, then analyzed using a Turner 10AU field fluorometer (Welschmeyer 1994; Fu et al. 2007).

For elemental analysis (particulate organic carbon and nitrogen, POC and PON), cells were filtered onto pre-combusted GF/F filters, dried, and analyzed on a Costech 4010 Elemental Analyzer (Fu et al. 2007). 

Samples for particulate domoic acid were filtered onto Supor 0.2 µm 47 mm PES filters. Samples were analyzed using LC-MS/MS on a Prominence UFLC system (Shimadzu, Kyoto, Japan) coupled to a SCIEX 4500 QTRAP mass spectrometer (AB Sciex, Framingham, MA, USA). Methods described in Wang et al. 2012.

Primary production was determined by measuring the uptake of radiolabeled bicar­bonate (Fu et al. 2008). 14C-bicarbonate was added to 45 mL sub-cultures at T24 h and incubated for 24 h (approximating net carbon fixation) under the respective experimental conditions. After the incubation period, cells were collected on GF/F filters and placed in a scintillation vial containing scintillation cocktail. Samples were stored for 24 h before being read on a Wallac System 1400 liquid scintillation counter.

pH measurements were made on a Mettler Toledo SevenCompact pH meter using a three-point calibration curve and total pH scale (Cooley and Yager 2006). Samples for total DIC analysis were collected at Tfinal. Seawater from undisturbed culture bottles was removed with a sterile syringe, ejected into pre-evacuated borosilicate Exetainers, and poisoned with 5% MgCl2. Total DIC was then measured using a Picarro cavity ring-down spectrophotometer according to Subhas et al. (2015).

For cell count samples (for normalizing cellular domoic acid), 1 mL of the final experimental culture was preserved with 40 ul glutaraldehyde and stored at 4°C in the dark. Cells were counted on a Olympus BX51 microscope using a Sedgewick Rafter Chamber.

Organism:
Pseudo-nitzschia australis, LSID (urn:lsid:marinespecies.org:taxname:246604)

Data was processed in using excel, which was used to calculate rates, averages, and standard deviations. Experimental seawater pCO2 and total alkalinity were calculated from measured DIC and pH using CO2SYS version 2.1 software (Lewis and Wallace, 1998).

Problems/Issues:

The following samples were removed from analysis, as there were issues during data collection or processing:

One DA sample was removed from analysis in the pre-industrial treatment in the CO2 single factor experiment, and from the NP=50, high nutrient, 19°C treatment in the N:P ratio experiment, due to a sampling and/or analytical error. In the cluster experiment, one sample from the extreme heatwave treatment was removed from primary production analyses due to an error made during the assay.

These data points were not included in this dataset.

* File cluster_experiment.csv was loaded into the BCO-DMO data system with missing identifier indicated by "NA".
** Missing data values are displayed differently based on the file format you download.  They are blank in csv files, "NaN" in MatLab files, etc.
* Column names adjusted to conform to BCO-DMO naming conventions designed to support broad re-use by a variety of research tools and scripting languages. [Only numbers, letters, and underscores.  Can not start with a number]

NSF Award Abstract:
The diatom Pseudo-nitzschia forms large, toxic harmful algal blooms along the U.S. West Coast, killing wildlife and harming valuable ocean fisheries. Understanding the causes of these blooms and predicting their occurrence, both now and under future changing climate conditions, is critical to coastal environmental and economic health. Puzzlingly, these blooms seem to happen during periods when coastal seawater upwelling results in cold, nutrient-rich, low pH sea surface conditions, and also during times when heat wave events cause warm, nutrient-poor, high pH conditions. These two extremes are forecast to get even more intense with climate change. This project is experimentally testing how Pseudo-nitzschia responds to upwelling and heat wave events using measurements of cell growth, toxin production, and gene expression. Broader impacts of this project include training the principal investigator in new gene expression methods, graduate and undergraduate research training, high school research mentoring experiences, and outreach and communications activities aimed at the commercial fishing industry. Societal benefits include obtaining a better understanding of the causes of damaging toxic algal blooms, and how they may change in the future coastal ocean.

The toxic diatom Pseudo-nitzschia causes annual harmful blooms along the US West Coast, a region where wind-driven upwelling brings rich nutrient supplies into the euphotic zone. However, this region is also experiencing unprecedented episodic ocean heatwave events linked to global warming. Thus, future climate trends in this region suggest an exaggeration of current physio-chemical extremes between colder, more nutrient-rich, low pH upwelling, and warmer, more nutrient-depleted, higher pH heatwaves. Surprisingly, toxic Pseudo-nitzschia spp. can bloom under both upwelling and heatwave conditions, despite opposite trends in key environmental controls like nutrients, temperature, and carbonate chemistry. This project is testing how this happens by first obtaining full response curves for each of the individual factors, temperature, pCO2, phosphorus, nitrogen, and silicon for two Pseudo-nitzschia isolates. Then, these variables are combined in holistic upwelling and heatwave scenario incubation experiments, to compare how growth and toxicity is affected in both cultures and natural blooms of Pseudo-nitzschia. The PI is assessing toxic diatom responses in these experiments using her existing expertise in algal physiology, as well as by expanding her professional horizons to develop new skills in transcriptome bioinformatics in partnership with Dr. Bethany Jenkins from the University of Rhode Island. Experiments are conducted to test the physiological responses of Pseudo-nitzschia to changes in nutrient concentrations, temperature and pCO2 during simulated upwelling or heatwave occurrences, and measure expression of key metabolic pathway genes such as toxin synthesis pathways. This project is helping to understand and interpret the surprising niche flexibility of toxic Pseudo-nitzschia in a changing ocean, and at the same time offers the PI a new avenue forward for her future career development.

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.