| Contributors | Affiliation | Role |
|---|---|---|
| Balch, William M. | Bigelow Laboratory for Ocean Sciences | Principal Investigator, Contact |
| Fields, David | Bigelow Laboratory for Ocean Sciences | Co-Principal Investigator |
| White, Meredith | Bigelow Laboratory for Ocean Sciences | Contact |
| Ake, Hannah | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
Growth cycle (14 day) culture dynamics of Pleurochrysis carterae (NCMA strain 645). It was isolated from 41.525 degrees North, 70.6736 degrees West (Woods Hole, Massachusetts USA), but has been maintained in culture since 1958.
Cultures: Pleurochrysis carterae cultures were maintained in exponential growth phase under axenic conditions in semi-continuous batch culture using L1-Si media prepared on 0.2 um-filtered, UV-sterilized, autoclaved seawater. Cultures were acclimated to one of three pCO2 treatments for > 9 generations before experiments were performed. Cultures were maintained in an incubator at 16.5 +/- 0.5 degrees C and 470 umol photons/m-2/s PAR on a 14-10 light-dark cycle where the lights turned on at 6 am and turned off at 8 pm.
pCO2 Treatments: Carbonate chemistry was manipulated by bubbling cultures and prepared media with 500 mL/min with 0.2 um-filtered 280, 380, or 750 ppm pCO2 air. The pCO2 levels of the treatment air were established using two mass flow controllers (Aalborg, Orangeburg, NY, USA) for each treatment to precisely mix in-house compressed air and pure CO2 (Maine Oxy, Auburn, ME, USA). The in-house compressed air was stripped of CO2 to less than 10 ppm CO2 using a Puregas VCD CO2 Adsorber (Puregas, LLC, Broomfield, CO, USA). The pCO2 of the gas mixtures was stable to +/- 8 ppm. pCO2 values of the cultures may be different than the target levels due to biological activity.
Growth Cycle (14 d) Culture Dynamics Monitoring: To understand the chemical and biological culture dynamics over an entire growth cycle (14 days, where days 1-9 represented exponential growth), the investigators took measurements of one culture from each pCO2 treatment every day at approximately the same time of day for one full growth cycle. Our pH electrode malfunctioned on days 7 and 8, and the total alkalinity sample from the 750 ppm treatment on day 8 was accidentally dropped and broken, so calculations of full carbonate chemistry parameters for days 7 and 8 were not possible.
pH Measurements: The pH of the cultures was measured using an OrionTM ROSSTM electrode connected to an Orion StarTM A211 Benchtop pH meter (ThermoFisher Scientific, Waltham, MA, USA), calibrated with NBS buffers (EK Industries, Inc., Joliet, IL, USA) and corrected to the total scale using weekly spectrophotometric pH measurements of culture samples. Spectrophotometric pH measurements of 0.2 um-filtered culture samples were made with 20 mM m-Cresol purple sodium salt indicator dye (Alfa Aesar, Ward Hill, MA, USA) using a Hitachi U-3010 spectrophotometer (Hitachi High-Technologies, Clarksburg, MD, USA) equipped with a water circulated cell holder connected to a VWR 1160 water bath (VWR, Radnor, PA, USA) set at 16.5 degrees C, holding a 1 cm quartz cell. The method followed the procedure described by Clayton and Byrne (1993) and Dickson et al. (2007), using the refit equation of Liu et al. (2011), resulting in a resolution of +/- 0.004 pH units.
Temperature: Temperature measurements were made with an OrionTM ROSSTM electrode connected to an Orion StarTM A211 Benchtop pH meter (ThermoFisher Scientific, Waltham, MA, USA).
Salinity: Salinity was measured using an Acorn SALT 6 handheld salinity meter (Oakton Instruments, Vernon Hills, IL, USA) with a resolution of +/- 0.1 ppt.
in vivo Fluorescence: Fluoresence was measured using a Turner 10-AU fluorometer (Turner Designs, Sunnyvale, CA, USA).
Cell density and cell diameter: Culture density and mean cell diameter were measured using a Moxi Z mini automated cell counter (ORFLO Technologies, Ketchum, ID, USA), which has a coefficient of variation of 4%.
Particulate Inorganic Carbon: Bulk culture PIC analyses followed the technique of Fernandez et al. (1993): 10 mL culture samples were filtered onto 0.4 um polycarbonate filters and rinsed with potassium borate buffer with the pH adjusted to 8.0 to remove seawater calcium chloride. Filters were carefully moved to trace-metal free centrifuge tubes and digested with 5 mL of 5% nitric acid. The calcium concentration was measured using a Jobin Yvon Ultima C inductively coupled plasma-atomic emission spectrometer (ICP-AES, HORIBA, Ltd., Kyoto, Japan). Bulk culture PIC measurements were corrected to PIC/cell using the corresponding cell density measurements.
Particulate Organic Carbon: To determine the bulk culture POC concentration, 10 mL of culture were filtered onto a pre-combusted Whatman GF/F filter, which was then fumed in 10 % HCl to remove inorganic carbonates. The dried filters were then analyzed on an ECS 4010 CHNSO Analyzer (Costech Analytical Technologies, Valencia, CA, USA) by Bigelow Analytical Services, East Boothbay, ME, USA. Bulk culture POC measurements were corrected to POC/cell using the corresponding cell density measurements.
Nutrients: Culture samples were filtered to 0.2 um to remove all algal cells and coccoliths, and samples were frozen prior to analysis. Total N (nitrate + nitrite), nitrite, phosphate, and silicate were measured by Continuous Flow Analysis by Bigelow Analytical Services using a SEAL AutoAnalyzer 3 HR (SEAL Analytical Inc., Mequon, WI, USA).
Total Alkalinity: Culture samples were filtered to 0.2 um to remove all algal cells and coccoliths. Total alkalinity was measured via titration with 0.01 N HCl using a Metrohm Titrando 888 controlled by Tiamo software (Metrohm, Riverview, FL, USA) to perform automated Gran titrations of 4 mL samples. Titrations were corrected to Certified Reference Materials (supplied by the laboratory of Andrew Dickson, Scripps Institution of Oceanography, La Jolla, CA, USA).
Nitrate: To determine the nitrate concentration of our cultures, Total N (nitrate+nitrite) and nitrite concentrations were measured and the nitrate concentration was calculated as:
Nitrate = Total N - Nitrite
Carbonate Chemistry Calculations: Using the measured values of pH (total scale), total alkalinity, temperature, salinity, phosphate, and silicate, the investigators used CO2SYS software (Pierrot et al. 2006) to calculate dissolved inorganic carbon (DIC), pCO2, [HCO3-], [CO32-], [CO2], and calcite using the first and second dissociation constants (K1 and K2) of carbonic acid in seawater from Mehrbach et al. (1973), refit by Dickson and Millero (1987); KHSO4 from Dickson (1990); and [B]T from Uppstrom et al. (1974). Full carbonate chemistry could not be calculated for days 7 and 8, since the pH meter malfunctioned.
DMO notes:
- added underscores and removed spaces and units from column names
- changed column names to comply with BCO-DMO standards.
- replaced all "na" with "nd"
| File |
|---|
14d_cultureDynamics.csv (Comma Separated Values (.csv), 9.07 KB) MD5:cca6aecb1e8dff0b7820eb15e4e72f3d Primary data file for dataset ID 660244 |
| Parameter | Description | Units |
| pCO2_treatment | The independent variable; one of three pCO2 levels (280 ppm, 380 ppm, or 750 ppm). These treatment levels are nominal values as they represent the target pCO2 for each treatment. Within each pCO2 treatment there are 14 days worth of culture measurements for the algal cultures. Measurements of the media (without algae) were made on days 0 and 14. Only chemical measurements were made on the media not biological measurements. | parts per million (ppm) |
| date | The date the measurement was taken; YYYY/mm/dd | unitless |
| day | The time elapsed since the beginning of the growth cycle. | unitless |
| time | The actual time that the measurements for that timepoint began; HH:MM | unitless |
| pH | pH measured on the total scale. | pH |
| temperature | Temperature | celsius |
| salinity | Salinity | practical salnity unit (PSU) |
| inVivo_fluorescence | fluorescence to indicate relative chlorophyll-a concentration. | relative fluorescence units |
| cell_density | cell density of the culture, as measured by the Moxi Z Automated Cell Counter. | cells per milliliter (cells/mL) |
| mean_cellDiameter | mean cell diameter as measured by the Moxi Z Automated Cell Counter. | microns (um) |
| fluorescencePerCellDensity | Fluorescence divided by cell density to give an estimate of fluorescence per cell. | cells per milliliter (cells/mL)-1 |
| PIC_ugCPerL | Particulate inorganic carbon concentration | ugC/L |
| PIC_pgCPerCell | Particulate inorganic carbon concentration with unit conversion to pg/mL and divided by the cell density to give pg C per cell. | pgC/cell |
| POC_ugCPermL | Particulate organic carbon concentration | ugC/mL |
| POC_pgCPerCell | Particulate organic carbon concentration with unit conversion to pg/mL and divided by the cell density to give pg C per cell. | pgC/cell |
| NO2 | Nitrite concentration. | umol/kg |
| NO3_NO2 | Nitrate + Nitrite concentration. | umol/kg |
| NO3 | Nitrate concentration calculated from the total N and nitrite measurements. | umol/kg |
| PO4 | Phosphate concentration. | umol/kg |
| SiO4 | Silicate concentration. | umol/kg |
| mean_SiO4 | Silicate concentration estimated as the average of the values from all the measured values. Biologically there should be no change in silicate concentration because coccolithophores do not take up silicate. However silicate values are needed to calculate the carbonate chemistry parameters in CO2sys so the average value was used for days when silicate concentrations were not measured. | umol/kg |
| TA | Total alkalinity | ueq/kg |
| DIC | Dissolved inorganic carbon concentration calculated using CO2sys. | umol/kg |
| pCO2 | Partial pressure of carbon dioxide in the water calculated using CO2sys. This calculated value more accurately represents the pCO2 of the culture than the nominal treatment value which represents the pCO2 of the air bubbled into the culture. | uatm |
| HCO3 | Bicarbonate concentration calculated using CO2sys. | umol/kg |
| CO3 | Carbonate concentration calculated using CO2sys. | umol/kg |
| CO2 | Carbon dioxide concentration calculated using CO2sys. | umol/kg |
| omega_calcite | Saturation state of calcium carbonate with respect to calcite calculated using CO2sys. | unitless |
| ISO_DateTime_UTC | Date/Time (UTC) ISO formatted; YYYY/mm/dd;HH:MM:SS | unitless |
| Dataset-specific Instrument Name | Moxi Z Automated Cell Counter |
| Generic Instrument Name | Automated Cell Counter |
| Dataset-specific Description | Measures culture density |
| Generic Instrument Description | An instrument that determines the numbers, types or viability of cells present in a sample. |
| Dataset-specific Instrument Name | Metrohm Titrando 888 |
| Generic Instrument Name | Automatic titrator |
| Dataset-specific Description | Automatic titrations controlled by Tiamo software (Metrohm, Riverview, FL, USA). |
| Generic Instrument Description | Instruments that incrementally add quantified aliquots of a reagent to a sample until the end-point of a chemical reaction is reached. |
| Dataset-specific Instrument Name | Puregas VCD CO2 Adsorber |
| Generic Instrument Name | CO2 Adsorber |
| Dataset-specific Description | Instrument stripped compressed air of CO2 |
| Generic Instrument Description | CO2 Adsorber - an instrument designed to remove CO2 and moisture from compressed air. |
| Dataset-specific Instrument Name | Jobin Yvon Ultima C |
| Generic Instrument Name | Inductively Coupled Plasma Optical Emission Spectrometer |
| Dataset-specific Description | Calcium concentration measured (ICP-AES, HORIBA, Ltd., Kyoto, Japan). |
| Generic Instrument Description | Also referred to as an Inductively coupled plasma atomic emission spectroscope (ICP-AES). These instruments pass nebulised samples into an inductively-coupled gas plasma (8-10000 K) where they are atomised and excited. The de-excitation optical emissions at characteristic wavelengths are spectroscopically analysed. It is often used in the detection of trace metals. |
| Dataset-specific Instrument Name | Aalborg Mass Flow Controller |
| Generic Instrument Name | Mass Flow Controller |
| Dataset-specific Description | Indicate and control set flow rates of gases. Manufactured in Orangeburg, NY USA. |
| Generic Instrument Description | Mass Flow Controller (MFC) - A device used to measure and control the flow of fluids and gases |
| Dataset-specific Instrument Name | SEAL AutoAnalyzer 3 HR |
| Generic Instrument Name | Nutrient Autoanalyzer |
| Dataset-specific Description | Continuous flow analysis performed by Bigelow Analytical Services (SEAL Analytical Inc., Mequon, WI, USA). |
| Generic Instrument Description | Nutrient Autoanalyzer is a generic term used when specific type, make and model were not specified. In general, a Nutrient Autoanalyzer is an automated flow-thru system for doing nutrient analysis (nitrate, ammonium, orthophosphate, and silicate) on seawater samples. |
| Dataset-specific Instrument Name | Orion ROSS electrode |
| Generic Instrument Name | pH Sensor |
| Dataset-specific Description | Orion ROSS electrode was connected to an Orion Star A211 Benchtop pH meter (ThermoFisher Scientific, Waltham, MA, USA) |
| Generic Instrument Description | An instrument that measures the hydrogen ion activity in solutions.
The overall concentration of hydrogen ions is inversely related to its pH. The pH scale ranges from 0 to 14 and indicates whether acidic (more H+) or basic (less H+). |
| Dataset-specific Instrument Name | Acorn SALT 6 handheld salinity meter |
| Generic Instrument Name | Salinity Sensor |
| Dataset-specific Description | Salinity measured using this instrument with a resolution of +/- 0.1 ppt (Oakton Instruments, Vernon Hills, IL, USA) |
| Generic Instrument Description | Category of instrument that simultaneously measures electrical conductivity and temperature in the water column to provide temperature and salinity data. |
| Dataset-specific Instrument Name | Hitachi U-3010 spectrophotometer |
| Generic Instrument Name | Spectrophotometer |
| Dataset-specific Description | Spectrophotometric pH measurements were taken of culture samples |
| Generic Instrument Description | An instrument used to measure the relative absorption of electromagnetic radiation of different wavelengths in the near infra-red, visible and ultraviolet wavebands by samples. |
| Website | |
| Platform | lab Bigelow |
| Start Date | 2013-07-07 |
| Description | Laboratory located at Bigelow Laboratory for Ocean Sciences |
(Extracted from the NSF award abstract)
Ocean acidification is one of the most pressing marine science issues of our time, with potential biological impacts spanning all marine phyla and potential societal impacts affecting man's relationship to the sea. Rising levels of atmospheric pCO2 are increasing the acidity of the world oceans. It is generally held that average surface ocean pH has already declined by 0.1 pH units relative to the pre-industrial level (Orr et al., 2005), and is projected to decrease 0.3 to 0.46 units by the end of this century, depending on CO2 emission scenarios (Caldeira and Wickett, 2005). The overall goal of this research is to parameterize how changes in pCO2 levels could alter the biological and alkalinity pumps of the world ocean. Specifically, the direct and indirect effects of ocean acidification will be examined within a simple, controlled predator/prey system containing a single prey phytoplankton species (the coccolithophore, Emiliania huxleyi) and a single predator (the oceanic metazoan grazer, Calanus finmarchicus). The experiments are designed to elucidate both direct effects (i.e. effects of ocean acidification on the individual organisms only) and interactive effects (i.e. effects on the combined predator/prey system). Interactive experiments with phytoplankton prey and zooplankton predator are a critical starting point for predicting the overall impact of ocean acidification in marine ecosystems. To meet these goals, a state-of-the-art facility will be constructed with growth chambers that are calibrated and have highly-controlled pH and alkalinity levels. The strength of this approach lies in meticulous calibration and redundant measurements that will be made to ensure that conditions within the chambers are well described and tightly monitored for DIC levels. Growth and calcification rates in coccolithophores and the developmental rates, morphological and behavioral effects on copepods will be measured. The PIC and POC in the algae and the excreted fecal pellets will be monitored for changes in the PIC/POC ratio, a key parameter for modeling feedback mechanisms for rising pCO2 levels. In addition, 14C experiments are planned to measure calcification rates in coccolithophores and dissolution rates as a result of grazing. These key experiments will verify closure in the mass balance of PIC, allowing the determination of actual dissolution rates of PIC within the guts of copepod grazers.
NSF Climate Research Investment (CRI) activities that were initiated in 2010 are now included under Science, Engineering and Education for Sustainability NSF-Wide Investment (SEES). SEES is a portfolio of activities that highlights NSF's unique role in helping society address the challenge(s) of achieving sustainability. Detailed information about the SEES program is available from NSF (https://www.nsf.gov/funding/pgm_summ.jsp?pims_id=504707).
In recognition of the need for basic research concerning the nature, extent and impact of ocean acidification on oceanic environments in the past, present and future, the goal of the SEES: OA program is to understand (a) the chemistry and physical chemistry of ocean acidification; (b) how ocean acidification interacts with processes at the organismal level; and (c) how the earth system history informs our understanding of the effects of ocean acidification on the present day and future ocean.
Solicitations issued under this program:
NSF 10-530, FY 2010-FY2011
NSF 12-500, FY 2012
NSF 12-600, FY 2013
NSF 13-586, FY 2014
NSF 13-586 was the final solicitation that will be released for this program.
PI Meetings:
1st U.S. Ocean Acidification PI Meeting(March 22-24, 2011, Woods Hole, MA)
2nd U.S. Ocean Acidification PI Meeting(Sept. 18-20, 2013, Washington, DC)
3rd U.S. Ocean Acidification PI Meeting (June 9-11, 2015, Woods Hole, MA – Tentative)
NSF media releases for the Ocean Acidification Program:
Press Release 10-186 NSF Awards Grants to Study Effects of Ocean Acidification
Discovery Blue Mussels "Hang On" Along Rocky Shores: For How Long?
Press Release 13-102 World Oceans Month Brings Mixed News for Oysters
| Funding Source | Award |
|---|---|
| NSF Division of Ocean Sciences (NSF OCE) |