http://lod.bco-dmo.org/id/dataset/744468
eng; USA
utf8
dataset
Highest level of data collection, from a common set of sensors or instrumentation, usually within the same research project
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
2018-08-22
ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data
ISO 19115-2:2009(E)
CSIA 15N AA data from phytoplankton, microzooplankton, and Calanus pacificus.
2018-08-23
publication
2018-08-23
revision
Marine Biological Laboratory/Woods Hole Oceanographic Institution Library (MBLWHOI DLA)
2019-06-10
publication
https://doi.org/10.1575/1912/bco-dmo.744468.1
Michael R. Landry
University of California-San Diego Scripps
principalInvestigator
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
publisher
Cite this dataset as: Landry, M. (2018) CSIA 15N AA data from phytoplankton, microzooplankton, and Calanus pacificus. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2018-08-23 [if applicable, indicate subset used]. doi:10.1575/1912/bco-dmo.744468.1 [access date]
15N AA from phytoplankton, microzooplankton and Calanus pacificus Dataset Description: <p>15N AA from phytoplankton, microzooplankton, and <em>Calanus pacificus.</em></p> Methods and Sampling: <p>Hydrolysis, derivatization and analyses of samples for CSIA –&nbsp; For Exps. 3-4, samples (1-2 mg dry weight) for CSIA-AA were added to 1 ml 6 N HPLC-grade HCl, then flushed with N2, capped with a Teflon-lined cap, and hydrolyzed at 150°C for 70 min.&nbsp; The resulting hydrolysate was evaporated to dryness under N2 at 55°C, redissolved in 1 ml 0.01 N HCl, purified by filtration (0.45-μm hydrophilic filter), washed with 1 ml of 0.01 N HCl, and further purified using cation-exchange chromatography with a 5-cm resin column (Dowex 50WX8-400) in a glass Pasteur pipette.&nbsp; AAs were eluted with 4 ml of 2 N NH4OH and evaporated to dryness under a stream of N2 at 80°C, then reacidified with 0.5 ml of 0.2 N HCl, flushed with N2, heated to 110°C for 5 min and evaporated to dryness under N2 at 55°C.&nbsp; Hydrolyzed samples were esterified with 2 ml of 4:1 isopropanol:acetyl chloride, flushed with N2 and heated to 110°C for 60 min.&nbsp; After drying at 60°C under N2, the samples were acylated by adding 1 ml of 3:1 methylene chloride:trifluoracetic anhydride (TFAA) and heated to 100°C for 15 min.&nbsp; The derivatized AAs were further purified by solvent extraction following (Ueda et al. 1989).&nbsp; The acylated AA esters were evaporated at room temperature under N2 and redissolved in 3 ml of 1:2 chloroform:P-buffer (KH2PO4 + Na2HPO4 in Milli-Q water, pH 7).&nbsp; Vigorous shaking ensured that the derivitized AAs were partitioned into chloroform while contaminants remained in the P-buffer.&nbsp; The solvents were separated by centrifugation (10 min at 600 g), the chloroform was transferred to a clean vial, and the solvent extraction process repeated.&nbsp; Finally, to ensure derivatization, the acylation step was repeated.&nbsp; Samples were stored at –20°C in 3:1 methylene chloride:TFAA for up to 2 weeks until analysis.&nbsp; AA derivatives were then analyzed by isotope monitoring gas chromatography-mass spectrometry.&nbsp; We used a Delta V Plus mass spectrometer interfaced with a Trace GC gas chromatograph through a GC-C III combustion furnace (980°C), reduction furnace (650°C), and liquid nitrogen cold trap.&nbsp; Internal reference compounds (aminoadipic acid and norleucine) of known nitrogen isotopic composition were co-injected with samples and used to normalize the measured δ15N values of unknown AAs, and a suite of eight AAs with known isotopic composition was analyzed every 3 injections for additional quality control.&nbsp; At least three injections per sample were analyzed.&nbsp; For Exps. 5 and 6, CSIA-AA samples were hydrolyzed in 6N HCl at 110°C for 20 h.&nbsp; The hydrolysate was evaporated to dryness at temperature under a continuous stream of N2 gas, then esterified with 4:1 isotopropanol:acetyl chloride at 110°C for 60 min, acylated in a 1:1 solution of methylene chloride:trifluoracetic anhydride (DCM:TFAA) for 10 min at 110°C.&nbsp; Samples were stored at -20°C in 1:1 DCM:TFAA for up to 3 months before isotope analysis using a Delta V Plus mass spectrometer (Thermo Scientific) interfaced through a Conflo IV to a GC 1310 gas chromatograph coupled to a GC Isolink combustion-reduction furnace (1000°C) and liquid nitrogen cold trap.&nbsp; All samples were injected (splitless injector) onto a forte BPx5 capillary column (60 m x 0.32 mm x 1.0-μm film thickness) at an injector temperature of 250°C with a constant helium flow rate of 1.4 ml min–1.&nbsp; The column was initially held at 50°C for 2 min and then increased to 125 °C at a rate of 15°C min-1.&nbsp; Once at 125°C, the temperature was increased at a rate of 3°C min-1 to 160°C and then to 190°C at a rate of 4°C min-1.&nbsp; The final temperature of 300°C was reached by ramping to 275°C at 6°C min-1 and then 15°C min-1 afterward.&nbsp; Samples were analyzed in triplicate and normalized to the known δ15N values of a suite of 14 AAs analyzed before and after each set of 3 samples.</p>
Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1260055 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1260055
completed
Michael R. Landry
University of California-San Diego Scripps
858-534-4702
9500 Gilman Drive Mail Code: 0227
La Jolla
CA
92093-0227
USA
mlandry@ucsd.edu
pointOfContact
asNeeded
Dataset Version: 1
Unknown
Experiment
Organism
stage
bulk_d15N
Ala_d15N
Asp_d15N
Glu_d15N
IsoL_d15N
Leu_d15N
Pro_d15N
Val_d15N
Gly_d15N
Lys_d15N
Phe_d15N
Ser_d15N
Thr_d15N
Note_Organism
Note_bulk_d15N
Note_Gly_d15N
Trace GC gas chromatograph, GC 1310 gas chromatograph
GC 1310
Delta V Plus
theme
None, User defined
experiment id
species scientific name (binomial)
experiment type
d15N
comments
featureType
BCO-DMO Standard Parameters
Gas Chromatograph
Gas Chromatograph
Mass Spectrometer
instrument
BCO-DMO Standard Instruments
otherRestrictions
otherRestrictions
Access Constraints: none. Use Constraints: Please follow guidelines at: http://www.bco-dmo.org/terms-use Distribution liability: Under no circumstances shall BCO-DMO be liable for any direct, incidental, special, consequential, indirect, or punitive damages that result from the use of, or the inability to use, the materials in this data submission. If you are dissatisfied with any materials in this data submission your sole and exclusive remedy is to discontinue use.
Resolving the trophic connection between protistan grazers and mesozooplankton in marine food webs using amino acid-specific stable isotope analyses
https://osprey.bco-dmo.org/project/556514
Resolving the trophic connection between protistan grazers and mesozooplankton in marine food webs using amino acid-specific stable isotope analyses
<p><em>Description from NSF award abstract:</em><br />
Energy dissipation and elemental cycling by protistan consumers in lower trophic levels of ocean food webs are of sufficient magnitude, based on global mean measures of the amount of primary production consumed, to strongly alter the efficiencies of material transfers to higher-level consumers and to export. We presently know very little about these microbial food web steps, how they vary regionally or temporally, or how they might be altered by climate change. Compound Specific Isotope Analysis of Amino Acids (CSIA-AA) offers an approach for advancing our understanding of microbial food web structure and trophic fluxes based on the trophic positions (TP) of mesozooplankton as temporal integrators of the fluxes from direct feeding on phytoplankton and indirect transfers via protistan microzooplankton. Preliminary laboratory experiments to test this idea have demonstrated that the standard application of the method, using labeled phenylalanine as the representative source AA for the primary producer baseline and labeled glutamic acid as the indicator AA for trophic enrichment, does not produce a measureable trophic-step signal for protistan grazers. However, the results have also shown that an alternative high-turnover AA, alanine, strongly enriches in protistan as well as metazoan consumers, and leads to substantially higher TP estimates of mesozooplankton in field-collected specimens than that based on labeled glutamic acid.</p>
<p>This research project will test the hypothesis that labeled alanine provides a quantifiable and consistent index of trophic enrichment for protistan steps in marine food webs. The research will involve three major elements. First, controlled laboratory experiments will be conducted with chemostat systems to compare 15N enrichments of alanine to other AAs for a representative suite of ciliate and flagellate grazers feeding on phytoplankton, and to evaluate the two-step enrichment from phytoplankton via a protistan grazer to a suspension-feeding copepod. Second, field-collected mesozooplankton from four distinct ecological regions of the Pacific Ocean will be analyzed by CSIA-AA to test the transfer of alanine enrichment through a metazoan trophic step (comparing suspension feeding species to primary carnivores) and to assess how the TP index differs with trophic structure over a broad range of ecological conditions. Last, CSIA-AA assessments of TP for size-structured zooplankton will be integrated into inverse models of nitrogen flows in the four regions (equatorial Pacific, subtropical North Pacific, California Current and Costa Rica Dome) as a major constraint for resolving and comparing fluxes through the microbial food web over the range of ecological conditions. A properly calibrated CSIA-AA assessment of mesozooplankton trophic position will provide a new and valuable approach for regional intercomparisons of lower-level food web structure, for assessing temporal and spatial trends in climate change, for ocean ecosystem model validation, and for better understanding of lower food-web energetic constraints on ocean fisheries.</p>
CSIA-AA Mesozooplankton TP
largerWorkCitation
project
eng; USA
oceans
-117.254046
-117.254046
32.868814
32.868814
2018-08-23
California Current, eastern Pacific Ocean
0
BCO-DMO catalogue of parameters from CSIA 15N AA data from phytoplankton, microzooplankton, and Calanus pacificus.
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
http://lod.bco-dmo.org/id/dataset-parameter/744528.rdf
Name: Experiment
Units: unitless
Description: <p>Experiment number</p>
http://lod.bco-dmo.org/id/dataset-parameter/744529.rdf
Name: Organism
Units: unitless
Description: <p>Species name or Particulate organic matter (POM)</p>
http://lod.bco-dmo.org/id/dataset-parameter/744530.rdf
Name: stage
Units: unitless
Description: <p>Refers to the experimental stage</p>
http://lod.bco-dmo.org/id/dataset-parameter/744531.rdf
Name: bulk_d15N
Units: per mil
Description: <p>15N of bulk material</p>
http://lod.bco-dmo.org/id/dataset-parameter/744532.rdf
Name: Ala_d15N
Units: per mil
Description: <p>15N of AA Alanine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744533.rdf
Name: Asp_d15N
Units: per mil
Description: <p>15N of AA Aspartic acid</p>
http://lod.bco-dmo.org/id/dataset-parameter/744534.rdf
Name: Glu_d15N
Units: per mil
Description: <p>15N of AA Glutamic acid</p>
http://lod.bco-dmo.org/id/dataset-parameter/744535.rdf
Name: IsoL_d15N
Units: per mil
Description: <p>15N of AA IsoLeucine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744536.rdf
Name: Leu_d15N
Units: per mil
Description: <p>15N of AA Leucine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744537.rdf
Name: Pro_d15N
Units: per mil
Description: <p>15N of AA Proline</p>
http://lod.bco-dmo.org/id/dataset-parameter/744538.rdf
Name: Val_d15N
Units: per mil
Description: <p>15N of AA Valine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744539.rdf
Name: Gly_d15N
Units: per mil
Description: <p>15N of AA Glycine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744540.rdf
Name: Lys_d15N
Units: per mil
Description: <p>15N of AA Lysine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744541.rdf
Name: Phe_d15N
Units: per mil
Description: <p>15N of AA Phenylalanine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744542.rdf
Name: Ser_d15N
Units: per mil
Description: <p>15N of AA Serine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744543.rdf
Name: Thr_d15N
Units: per mil
Description: <p>15N of AA Treonine</p>
http://lod.bco-dmo.org/id/dataset-parameter/744544.rdf
Name: Note_Organism
Units: unitless
Description: <p>Each note corresponds to the cell in this row in the Organism column</p>
http://lod.bco-dmo.org/id/dataset-parameter/744545.rdf
Name: Note_bulk_d15N
Units: unitless
Description: <p>Each note corresponds to the cell in this row in the bulk d15N column</p>
http://lod.bco-dmo.org/id/dataset-parameter/744546.rdf
Name: Note_Gly_d15N
Units: unitless
Description: <p>Each note corresponds to the cell in this row in the Gly d15N column</p>
GB/NERC/BODC > British Oceanographic Data Centre, Natural Environment Research Council, United Kingdom
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
3493
https://darchive.mblwhoilibrary.org/bitstream/1912/24237/1/dataset-744468_zooplanktonphytoplankton-csia-aa__v1.tsv
download
https://doi.org/10.1575/1912/bco-dmo.744468.1
download
onLine
dataset
<p>Hydrolysis, derivatization and analyses of samples for CSIA –&nbsp; For Exps. 3-4, samples (1-2 mg dry weight) for CSIA-AA were added to 1 ml 6 N HPLC-grade HCl, then flushed with N2, capped with a Teflon-lined cap, and hydrolyzed at 150°C for 70 min.&nbsp; The resulting hydrolysate was evaporated to dryness under N2 at 55°C, redissolved in 1 ml 0.01 N HCl, purified by filtration (0.45-μm hydrophilic filter), washed with 1 ml of 0.01 N HCl, and further purified using cation-exchange chromatography with a 5-cm resin column (Dowex 50WX8-400) in a glass Pasteur pipette.&nbsp; AAs were eluted with 4 ml of 2 N NH4OH and evaporated to dryness under a stream of N2 at 80°C, then reacidified with 0.5 ml of 0.2 N HCl, flushed with N2, heated to 110°C for 5 min and evaporated to dryness under N2 at 55°C.&nbsp; Hydrolyzed samples were esterified with 2 ml of 4:1 isopropanol:acetyl chloride, flushed with N2 and heated to 110°C for 60 min.&nbsp; After drying at 60°C under N2, the samples were acylated by adding 1 ml of 3:1 methylene chloride:trifluoracetic anhydride (TFAA) and heated to 100°C for 15 min.&nbsp; The derivatized AAs were further purified by solvent extraction following (Ueda et al. 1989).&nbsp; The acylated AA esters were evaporated at room temperature under N2 and redissolved in 3 ml of 1:2 chloroform:P-buffer (KH2PO4 + Na2HPO4 in Milli-Q water, pH 7).&nbsp; Vigorous shaking ensured that the derivitized AAs were partitioned into chloroform while contaminants remained in the P-buffer.&nbsp; The solvents were separated by centrifugation (10 min at 600 g), the chloroform was transferred to a clean vial, and the solvent extraction process repeated.&nbsp; Finally, to ensure derivatization, the acylation step was repeated.&nbsp; Samples were stored at –20°C in 3:1 methylene chloride:TFAA for up to 2 weeks until analysis.&nbsp; AA derivatives were then analyzed by isotope monitoring gas chromatography-mass spectrometry.&nbsp; We used a Delta V Plus mass spectrometer interfaced with a Trace GC gas chromatograph through a GC-C III combustion furnace (980°C), reduction furnace (650°C), and liquid nitrogen cold trap.&nbsp; Internal reference compounds (aminoadipic acid and norleucine) of known nitrogen isotopic composition were co-injected with samples and used to normalize the measured δ15N values of unknown AAs, and a suite of eight AAs with known isotopic composition was analyzed every 3 injections for additional quality control.&nbsp; At least three injections per sample were analyzed.&nbsp; For Exps. 5 and 6, CSIA-AA samples were hydrolyzed in 6N HCl at 110°C for 20 h.&nbsp; The hydrolysate was evaporated to dryness at temperature under a continuous stream of N2 gas, then esterified with 4:1 isotopropanol:acetyl chloride at 110°C for 60 min, acylated in a 1:1 solution of methylene chloride:trifluoracetic anhydride (DCM:TFAA) for 10 min at 110°C.&nbsp; Samples were stored at -20°C in 1:1 DCM:TFAA for up to 3 months before isotope analysis using a Delta V Plus mass spectrometer (Thermo Scientific) interfaced through a Conflo IV to a GC 1310 gas chromatograph coupled to a GC Isolink combustion-reduction furnace (1000°C) and liquid nitrogen cold trap.&nbsp; All samples were injected (splitless injector) onto a forte BPx5 capillary column (60 m x 0.32 mm x 1.0-μm film thickness) at an injector temperature of 250°C with a constant helium flow rate of 1.4 ml min–1.&nbsp; The column was initially held at 50°C for 2 min and then increased to 125 °C at a rate of 15°C min-1.&nbsp; Once at 125°C, the temperature was increased at a rate of 3°C min-1 to 160°C and then to 190°C at a rate of 4°C min-1.&nbsp; The final temperature of 300°C was reached by ramping to 275°C at 6°C min-1 and then 15°C min-1 afterward.&nbsp; Samples were analyzed in triplicate and normalized to the known δ15N values of a suite of 14 AAs analyzed before and after each set of 3 samples.</p>
Specified by the Principal Investigator(s)
<p>Data were processed using R software to correct sample values using a standard curve of known versus measured δ15N values.&nbsp; The standard deviations of measured δ15N values of standard AAs ranged from 0.1 to 1.7‰.&nbsp; Analyses of replicate samples are noted in data&nbsp;table.&nbsp;&nbsp;</p>
<p><strong>BCO-DMO Processing:</strong><br />
- modified parameter names to conform with BCO-DMO naming conventions;<br />
- replaced spaces with underscores in organism and stage columns;<br />
- replaced missing data with 'nd'</p>
Specified by the Principal Investigator(s)
asNeeded
7.x-1.1
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
Trace GC gas chromatograph, GC 1310 gas chromatograph
Trace GC gas chromatograph, GC 1310 gas chromatograph
PI Supplied Instrument Name: Trace GC gas chromatograph, GC 1310 gas chromatograph PI Supplied Instrument Description:Used with mass spectrometer Instrument Name: Gas Chromatograph Instrument Short Name:Gas Chromatograph Instrument Description: Instrument separating gases, volatile substances, or substances dissolved in a volatile solvent by transporting an inert gas through a column packed with a sorbent to a detector for assay. (from SeaDataNet, BODC) Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB02/
GC 1310
GC 1310
PI Supplied Instrument Name: GC 1310 PI Supplied Instrument Description:Coupled to a GC Isolink combustion-reduction furnace (1000 deg C) and liquid nitrogen cold trap. Instrument Name: Gas Chromatograph Instrument Short Name:Gas Chromatograph Instrument Description: Instrument separating gases, volatile substances, or substances dissolved in a volatile solvent by transporting an inert gas through a column packed with a sorbent to a detector for assay. (from SeaDataNet, BODC) Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB02/
Delta V Plus
Delta V Plus
PI Supplied Instrument Name: Delta V Plus PI Supplied Instrument Description:Used for sampling Instrument Name: Mass Spectrometer Instrument Short Name:Mass Spec Instrument Description: General term for instruments used to measure the mass-to-charge ratio of ions; generally used to find the composition of a sample by generating a mass spectrum representing the masses of sample components. Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB16/