http://lod.bco-dmo.org/id/dataset/963872 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 2025-06-05 ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data ISO 19115-2:2009(E) Effects of sponge excurrent seawater on coral reef picoplankton composition in the Florida Keys January 2022 2025-06-17 publication 2025-06-17 revision Marine Biological Laboratory/Woods Hole Oceanographic Institution Library (MBLWHOI DLA) 2025-06-20 publication https://doi.org/10.26008/1912/bco-dmo.963872.1 Amy Apprill Woods Hole Oceanographic Institution principalInvestigator Cole G. Easson Middle Tennessee State University principalInvestigator Cara L. Fiore Appalachian State University principalInvestigator Alicia M. Reigel Appalachian State University 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: Apprill, A., Easson, C. G., Fiore, C. L., Reigel, A. M. (2025) Effects of sponge excurrent seawater on coral reef picoplankton composition in the Florida Keys January 2022. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2025-06-17 [if applicable, indicate subset used]. doi:10.26008/1912/bco-dmo.963872.1 [access date] Inorganic nutrients, targeted metabolomics and flow cytometry Dataset Description: Methods and Sampling: &lt;p&gt;Incubation experiments were set up with sponge excurrent water, coral exudates, sponge excurrent and coral exudates, or surface reef water as media and with reef surface picoplankton as inoculum to compare microbial growth over 48 hours. Excurrent seawater samples of three sponge species (Verongula rigida, Xestospongia muta, and Niphates digitalis) were collected at Looe Key Reef in the Florida Keys, U.S.A. at approximately 10m depth (24.54605, -81.40610). The exhalent seawater was collected using acid-rinsed glass syringes with a three-way valve and PharMed BPT tubing (Cole-Parmer, USA) connected to 1L FlexFoil Plus sampling bags (SKC, Pittsburgh, PA, USA). Water was drawn in at about 2 mL per minute, which is slower than the pumping rate of the sponges. The exhalent seawater was collected the day before the experiment and filtered (0.2 µm, Omnipore, Millipore Sigma, USA) prior to storage at 4°C. Coral exudates were collected by incubating four fragments each of Acropora cervicornis and Orbicella faveolata were incubated in aquaria containing 0.2 µm filtered (Omnipore) seawater from Mote Marine laboratory seawater intake which pulls from the nearby seawater canal. The aquaria was placed in flowthrough seawater raceway in the sunlight under one layer of shade cloth (~23 – 262 µE s-1 with a mix of sun and clouds) for 6.5 hrs. The coral exudate seawater was then filtered 0.2 µm filtered and stored at 4°C.&lt;/p&gt; &lt;p&gt;Surface seawater over Looe Key Reef was collected the morning of the experimental setup and was used for both the inoculum of bacterioplankton and for background seawater control medium (0.2 µm filtered to create the ambient seawater medium). The sponge excurrent seawater was combined in a 1:1:1 ratio from the three sponge species. Then four different mediums were created: coral exudate water, sponge exhalant water, sponge exhalant water plus coral exudates (1:1 ratio of each medium), and ambient seawater. Coral reef surface picoplankton, served as inoculum and was added to the media in a 1:3 ratio of inoculum to medium. An initial ‘soup’ was made for each treatment in 5L acid-cleaned polycarbonate food grade containers and the total volume of the initial soup was 3900 mL for treatments with microbial inoculum and 2900 mL for control treatments without inoculum. From the initial soup, three T0 samples of seawater (1400 mL) were taken for chemical and microbial analyses. The rest of the experimental seawater with inoculum was then distributed into 2L acid-cleaned polycarbonate bottles (1L per bottle) and incubated in a flow through seawater table in the dark for 48 hours. There were five replicate bottles for each treatment (coral, sponge, sponge + coral, ambient seawater) and three replicate bottles for each no-inoculum control corresponding to each treatment. Nutrients and the bacterioplankton community composition and abundance were assessed at the start of the experiment (T0) and after 48hrs (T48).&lt;/p&gt; &lt;p&gt;Seawater sampled from the incubation ‘soup’ or bottles was used for multiple nutrient analyses and DNA extraction. Prior to filtration, 1 mL of seawater was preserved in&amp;nbsp;500 µL of paraformaldehyde (0.5% final concentration), which were stored in the refrigerator for 1-2 hours and then stored at -80°C until shipped to the University of Hawai’i for flow cytometry analysis. Also prior to filtration, approximately 30 mL of sample was poured into acid and milliQ water rinsed and combusted glass amber EPA vials with acid cleaned septa and lids. These 30 mL samples were acidified to pH ~2 with concentrated HCl and stored at room temperature until analysis for total organic carbon (TOC) and total nitrogen (TN) at Woods Hole Oceanographic Institution. The rest of the seawater was filtered through an Omnipore 0.2 um filter using a peristaltic pump and acid and milliQ water rinsed Pharmed BP tubing and Teflon filter holders. The filter was stored at -80C until DNA extraction, while the filtrate was poured into acid and milliQ water rinsed: 1) HDPE bottles (~20 mL) for inorganic nutrient analysis at the University of Oregon, 2) polycarbonate bottles (~500 mL) for extraction of dissolved organic matter (DOM).&amp;nbsp; Filtered seawater was processed using PPL solid phase extraction following the protocol by Dittmar et al. 2008. Extracts were dried to nearly completeness, leaving a small viscous drop in the vial. These extracts were then shipped to WHOI for metabolomics analysis.&lt;/p&gt; &lt;p&gt;DNA from the Omnipore filters was extracted using a commercial kit and the 16S rRNA gene was amplified using the modified earth microbiome primer set for the V4 region&amp;nbsp;(515F and 806R, Apprill et al. 2015). PCRs were sent to Middle Tennessee State University for library construction and sequencing on an Illumina MiSeq, producing FASTQ files as output.&lt;/p&gt; &lt;p&gt;Targeted Metabolite Analysis by UPLC-MS:&lt;/p&gt; &lt;p&gt;DOM extracts were reconstituted in 200 μl MilliQ water with 50 ng/ml isotopically-labeled injection standards d2 biotin, d6 succinic acid, d4 cholic acid, and d7 indole 3 acetic acid. We used ultra-performance liquid chromatography (Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific) coupled to a heated electrospray ionization source (H-ESI) and a triple quadrupole mass spectrometer (TSQ Vantage, Thermo Scientific) operated under selected reaction monitoring (SRM) mode. We performed chromatographic separation with a Waters Acquity HSS T3 column (2.1 × 100 mm, 1.8 μm) equipped with a Vanguard pre-column and maintained at 40°C. We eluted the metabolites from the column with (A) 0.1% formic acid in water and (B) 0.1% formic acid in acetonitrile at a flow rate of 0.5 mL min-1, according to the gradient: 0 min, 1% B; 1 min, 1%B; 3 min, 15%B; 6 min, 50%B; 9 min, 95%B; 10 min, 95%B; 10.2 min, 1%B; 12 min, 1%B (total run time = 12 min). Settings for source gases were 55 (sheath), 20 (auxiliary) and 0 (sweep), and these settings are presented in arbitrary units. The heated capillary temperature was 375 °C and the vaporizer temperature was 400 °C. For positive and negative modes, we performed separate autosampler injections of 5 μL each.&lt;/p&gt; &lt;p&gt;Inorganic nutrients:&amp;nbsp;&lt;/p&gt; &lt;p&gt;Inorganic nutrients included phosphate, nitrate+nitrite, nitrite, ammonia, and silicic acid. The phosphate method is a modification of the molybdenum blue procedure of&lt;/p&gt; &lt;p&gt;Bernhardt and Wilhelms (1967), in which phosphate is determined as reduced phosphomolybdic acid employing hydrazine as the reductant. The nitrate + nitrite analysis uses the basic method of Armstrong et al. (1967), with modifications to improve the precision and ease of operation.&amp;nbsp; Sulfanilamide and N-(1-Napthyl)ethylenediamine dihydrochloride react with nitrite to form a colored diazo compound. For the nitrate + nitrite analysis, nitrate is first reduced to nitrite using an OTCR and imidazole buffer as described by Patton (1983). Nitrite analysis is performed on a separate channel, omitting the cadmium reductor and the buffer. The method is based on that of Armstrong et al. (1967) as adapted by Atlas et al. (1971).&amp;nbsp; Addition of an acidic molybdate reagent forms silicomolybdic acid which is then reduced by stannous chloride. This indophenol blue method is modified from ALPKEM RFA methodology which references Methods for Chemical Analysis of Water and Wastes, March 1984, EPA-600/4-79-020, &quot;Nitrogen Ammonia&quot;, Method 350.1 (Colorimetric, Automated Phenate) A detailed description of the continuous segmented flow procedures used can be found in Gordon et. al.&amp;nbsp; (1994).&amp;nbsp;&lt;/p&gt; &lt;p&gt;Flow cytometry:&amp;nbsp;&lt;/p&gt; &lt;p&gt;Samples were preserved and stored at -80°C until later batch analysis at the University of Hawaii SOEST Flow Cytometry Facility (www.soest.hawaii.edu/sfcf). Microbial cells were enumerated using flow cytometry (Selph, 2021). In brief, samples (0.1 mL) were thawed in batches, stained with the DNA dye Hoechst 34580 (1 µg/mL final), then run at 30 µL min-1 on a Beckman-Coulter CytoFlex S flow cytometer, using lasers emitting at 375 nm (to detect Hoechst), 488 nm (for scatter and chlorophyll parameters), and 561 nm (for phycoerythrin). Resulting listmode files (FCS 3.0) were analyzed using FlowJo software (Becton Dickinson, v. 10.8.2) to distinguish microbial populations based on their fluorescence signals (chlorophyll, phycoerythrin, DNA), as well as forward and right-angle light scatter.&amp;nbsp; Heterotrophic bacteria were distinguished from phytoplankton by their DNA signature and absence of pigment.&amp;nbsp; Prochlorococcus and Synechococcus were separated from larger eukaryotic phytoplankton by their light scatter signatures, as well as their characteristic pigment and DNA signatures.&amp;nbsp; Other phytoplankton (eukaryotes, mostly 2-20 µm pico- and nano-sized cells given the small volume analyzed) had higher light scatter and more chlorophyll fluorescence per cell. &amp;gt;&lt;/p&gt; Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1924540 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1924540 Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1923962 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1923962 completed Amy Apprill Woods Hole Oceanographic Institution 508-289-2649 Marine Chemistry and Geochemistry Department 266 Woods Hole Road, MS# 4 Woods Hole MA 02543 USA aapprill@whoi.edu pointOfContact Cole G. Easson Middle Tennessee State University 615-898-5822 1672 Greenland Drive SCI 2044 Murfreesboro TN 37132 USA Cole.Easson@mtsu.edu pointOfContact Cara L. Fiore Appalachian State University 828-262-7548 572 Rivers St Boone NC 28607 USA fiorec@appstate.edu pointOfContact Alicia M. Reigel Appalachian State University Lexington VA USA areigel@wlu.edu pointOfContact asNeeded Dataset Version: 1 Unknown SampleID latitude longitude sampling_date Trtmt Time PO4 NOx Silicate NH4 TOC TON metabolite_2_3_dihydroxypropane_1_sulfonate metabolite_2_3_dihydroxybenzoic_acid metabolite_2_deoxycytidine metabolite_2_methyl_4_oxopentanoic metabolite_2_Keto_3_deoxy_6_phosphogluconate metabolite_3_methyl_2_oxobutanoic_acid metabolite_4_aminobenzoic_acid metabolite_4_methyl_2_oxopentanoic_acid metabolite_6_hydroxymelatonin metabolite_adenine metabolite_adenosine_5_monophosphate metabolite_adenosine metabolite_alanine metabolite_4_amino_5_aminomethyl_2_methylpyrimidine metabolite_anthranilate metabolite_arginine metabolite_argininosuccinic_acid metabolite_caffeine metabolite_carnitine metabolite_choline metabolite_citric_acid metabolite_cyanocobalamin metabolite_cysteine metabolite_cytidine metabolite_cytosine metabolite_desthiobiotin metabolite_chitobiose metabolite_dimethylsulfoniopropionate metabolite_D_fructose_1_6_bisphosphate metabolite_fructose_6_phosphate metabolite_fumaric_acid metabolite_x001a_aminobutyric_acid metabolite_D_glucosamine metabolite_glucose_6_phosphate metabolite_glutamic_acid metabolite_glutamine metabolite_glutathione metabolite_sn_glycerol_3_phosphate metabolite_glycine metabolite_glycine_betaine metabolite_guanine metabolite_guanosine metabolite_4_methyl_5_thiazoleethanol metabolite_histadine metabolite_histidinol metabolite_homocysteine metabolite_hydroxocobalamin metabolite_hydroxyproline metabolite_hypoxanthine metabolite_inosine metabolite_inosine_5_monophosphate metabolite_isoleucine metabolite_kynurenine metabolite_leucine metabolite_lumichrome metabolite_malic_acid metabolite_methionine metabolite_methylmalonic_acid metabolite_5_methylthioadenosine metabolite_n_acetyl_glucosamine metabolite_N_acetyl_D_glucosamine_6_phosphate metabolite_n_acetyl_glutamic_acid metabolite_n_acetyl_muramic_acid metabolite_phosphoserine metabolite_ornithine metabolite_pantothenic_acid metabolite_phenylalanine metabolite_proline metabolite_riboflavin metabolite_D_Ribose_5_phosphate metabolite_serine metabolite_taurocholic_acid metabolite_spermidine metabolite_succinic_acid metabolite_sucrose metabolite_syringic_acid metabolite_thiamine metabolite_threonine_homoserine metabolite_thymidine metabolite_chitotriose metabolite_tryptophan metabolite_tyrosine metabolite_uric_acid metabolite_uridine metabolite_uridine_5_monophosphate metabolite_valine metabolite_xanthine metabolite_xanthosine Proch_conc Syn_conc Picoeuk_conc HetBac_conc Beckman-Coulter CytoFLEX S Horiba Aqualog scanning fluorometer TSQ Vantage, Thermo Scientific Technicon AutoAnalyzer II™ and Alpkem RFA 300™ Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific theme None, User defined sample identification latitude longitude date treatment time of day reactive phosphorus (PO4) nitrate plus nitrite Silicate, Si(OH)4, silicic acid Ammonium total organic Carbon total organic Nitrogen metabolite prochlorococcus abundance prochlorococcus abundance pico_euks heterotrophic bacteria abundance featureType BCO-DMO Standard Parameters Flow Cytometer Fluorometer Quadrupole Mass Spectrometer Technicon AutoAnalyzer II Ultra-high-performance liquid chromatography 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. Collaborative Research: The Influence of Sponge Holobiont Metabolism on Coral Reef Dissolved Organic Matter and Reef Microorganisms https://osprey.bco-dmo.org/project/822468 Collaborative Research: The Influence of Sponge Holobiont Metabolism on Coral Reef Dissolved Organic Matter and Reef Microorganisms <p><em>NSF Award Abstract:</em><br /> The seawater around coral reefs is typically low in nutrients, yet coral reefs are teeming with life and are often compared to oases in a desert. Life exists in these 'marine deserts' in large part, due to symbiotic associations between single-celled microbes and invertebrates such as corals and sponges. The concentration and type of dissolved organic matter (DOM), a complex pool of organic nutrients such as amino acids, vitamins, and other diverse compounds, also affects the health of coral reefs. The composition of DOM on coral reefs is linked to both the composition of free-living microbes in the seawater and to the nutrition of filter-feeding organisms, such as corals and sponges. However, the factors that influence the composition of DOM on coral reefs and the consequences of how it changes are not well understood. Recent work suggests that sponges could have a significant impact on the composition of reef dissolved organic nutrients, depending on sponge species due to differences in filtration capacity and in their symbiotic microbial communities. This project characterizes how diverse sponge species process DOM on coral reefs and determines the impacts of this processing on the free-living microbial community. Seawater is collected from sponges (pre- and post- sponge filtration) on coral reefs in the relatively pristine region of Curacao, and incubation experiments measure the impact of sponge filtration on the growth of the free-living microbial community. The organic nutrients of seawater samples are analyzed using cutting-edge techniques to distinguish the types of nutrients that are processed by sponges. The incubation experiments, using free-living microbes collected from the coral reef, quantify the impact of sponge filtration on the growth and composition of this community. This project provides fundamental understanding of how sponges contribute to the base of the coral reef food web. As the human-driven impacts continue to alter the composition of organisms on reefs, this understanding is necessary to predict changes to reef microbial food webs and is thus essential for scientists, reef managers, and policy decision makers. This project trains undergraduate students and a postdoctoral scholar and contributes to undergraduate and K-12 education through development of sponge-centric lessons that focus on local U.S. east coast aquatic environments as well as coral reef ecosystems.</p> <p>Sponges vary in their capacity to filter seawater and in their associated microbial communities, leading to diverse metabolic strategies that often coexist in one habitat. While it is well-established that sponges are important in processing dissolved organic matter (DOM), an important reservoir of reduced carbon compounds, and transferring this energy to benthic food webs, there has been limited work to understand the consequences of sponge processing on the composition of coral reef DOM and on pelagic food webs. Specifically, while studies have shown that exudates of corals and algae select for specific groups of picoplankton (autotrophic and heterotrophic, respectively), similar data for sponges are required to understand the multiple factors that shape the composition of DOM and of the picoplankton community on coral reefs. Thus, this project is aimed at addressing a major knowledge gap of the role of sponge-derived DOM (sponge exometabolome) in coral reef biogeochemistry. An in situ sampling design targeting prominent Caribbean sponges and picoplankton incubation experiments is coupled to address both the composition of sponge exometabolomes and delineate shifts in the picoplankton community derived from sponge exometabolomes. Molecular-level changes to seawater DOM by sponge processing and the impact of these changes on the overall coral reef DOM profile is assessed with two DOM analysis techniques: a commonly used fluorometry technique (fDOM analysis) and with high-resolution mass spectrometry (LC-MS/MS). Additionally, microbiome and functional gene profiling, growth metrics, and nutrient analyses are employed to assess changes in the picoplankton community in response to sponge exometabolomes. Advanced data analysis techniques then synthesize data generated by each approach to provide novel insight on a poorly uncharacterized biogeochemical pathway on coral reefs. The work outlined here represents entirely novel information on the impact of sponge metabolism on the composition of DOM, sheds light on biologically important molecules involved in benthic-pelagic coupling, and importantly, generates data using standardized methods, thus facilitating comparison to previous and future DOM datasets.</p> <p>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.</p> Sponge Holobiont DOM largerWorkCitation project eng; USA oceans -81.40871111 -81.40871111 24.562002777778 24.562002777778 2022-01-06 2022-01-08 Caribbean Sea 0 BCO-DMO catalogue of parameters from Effects of sponge excurrent seawater on coral reef picoplankton composition in the Florida Keys January 2022 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/964048.rdf Name: SampleID Units: unitless Description: <p>Unique Sample Identity as Treatment_Time_TreatmentNumber_RreplicateNumber_</p> http://lod.bco-dmo.org/id/dataset-parameter/964049.rdf Name: latitude Units: decimal degrees Description: <p>Latitude of sampling location, south is negative</p> http://lod.bco-dmo.org/id/dataset-parameter/964050.rdf Name: longitude Units: decimal degrees Description: <p>Longitude of sampling location, west is negative</p> http://lod.bco-dmo.org/id/dataset-parameter/965037.rdf Name: sampling_date Units: unknown Description: <p>Sampling date</p> http://lod.bco-dmo.org/id/dataset-parameter/964051.rdf Name: Trtmt Units: unitless Description: <p>Treatment of sponge exhalent media, coral exhalent media, reef ambient seawater media, or sponge exhalant + coral exhalent media, each with and without picoplankton inoculum. SpongeM: Treatment 1, Sponge exhalant (0.2µm filtered) inoculated with picoplankton (1.6µm filtered surface water); SpongeCtl: Treatment 2, Sponge exhalant (0.2µm filtered) without picoplankton inoculum; SpongeCoralM: Treatment 3, Sponge exhalant + Coral exhalant media (0.2 µm filtered) inoculated with picoplankton (1.6 µm filtered surface water); SpongeCoralCtl: Treatment 4, Sponge exhalant + Coral exhalant media (0.2 µm filtered) without picoplankton inoculum; FswM: Treatment 5, reef ambient seawater media (0.2 µm filtered) inoculated with picoplankton (1.6 µm filtered surface water); FswCtl: Treatment 6, reef ambient seawater media (0.2 µm filtered) without picoplankton inoculum; CoralM: Treatment 7, Coral exhalant media (0.2 µm filtered) inoculated with picoplankton (1.6 µm filtered surface water); CoralCtl: Treatment 8, Coral exhalant media (0.2 µm filtered) without picolanton inoculum</p> http://lod.bco-dmo.org/id/dataset-parameter/964052.rdf Name: Time Units: hours Description: <p>Time of sampling in hours with the start of the experiment as T0</p> http://lod.bco-dmo.org/id/dataset-parameter/964053.rdf Name: PO4 Units: micromolar per liter (µM/L) Description: <p>Concentration of phosphate per liter of surface water sample</p> http://lod.bco-dmo.org/id/dataset-parameter/964054.rdf Name: NOx Units: micromolar per liter (µM/L) Description: <p>Concentration of nitrite + nitrate</p> http://lod.bco-dmo.org/id/dataset-parameter/964055.rdf Name: Silicate Units: micromoles per liter (µM/L) Description: <p>Concentration of dissolved silicate in micromoles per liter of sample water</p> http://lod.bco-dmo.org/id/dataset-parameter/964056.rdf Name: NH4 Units: micromolar per liter (µM/L) Description: <p>Concentration of ammonium per liter of surface water sample</p> http://lod.bco-dmo.org/id/dataset-parameter/964057.rdf Name: TOC Units: micromolar per liter (µM)/L Description: <p>Concentration of dissolved organic carbon</p> http://lod.bco-dmo.org/id/dataset-parameter/964058.rdf Name: TON Units: micromolar per liter (µM/L) Description: <p>Concentration of total organic nitrogen (derived from TOC - dissolved inorganic nitrogen)</p> http://lod.bco-dmo.org/id/dataset-parameter/964059.rdf Name: metabolite_2_3_dihydroxypropane_1_sulfonate Units: picomolar (pM) Description: <p>2,3-dihydroxypropane-1-sulfonate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964060.rdf Name: metabolite_2_3_dihydroxybenzoic_acid Units: picomolar (pM) Description: <p>2,3-dihydroxybenzoic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964061.rdf Name: metabolite_2_deoxycytidine Units: picomolar (pM) Description: <p>2&#039;-deoxycytidine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964062.rdf Name: metabolite_2_methyl_4_oxopentanoic Units: picomolar (pM) Description: <p>2-methyl-4-oxopentanoic metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964063.rdf Name: metabolite_2_Keto_3_deoxy_6_phosphogluconate Units: picomolar (pM) Description: <p>2-Keto-3-deoxy-6-phosphogluconate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964064.rdf Name: metabolite_3_methyl_2_oxobutanoic_acid Units: picomolar (pM) Description: <p>3-methyl-2-oxobutanoic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964065.rdf Name: metabolite_4_aminobenzoic_acid Units: picomolar (pM) Description: <p>4-aminobenzoic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964066.rdf Name: metabolite_4_methyl_2_oxopentanoic_acid Units: picomolar (pM) Description: <p>4-methyl-2-oxopentanoic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964067.rdf Name: metabolite_6_hydroxymelatonin Units: picomolar (pM) Description: <p>6-hydroxymelatonin metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964068.rdf Name: metabolite_adenine Units: picomolar (pM) Description: <p>Adenine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964069.rdf Name: metabolite_adenosine_5_monophosphate Units: picomolar (pM) Description: <p>Adenosine 5&#039;-monophosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964070.rdf Name: metabolite_adenosine Units: picomolar (pM) Description: <p>Adenosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964071.rdf Name: metabolite_alanine Units: picomolar (pM) Description: <p>Alanine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964072.rdf Name: metabolite_4_amino_5_aminomethyl_2_methylpyrimidine Units: picomolar (pM) Description: <p>4-amino-5-aminomethyl-2-methylpyrimidine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964073.rdf Name: metabolite_anthranilate Units: picomolar (pM) Description: <p>Anthranilate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964074.rdf Name: metabolite_arginine Units: picomolar (pM) Description: <p>Arginine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964075.rdf Name: metabolite_argininosuccinic_acid Units: picomolar (pM) Description: <p>Argininosuccinic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964076.rdf Name: metabolite_caffeine Units: picomolar (pM) Description: <p>Caffeine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964077.rdf Name: metabolite_carnitine Units: picomolar (pM) Description: <p>Carnitine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964078.rdf Name: metabolite_choline Units: picomolar (pM) Description: <p>Choline metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964079.rdf Name: metabolite_citric_acid Units: picomolar (pM) Description: <p>Citric acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964080.rdf Name: metabolite_cyanocobalamin Units: picomolar (pM) Description: <p>Cyanocobalamin metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964081.rdf Name: metabolite_cysteine Units: picomolar (pM) Description: <p>Cysteine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964082.rdf Name: metabolite_cytidine Units: picomolar (pM) Description: <p>Cytidine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964083.rdf Name: metabolite_cytosine Units: picomolar (pM) Description: <p>Cytosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964084.rdf Name: metabolite_desthiobiotin Units: picomolar (pM) Description: <p>Desthiobiotin metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964085.rdf Name: metabolite_chitobiose Units: picomolar (pM) Description: <p>Chitobiose metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964086.rdf Name: metabolite_dimethylsulfoniopropionate Units: picomolar (pM) Description: <p>Dimethylsulfoniopropionate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964087.rdf Name: metabolite_D_fructose_1_6_bisphosphate Units: picomolar (pM) Description: <p>D-fructose 1,6-bisphosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964088.rdf Name: metabolite_fructose_6_phosphate Units: picomolar (pM) Description: <p>Fructose 6-phosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964089.rdf Name: metabolite_fumaric_acid Units: picomolar (pM) Description: <p>Fumaric acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964090.rdf Name: metabolite_x001a_aminobutyric_acid Units: picomolar (pM) Description: <p>-aminobutyric acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964091.rdf Name: metabolite_D_glucosamine Units: picomolar (pM) Description: <p>D-glucosamine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964092.rdf Name: metabolite_glucose_6_phosphate Units: picomolar (pM) Description: <p>Glucose 6-phosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964093.rdf Name: metabolite_glutamic_acid Units: picomolar (pM) Description: <p>Glutamic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964094.rdf Name: metabolite_glutamine Units: picomolar (pM) Description: <p>Glutamine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964095.rdf Name: metabolite_glutathione Units: picomolar (pM) Description: <p>Glutathione metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964096.rdf Name: metabolite_sn_glycerol_3_phosphate Units: picomolar (pM) Description: <p>sn-glycerol 3-phosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964097.rdf Name: metabolite_glycine Units: picomolar (pM) Description: <p>Glycine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964098.rdf Name: metabolite_glycine_betaine Units: picomolar (pM) Description: <p>Glycine betaine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964099.rdf Name: metabolite_guanine Units: picomolar (pM) Description: <p>Guanine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964100.rdf Name: metabolite_guanosine Units: picomolar (pM) Description: <p>Guanosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964101.rdf Name: metabolite_4_methyl_5_thiazoleethanol Units: picomolar (pM) Description: <p>4-methyl-5-thiazoleethanol metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964102.rdf Name: metabolite_histadine Units: picomolar (pM) Description: <p>Histadine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964103.rdf Name: metabolite_histidinol Units: picomolar (pM) Description: <p>Histidinol metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964104.rdf Name: metabolite_homocysteine Units: picomolar (pM) Description: <p>Homocysteine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964105.rdf Name: metabolite_hydroxocobalamin Units: picomolar (pM) Description: <p>Hydroxocobalamin metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964106.rdf Name: metabolite_hydroxyproline Units: picomolar (pM) Description: <p>Hydroxyproline metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964107.rdf Name: metabolite_hypoxanthine Units: picomolar (pM) Description: <p>Hypoxanthine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964108.rdf Name: metabolite_inosine Units: picomolar (pM) Description: <p>Inosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964109.rdf Name: metabolite_inosine_5_monophosphate Units: picomolar (pM) Description: <p>Inosine 5&#039;-monophosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964110.rdf Name: metabolite_isoleucine Units: picomolar (pM) Description: <p>Isoleucine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964111.rdf Name: metabolite_kynurenine Units: picomolar (pM) Description: <p>Kynurenine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964112.rdf Name: metabolite_leucine Units: picomolar (pM) Description: <p>Leucine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964113.rdf Name: metabolite_lumichrome Units: picomolar (pM) Description: <p>Lumichrome metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964114.rdf Name: metabolite_malic_acid Units: picomolar (pM) Description: <p>Malic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964115.rdf Name: metabolite_methionine Units: picomolar (pM) Description: <p>Methionine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964116.rdf Name: metabolite_methylmalonic_acid Units: picomolar (pM) Description: <p>Methylmalonic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964117.rdf Name: metabolite_5_methylthioadenosine Units: picomolar (pM) Description: <p>5&#039;-methylthioadenosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964118.rdf Name: metabolite_n_acetyl_glucosamine Units: picomolar (pM) Description: <p>n-acetyl glucosamine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964119.rdf Name: metabolite_N_acetyl_D_glucosamine_6_phosphate Units: picomolar (pM) Description: <p>N-acetyl-D-glucosamine 6-phosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964120.rdf Name: metabolite_n_acetyl_glutamic_acid Units: picomolar (pM) Description: <p>n-acetyl glutamic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964121.rdf Name: metabolite_n_acetyl_muramic_acid Units: picomolar (pM) Description: <p>n-acetyl muramic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964122.rdf Name: metabolite_phosphoserine Units: picomolar (pM) Description: <p>Phosphoserine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964123.rdf Name: metabolite_ornithine Units: picomolar (pM) Description: <p>Ornithine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964124.rdf Name: metabolite_pantothenic_acid Units: picomolar (pM) Description: <p>Pantothenic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964125.rdf Name: metabolite_phenylalanine Units: picomolar (pM) Description: <p>Phenylalanine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964126.rdf Name: metabolite_proline Units: picomolar (pM) Description: <p>Proline metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964127.rdf Name: metabolite_riboflavin Units: picomolar (pM) Description: <p>Riboflavin metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964128.rdf Name: metabolite_D_Ribose_5_phosphate Units: picomolar (pM) Description: <p>D-Ribose 5-phosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964129.rdf Name: metabolite_serine Units: picomolar (pM) Description: <p>Serine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964130.rdf Name: metabolite_taurocholic_acid Units: picomolar (pM) Description: <p>Taurocholic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964131.rdf Name: metabolite_spermidine Units: picomolar (pM) Description: <p>Spermidine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964132.rdf Name: metabolite_succinic_acid Units: picomolar (pM) Description: <p>Succinic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964133.rdf Name: metabolite_sucrose Units: picomolar (pM) Description: <p>Sucrose metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964134.rdf Name: metabolite_syringic_acid Units: picomolar (pM) Description: <p>Syringic acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964135.rdf Name: metabolite_thiamine Units: picomolar (pM) Description: <p>Thiamine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964136.rdf Name: metabolite_threonine_homoserine Units: picomolar (pM) Description: <p>Threonine / Homoserine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964137.rdf Name: metabolite_thymidine Units: picomolar (pM) Description: <p>Thymidine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964138.rdf Name: metabolite_chitotriose Units: picomolar (pM) Description: <p>Chitotriosemetabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964139.rdf Name: metabolite_tryptophan Units: picomolar (pM) Description: <p>Tryptophan metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964140.rdf Name: metabolite_tyrosine Units: picomolar (pM) Description: <p>Tyrosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964141.rdf Name: metabolite_uric_acid Units: picomolar (pM) Description: <p>Uric acid metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964142.rdf Name: metabolite_uridine Units: picomolar (pM) Description: <p>Uridine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964143.rdf Name: metabolite_uridine_5_monophosphate Units: picomolar (pM) Description: <p>Uridine 5&#039;-monophosphate metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964144.rdf Name: metabolite_valine Units: picomolar (pM) Description: <p>Valine metabolic concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964145.rdf Name: metabolite_xanthine Units: picomolar (pM) Description: <p>Xanthine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964146.rdf Name: metabolite_xanthosine Units: picomolar (pM) Description: <p>Xanthosine metabolite concentration</p> http://lod.bco-dmo.org/id/dataset-parameter/964147.rdf Name: Proch_conc Units: cells per millliliter (mL-1) Description: <p>Prochlorococcus concentration (mL-1), very low abndances (&lt; 100 cells/mL) may represent noise</p> http://lod.bco-dmo.org/id/dataset-parameter/964148.rdf Name: Syn_conc Units: cells per millliliter (mL-1) Description: <p>Synechococcus concentration (mL-1), very low abundances (&lt; 100 cells/mL) may represent noise</p> http://lod.bco-dmo.org/id/dataset-parameter/964149.rdf Name: Picoeuk_conc Units: cells per millliliter (mL-1) Description: <p>Picoeukaryote (photosynthetic) concentration (mL-1)</p> http://lod.bco-dmo.org/id/dataset-parameter/964150.rdf Name: HetBac_conc Units: cells per millliliter (mL-1) Description: <p>Heterotrophic (nonpigmented) bacteria concentration (mL-1)</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 121262 https://darchive.mblwhoilibrary.org/server/api/core/bitstreams/c3a5cce2-8cd7-4e9c-825a-c9a2459190c7/content download https://doi.org/10.26008/1912/bco-dmo.963872.1 download onLine dataset &lt;p&gt;Incubation experiments were set up with sponge excurrent water, coral exudates, sponge excurrent and coral exudates, or surface reef water as media and with reef surface picoplankton as inoculum to compare microbial growth over 48 hours. Excurrent seawater samples of three sponge species (Verongula rigida, Xestospongia muta, and Niphates digitalis) were collected at Looe Key Reef in the Florida Keys, U.S.A. at approximately 10m depth (24.54605, -81.40610). The exhalent seawater was collected using acid-rinsed glass syringes with a three-way valve and PharMed BPT tubing (Cole-Parmer, USA) connected to 1L FlexFoil Plus sampling bags (SKC, Pittsburgh, PA, USA). Water was drawn in at about 2 mL per minute, which is slower than the pumping rate of the sponges. The exhalent seawater was collected the day before the experiment and filtered (0.2 µm, Omnipore, Millipore Sigma, USA) prior to storage at 4°C. Coral exudates were collected by incubating four fragments each of Acropora cervicornis and Orbicella faveolata were incubated in aquaria containing 0.2 µm filtered (Omnipore) seawater from Mote Marine laboratory seawater intake which pulls from the nearby seawater canal. The aquaria was placed in flowthrough seawater raceway in the sunlight under one layer of shade cloth (~23 – 262 µE s-1 with a mix of sun and clouds) for 6.5 hrs. The coral exudate seawater was then filtered 0.2 µm filtered and stored at 4°C.&lt;/p&gt; &lt;p&gt;Surface seawater over Looe Key Reef was collected the morning of the experimental setup and was used for both the inoculum of bacterioplankton and for background seawater control medium (0.2 µm filtered to create the ambient seawater medium). The sponge excurrent seawater was combined in a 1:1:1 ratio from the three sponge species. Then four different mediums were created: coral exudate water, sponge exhalant water, sponge exhalant water plus coral exudates (1:1 ratio of each medium), and ambient seawater. Coral reef surface picoplankton, served as inoculum and was added to the media in a 1:3 ratio of inoculum to medium. An initial ‘soup’ was made for each treatment in 5L acid-cleaned polycarbonate food grade containers and the total volume of the initial soup was 3900 mL for treatments with microbial inoculum and 2900 mL for control treatments without inoculum. From the initial soup, three T0 samples of seawater (1400 mL) were taken for chemical and microbial analyses. The rest of the experimental seawater with inoculum was then distributed into 2L acid-cleaned polycarbonate bottles (1L per bottle) and incubated in a flow through seawater table in the dark for 48 hours. There were five replicate bottles for each treatment (coral, sponge, sponge + coral, ambient seawater) and three replicate bottles for each no-inoculum control corresponding to each treatment. Nutrients and the bacterioplankton community composition and abundance were assessed at the start of the experiment (T0) and after 48hrs (T48).&lt;/p&gt; &lt;p&gt;Seawater sampled from the incubation ‘soup’ or bottles was used for multiple nutrient analyses and DNA extraction. Prior to filtration, 1 mL of seawater was preserved in&amp;nbsp;500 µL of paraformaldehyde (0.5% final concentration), which were stored in the refrigerator for 1-2 hours and then stored at -80°C until shipped to the University of Hawai’i for flow cytometry analysis. Also prior to filtration, approximately 30 mL of sample was poured into acid and milliQ water rinsed and combusted glass amber EPA vials with acid cleaned septa and lids. These 30 mL samples were acidified to pH ~2 with concentrated HCl and stored at room temperature until analysis for total organic carbon (TOC) and total nitrogen (TN) at Woods Hole Oceanographic Institution. The rest of the seawater was filtered through an Omnipore 0.2 um filter using a peristaltic pump and acid and milliQ water rinsed Pharmed BP tubing and Teflon filter holders. The filter was stored at -80C until DNA extraction, while the filtrate was poured into acid and milliQ water rinsed: 1) HDPE bottles (~20 mL) for inorganic nutrient analysis at the University of Oregon, 2) polycarbonate bottles (~500 mL) for extraction of dissolved organic matter (DOM).&amp;nbsp; Filtered seawater was processed using PPL solid phase extraction following the protocol by Dittmar et al. 2008. Extracts were dried to nearly completeness, leaving a small viscous drop in the vial. These extracts were then shipped to WHOI for metabolomics analysis.&lt;/p&gt; &lt;p&gt;DNA from the Omnipore filters was extracted using a commercial kit and the 16S rRNA gene was amplified using the modified earth microbiome primer set for the V4 region&amp;nbsp;(515F and 806R, Apprill et al. 2015). PCRs were sent to Middle Tennessee State University for library construction and sequencing on an Illumina MiSeq, producing FASTQ files as output.&lt;/p&gt; &lt;p&gt;Targeted Metabolite Analysis by UPLC-MS:&lt;/p&gt; &lt;p&gt;DOM extracts were reconstituted in 200 μl MilliQ water with 50 ng/ml isotopically-labeled injection standards d2 biotin, d6 succinic acid, d4 cholic acid, and d7 indole 3 acetic acid. We used ultra-performance liquid chromatography (Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific) coupled to a heated electrospray ionization source (H-ESI) and a triple quadrupole mass spectrometer (TSQ Vantage, Thermo Scientific) operated under selected reaction monitoring (SRM) mode. We performed chromatographic separation with a Waters Acquity HSS T3 column (2.1 × 100 mm, 1.8 μm) equipped with a Vanguard pre-column and maintained at 40°C. We eluted the metabolites from the column with (A) 0.1% formic acid in water and (B) 0.1% formic acid in acetonitrile at a flow rate of 0.5 mL min-1, according to the gradient: 0 min, 1% B; 1 min, 1%B; 3 min, 15%B; 6 min, 50%B; 9 min, 95%B; 10 min, 95%B; 10.2 min, 1%B; 12 min, 1%B (total run time = 12 min). Settings for source gases were 55 (sheath), 20 (auxiliary) and 0 (sweep), and these settings are presented in arbitrary units. The heated capillary temperature was 375 °C and the vaporizer temperature was 400 °C. For positive and negative modes, we performed separate autosampler injections of 5 μL each.&lt;/p&gt; &lt;p&gt;Inorganic nutrients:&amp;nbsp;&lt;/p&gt; &lt;p&gt;Inorganic nutrients included phosphate, nitrate+nitrite, nitrite, ammonia, and silicic acid. The phosphate method is a modification of the molybdenum blue procedure of&lt;/p&gt; &lt;p&gt;Bernhardt and Wilhelms (1967), in which phosphate is determined as reduced phosphomolybdic acid employing hydrazine as the reductant. The nitrate + nitrite analysis uses the basic method of Armstrong et al. (1967), with modifications to improve the precision and ease of operation.&amp;nbsp; Sulfanilamide and N-(1-Napthyl)ethylenediamine dihydrochloride react with nitrite to form a colored diazo compound. For the nitrate + nitrite analysis, nitrate is first reduced to nitrite using an OTCR and imidazole buffer as described by Patton (1983). Nitrite analysis is performed on a separate channel, omitting the cadmium reductor and the buffer. The method is based on that of Armstrong et al. (1967) as adapted by Atlas et al. (1971).&amp;nbsp; Addition of an acidic molybdate reagent forms silicomolybdic acid which is then reduced by stannous chloride. This indophenol blue method is modified from ALPKEM RFA methodology which references Methods for Chemical Analysis of Water and Wastes, March 1984, EPA-600/4-79-020, &quot;Nitrogen Ammonia&quot;, Method 350.1 (Colorimetric, Automated Phenate) A detailed description of the continuous segmented flow procedures used can be found in Gordon et. al.&amp;nbsp; (1994).&amp;nbsp;&lt;/p&gt; &lt;p&gt;Flow cytometry:&amp;nbsp;&lt;/p&gt; &lt;p&gt;Samples were preserved and stored at -80°C until later batch analysis at the University of Hawaii SOEST Flow Cytometry Facility (www.soest.hawaii.edu/sfcf). Microbial cells were enumerated using flow cytometry (Selph, 2021). In brief, samples (0.1 mL) were thawed in batches, stained with the DNA dye Hoechst 34580 (1 µg/mL final), then run at 30 µL min-1 on a Beckman-Coulter CytoFlex S flow cytometer, using lasers emitting at 375 nm (to detect Hoechst), 488 nm (for scatter and chlorophyll parameters), and 561 nm (for phycoerythrin). Resulting listmode files (FCS 3.0) were analyzed using FlowJo software (Becton Dickinson, v. 10.8.2) to distinguish microbial populations based on their fluorescence signals (chlorophyll, phycoerythrin, DNA), as well as forward and right-angle light scatter.&amp;nbsp; Heterotrophic bacteria were distinguished from phytoplankton by their DNA signature and absence of pigment.&amp;nbsp; Prochlorococcus and Synechococcus were separated from larger eukaryotic phytoplankton by their light scatter signatures, as well as their characteristic pigment and DNA signatures.&amp;nbsp; Other phytoplankton (eukaryotes, mostly 2-20 µm pico- and nano-sized cells given the small volume analyzed) had higher light scatter and more chlorophyll fluorescence per cell. &amp;gt;&lt;/p&gt; Specified by the Principal Investigator(s) &lt;p&gt;Targeted Metabolomics:&lt;/p&gt; &lt;p&gt;Samples were analyzed in random order and injected pooled samples at regular intervals (every 8 samples). We monitored two SRM transitions per compound for quantification and confirmation; these transitions were optimized previously using authentic standards. We generated 8-point external calibration curves based on peak area for each compound. We converted raw data files from proprietary Thermo (.RAW) format to mzML using the msConvert tool (Chambers et al., 2012) prior to processing with El-MAVEN (Agrawal et al., 2019). Metabolite concentrations were provided from WHOI as raw data in ng per ml in the 200 μl extract. Extraction efficiency values in seawater were used from Johnson et al. 2017. For any metabolites with extraction efficiency below 1%, these were removed from the analysis. For any metabolites with extraction efficiency of 30% or higher, the values were corrected based on the extraction efficiency (e.g., if the extraction efficiency was 50% then the value was multiplied by 2). We then converted the ng/ml values into total nanograms by multiplying by 0.2 then divided by the sample volume in liters to produce ng/L concentrations. These concentrations were converted to picomolar concentrations using the formula weight for each metabolite.&lt;/p&gt; Specified by the Principal Investigator(s) * adjusted names to comply with database requirements * Added sampling lat/lon and dates where missing * Specified by BCO-DMO Data Managers 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 Beckman-Coulter CytoFLEX S Beckman-Coulter CytoFLEX S PI Supplied Instrument Name: Beckman-Coulter CytoFLEX S PI Supplied Instrument Description:Flow Cytometry: Preserved samples were analyzed using a Beckman-Coulter CytoFLEX S flow cytometer with 375 nm, 405 nm, 488 nm, and 561 nm lasers activated. Instrument Name: Flow Cytometer Instrument Short Name:Flow Cytometer Instrument Description: Flow cytometers (FC or FCM) are automated instruments that quantitate properties of single cells, one cell at a time. They can measure cell size, cell granularity, the amounts of cell components such as total DNA, newly synthesized DNA, gene expression as the amount messenger RNA for a particular gene, amounts of specific surface receptors, amounts of intracellular proteins, or transient signalling events in living cells. (from: http://www.bio.umass.edu/micro/immunology/facs542/facswhat.htm) Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB37/ Horiba Aqualog scanning fluorometer Horiba Aqualog scanning fluorometer PI Supplied Instrument Name: Horiba Aqualog scanning fluorometer PI Supplied Instrument Description:Fluorometer was used to measure fDOM Instrument Name: Fluorometer Instrument Short Name:Fluorometer Instrument Description: A fluorometer or fluorimeter is a device used to measure parameters of fluorescence: its intensity and wavelength distribution of emission spectrum after excitation by a certain spectrum of light. The instrument is designed to measure the amount of stimulated electromagnetic radiation produced by pulses of electromagnetic radiation emitted into a water sample or in situ. Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/113/ TSQ Vantage, Thermo Scientific TSQ Vantage, Thermo Scientific PI Supplied Instrument Name: TSQ Vantage, Thermo Scientific PI Supplied Instrument Description:Targeted metabolomics were measured using the ultra-performance liquid chromatography (Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific) coupled to a heated electrospray ionization source (H-ESI) and a triple quadrupole mass spectrometer (TSQ Vantage, Thermo Scientific) operated under selected reaction monitoring (SRM) mode at the Woods Hole Oceanographic Institution (WHOI).  Instrument Name: Quadrupole Mass Spectrometer Instrument Short Name:QMS Instrument Description: A piece of apparatus that consists of an ion source, a mass-to-charge analyser, a detector and a vacuum system and is used to measure mass spectra. The detector is a quadrupole mass-to-charge analyser, which holds the ions in a stable orbit by an electric field generated by four parallel electrodes.   Technicon AutoAnalyzer II™ and Alpkem RFA 300™ Technicon AutoAnalyzer II™ and Alpkem RFA 300™ PI Supplied Instrument Name: Technicon AutoAnalyzer II™ and Alpkem RFA 300™ PI Supplied Instrument Description:Inorganic nutrients measured using Technicon AutoAnalyzer II™ components were used to measure phosphate and ammonium; and Alpkem RFA 300™ components were used for silicic acid, nitrate plus nitrite, and nitrite.> Instrument Name: Technicon AutoAnalyzer II Instrument Short Name: Instrument Description: A rapid flow analyzer that may be used to measure nutrient concentrations in seawater. It is a continuous segmented flow instrument consisting of a sampler, peristaltic pump, analytical cartridge, heating bath, and colorimeter. See more information about this instrument from the manufacturer. Community Standard Description: http://vocab.nerc.ac.uk/collection/L22/current/TOOL0291/ Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific PI Supplied Instrument Name: Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific PI Supplied Instrument Description:Targeted metabolomics were measured using the ultra-performance liquid chromatography (Accela Open Autosampler and Accela 1250 Pump, Thermo Scientific) coupled to a heated electrospray ionization source (H-ESI) and a triple quadrupole mass spectrometer (TSQ Vantage, Thermo Scientific) operated under selected reaction monitoring (SRM) mode at the Woods Hole Oceanographic Institution (WHOI).  Instrument Name: Ultra-high-performance liquid chromatography Instrument Short Name:UHPLC Instrument Description: Ultra high-performance liquid chromatography: Column chromatography where the mobile phase is a liquid, the stationary phase consists of very small (< 2 microm) particles and the inlet pressure is relatively high.