http://lod.bco-dmo.org/id/dataset/889857 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 2023-02-14 ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data ISO 19115-2:2009(E) Isotopic analysis of ¹³C and ¹⁵N for sponges, coral, and zooxanthellae (family Symbiodiniaceae) used in a 'pulse-chase' experiment to examine the uptake of sponge-derived nutrients by the coral holobiont 2023-02-16 publication 2023-02-16 revision Marine Biological Laboratory/Woods Hole Oceanographic Institution Library (MBLWHOI DLA) 2023-02-24 publication https://doi.org/10.26008/1912/bco-dmo.889857.1 Cara L. Fiore Appalachian State University principalInvestigator Amy Apprill Woods Hole Oceanographic Institution principalInvestigator Cole G. Easson Middle Tennessee State University principalInvestigator Christopher J. Freeman College of Charleston 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: Reigel, A. M., Easson, C. G., Apprill, A., Freeman, C. J., Bartley, M. M., Fiore, C. L. (2023) Isotopic analysis of ¹³C and ¹⁵N for sponges, coral, and zooxanthellae (family Symbiodiniaceae) used in a 'pulse-chase' experiment to examine the uptake of sponge-derived nutrients by the coral holobiont. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2023-02-16 [if applicable, indicate subset used]. doi:10.26008/1912/bco-dmo.889857.1 [access date] Stable Isotope 13C and 15N Dataset Description: Methods and Sampling: &lt;p&gt;&lt;strong&gt;Sample Collection and Maintenance&lt;/strong&gt;&lt;br /&gt; Three coral species, 1 octocoral and 2 scleractinians, were utilized in this experiment. The octocoral, &lt;em&gt;Eunicea flexuosa&lt;/em&gt;, was collected from Wonderland Reef (24.558694, -81.503528) within the Florida Keys National Marine Sanctuary (FKNMS) under the state of Florida (FL) saltwater fishing permit (Permit number: I-H1R76333834 held by A.M. Reigel). Three axial branch tips were clipped from each of 10 healthy &lt;em&gt;E. flexuosa&lt;/em&gt; colonies located at depths of ~5-8 meters (m). Branch tips were kept in seawater and immediately transported to the outdoor land-based nursery (Climate and Acidification Ocean Simulator (CAOS)) at Mote Marine Laboratory at Summerland Key, FL, USA where they were placed into a shaded, temperature- and pH-maintained flow-through tank and allowed to acclimate for ~24 hours. The two hard coral species, &lt;em&gt;Acropora cervicornis&lt;/em&gt; and &lt;em&gt;Orbicella faveolata&lt;/em&gt; were provided by Mote Marine Laboratory’s field (&lt;em&gt;A. cervicornis; &lt;/em&gt;Coordinates: 24.562747, -81.400455) and land-based (&lt;em&gt;O. faveolata&lt;/em&gt;) nurseries as permitted under the FKNMS-2015-163-A3. All hard coral fragments were placed in the same flow-through CAOS tanks as the &lt;em&gt;E. Flexuosa&lt;/em&gt; samples and allowed to acclimate for ~24 hours.&lt;/p&gt; &lt;p&gt;To develop a representative sponge community for the Florida Keys reefs, we collected 5 individuals of each of 6 sponge species (&lt;em&gt;Niphates digitalis&lt;/em&gt;, Verongula rigida, &lt;em&gt;Aplysina fulva&lt;/em&gt;, &lt;em&gt;Aplysina cauliformis&lt;/em&gt;, &lt;em&gt;Xestospongia muta&lt;/em&gt;, &lt;em&gt;Callyspongia aculeata&lt;/em&gt;) from Wonderland Reef under FL saltwater fishing permit (Permit #: I-H1R76333834 held by A.M. Reigel). Sponges were kept in seawater and immediately transported to the lab where they were placed in a shaded CAOS flow-through tank to acclimate for ~24 hours. Corals and sponges were not in the same CAOS tanks during the acclimation period.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Stable Isotope Pulse-Chase Incubations&lt;/strong&gt;&lt;br /&gt; Following 24 hours of acclimation time, sponges (n=2 per species) and coral fragments (n=4-5 per species) were removed from their respective acclimation tanks and immediately placed into labeled, sterile WhirlPak© bags and stored in the -20º Celsius (C) freezer (i.e., initial, or T₀, samples). The remaining sponges were moved from the acclimation tank to a 'pulse' tank for a 3-hour incubation. The 'pulse' tank was pre-filled with 84 liters of 0.22 micrometer (µm) filtered seawater spiked with isotopically labeled inorganic compounds in the following final concentrations: 0.1 grams per liter (g L⁻¹) sodium bicarbonate (NaH¹³CO₃), 0.01g L⁻¹ sodium nitrate (Na¹⁵NO₃), and 0.01g L⁻¹ ammonium chloride (¹⁵NH₄Cl). At the end of the 3-hour 'pulse', a subset of sponges ('pulse' samples) was destructively sampled following the same procedure detailed above. The remaining enriched sponges (n=5 per species) were transferred to five separate flow-through tanks, such that each tank contained one individual of each sponge species, for a 1-hour 'rinse' period to flush away any labeled compounds that were not incorporated into the sponge tissue. At the end of the rinse, two fragments from each coral species were placed into each of eight experimental tanks, five enriched sponge-containing tanks, and three no-sponge control tanks, for the 6-hr 'chase'. During the duration of the 'chase', the flow-through system was turned off and a submersible aquarium pump was added to each tank to maintain circulation and gas exchange. A single fragment from each coral species was destructively sampled from each experimental tank twice during the 'chase': at the half point (T₃) and at the end (T₆). All enriched sponges were also destructively sampled at the end of the 'chase'.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Coral and Zooxanthellae Separations&lt;/strong&gt;&lt;br /&gt; To prepare for downstream analyses the coral fractions, host and zooxanthellae, were manually separated. Scleractinian fragments were thawed and airbrushed with an aerosolized jet of 0.22 micrometer (µm) filtered seawater to physically separate the coral tissue and skeleton and suspend the coral tissue material into a homogenate. To separate host tissue from zooxanthellae cells, the homogenate was centrifuged at 2000g for 3-5 minutes. Centrifugation formed a pellet comprised of zooxanthellae cells and a homogenate of host material. The host homogenate was pipetted into a separate sterile 50 milliliter (ml) Falcon tube. The homogenate, zooxanthellae pellet, and skeletal fragments were frozen and transported to Appalachian State University where they were stored at -20 degrees Fahrenheit (F) until further processing. At Appalachian State, host homogenates and zooxanthellae pellets were thawed and checked for purity. Impure fractions were combined, homogenized with a tissue homogenizer (maximum speed for ~15 seconds) to physically separate zooxanthellae cells from host tissue, and centrifuged (3000 x g, 6 minutes) to pellet the zooxanthellae cells. Separated fractions were combined with original fractions each time and checked for purity under the microscope. The process was repeated until at least 80% purity was reached.&lt;/p&gt; &lt;p&gt;&lt;em&gt;E. flexuosa&lt;/em&gt; fractions were separated using a different process. First, the frozen coral branches were lyophilized (Labconco™ FreeZone™ Bulk Tray Dryer) for 22-24 hours, until they were completely dry. Following lyophilization, the axial skeleton was removed and the tissue was ground up using a mortar and pestle (note: separate mortar and pestle sets were used for control and enriched samples). The ground tissue was weighed and then rehydrated in 10ml of MilliQ water in a sterile 15ml Falcon tube. Very quickly following rehydration, the sclerites (skeletal fragments) sank to the bottom of the tube and the remaining host homogenate was pipetted into a new tube taking care not to transfer the sclerites. The homogenate was homogenized using a tissue homogenizer for ~15 seconds at maximum speed and centrifuged at 4000g for 5 minutes to separate the fractions. The centrifugation step was repeated as necessary until the host homogenate and zooxanthellae pellets were pure. Following both octocoral and scleractinian fraction separations, 50 microliters (µl) of pure zooxanthellae from each sample was transferred to a cryovial with 50µl of 10% paraformaldehyde (PFA) to fix the cells and stored in the refrigerator for future zooxanthellae counts. The pure host homogenates and remaining zooxanthellae pellets were stored in the -20F freezer.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Stable Isotope Analysis&lt;/strong&gt;&lt;br /&gt; All sponge and zooxanthellae tissues and the octocoral host homogenate were lyophilized, weighed, and the dried material was homogenized with a mortar and pestle. Homogenized samples were decalcified via exposure to 12M HCL in desiccator chambers. Subsamples of ~1-2.5 milligrams (mg) of decalcified tissue from each sample were packed into silver cups. Scleractinian coral homogenates, made as previously detailed, contained filtered seawater, and if lyophilized, the salts could contaminate the host tissue. To remove the salts, host homogenates were passed through a sterile pre-weighed 1.6 µm GF/F filter that captured host material while seawater passed through and was discarded. The filters containing host material were frozen, dried overnight in a drying oven, weighed, decalcified as detailed above, and sent to the Marine Biological Laboratory (MBL) Stable Isotope Laboratory where they were also packed into silver cups.&lt;/p&gt; &lt;p&gt;Packed samples were analyzed at the MBL Stable Isotope Laboratory for δ¹⁵N and δ¹³C using a Europa 20-20 continuous-flow isotope ratio mass spectrometer interfaced with a Europa ANCA-SL elemental analyzer. The analytical precision based on replicate analyses of isotopically homogeneous international standards is +/- 0.1 ‰ for both δ¹⁵N and δ¹³C measurements.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Sampling and Analysis Dates&lt;/strong&gt;&lt;br /&gt; Following the 08-Dec-2020 sampling, all field-collected samples were acclimated in the aquarium system for 24 hours and the stable isotope 'pulse-chase' experiment was conducted over 11 hours on 09-Dec-2020. The host and zooxanthellae fractions were frozen and then the separations of the fractions from &lt;em&gt;A. cervicornis &lt;/em&gt;and &lt;em&gt;O. faveolata&lt;/em&gt; happened on 17-Dec-2020 and both fractions were frozen again. The other experimental steps occurred over several months in Spring 2021.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Known Issues/Problems&lt;/strong&gt;&lt;br /&gt; Four control (i.e., non-enriched) coral samples (Of_T6_7, AC_T3_3, AC_T6_1, and EF_T6_1) and their corresponding zooxanthellae have δ¹⁵N and δ¹³C values that suggest they may have been contaminated by the labeled isotopes at some point during the sample preparation process. These samples are found at the bottom of the data file and are identified as 'Control-Contaminated' in the Treatment column. These samples should be excluded or used with caution.&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 Cara L. Fiore Appalachian State University 828-262-7548 572 Rivers St Boone NC 28607 USA fiorec@appstate.edu pointOfContact 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 Christopher J. Freeman College of Charleston 817-707-5840 66 George Street Department of Biology Charleston SC 29424 USA freemancj@cofc.edu pointOfContact Alicia M. Reigel Appalachian State University Lexington VA USA areigel@wlu.edu pointOfContact asNeeded Dataset Version: 1 Unknown Sample_ID Tissue_Type Species Treatment TimePoint Field_Sampling_Date Latitude Longitude Tissue_Weight_mg d13C Carbon_umol d15N Nitrogen_umol Molar_C_N_ratio Europa ANCA-SL elemental analyzer Europa 20-20 continuous-flow isotope ratio mass spectrometer Labconco FreeZone Bulk Tray Dryer theme None, User defined sample identification sample description species scientific name (binomial) treatment time_point date latitude longitude weight d13C Elemental carbon (C) d15N Nitrogen ratio featureType BCO-DMO Standard Parameters Elemental Analyzer Isotope-ratio Mass Spectrometer Lyophilizer 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 -81.503528 -81.400455 24.558694 24.562747 2020-12-08 2021-05-31 Caribbean Sea 0 BCO-DMO catalogue of parameters from Isotopic analysis of ¹³C and ¹⁵N for sponges, coral, and zooxanthellae (family Symbiodiniaceae) used in a 'pulse-chase' experiment to examine the uptake of sponge-derived nutrients by the coral holobiont 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/889887.rdf Name: Sample_ID Units: unitless Description: <p>Sample ID number</p> http://lod.bco-dmo.org/id/dataset-parameter/889888.rdf Name: Tissue_Type Units: unitless Description: <p>Type of organism the tissue sample was obtained from: Sponge = tissue from a sponge; Zoox = tissue of coral-associated dinoflagellates obtained by centrifuge separation from coral host tissue; coral = coral tissue (includes material from coral host and + associated microbial community); blank = experimental control samples.</p> http://lod.bco-dmo.org/id/dataset-parameter/889889.rdf Name: Species Units: unitless Description: <p>Species name</p> http://lod.bco-dmo.org/id/dataset-parameter/889890.rdf Name: Treatment Units: unitless Description: <p>Control or enriched with stable isotopes. Those labeled with Control_Contaminated = control samples that appear to have been contained with enriched isotopes during the sample preparation process and should be removed or cautiously utilized by other researchers.</p> http://lod.bco-dmo.org/id/dataset-parameter/889891.rdf Name: TimePoint Units: unitless Description: <p>Time point. T0 = initial; pulse = at the end of the 3-hr pulse; T3= 3 hrs into the chase; T6 = 6 hrs into the chase.</p> http://lod.bco-dmo.org/id/dataset-parameter/889892.rdf Name: Field_Sampling_Date Units: unitless Description: <p>Date when the samples were collected from in-situ nursery (Acropora cervicornis samples) or natural reef habitat (Eunicea flexuosa and all sponge samples).</p> http://lod.bco-dmo.org/id/dataset-parameter/889893.rdf Name: Latitude Units: decimal degrees North Description: <p>Latitude coordinate of in-situ collection location</p> http://lod.bco-dmo.org/id/dataset-parameter/889894.rdf Name: Longitude Units: decimal degrees North Description: <p>Longitude coordinate of in-situ collection location</p> http://lod.bco-dmo.org/id/dataset-parameter/889895.rdf Name: Tissue_Weight_mg Units: milligrams (mg) Description: <p>Weight of tissue used in analysis of isotope content</p> http://lod.bco-dmo.org/id/dataset-parameter/889896.rdf Name: d13C Units: parts per thousand (per mil, o/oo) Description: <p>Measure of the ratio of 13C to 12C in the sample as compared to the same ratio in the standard Vienna Pee Dee Belemnite (PDB) reference material PDB</p> http://lod.bco-dmo.org/id/dataset-parameter/889897.rdf Name: Carbon_umol Units: micromoles (umol) Description: <p>Total carbon content in the sample</p> http://lod.bco-dmo.org/id/dataset-parameter/889898.rdf Name: d15N Units: parts per thousand (per mil, o/oo) Description: <p>Measure of the ratio of 15N to 14N in the sample as compared to the same ratio in the standard Atmosphere (AIR) reference material</p> http://lod.bco-dmo.org/id/dataset-parameter/889899.rdf Name: Nitrogen_umol Units: micromoles (umol) Description: <p>Total nitrogen content in the sample</p> http://lod.bco-dmo.org/id/dataset-parameter/889900.rdf Name: Molar_C_N_ratio Units: unitless Description: <p>Molar ratio of C:N in the sample</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 18314 https://darchive.mblwhoilibrary.org/server/api/core/bitstreams/bc3907a9-cd72-50f9-ba51-baeee857655d/content download https://doi.org/10.26008/1912/bco-dmo.889857.1 download onLine dataset &lt;p&gt;&lt;strong&gt;Sample Collection and Maintenance&lt;/strong&gt;&lt;br /&gt; Three coral species, 1 octocoral and 2 scleractinians, were utilized in this experiment. The octocoral, &lt;em&gt;Eunicea flexuosa&lt;/em&gt;, was collected from Wonderland Reef (24.558694, -81.503528) within the Florida Keys National Marine Sanctuary (FKNMS) under the state of Florida (FL) saltwater fishing permit (Permit number: I-H1R76333834 held by A.M. Reigel). Three axial branch tips were clipped from each of 10 healthy &lt;em&gt;E. flexuosa&lt;/em&gt; colonies located at depths of ~5-8 meters (m). Branch tips were kept in seawater and immediately transported to the outdoor land-based nursery (Climate and Acidification Ocean Simulator (CAOS)) at Mote Marine Laboratory at Summerland Key, FL, USA where they were placed into a shaded, temperature- and pH-maintained flow-through tank and allowed to acclimate for ~24 hours. The two hard coral species, &lt;em&gt;Acropora cervicornis&lt;/em&gt; and &lt;em&gt;Orbicella faveolata&lt;/em&gt; were provided by Mote Marine Laboratory’s field (&lt;em&gt;A. cervicornis; &lt;/em&gt;Coordinates: 24.562747, -81.400455) and land-based (&lt;em&gt;O. faveolata&lt;/em&gt;) nurseries as permitted under the FKNMS-2015-163-A3. All hard coral fragments were placed in the same flow-through CAOS tanks as the &lt;em&gt;E. Flexuosa&lt;/em&gt; samples and allowed to acclimate for ~24 hours.&lt;/p&gt; &lt;p&gt;To develop a representative sponge community for the Florida Keys reefs, we collected 5 individuals of each of 6 sponge species (&lt;em&gt;Niphates digitalis&lt;/em&gt;, Verongula rigida, &lt;em&gt;Aplysina fulva&lt;/em&gt;, &lt;em&gt;Aplysina cauliformis&lt;/em&gt;, &lt;em&gt;Xestospongia muta&lt;/em&gt;, &lt;em&gt;Callyspongia aculeata&lt;/em&gt;) from Wonderland Reef under FL saltwater fishing permit (Permit #: I-H1R76333834 held by A.M. Reigel). Sponges were kept in seawater and immediately transported to the lab where they were placed in a shaded CAOS flow-through tank to acclimate for ~24 hours. Corals and sponges were not in the same CAOS tanks during the acclimation period.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Stable Isotope Pulse-Chase Incubations&lt;/strong&gt;&lt;br /&gt; Following 24 hours of acclimation time, sponges (n=2 per species) and coral fragments (n=4-5 per species) were removed from their respective acclimation tanks and immediately placed into labeled, sterile WhirlPak© bags and stored in the -20º Celsius (C) freezer (i.e., initial, or T₀, samples). The remaining sponges were moved from the acclimation tank to a 'pulse' tank for a 3-hour incubation. The 'pulse' tank was pre-filled with 84 liters of 0.22 micrometer (µm) filtered seawater spiked with isotopically labeled inorganic compounds in the following final concentrations: 0.1 grams per liter (g L⁻¹) sodium bicarbonate (NaH¹³CO₃), 0.01g L⁻¹ sodium nitrate (Na¹⁵NO₃), and 0.01g L⁻¹ ammonium chloride (¹⁵NH₄Cl). At the end of the 3-hour 'pulse', a subset of sponges ('pulse' samples) was destructively sampled following the same procedure detailed above. The remaining enriched sponges (n=5 per species) were transferred to five separate flow-through tanks, such that each tank contained one individual of each sponge species, for a 1-hour 'rinse' period to flush away any labeled compounds that were not incorporated into the sponge tissue. At the end of the rinse, two fragments from each coral species were placed into each of eight experimental tanks, five enriched sponge-containing tanks, and three no-sponge control tanks, for the 6-hr 'chase'. During the duration of the 'chase', the flow-through system was turned off and a submersible aquarium pump was added to each tank to maintain circulation and gas exchange. A single fragment from each coral species was destructively sampled from each experimental tank twice during the 'chase': at the half point (T₃) and at the end (T₆). All enriched sponges were also destructively sampled at the end of the 'chase'.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Coral and Zooxanthellae Separations&lt;/strong&gt;&lt;br /&gt; To prepare for downstream analyses the coral fractions, host and zooxanthellae, were manually separated. Scleractinian fragments were thawed and airbrushed with an aerosolized jet of 0.22 micrometer (µm) filtered seawater to physically separate the coral tissue and skeleton and suspend the coral tissue material into a homogenate. To separate host tissue from zooxanthellae cells, the homogenate was centrifuged at 2000g for 3-5 minutes. Centrifugation formed a pellet comprised of zooxanthellae cells and a homogenate of host material. The host homogenate was pipetted into a separate sterile 50 milliliter (ml) Falcon tube. The homogenate, zooxanthellae pellet, and skeletal fragments were frozen and transported to Appalachian State University where they were stored at -20 degrees Fahrenheit (F) until further processing. At Appalachian State, host homogenates and zooxanthellae pellets were thawed and checked for purity. Impure fractions were combined, homogenized with a tissue homogenizer (maximum speed for ~15 seconds) to physically separate zooxanthellae cells from host tissue, and centrifuged (3000 x g, 6 minutes) to pellet the zooxanthellae cells. Separated fractions were combined with original fractions each time and checked for purity under the microscope. The process was repeated until at least 80% purity was reached.&lt;/p&gt; &lt;p&gt;&lt;em&gt;E. flexuosa&lt;/em&gt; fractions were separated using a different process. First, the frozen coral branches were lyophilized (Labconco™ FreeZone™ Bulk Tray Dryer) for 22-24 hours, until they were completely dry. Following lyophilization, the axial skeleton was removed and the tissue was ground up using a mortar and pestle (note: separate mortar and pestle sets were used for control and enriched samples). The ground tissue was weighed and then rehydrated in 10ml of MilliQ water in a sterile 15ml Falcon tube. Very quickly following rehydration, the sclerites (skeletal fragments) sank to the bottom of the tube and the remaining host homogenate was pipetted into a new tube taking care not to transfer the sclerites. The homogenate was homogenized using a tissue homogenizer for ~15 seconds at maximum speed and centrifuged at 4000g for 5 minutes to separate the fractions. The centrifugation step was repeated as necessary until the host homogenate and zooxanthellae pellets were pure. Following both octocoral and scleractinian fraction separations, 50 microliters (µl) of pure zooxanthellae from each sample was transferred to a cryovial with 50µl of 10% paraformaldehyde (PFA) to fix the cells and stored in the refrigerator for future zooxanthellae counts. The pure host homogenates and remaining zooxanthellae pellets were stored in the -20F freezer.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Stable Isotope Analysis&lt;/strong&gt;&lt;br /&gt; All sponge and zooxanthellae tissues and the octocoral host homogenate were lyophilized, weighed, and the dried material was homogenized with a mortar and pestle. Homogenized samples were decalcified via exposure to 12M HCL in desiccator chambers. Subsamples of ~1-2.5 milligrams (mg) of decalcified tissue from each sample were packed into silver cups. Scleractinian coral homogenates, made as previously detailed, contained filtered seawater, and if lyophilized, the salts could contaminate the host tissue. To remove the salts, host homogenates were passed through a sterile pre-weighed 1.6 µm GF/F filter that captured host material while seawater passed through and was discarded. The filters containing host material were frozen, dried overnight in a drying oven, weighed, decalcified as detailed above, and sent to the Marine Biological Laboratory (MBL) Stable Isotope Laboratory where they were also packed into silver cups.&lt;/p&gt; &lt;p&gt;Packed samples were analyzed at the MBL Stable Isotope Laboratory for δ¹⁵N and δ¹³C using a Europa 20-20 continuous-flow isotope ratio mass spectrometer interfaced with a Europa ANCA-SL elemental analyzer. The analytical precision based on replicate analyses of isotopically homogeneous international standards is +/- 0.1 ‰ for both δ¹⁵N and δ¹³C measurements.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Sampling and Analysis Dates&lt;/strong&gt;&lt;br /&gt; Following the 08-Dec-2020 sampling, all field-collected samples were acclimated in the aquarium system for 24 hours and the stable isotope 'pulse-chase' experiment was conducted over 11 hours on 09-Dec-2020. The host and zooxanthellae fractions were frozen and then the separations of the fractions from &lt;em&gt;A. cervicornis &lt;/em&gt;and &lt;em&gt;O. faveolata&lt;/em&gt; happened on 17-Dec-2020 and both fractions were frozen again. The other experimental steps occurred over several months in Spring 2021.&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Known Issues/Problems&lt;/strong&gt;&lt;br /&gt; Four control (i.e., non-enriched) coral samples (Of_T6_7, AC_T3_3, AC_T6_1, and EF_T6_1) and their corresponding zooxanthellae have δ¹⁵N and δ¹³C values that suggest they may have been contaminated by the labeled isotopes at some point during the sample preparation process. These samples are found at the bottom of the data file and are identified as 'Control-Contaminated' in the Treatment column. These samples should be excluded or used with caution.&lt;/p&gt; Specified by the Principal Investigator(s) &lt;p&gt;&lt;strong&gt;BCO-DMO Processing:&lt;/strong&gt;&lt;br /&gt; - converted dates to YYYY-MM-DD format;&lt;br /&gt; - removed spaces and special characters from parameter names;&lt;br /&gt; - created separate columns for latitude and longitude values.&lt;/p&gt; 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 Europa ANCA-SL elemental analyzer Europa ANCA-SL elemental analyzer PI Supplied Instrument Name: Europa ANCA-SL elemental analyzer Instrument Name: Elemental Analyzer Instrument Short Name: Instrument Description: Instruments that quantify carbon, nitrogen and sometimes other elements by combusting the sample at very high temperature and assaying the resulting gaseous oxides. Usually used for samples including organic material. Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB01/ Europa 20-20 continuous-flow isotope ratio mass spectrometer Europa 20-20 continuous-flow isotope ratio mass spectrometer PI Supplied Instrument Name: Europa 20-20 continuous-flow isotope ratio mass spectrometer Instrument Name: Isotope-ratio Mass Spectrometer Instrument Short Name:IR Mass Spec; IRMS Instrument Description: The Isotope-ratio Mass Spectrometer is a particular type of mass spectrometer used to measure the relative abundance of isotopes in a given sample (e.g. VG Prism II Isotope Ratio Mass-Spectrometer). Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB16/ Labconco FreeZone Bulk Tray Dryer Labconco FreeZone Bulk Tray Dryer PI Supplied Instrument Name: Labconco FreeZone Bulk Tray Dryer Instrument Name: Lyophilizer Instrument Short Name:freeze dryer Instrument Description: A lyophilizer, also known as freeze dryer or liofilizador, is a device that is used to freeze-dry material.