| Contributors | Affiliation | Role |
|---|---|---|
| Bahr, Keisha D. | Texas A&M, Corpus Christi (TAMU-CC) | Principal Investigator |
| McNicholl, Conall | Hawaii Institute of Marine Biology (HIMB) | Scientist |
| Armstrong, David A. | Texas A&M, Corpus Christi (TAMU-CC) | Student |
| Bretzing-Tungate, Robert | Texas A&M, Corpus Christi (TAMU-CC) | Data Manager |
| York, Amber D. | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
Note: This section includes a description of coral biological data not included in this dataset. See "Related Datasets" section for additional data from this experiment.
Overview
Seasonal mesocosm incubations of Montipora capitata and Pocillopora acuta were conducted over a two-year period (June 2022–March 2024) at the Hawai‘i Institute of Marine Biology (HIMB). Corals were collected from fringe reefs located around the institute. Experiments were carried out during winter (January 10–March 17) and summer (June 10–August 17) in outdoor flow-through mesocosms at the Hawaii Institute of Marine Biology in Kane`ohe, Hawai`i. Due to facility constraints, all treatments could not run simultaneously. Therefore, each seasonal experiment consisted of two consecutive 30-day exposures, each using newly collected coral colonies. All procedures were standardized across replicates, except for adjustments in total alkalinity (TA) and pH required to create the treatments.
Coral Collection and Pre-Experiment Holding
Coral colonies of Montipora capitata and Pocillopora acuta were collected by hand while snorkeling from shallow (1 m) fringing reef habitat surrounding HIMB in Kāne‘ohe Bay, Hawai‘i. Colonies were transported immediately to the mesocosm facility and held for one week in flow-through tanks that were identical in design to the experimental mesocosms.
One day prior to placement in experimental mesocosms, all colonies were stained for eight hours with alizarin red to mark the initial skeletal growth band (reference: Jokiel and Morrissey 1993).
At the start of each 30-day experiment, colonies were randomly assigned to mesocosms, buoyant weighed following Jokiel (1978), and placed into a standardized grid layout. Each mesocosm contained 20 colonies of each species arranged in alternating species order to avoid species-level clustering effects.
Mesocosm Facility Design
The flow-through mesocosm facility (Jokiel et al. 2014) consisted of twelve 450 L fiberglass tanks operating on continuous seawater flow. Tanks were arranged in four rows of three mesocosms, with each row supplied by a dedicated header tank. Water was pumped at ~3 m depth from the adjacent bay, supplying unfiltered seawater that preserved natural diel and seasonal variability.
Each header tank fed seawater into a 100 L mixing reservoir positioned above the system to facilitate chemical manipulation before seawater entered the mesocosms. Mesocosm residence time was greater than one hour. Each mesocosm contained two submersible circulation pumps and an airstone that ran continuously to maintain water movement and oxygenation.
Treatment Conditions and Carbonate Chemistry Manipulation
Four carbonate chemistry conditions were generated across consecutive 30-day exposures by altering total alkalinity (TA), dissolved inorganic carbon (DIC), and pH. Target conditions included:
1. Ambient control
2. Low pH (ΔpH ~ -0.3 from ambient)
3. High or low TA (±100 µmol kg-1 from ambient)
4. Combined low pH and TA alteration
TA adjustments were made using a peristaltic pump delivering either 1.0 M HCl (for low TA) at ~3 mL min-1 or 1.0 M Na2CO3 (for high TA) at ~2 mL min-1 directly into each row’s mixing tank.
pH reductions were achieved by bubbling pure CO2 or a CO2-air mixture directly into designated mesocosms using a Maxi-Jet 1600 pump-driven venturi injector.
Because each row was supplied by a single header tank, only one direction of TA manipulation (increase or decrease) could be applied per row in a given 30-day experiment. The subsequent 30-day experiment performed the opposite TA manipulation, ensuring all conditions were tested each season.
Seawater Sampling and Measurements
Daily Parameters
Temperature, salinity, dissolved oxygen, and pHNBS were measured daily at mid-day in all mesocosms and header tanks using a YSI multimeter (YSI ProDSS or YSI 556 MPS).
pH electrodes were calibrated daily using NIST-traceable pH 4, 7, and 10 buffers and corrected to the total scale using Tris buffer from A. Dickson (Scripps Institution of Oceanography). Dissolved oxygen calibrations followed manufacturer water-saturated air calibration protocols.
Total Alkalinity Sampling and Storage
Discrete seawater samples (100 mL) for total alkalinity (TA) were collected twice weekly.
Sampling procedures followed best practices described in Dickson et al. (2007):
Rinsed sample bottles three times with sample water.
Collected seawater in acid-cleaned 100 mL borosilicate bottles.
No headspace was left to prevent gas exchange.
Samples were analyzed within 12 hours of collection and stored at ambient temperature in the dark until analysis.
TA Determination
TA was determined using open-cell potentiometric titration on a Metrohm 877 Titrino Plus equipped with a Metrohm 9101 Herisau glass pH electrode.
All titrations were standardized using certified reference materials (CRM, batch number provided by Dickson Laboratory). Electrode slope, offset, and drift were checked daily.
Carbonate system variables (DIC, pCO2, HCO3-, CO32-, and Omega aragonite) were calculated using the R package seacarb.
Calcification Measurements
Calcification rates were quantified using buoyant weighing following Jokiel (1978). Colonies were weighed on day 0 and day 30. Calcification rate (G) was calculated as:
G (g CaCO3 d-1) = 1.54 × (Wf – Wi) / d
where 1.54 g cm-3 is the density of aragonite, Wf and Wi are final and initial buoyant weights, and d is the exposure duration in days.
Three colonies per species per mesocosm were randomly selected for further biometric analysis. Three branch tips per colony were sampled immediately before and after the exposure period. Collected fragments were frozen at -20 C until post-processing.
Mortality was low and did not differ across treatments; dead colonies were excluded from analysis. All corals were returned to the reef at the conclusion of experiments.
Post-Hoc Processing of Coral Samples
At the end of each experimental season, all frozen branch tips were transported to Texas A&M University–Corpus Christi for tissue, symbiont, pigment, and skeletal analysis.
Tissue Removal and Homogenization
Branch tips were thawed and sprayed with phosphate-buffered saline (PBS) using a Paasche airbrush.
Tissue slurry (25 mL) was sonicated for 20 seconds (Qsonica ultrasonic processor).
Samples were vortexed and centrifuged (VWR International centrifuge) to separate host tissue and symbionts.
Host Tissue Analyses
Aliquots of the host fraction were used to quantify protein concentration using spectrophotometry (SpectraMax M3). PBS was used as a blank.
Symbiont Analyses
Symbiont pellets were resuspended and counted using a Bright-Line hemocytometer (Hausser Scientific) under 10× magnification on a Leica ICC50W microscope.
Pigment Analyses
Chlorophyll a and c were extracted using 100 percent acetone. Absorbance was measured on a SpectraMax M3 spectrophotometer, with acetone blanks. Calculations followed Parsons et al. (1984).
Skeletal Preparation and Biometric Measurements
After tissue removal:
Skeletons were soaked in 10 percent bleach to remove remaining tissue.
Samples were dried for 4 hours at 60 C (DX302C drying oven).
Dry weights were recorded using a VWR-4002B2 analytical balance.
Skeletal volume was measured by water displacement in a 100 mL graduated cylinder.
Three-dimensional skeletal scans were generated using an Einscan-SE scanner and processed in MeshLab to estimate total surface area.
Note: This section includes a description of coral biological data not included in this dataset. See "Related Datasets" section for additional data from this experiment.
Overview
Seasonal mesocosm incubations of Montipora capitata and Pocillopora acuta were conducted over a two-year period (June 2022–March 2024) at the Hawai‘i Institute of Marine Biology (HIMB). Corals were collected from fringe reefs located around the institute. Experiments were carried out during winter (January 10–March 17) and summer (June 10–August 17) in outdoor flow-through mesocosms at the Hawaii Institute of Marine Biology in Kane`ohe, Hawai`i. Due to facility constraints, all treatments could not run simultaneously. Therefore, each seasonal experiment consisted of two consecutive 30-day exposures, each using newly collected coral colonies. All procedures were standardized across replicates, except for adjustments in total alkalinity (TA) and pH required to create the treatments.
Coral Collection and Pre-Experiment Holding
Coral colonies of Montipora capitata and Pocillopora acuta were collected by hand while snorkeling from shallow (1 m) fringing reef habitat surrounding HIMB in Kāne‘ohe Bay, Hawai‘i. Colonies were transported immediately to the mesocosm facility and held for one week in flow-through tanks that were identical in design to the experimental mesocosms.
One day prior to placement in experimental mesocosms, all colonies were stained for eight hours with alizarin red to mark the initial skeletal growth band (reference: Jokiel and Morrissey 1993).
At the start of each 30-day experiment, colonies were randomly assigned to mesocosms, buoyant weighed following Jokiel (1978), and placed into a standardized grid layout. Each mesocosm contained 20 colonies of each species arranged in alternating species order to avoid species-level clustering effects.
Mesocosm Facility Design
The flow-through mesocosm facility (Jokiel et al. 2014) consisted of twelve 450 L fiberglass tanks operating on continuous seawater flow. Tanks were arranged in four rows of three mesocosms, with each row supplied by a dedicated header tank. Water was pumped at ~3 m depth from the adjacent bay, supplying unfiltered seawater that preserved natural diel and seasonal variability.
Each header tank fed seawater into a 100 L mixing reservoir positioned above the system to facilitate chemical manipulation before seawater entered the mesocosms. Mesocosm residence time was greater than one hour. Each mesocosm contained two submersible circulation pumps and an airstone that ran continuously to maintain water movement and oxygenation.
Treatment Conditions and Carbonate Chemistry Manipulation
Four carbonate chemistry conditions were generated across consecutive 30-day exposures by altering total alkalinity (TA), dissolved inorganic carbon (DIC), and pH. Target conditions included:
1. Ambient control
2. Low pH (ΔpH ~ -0.3 from ambient)
3. High or low TA (±100 µmol kg-1 from ambient)
4. Combined low pH and TA alteration
TA adjustments were made using a peristaltic pump delivering either 1.0 M HCl (for low TA) at ~3 mL min-1 or 1.0 M Na2CO3 (for high TA) at ~2 mL min-1 directly into each row’s mixing tank.
pH reductions were achieved by bubbling pure CO2 or a CO2-air mixture directly into designated mesocosms using a Maxi-Jet 1600 pump-driven venturi injector.
Because each row was supplied by a single header tank, only one direction of TA manipulation (increase or decrease) could be applied per row in a given 30-day experiment. The subsequent 30-day experiment performed the opposite TA manipulation, ensuring all conditions were tested each season.
Seawater Sampling and Measurements
Daily Parameters
Temperature, salinity, dissolved oxygen, and pHNBS were measured daily at mid-day in all mesocosms and header tanks using a YSI multimeter (YSI ProDSS or YSI 556 MPS).
pH electrodes were calibrated daily using NIST-traceable pH 4, 7, and 10 buffers and corrected to the total scale using Tris buffer from A. Dickson (Scripps Institution of Oceanography). Dissolved oxygen calibrations followed manufacturer water-saturated air calibration protocols.
Total Alkalinity Sampling and Storage
Discrete seawater samples (100 mL) for total alkalinity (TA) were collected twice weekly.
Sampling procedures followed best practices described in Dickson et al. (2007):
Rinsed sample bottles three times with sample water.
Collected seawater in acid-cleaned 100 mL borosilicate bottles.
No headspace was left to prevent gas exchange.
Samples were analyzed within 12 hours of collection and stored at ambient temperature in the dark until analysis.
TA Determination
TA was determined using open-cell potentiometric titration on a Metrohm 877 Titrino Plus equipped with a Metrohm 9101 Herisau glass pH electrode.
All titrations were standardized using certified reference materials (CRM, batch number provided by Dickson Laboratory). Electrode slope, offset, and drift were checked daily.
Carbonate system variables (DIC, pCO2, HCO3-, CO32-, and Omega aragonite) were calculated using the R package seacarb.
Calcification Measurements
Calcification rates were quantified using buoyant weighing following Jokiel (1978). Colonies were weighed on day 0 and day 30. Calcification rate (G) was calculated as:
G (g CaCO3 d-1) = 1.54 × (Wf – Wi) / d
where 1.54 g cm-3 is the density of aragonite, Wf and Wi are final and initial buoyant weights, and d is the exposure duration in days.
Three colonies per species per mesocosm were randomly selected for further biometric analysis. Three branch tips per colony were sampled immediately before and after the exposure period. Collected fragments were frozen at -20 C until post-processing.
Mortality was low and did not differ across treatments; dead colonies were excluded from analysis. All corals were returned to the reef at the conclusion of experiments.
Post-Hoc Processing of Coral Samples
At the end of each experimental season, all frozen branch tips were transported to Texas A&M University–Corpus Christi for tissue, symbiont, pigment, and skeletal analysis.
Tissue Removal and Homogenization
Branch tips were thawed and sprayed with phosphate-buffered saline (PBS) using a Paasche airbrush.
Tissue slurry (25 mL) was sonicated for 20 seconds (Qsonica ultrasonic processor).
Samples were vortexed and centrifuged (VWR International centrifuge) to separate host tissue and symbionts.
Host Tissue Analyses
Aliquots of the host fraction were used to quantify protein concentration using spectrophotometry (SpectraMax M3). PBS was used as a blank.
Symbiont Analyses
Symbiont pellets were resuspended and counted using a Bright-Line hemocytometer (Hausser Scientific) under 10× magnification on a Leica ICC50W microscope.
Pigment Analyses
Chlorophyll a and c were extracted using 100 percent acetone. Absorbance was measured on a SpectraMax M3 spectrophotometer, with acetone blanks. Calculations followed Parsons et al. (1984).
Skeletal Preparation and Biometric Measurements
After tissue removal:
Skeletons were soaked in 10 percent bleach to remove remaining tissue.
Samples were dried for 4 hours at 60 C (DX302C drying oven).
Dry weights were recorded using a VWR-4002B2 analytical balance.
Skeletal volume was measured by water displacement in a 100 mL graduated cylinder.
Three-dimensional skeletal scans were generated using an Einscan-SE scanner and processed in MeshLab to estimate total surface area.
Primary Data File:
- Loaded "biomass.csv" as resource "995474_v1_buoyant-wgt" in CSV format with header row 1; treated empty strings, "nd", and "NA" as missing values
- Converted field "date" from format "%m/%d/%Y" to ISO 8601 date format "%Y-%m-%d" (output type: date)
- Set field types: "bwt" as number; "coral", "experiment", "mesocosm", "tp" as integer; "date" as date ("%Y-%m-%d"); "season", "species", "treatment" as string
- Applied BCO-DMO metadata to all fields: "bwt" (buoyant weight, grams), "coral" (coral ID number), "date" (sampling date, ISO 8601), "experiment" (experiment ID), "mesocosm" (tank ID), "season" (season of measurement; smr=summer, wntr=winter), "species" (coral species; mc=Montipora capitata, pa=Pocillopora acuta), "tp" (timepoint sampling day since experiment start), "treatment" (treatment ID; amb=control, co2=elevated pCO2, lta=lowered total alkalinity, hta=elevated total alkalinity)
- Updated package-level metadata with field-level statistics (count, min, max) for all 9 fields across 7,680 rows
- Output written to "995474_v1_buoyant-wgt.csv"
Tables added as supplemental files (same files attached from the related biological data mesocosm dataset):
- Loaded three CSV files (env_data.csv, sw_chem.csv, crm_tris_std.csv) using filenames as resource names, with missing values defined as "", "nd", and "NA"
- Renamed column "experiment (#)" to "experiment_num" in crm_tris_std
- Converted date col from %m/%d/%Y to ISO 8601 format (%Y-%m-%d) in crm_tris_std, env_data, and sw_chem; the tod column in sw_chem contains "day"/"night" text values, not actual times, so no ISO datetime conversion was applied to it
- Applied find/replace on tod_hr in env_data to add missing colons to time values in HHMM/HMM format (e.g., "2300" → "23:00")
- Applied second find/replace on tod_hr in env_data to standardize format to HH:MM by truncating seconds and zero-padding hours; values already containing colons (e.g., "23:00") were left unchanged
- Set col types for all four resources: numeric fields as number or integer, date fields as date with %Y-%m-%d format, tod_hr in env_data as time with %H:%M format, and various categorical fields as string
- Output written to env_data.csv, sw_chem.csv, crm_tris_std.csv, and 995164_v1_coral-biological-data.csv
| Parameter | Description | Units |
| date | the date of sampling (ISO 8601 format yyyy-mm-dd) | unitless |
| experiment | the experiment ID number | unitless |
| tp | Timepoint indicator describing sampling day within the experiment since the start (0). (e.g., Day 0 or Day 30). | unitless |
| season | the season in which measurements were conducted (smr = summer and wntr = winter) | unitless |
| treatment | treatment identification (amb = control, co2 = elevated pCO2, lta = lowered total alkalinity, hta = elevated total alkalinity) | unitless |
| mesocosm | tank identification number | unitless |
| species | coral species [mc = Montipora capitata (urn:lsid:marinespecies.org:taxname:287697), pa = Pocillopora acuta (urn:lsid:marinespecies.org:taxname:759099)] | unitless |
| coral | the coral identification number | unitless |
| bwt | buoyant weight | grams (g) |
| Dataset-specific Instrument Name | Flow-through Mesocosm System |
| Generic Instrument Name | Aquarium |
| Dataset-specific Description | Reference: Jokiel et al. 2014
Description: Outdoor continuous-flow mesocosm facility at HIMB consisting of twelve 450 L fiberglass tanks (1 m x 1 m x 0.5 m) supplied by seawater pumped from ~3 m depth in Kāneʻohe Bay.
Flow: Adjustable head box delivering ~50 min-1 seawater residence time.
Notes: All mesocosms contained two submersible pumps and an airstone for continuous mixing. |
| Generic Instrument Description | Aquarium - a vivarium consisting of at least one transparent side in which water-dwelling plants or animals are kept |
| Dataset-specific Instrument Name | |
| Generic Instrument Name | Diving Mask and Snorkel |
| Dataset-specific Description | Snorkel-based Coral Collection Gear
Use: Hand-collection of coral colonies at 1 m depth surrounding HIMB.
Notes: Standard snorkeling gear (mask, snorkel, fins) and hand tools (bone cutters, chisels) were used; no specialized electronic instrumentation. |
| Generic Instrument Description | A diving mask (also half mask, dive mask or scuba mask) is an item of diving equipment that allows underwater divers, including, scuba divers, free-divers, and snorkelers to see clearly underwater.
Snorkel: A breathing apparatus for swimmers and surface divers that allows swimming or continuous use of a face mask without lifting the head to breathe, consisting of a tube that curves out of the mouth and extends above the surface of the water. |
| Dataset-specific Instrument Name | CO2 Gas Delivery System |
| Generic Instrument Name | no_bcodmo_term |
| Dataset-specific Description | Components: CO2 cylinder, regulator, and gas injection line.
Use: Manipulation of mesocosm pH by bubbling pure CO2 or CO2-air mixtures directly into individual mesocosms.
Calibration: Flow checked daily using bubble-rate timing; regulator pressures recorded each morning. |
| Generic Instrument Description | No relevant match in BCO-DMO instrument vocabulary. |
| Dataset-specific Instrument Name | |
| Generic Instrument Name | Pump |
| Dataset-specific Description | Submersible Pumps
Manufacturer: Maxi-Jet
Model: Maxi-Jet 1600
Use: Internal water circulation and CO2 injection mixing inside mesocosms.
Calibration: Flow rate verified twice per season using volumetric bucket timing. |
| Generic Instrument Description | A pump is a device that moves fluids (liquids or gases), or sometimes slurries, by mechanical action. Pumps can be classified into three major groups according to the method they use to move the fluid: direct lift, displacement, and gravity pumps |
| Dataset-specific Instrument Name | Peristaltic Pumps for TA Manipulation |
| Generic Instrument Name | Pump |
| Dataset-specific Description | Manufacturer: To be filled by user (brand not provided)
Flow Rate: 2–3 mL min-1
Use: Delivery of 1.0 M HCl or 1.0 M Na2CO3 to achieve target alkalinity offsets.
Calibration: Flow rate calibrated at the start of each 30-day experiment by measuring timed output volumes. |
| Generic Instrument Description | A pump is a device that moves fluids (liquids or gases), or sometimes slurries, by mechanical action. Pumps can be classified into three major groups according to the method they use to move the fluid: direct lift, displacement, and gravity pumps |
| Dataset-specific Instrument Name | Buoyant Weighing Apparatus |
| Generic Instrument Name | scale or balance |
| Dataset-specific Description | Use: Measurement of initial and final coral colony mass for calcification estimates.
Components:
Electronic balance
Custom platform submerged in seawater for buoyant weight readings
Calibration:
Balance calibrated daily using known-weight standards.
System checked for drift by repeated weighing of a reference coral fragment. |
| Generic Instrument Description | Devices that determine the mass or weight of a sample. |
| Dataset-specific Instrument Name | |
| Generic Instrument Name | Titrator |
| Dataset-specific Description | Open-cell Potentiometric Titrator for Total Alkalinity
Manufacturer: Metrohm
Model: 877 Titrino Plus with 9101 Herisau glass pH electrode
Use: Measurement of TA from discrete 100 mL seawater samples twice weekly.
Calibration:
Daily electrode slope and offset verification.
Weekly CRM calibration using certified reference materials from A. Dickson (batch number supplied by user).
Notes: All titrations performed at constant temperature. |
| Generic Instrument Description | Titrators are instruments that incrementally add quantified aliquots of a reagent to a sample until the end-point of a chemical reaction is reached. |
| Dataset-specific Instrument Name | YSI Multiparameter Water Quality Meters |
| Generic Instrument Name | Water Quality Multiprobe |
| Dataset-specific Description | YSI ProDSS
YSI 556 MPS
Manufacturer: YSI Inc.
Parameters Measured: Temperature, salinity, dissolved oxygen, and pHNBS.
Calibration:
DO probe calibrated daily using water-saturated air.
pH electrode calibrated daily using pH 4, 7, and 10 NIST buffers; corrected to total scale using Tris buffer from Dickson Laboratory.
Conductivity and temperature routinely checked using certified standards. |
| Generic Instrument Description | An instrument which measures multiple water quality parameters based on the sensor configuration. |
NSF Award Abstract:
Corals build calcium carbonate skeletons to maintain the three-dimensional structure of a coral reef, which provides habitat for many organisms and protects shorelines from bioerosion and storm damage. However, changes in ocean chemistry threaten the ability of corals to build and sustain these ecological important structures. To further the understanding of how climate change impacts coral reefs, this project investigates how changes in ocean carbonate chemistry directly influence coral calcification. The researchers are conducting a series of experiments on corals grown in seawater tanks to study corals responses to seawater chemistry in a changing ocean. Broader impacts of the project include student research opportunities, science-inquiry labs, and virtual learning. This project supports the training of several early career researchers, Ph.D. students, undergraduates, and high school students in the disciplines of chemistry, engineering, and marine ecology. Researchers partner with the Texas State Aquarium to communicate with the general public through a virtual research expedition series that will focus on coral reef health. This series includes interviews, behind the scene tours, and virtual dives on coral reefs in Hawaii.
This project examines the fundamental connections between seawater chemistry and coral physiology by investigating the modulation of seawater chemistry in the microenvironment surrounding corals. Specifically, this project 1) examines the response of corals to differing carbonate chemistry and 2) characterizes the proton gradient across the corals' boundary layers under differing ocean acidification conditions. Results of this work isolate whether carbonate ions or hydrogen ions have a stronger influence on calcification rates. This work utilizes a state-of-the-art experimental mesocosm facility that combines an automated systems to simultaneously and independently control both total alkalinity and carbon dioxide in the tanks to examine coral response under different carbon chemistry scenarios. Small-scale gradients in carbon chemistry surrounding the corals are being characterized using an innovative solid-state, reagentless sensor capable of making simultaneous measurements of two critical carbon system parameters. Coral biological response variables are quantified during short-term incubations and long-term mesocosm manipulations to understand physiological implications across multiple scales (i.e., individual and community scales) and across boundary layers.
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.
| Funding Source | Award |
|---|---|
| NSF Division of Ocean Sciences (NSF OCE) | |
| NSF Division of Ocean Sciences (NSF OCE) |