DOI: 10.7280/d1p10v
title: Factorial field manipulation reveals multi-stressor effects on a critical habitat-forming shellfish

description: Ocean acidification and warming are expected to have negative impacts on
 marine organisms, and particularly on shell-building species.
 Acidification and warming may operate independently or interactively,
 amplifying or mitigating impacts on individuals. Previous results have
 primarily come from lab studies of single species, yet these climate
 stressors are occurring within naturally dynamic systems with high abiotic
 and biotic variability. As a result, the impacts of these stressors in
 situ remains poorly understood. We conducted a 6-month field manipulation
 to determine the effects of ocean acidification and warming on a
 habitat-forming shellfish, the Pacific blue mussel (Mytilus trossulus), in
 a dynamic coastal system. Twenty tide pools were factorially manipulated,
 including unmanipulated control, CO2 added, warmed, and combined CO2 added
 and warmed treatments. We measured mussel shell thickness, strength, and
 corrosion at 0, 3, and 6 months of exposure to treatment conditions. CO2
 addition led to a decrease in shell thickness and strength and an increase
 in shell corrosion. However, we also detected an increase in shell
 strength for mussels exposed to both CO2 addition and warming. These
 findings indicate that ocean acidification negatively impacted shellfish
 in situ and that these effects might be mitigated when exposed
 concurrently to moderate warming, leading to an interactive effect of
 acidification and warming on this critical habitat-forming shellfish.
description: Temperature Data:<br> Temperature data was collected at the
 tide pool level using Onset ® HOBO TidbiT temperature loggers (±0.2
 accuracy) that recorded temperature consecutively every 5 min from March
 to September 2019. Temperature data were summarized for the 28-day periods
 preceding each collection period (March, July, and September). For each
 tidepool, we calculated the following thermal parameters: minimum, mean
 daily minimum, 10th percentile, median, average, range, mean daily range,
 mean daily average, maximum, mean daily maximum, 90th percentile, mean
 daily 90th percentile, 95th percentile, 99th percentile, mean daily 90th
 percentile temperatures, mean daily 95th percentile temperatures, mean
 daily 99th percentile temperatures. All temperature data was separated
 into low day and nighttime tides based on known shore heights and observed
 tide data from  Sitka Harbor, Baranof Island, Sitka, Alaska (tbone tides;
 https://tide.arthroinfo.org/). Environmental Tide Pool Data: <br> Tide pool parameters were collected using handheld instruments and total alkalinity was determined at UCSC.<br> <br> Shell Trait Data:<br> All shell trait data were measured within the laboratory. Carbonate Chemistry Variables:<br> pCO<sub>2</sub>, DIC, and saturation states of calcite and aragonite were calculated using the R package <em>seacarb.</em>
description: # Factorial field manipulation reveals multi-stressor effects on a
 critical habitat-forming shellfish This file contains the information
 about the datasets used in Rangel et al. 2025 where we conducted a 6-month
 long field manipulation of pH and temperature in 20 tide pools in Sitka,
 Alaska and measured effects on the mussel *Mytilus trossulus* shell traits
 and function. We measured mussel shell thickness, strength, and corrosion.
 **File names:** DayCarbChem_Mussel.csv * Sample ID: Number of sample in
 collection sequence * Date: Date sample was collected * Month: Month
 sample was collected * Year: Year sample was collected * DayNight: Whether
 the sample was collected during daytime or nighttime * OnOff: On means
 climate manipulations were on and off means when climate manipulations
 were turned off * Timepoint: Time point during each tide when sample was
 taken (1-3) with 3 being the time point after the longest time of
 separation from the ocean * Pool: Tide pool number where sample was
 collected * Treatment: Climate manipulation treatment name * Heat:
 Indicates whether sample underwent a warmed treatment * CO2YN: Indicates
 whether sample underwent a CO2 added treatment (Y = Yes ; N = No) *
 Time.water.collected: The actual time sample was collected from tide pools
 * Temp: Handheld temperature data (°C) * Hanna pH: uncorrected handled
 tide pool pH (mV) * Salinity: salinity of tide pool (ppt) * DO: Dissolved
 oxygen of tide pool (mg/L) * Light: Value of light from hand held
 instrument (umol/m/s) * HannapH_tot: pH of tide pool corrected (total pH
 units) * SpecpH_tot: Raw Spec value for pH (total units) * Raw_TA:
 Uncorrected total alkalinity values (mmol kg-1) * Corrected_TA: Salinity
 corrected Total alkalinity (mmol kg-1) * Calcite: Saturation state of
 calcite from CO2sys * Aragonite: Saturation state of aragonite from CO2sys
 * Volume: Volume of water in the pool (L) * seacarbCO2: CO2 concentration
 calculated by seacarb packing in R (mmol kg-1) *
 fCO2: "potential" fCO2, CO2 fugacity computed at potential
 temperature and atmospheric pressure (μatm) * pCO2:"in situ"
 pCO2, CO2 partial pressure computed at in situ temperature and total
 pressure (atm + hydrostatic) (μatm) * fCO2pot: "in situ" fCO2,
 CO2 fugacity computed at in situ temperature and total pressure (atm +
 hydrostatic) (μatm) * HCO3: HCO3 concentration (mmol kg-1) * CO3: CO3
 concentration (mmol kg-1) * DIC: Dissolved inorganic carbon concentration
 (mmol kg-1) * ALK: total alkalinity from seacarb (mmol kg-1) *
 OmegaAragonite: seacarb saturation state of aragonite * OmegaCalcite:
 seacarb saturation state of calcite **File Name:**
 NightCarbChem_Mussel.csv * Sample: Number of sample in collection sequence
 (ID) * Date: Date sample was collected * Month: Month sample was collected
 * Year: Year sample was collected * DayNight: Whether the sample was
 collected during daytime or nighttime * OnOff: On means climate
 manipulations were on and off means when climate manipulations were turned
 off * Timepoint: Time point during each tide when sample was taken (1-3)
 with 3 being the time point after the longest time of separation from the
 ocean * Completed: If there is a T1, T2, T3 water sample Y = Yes, N = No *
 MissingT: Indicated is there was a missing Timepoint (T1, T2, T3) * Pool:
 Tide pool number where sample was collected * Treatment: Climate
 manipulation treatment name * Heat: Indicates whether sample underwent a
 warmed treatment * CO2YN: Indicates whether sample underwent a CO2 added
 treatment (Y = Yes ; N = No) * SA: Surface area calculated for the pools
 (m2) * Volume: Volume of water in tide pools (L) * DeltaVolume: Change in
 pool volume with each water collection timepoint through low tide (L) *
 Time.water.collected: The actual time sample was collected from tide pools
 * Time.water.collected2: The actual time sample was collected from tide
 pools * Temp: Handheld temperature data (°C) * Hanna pH raw: uncorrected
 handled tide pool pH (mV) * Calcite: Saturation state of calcite from
 CO2sys * Aragonite: Saturation state of aragonite from CO2sys * Salinity:
 salinity of tide pool (ppt) * DO: Dissolved oxygen of tide pool (mg/L) *
 Light: Value of light from hand held instrument (umol/m/s) * Raw_TA:
 Uncorrected total alkalinity values (mmol kg-1) * Corrected_TA: Salinity
 corrected Total alkalinity (mmol kg-1) * HannapH: pH of tide pool
 corrected (total pH units) * SpecpH_tot: Raw Spec value for pH (total
 units) * Raw.TA: Uncorrected total alkalinity values (mmol kg-1) *
 CorrectedTA: Salinity corrected Total alkalinity (mmol kg-1) *
 CorrectedNN: Corrected nutrients (umol/L) * CorrectedNitrite: Corrected
 nitrite (umol/L) * CalculatedNitrate: Calculated nitrate (umol/L) *
 Phosphate: Phosphate value (umol/L) * NH4: Ammonium (umol/L) * Date_Time:
 Date and time sample was collected * TADeltaTime: Change in TA over the
 sampling period (mmol kg-1) * DeltaTime: Change in time over the sampling
 period * DeltaNN: Change in corrected nutrients over time (umol/L) *
 Phosphate: Change in phosphate value over time (umol/L) * DeltaNH4: Change
 in ammonium value over time (umol/L) * DeltaTA_N_Norm: Change in total
 alkalinity nutrient corrected over time that is normalized (mmol kg-1) *
 NEC.mmol.m2.hr: Net ecosystem calcification (mmol m2 hr) * seacarbCO2: CO2
 concentration calculated by seacarb packing in R (mmol kg-1) *
 fCO2: "potential" fCO2, CO2 fugacity computed at potential
 temperature and atmospheric pressure (μatm) * pCO2: "in situ"
 pCO2, CO2 partial pressure computed at in situ temperature and total
 pressure (atm + hydrostatic) (μatm) * fCO2pot: "in situ" fCO2,
 CO2 fugacity computed at in situ temperature and total pressure (atm +
 hydrostatic) (μatm) * HCO3: HCO3 concentration (mmol kg-1) * CO3: CO3
 concentration (mmol kg-1) * DIC: Dissolved inorganic carbon concentration
 (mmol kg-1) * ALK: total alkalinity from seacarb (mmol kg-1) *
 OmegaAragonite: seacarb saturation state of aragonite * OmegaCalcite:
 seacarb saturation state of calcite **File names:**
 DayCarbChemProjected.csv and NightCarbChemProjected.csv * SampleID: Number
 of sample in collection sequence (ID) * Date: Date sample was collected *
 Month: Month sample was collected * Year: Year sample was collected *
 DayNight: Whether the sample was collected during daytime or nighttime *
 OnOff: On means climate manipulations were on and off means when climate
 manipulations were turned off * Timepoint: Time point during each tide
 when sample was taken (1-3) with 3 being the time point after the longest
 time of separation from the ocean] * Completed: If there is a T1, T2, T3
 water sample Y = Yes, N = No * MissingT: Indicated is there was a missing
 Timepoint (T1, T2, T3) * Pool: Tide pool number where sample was collected
 * Treatment: Climate manipulation treatment name * Heat: Indicates whether
 sample underwent a warmed treatment * CO2YN: Indicates whether sample
 underwent a CO2 added treatment (Y = Yes ; N = No) * SA: Surface area
 calculated for the pools (m2) * Volume: Volume of water in tide pools (L)
 * DeltaVolume: Change in pool volume with each water collection timepoint
 through low tide (L) * Time.water.collected: The actual time sample was
 collected from tide pools * Time.water.collected2: The actual time sample
 was collected from tide pools * Temp: Handheld temperature data (°C) *
 Hanna pH raw: uncorrected handled tide pool pH (mV) * Salinity: salinity
 of tide pool (ppt) * DO: Dissolved oxygen of tide pool (mg/L) * Light:
 Value of light from hand held instrument (umol/m/s) * HannapH: pH of tide
 pool corrected (total pH units) * Raw.TA: Uncorrected total alkalinity
 values (mmol kg-1) * CorrectedTA: Salinity corrected Total alkalinity
 (mmol kg-1) * CorrectedNN: Corrected nutrients (umol/L) *
 CorrectedNitrite: Corrected nitrite (umol/L) * CalculatedNitrate:
 Calculated nitrate (umol/L) * Phosphate: Phosphate value (umol/L) * NH4:
 Ammonium (umol/L) * pH: pH calculated from seacarb (total units) *
 seacarbCO2: CO2 concentration calculated by seacarb packing in R (mmol
 kg-1) * fCO2: "potential" fCO2, CO2 fugacity computed at
 potential temperature and atmospheric pressure (μatm) * HCO3: HCO3
 concentration (mmol kg-1) * CO3: CO3 concentration (mmol kg-1) *
 DIC: Dissolved inorganic carbon concentration (mmol kg-1) * ALK: total
 alkalinity from seacarb (mmol kg-1) * OmegaAragonite: seacarb saturation
 state of aragonite * OmegaCalcite: seacarb saturation state of calcite
 **File Name:** DetidedTemp.csv * Month: Month sample was collected *
 Timepoint: Timepoint of exposure to climate manipulations when mussels
 were collected (0, 3, 6) * Pool: Tide pool number where sample was
 collected * Treatment:  Climate manipulation treatment name * Heat: If the
 pool had a warming treatment (H = Heat, N = No Heat) * CO2: If the pool
 had a CO2 treatment (C = CO2, N = No CO2) * DayNight: Whether the sample
 was collected during daytime or nighttime * Minimum: Minimum of pool
 temperatures (°C) * Maximum: Maximum of pool temperatures (°C) * Range:
 Range of pool temperatures (°C) * Median: Median of pool temperatures (°C)
 * Average: Average of pool temperatures (°C) * DailyAverage: Daily average
 of pool temperatures (°C) * Max90Percentile: 90th percentile maximum of
 pool temperatures (°C) * Max95Percentile: 95th percentile maximum of pool
 temperatures (°C) * Max99Percentile: 99th percentile maximum of pool
 temperatures (°C) * Min10Percentile: 10th percentile minimum of pool
 temperatures (°C) * DailyMaximum: Daily maximum of pool temperatures (°C)
 * Daily90MaxPercentile: Daily 90th percentile maximum of pool temperatures
 (°C) * Daily95MaxPercentile: Daily 95th percentile maximum of pool
 temperatures (°C) * Daily99MaxPercentile: Daily 99th percentile maximum of
 pool temperatures (°C) * DailyMaximumPercentile: Daily Maximum of pool
 temperatures (°C) * DailyMinimum: Daily minimum of pool temperatures (°C)
 * DailyRange: Daily range of pool temperatures (°C) * Volume: Volume of
 water in pools (L) **File Name:** MS_AvgShellCorr_Inner_update.csv and
 MS_AvgShellCorr_Outer_update.csv * Month: Month sample was collected *
 Timepoint: Timepoint of exposure to climate manipulations when mussels
 were collected (0, 3, 6) * Treatment: Climate manipulation treatment name
 * mean: Mean of shell corrosion for inner or outer for each treatment * n:
 Number of shells used in the calculation * sd: Standard deviation of mean
 inner and outer shell corrosion * se: Standard error of mean inner and
 outer shell corrosion **File Name:** Mussel_Biomass_Jeb.csv * Month: Month
 sample was collected * Timepoint: Timepoint of exposure to climate
 manipulations when mussels were collected (0, 3, 6) * Pool: Tide pool
 number where sample was collected * Treatment:  Climate manipulation
 treatment name * Pool Volume:  Volume of water in pools (L) * Percent
 Cover: Percent cover of mussels in the pools (%) * Surface Area: Surface
 area of the pools (m2) * WetBiomass: g L-1 * Norm Biomass: g L-1 **File
 Name:** pH_Meta.csv * Sample ID: Number of sample in collection sequence *
 Date: Date sample was collected * Month: Month sample was collected *
 Year: Year sample was collected * DayNight: Whether the sample was
 collected during daytime or nighttime * OnOff: On means climate
 manipulations were on and off means when climate manipulations were turned
 off * Timepoint: Time point during each tide when sample was taken (1-3)
 with 3 being the time point after the longest time of separation from the
 ocean * Pool: Tide pool number where sample was collected * Treatment:
 Climate manipulation treatment name * Heat: If the pool had a warming
 treatment (H = Heat, N = No Heat) * CO2: If the pool had a CO2 treatment
 (C = CO2, N = No CO2) * Time.water.collected: The actual time sample was
 collected from tide pools * Temp: Handheld temperature data (°C) * Hanna
 pH: uncorrected hanna tide pool pH (mV) * Salinity: salinity of tide pool
 (ppt) * DO: Dissolved oxygen of tide pool (mg/L) * Light: Value of light
 from hand held instrument (umol/m/s) * HannapH_tot: pH of tide pool
 corrected (total pH units) * Raw_TA: Uncorrected total alkalinity values
 (mmol kg-1) * Corrected_TA: Salinity corrected Total alkalinity (mmol
 kg-1) * Volume: Volume of water in tide pools (L) **File
 Name:** Sitka_Mussel_CLShells.csv * Date_Measured: Date shell trait
 measured * Month: Month sample was collected * Timepoint: Timepoint of
 exposure to climate manipulations when mussels were collected (0, 3, 6) *
 Species: Type of species that was collected (mussel) * Mussel_ID:
 Identification of mussel number * Pool: Tide pool number where sample was
 collected * Treatment: Climate manipulation treatment name * Heat: If the
 pool had a warming treatment (H = Heat, N = No Heat) * CO2: If the pool
 had a CO2 treatment (C = CO2, N = No CO2) * Total_L: total length of
 mussel shell from lip to base (mm) * Side: Either left or right valve that
 was measured (1 = right, 2=left) * Final_LipA: Averaged shell thickness
 measurements at the lip of shell (mm) * Final_MidB: Averaged shell
 thickness measurements at middle point of shell (mm) * Final_BaseC:
 Averaged shell thickness measurements at base of shell (mm) * StandardA:
 Total length standardized lip shell thickness (mm) * StandardB: Total
 length standardized middle shell thickness (mm) * Standard C: Total length
 standardized base shell thickness (mm) * ThreeTrials: Y is mussels were
 collected from these pool at all three timepoints - N if mussels were not
 collected at all 3 timepoints * Volume: Volume of water in tide pools (L)
 **File Name:** ShellStrength_CL.csv * ID: Mussel Identification *
 Pool: Tide pool number where sample was collected * Month: Month sample
 was collected * Timepoint: Time point during each tide when sample was
 taken (1-3) with 3 being the time point after the longest time of
 separation from the ocean * Treatment: Climate manipulation treatment name
 * Heat: If the pool had a warming treatment (H = Heat, N = No Heat) * CO2:
 If the pool had a CO2 treatment (C = CO2, N = No CO2) * Shell_ID: Shell
 identification * Depth: Depth of mussel shell measured (mm) * Length:
 Total length of mussel shell (lip to base) measured (mm) * Width: Width of
 mussel shell measured (mm) * Mass: Weight of dried mussel shell (g) *
 Strength: Raw strength value measured from Instron force test (N) *
 Strength.Stand: Total length standardized strength value (N) * Valve:
 Right (R) or left (L) value was used in strength test * ThreeTrials:
 Whether or not this pool was collected at 3 timepoints (Y = yes, N = No) *
 Volume: Volume of water in tide pools (L) **File
 Name:** Shell_Corrosion_Inner.csv * Month: Month sample was collected *
 Timepoint: Time point during each tide when sample was taken (1-3) with 3
 being the time point after the longest time of separation from the ocean *
 Mussel.ID: Mussel Identification * Pool: Tide pool number where sample was
 collected * Treatment: Climate manipulation treatment name * Heat: If the
 pool had a warming treatment (H = Heat, N = No Heat) * CO2: If the pool
 had a CO2 treatment (C = CO2, N = No CO2) * Valve: Either left (L) or
 right (R) valve used in measurement * Side: Either inner (In) or outer
 (Out) shell measured * ThreeTrials: Whether or not this pool was collected
 at 3 timepoints (Y = yes, N = No) * Outer_TA: Total outer shell area
 measured (mm) * Corroded_Area: Total corroded shell area (mm) *
 Uncorroded_Area: Total uncorroded shell area (mm) * Per_Corroded: Percent
 of corroded shell area calculated (%) * Per_Uncorroded: Percent of
 uncorroded shell area (%) * SL: Total length of mussel shell valve (mm) *
 Gam_Corr: Gamma corrected mussel percent corroded (%) * Volume: Volume of
 water in tide pools (L) **File Name:** Shell_Corrosion_Outer.csv *
 Month: Month sample was collected * Timepoint: Time point during each tide
 when sample was taken (1-3) with 3 being the time point after the longest
 time of separation from the ocean. * Mussel.ID: Mussel Identification *
 Pool: Tide pool number where sample was collected * Treatment: Climate
 manipulation treatment name * Heat: If the pool had a warming treatment (H
 = Heat, N = No Heat) * CO2: If the pool had a CO2 treatment (C = CO2, N =
 No CO2) * Valve: Either left (L) or right (R) valve used in measurement *
 Side: Either inner (In) or outer (Out) shell measured * ThreeTrials:
 Whether or not this pool was collected at 3 timepoints (Y = yes, N = No) *
 Outer_TA: Total outer shell area measured (mm) * Corroded_Area: Total
 corroded shell area (mm) * Uncorroded_Area: Total uncorroded shell area
 (mm) * Per_Corroded: Percent of corroded shell area calculated (%) *
 Per_Uncorroded: Percent of uncorroded shell area (%) * SL: Total length of
 mussel shell valve (mm) * Volume: Volume of water in tide pools (L) **File
 Name:** TidePool_Propertities_Mussels.csv * Pool: Tide pool number where
 sample was collected * Treatment: Climate manipulation treatment name *
 Max Depth: Depth of the pool (cm) * Perimeter: Perimeter of the pool (cm)
 * Volume: Volume of water in tide pools (L) * Interior surface area:
 Number of squares that were counted to calculate inner pool surface area
 (squares) * Tide Height: Location of the pool on the shoreline from
 waterline to high tide (m) * A: Surface area calculated for the pools (m2)
 **File Name:** WaterMeta_DIC_Musselsl.csv * Sample: Number of sample in
 collection sequence (ID) * Date: Date sample was collected * Month: Month
 sample was collected * Year: Year sample was collected * DayNight: Whether
 the sample was collected during daytime or nighttime * OnOff: On means
 climate manipulations were on and off means when climate manipulations
 were turned off * Timepoint: Time point during each tide when sample was
 taken (1-3) with 3 being the time point after the longest time of
 separation from the ocean * Completed: If there is a T1, T2, T3 water
 sample Y = Yes, N = No * MissingT: Indicated is there was a missing
 Timepoint (T1, T2, T3) * Pool: Tide pool number where sample was collected
 * Treatment: Climate manipulation treatment name * Heat: Indicates whether
 sample underwent a warmed treatment * CO2YN: Indicates whether sample
 underwent a CO2 added treatment (Y = Yes ; N = No) * SA: Surface area
 calculated for the pools (m2) * Volume: Volume of water in tide pools (L)
 * DeltaVolume: Change in pool volume with each water collection timepoint
 through low tide (L) * Time.water.collected: The actual time sample was
 collected from tide pools * Temp: Handheld temperature data (°C) * Hanna
 pH raw: uncorrected handled tide pool pH (mV) * Salinity: salinity of tide
 pool (ppt) * DO: Dissolved oxygen of tide pool (mg/L) * Light: Value of
 light from hand held instrument (umol/m/s) * HannapH: pH of tide pool
 corrected (total pH units) * Raw.TA: Uncorrected total alkalinity values
 (mmol kg-1) * CorrectedTA: Salinity corrected Total alkalinity (mmol kg-1)
 * CorrectedNN: Corrected nutrients (umol/L) * CorrectedNitrite: Corrected
 nitrite (umol/L) * CalculatedNitrate: Calculated nitrate (umol/L) *
 Phosphate: Phosphate value (umol/L) * NH4: Ammonium (umol/L) Datasets
 Include: * DayCarbChem.csv * DayCarbChemProjected.csv * DetidedTemp.csv *
 MS_AvgShellCorr_Inner_update.csv * MS_AvgShellCorr_Outer_update.csv *
 Mussel_Biomass_Jeb.csv * NightCarbChem_Mussel.csv *
 NightCarbChemProjected.csv * pH_Meta.csv * Shell_Corrosion_Inner.csv *
 Shell_Corrosion_Outer.csv * ShellStrength_CL.csv *
 Sitka_Mussel_CLShells.csv * TidePool_Propertities_Mussels.csv *
 WaterMeta_DIC_Musselsl.csv