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            <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/dataset/925689.rdf" xlink:actuate="onRequest">Allometric scaling of calcification data for Mytilus californianus from 2021-2022 (OA decoupling project)</gmx:Anchor>
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            <gco:CharacterString>Cite this dataset as: Gaylord, B. (2024) Allometric scaling of calcification data for Mytilus californianus from 2021-2022 (OA decoupling project). Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2024-05-06 [if applicable, indicate subset used]. doi:10.26008/1912/bco-dmo.925689.1 [access date]</gco:CharacterString>
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        <gco:CharacterString>Methods and Sampling: &amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Sampling collection details&amp;lt;/strong&amp;gt;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;We gathered naturally settled, adult California mussels (&amp;lt;em&amp;gt;M. californianus&amp;lt;/em&amp;gt;&amp;amp;nbsp;between 30 and 80 mm in maximum shell length) by hand from the mid-intertidal zone of Carmet Beach, along the northern California coast. We cleaned mussels of all epibionts and external byssal threads, then transported them in buckets (&amp;amp;lt; 0.5 hr transit time) to Bodega Marine Laboratory, where we acclimated individuals for seven days in flow-through seawater tables prior to subsequent experiments.&amp;amp;nbsp;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Experiment details&amp;amp;nbsp;&amp;lt;/strong&amp;gt;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;To account for calcification scaling allometrically with animal size, we determined the relationship between mussel tissue mass and calcification rate by incubating 26 different mussels spanning 24 mm to 67 mm in shell length and 0.07 g to 2.55 g of dry tissue in ambient, unmanipulated seawater (Supplementary Figure S3 in Romano de Orte et al. (2021), F&amp;lt;sub&amp;gt;1,24&amp;lt;/sub&amp;gt;&amp;amp;nbsp;= 61.76, R&amp;lt;sup&amp;gt;2&amp;lt;/sup&amp;gt;&amp;amp;nbsp;= 0.71, p &amp;amp;lt;0.001). This procedure yielded a scaling exponent of 0.72 and we therefore divided calcification rates of the experimental mussels by g&amp;lt;sup&amp;gt;0.72&amp;lt;/sup&amp;gt;.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;We calculated net calcification rates with the ammonia-corrected alkalinity anomaly technique&amp;amp;nbsp;(Gazeau et al&amp;amp;nbsp; 2015), divided by incubation duration and mussel dry tissue mass raised by a factor of 0.72 (see the Related Datasets section of this metadata page for this incubation data). The alkalinity anomaly technique builds on the observation that precipitation of CaCO&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt;&amp;amp;nbsp;results in an equivalent reduction in seawater [CO&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt;&amp;lt;sup&amp;gt;2-&amp;lt;/sup&amp;gt;] (or reduction of [HCO&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt;&amp;lt;sup&amp;gt;-&amp;lt;/sup&amp;gt;] followed by an increase in [H&amp;lt;sup&amp;gt;+&amp;lt;/sup&amp;gt;]) which contributes two equivalents of total alkalinity—simultaneous production of ammonia is the major metabolic process in mussels that can obscure this and its signal must be removed&amp;amp;nbsp;(Gazeau et al&amp;amp;nbsp; 2015). Following the incubation, we dissected each mussel and dried it at 60 °C for at least 24 hours to obtain the dry tissue mass (excluding byssal threads) and dry shell mass of each individual mussel.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;We conducted additional incubations (n=87, between 3 and 9 per experiment day) without mussels throughout the trials as experimental blanks to determine background changes in alkalinity. We excluded from our analysis any experimental days where background alkalinity changes exceeded 5 µmol kg&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;The mean of the absolute values of alkalinity change during the incubations of these experimental blanks was 1.3 ± 1.2 µmol kg&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;(n = 72).&amp;amp;nbsp;&amp;lt;/p&amp;gt;</gco:CharacterString>
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        <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/award/886710.rdf" xlink:title="OCE-2129942" xlink:actuate="onRequest">Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-2129942 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=2129942</gmx:Anchor>
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	Name: species
	Units: unitless
	Description: &lt;p&gt;Mussel species used in incubation.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/926762.rdf
	Name: AphiaID
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http://lod.bco-dmo.org/id/dataset-parameter/926763.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926764.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926765.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926766.rdf
	Name: ISO_Start_DateTime_UTC
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http://lod.bco-dmo.org/id/dataset-parameter/926767.rdf
	Name: duration
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http://lod.bco-dmo.org/id/dataset-parameter/926768.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926769.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926770.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926771.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926772.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926773.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926774.rdf
	Name: TA
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	Description: &lt;p&gt;Mean alkalinity during incubation.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/926775.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926776.rdf
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	Description: &lt;p&gt;Mean dissolved oxygen concentration during the incubation.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/926777.rdf
	Name: ci
	Units: umol kg-1
	Description: &lt;p&gt;Mussel condition index, dry tissue mass divided by total dry mass.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/926778.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926779.rdf
	Name: byssal_threads
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http://lod.bco-dmo.org/id/dataset-parameter/926780.rdf
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http://lod.bco-dmo.org/id/dataset-parameter/926781.rdf
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	Units: umol kg-1
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                <gco:CharacterString>&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Sampling collection details&amp;lt;/strong&amp;gt;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;We gathered naturally settled, adult California mussels (&amp;lt;em&amp;gt;M. californianus&amp;lt;/em&amp;gt;&amp;amp;nbsp;between 30 and 80 mm in maximum shell length) by hand from the mid-intertidal zone of Carmet Beach, along the northern California coast. We cleaned mussels of all epibionts and external byssal threads, then transported them in buckets (&amp;amp;lt; 0.5 hr transit time) to Bodega Marine Laboratory, where we acclimated individuals for seven days in flow-through seawater tables prior to subsequent experiments.&amp;amp;nbsp;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Experiment details&amp;amp;nbsp;&amp;lt;/strong&amp;gt;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;To account for calcification scaling allometrically with animal size, we determined the relationship between mussel tissue mass and calcification rate by incubating 26 different mussels spanning 24 mm to 67 mm in shell length and 0.07 g to 2.55 g of dry tissue in ambient, unmanipulated seawater (Supplementary Figure S3 in Romano de Orte et al. (2021), F&amp;lt;sub&amp;gt;1,24&amp;lt;/sub&amp;gt;&amp;amp;nbsp;= 61.76, R&amp;lt;sup&amp;gt;2&amp;lt;/sup&amp;gt;&amp;amp;nbsp;= 0.71, p &amp;amp;lt;0.001). This procedure yielded a scaling exponent of 0.72 and we therefore divided calcification rates of the experimental mussels by g&amp;lt;sup&amp;gt;0.72&amp;lt;/sup&amp;gt;.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;We calculated net calcification rates with the ammonia-corrected alkalinity anomaly technique&amp;amp;nbsp;(Gazeau et al&amp;amp;nbsp; 2015), divided by incubation duration and mussel dry tissue mass raised by a factor of 0.72 (see the Related Datasets section of this metadata page for this incubation data). The alkalinity anomaly technique builds on the observation that precipitation of CaCO&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt;&amp;amp;nbsp;results in an equivalent reduction in seawater [CO&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt;&amp;lt;sup&amp;gt;2-&amp;lt;/sup&amp;gt;] (or reduction of [HCO&amp;lt;sub&amp;gt;3&amp;lt;/sub&amp;gt;&amp;lt;sup&amp;gt;-&amp;lt;/sup&amp;gt;] followed by an increase in [H&amp;lt;sup&amp;gt;+&amp;lt;/sup&amp;gt;]) which contributes two equivalents of total alkalinity—simultaneous production of ammonia is the major metabolic process in mussels that can obscure this and its signal must be removed&amp;amp;nbsp;(Gazeau et al&amp;amp;nbsp; 2015). Following the incubation, we dissected each mussel and dried it at 60 °C for at least 24 hours to obtain the dry tissue mass (excluding byssal threads) and dry shell mass of each individual mussel.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;We conducted additional incubations (n=87, between 3 and 9 per experiment day) without mussels throughout the trials as experimental blanks to determine background changes in alkalinity. We excluded from our analysis any experimental days where background alkalinity changes exceeded 5 µmol kg&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;The mean of the absolute values of alkalinity change during the incubations of these experimental blanks was 1.3 ± 1.2 µmol kg&amp;lt;sup&amp;gt;-1&amp;lt;/sup&amp;gt;&amp;amp;nbsp;(n = 72).&amp;amp;nbsp;&amp;lt;/p&amp;gt;</gco:CharacterString>
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              <gmd:description>
                <gco:CharacterString>&amp;lt;p&amp;gt;All computations were performed with R statistical software, version 4.1.0. We performed carbonate system calculations using the package&amp;amp;nbsp;&amp;lt;em&amp;gt;seacarb&amp;lt;/em&amp;gt;, using equilibrium constants from Lueker&amp;amp;nbsp;&amp;lt;em&amp;gt;et al&amp;lt;/em&amp;gt;. We computed linear mixed models using the&amp;amp;nbsp;&amp;lt;em&amp;gt;lmer&amp;lt;/em&amp;gt;&amp;amp;nbsp;function in in the&amp;amp;nbsp;&amp;lt;em&amp;gt;lmertest&amp;lt;/em&amp;gt;&amp;lt;em&amp;gt;&amp;amp;nbsp;&amp;lt;/em&amp;gt;package in R and focused on assessing likely candidate parameters as fixed factors, and mussel collection date as a random intercept to account for natural seasonal differences between cohorts. Conditional R&amp;lt;sup&amp;gt;2&amp;lt;/sup&amp;gt;&amp;amp;nbsp;was calculated with the package&amp;amp;nbsp;&amp;lt;em&amp;gt;MuMIn&amp;lt;/em&amp;gt;. We determined parameters for non-linear fits employed to model dissolution rates by minimizing the sum of squares of model residuals using the&amp;amp;nbsp;&amp;lt;em&amp;gt;optim&amp;lt;/em&amp;gt;&amp;amp;nbsp;function.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;Colors for plots were chosen from color palettes in the&amp;amp;nbsp;&amp;lt;em&amp;gt;cmocean&amp;lt;/em&amp;gt;&amp;amp;nbsp;package in&amp;amp;nbsp;&amp;lt;em&amp;gt;R&amp;lt;/em&amp;gt;.&amp;lt;/p&amp;gt;</gco:CharacterString>
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                        <gco:CharacterString>Specified by the Principal Investigator(s)</gco:CharacterString>
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          </gmd:processStep>
        <gmd:processStep xlink:title="BCO-DMO Data Processing Description">
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                <gmd:description>
                  <gco:CharacterString>- Removed special characters (e.g., periods) from column names and replaced with underscores
- Added a datetime field (&amp;quot;start_datetime)from the date and start time fields
- Changed the presentation of species values from &amp;quot;mytilus_californianus&amp;quot; to &amp;quot;Mytilus californianus&amp;quot; and added AphiaID and LSID to the data file
- All numeric float fields rounded to 2 degrees of precision</gco:CharacterString>
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                        <gmd:title>
                          <gco:CharacterString>Specified by BCO-DMO Data Managers</gco:CharacterString>
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                        <gmd:date gco:nilReason="unknown"/>
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    <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/affiliation/191.rdf" xlink:actuate="onRequest">Biological and Chemical Oceanography Data Management Office (BCO-DMO)</gmx:Anchor>
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