Larval respiration was measured in a confined respirometer use a Ruthenium-based optrode (FOXY-R, 1.58 diameter, Ocean Optics) connected to a spectrophotometer (USB2000, Ocean Optics) and interfaced with a computer running the manufacturers software (OOISensor, version 1.00.08). The optrode was calibrated using a zero solution (0.01 M Na2B4O7·10H2O supersaturated with Na2SO3) and 100% air saturation usingwater-saturated air at the treatment temperature. Tomeasure respiration, 6 larvae were removed from the tubs in the treatment tanks and placed into 2-mL glass Wheaton vials filled with filtered seawater from the same treatment tank and sealed with Parafilm. A study conducted concurrently with the present analysis demonstrated that respiration of P. damicornis larvae in identical vials could be measured accurately with 5 larvae vial-1 (Edmunds et al., 2011).
Initial O2 concentration in the seawater was determined before the vials were sealed, and vials without larvae used as controls. Larvae in the sealed vials were incubated at their temperature treatments for 1.5-2 h in the dark using water baths set to the temperature of the respective treatments (±0.1 °C, Hipoint, models LC-06 and LC-10); vials were carefully moved throughout the incubations to ensure the seawater remained mixed. Incubation timeswere selected to ensure O2 concentrations remained >75% saturation. On completion of the incubations, vials were removed from the water baths, gently inverted to mix the seawater, and analyzed for O2 saturation. O2 saturation was converted to concentration using tabulated values of O2 solubility (N. Ramsing and J. Gundersen at http://www.unisense.com [based on Garcia and Gordon, 1992]) and the temperature and salinity of the seawater, and the change in O2 concentration used to calculate O2 flux after adjusting for controls. The respiration rates are expressed as nmol O2 mg protein-1 min-1.
The 'ambient' and 'high' pCO2 levels: 49.4 Pa versus 86.2 Pa
The 'ambient' and 'high' temperatures: 24.00 °C [ambient] versus 30.49 °C
Data also available from PANGAEA: doi:10.1594/PANGAEA.823582