The basic observed water characteristics (pH, temperature, salinity and total alkalinity (TA)) and calculated water carbonate chemistry in the field (22.44°N, 113.95°E, Pak Nai, Hong Kong) and laboratory (22.28°N, 114.14°E, the University of Hong Kong, Hong Kong) during a two-factor orthogonal experiment. To be specific, we designed a two-factor (low pH, as a proxy for OA, and Vibrio infection) orthogonal experiment to examine the immunological and molecular responses in the ecologically and economically important edible Hong Kong oyster, Crassostrea hongkongensis. Wild oyster population was collected from their native coastal area (22.44°N, 113.95°E, Pak Nai, Hong Kong). Vibrio parahaemolyticus, as a major global concern of foodborne human-related disease, was used in this study to investigate the antimicrobial characteristics of edible commercial oysters in the context of future OA. pH and temperature were monitored daily with pH meter (Mettler Toledo InLab®, Switzerland) calibrated with NBS standards. Salinity was monitored using a dual-scale refractometer (MingRui, China). The seawater was utterly changed every two days and 50 mL seawater was preserved with HgCl2 every week for total alkalinity (TA) measurement. The TA was measured using an alkalinity titrator (Mettler Toledo G20S Compact Titrator, Switzerland), and pCO2, CO32-, saturation states of calcite and aragonite were calculated by CO2SYS program with equilibrium constants K2 and KSO4 (Millero, F. J., Graham, T. B., Huang, F., Bustos-Serrano, H., & Pierrot, D. (2006). Dissociation constants of carbonic acid in seawater as a function of salinity and temperature. Marine Chemistry, 100(1), 80-94. https://doi.org/10.1016/j.marchem.2005.12.001).
