Dataset: Palau temperature experiment 2014-2015

Sampling and analytical procedures:

Coral Collection: A total of 8 colonies of each species were collected at each site at a depth between 5–10 meters (offshore) or 1–5 meters (Inshore) and at least 10 meters apart. Differences in collection depth were necessary due to the natural distribution of these species at each site and in order to ensure all colonies were collected from similar light conditions (maximal mid-day in situ light of 800-1000-μmol quanta m-2 s-1). Colonies were transported back to the Palau International Coral Research Center (PICRC) and fragmented into five replicate nubbins and placed into a 1200L flow-through aquarium and held at 27.5°C. Seawater was collected directly off of a nearby pier at a depth of 3 m and then passed through a pressurized sand filter and aquarium filter pads prior to use in flow-through and experimental treatment systems. Coral nubbins were attached to 2-inch square PVC tiles with marine epoxy (Splash zone compound A-788) and held at ambient conditions in flow-through bins as described above for 12-16 days prior to the start of the experiment. Control and experimental bins (see below) were maintained outdoors underneath clear plastic film (Sun Selector, Ginegar Plastic Products) to block periodic rainfall and a 60% shade cloth providing a peak midday light intensity of 800 μmol quanta m-2 s-1, as measured with a light sensor (LiCor LI-192).

Experimental System: Each treatment system consisted of 7–12 (56 L) plastic treatment bins connected to a central (~1200 L) sump. Seawater within each sump was set to either heated or ambient temperature conditions prior to being sent to the treatment bins. Ambient temperature was maintained via a chiller system and a series of titanium heating elements were used for high temperature treatments. For each treatment, three replicate fragments from each colony were placed within separate treatment bins. For the heated treatment, the temperature was gradually ramped from 27.5°C to 32°C over 4 days, and then maintained at 32°C for an additional 10 days for a total of 14 days of heating. Temperature within the control treatment was maintained at 27.5°C throughout the 14-day experiment. Treatment bins and PVC tiles were cleaned every other day to prevent algal fouling, and coral fragments were rotated within their respective bins every other day to ensure a uniform light exposure and minimize possible tank effects.

At the start of the experiment (day 0), one fragment from each coral colony was removed from control and treatment tanks and processed for symbiont photo-physiology and biomass metrics (described below). Additional fragments were then sampled on days 9 (4 days of temperature ramping + 5 days at 32°C) and 14 (4 days of temperature ramping and 10 days at 32°C). Coral tissue was removed by airbrush (100 psi) with filtered (0.22 m) seawater. The resulting slurry was homogenized with a Tissue Tearor (BioSpec products, Inc), and then divided into 2 mL aliquots. One aliquot was preserved with 1% glutaraldehyde for cell enumeration and stored at 4°C. All other aliquots were centrifuged for 2 minutes (5,000×g) and the supernatant was discarded. Algal subsamples from each colony were suspended in DNA preservation buffer (Seutin et al. 1991) and stored at 4°C. The remaining algal samples were immediately frozen (-20°C) and shipped back to the United States and stored at -20°C until further analyses.

Remaining information on procedures can be found in Hoadley et al., 2019.