Meredith Barnard and Group 4 Coral Bleaching Meredith Barnard and Group 4

Protein Assay Measure of fitness Amount of protein the the polyps More stress causes increased protein production

Protein Assay Procedure Gather the series of dilutions provided (.24 mg/mL, .50 mg/mL, .75 mg/mL, 1.00 mg/mL, and 1.25 mg/mL) Measure the absorbance for each dilution More concentrated  Higher absorbance

Results Trail 2 Trial 1 Concentration (mg/mL) Absorbance .25 0.259 .50 0.58 .75 0.786 1.00 0.875 1.25 1.24 Concentration (mg/mL) Absorbance .25 0.24 .50 0.627 .75 0.755 1.00 0.986 1.25 1.22

Results cont… Concentration (mg/mL) Average Absorbance .25 0.2495 .50 0.6035 .75 0.7705 1.00 0.9305 1.25 1.23 Homogenate (100 µL) Absorbance Trial 1 .129 Trial 2 .132 Avg .1305 Y = mx + b Abs = slope * X Y = .9554x + .0335 Avg Abs = (.129 + .132)/ 2 .1305 = .9554x X = .13659 mg/mL of proteins

Zooxanthellae Density Procedure 3 Xenia polyps were gathered and weighed (.0135 g) They were placed in 2 mL of seawater and homogenized for about 20 seconds A sample of the homogenate was placed on a counting chamber and the number of zoox present was counted

Number of Zoox Count 1 Count 2 Cell # 1 8 13 17 21 Top 9 14 Bottom 7 6 11 Cell # 4 6 10 15 23 Top 11 7 Bottom 8 3 Top Bottom Count 1 49 39 Count 2 36 31 Avg between count 1 and count 2 42.5 35 Avg of top and bottom 38.75

Zooxanthellae Pictures!!!

Calculations 38.75 / 5 = 7.75 zoox/ box 7.75 / (.004*10-3) = 1,937,500 zoox/mL (1,937,500*2)/.0135g= 2.87*10^9 zoox/g

Chlorophyll Extraction and Measurement Procedure Dilute 1 mL of the homogenate with 9 mL of acetone. Vortex and extract for 10 minutes Centrifuge at high speed for 5 minutes Measure absorbance at 665, 647, and 630 nm. Calculate amt of Chlorophyll using : Ca = (11.85*A665)- (1.54*A647) – (.08*A630) - Calculate the total amount of chlorophyll a in the sample

Results & Calculations A665 = .024 A647 = .003 A630 = .004 Ca = (11.85*.024)- (1.54*.003) – (.08*.004) Ca = .27946 mg/mL

Results and Calculations Continued… Chlorophyll a (µ/mL) = [V(Ca)] / l Ca = .27946 mg/mL l = path length of cuvette (1.6 cm) V= total volume of homogenate Chlorophyll a(µ/mL)= [2 mL*.27496 mg/ mL] / 1.6 cm Chlorophyll a = .174663 µ/mL

What’s to Come?!? Temperature effects on Xenia corals

How will Changes in Temperature Affect Corals?!? Experiment: 3 trials 2 tanks Control Tank Constant temperature ~ 28º C Group 4 Change in temperature by one degree each week