1. Meredith Barnard and Group 4
Coral Bleaching
Meredith Barnard and Group 4

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

3. 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

4. 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

5. 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 Avg Abs = ( )/ 2
.1305 = .9554x
X = mg/mL of proteins

6. 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

7. 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

8. Zooxanthellae Pictures!!!

9. 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

10. 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

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

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

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

14. 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