1. Coral Bleaching Presentation 4
Presented by: Selima
Group 2: Lacey, Lauren, Kelli,
Selima, & Randy

2. Methods
Last week, we repeated previous steps of homogenation, vortexing, centrifugation, and absorbency readings to obtain control data for Aiptasia and create another Bradford Assay Curve

3. Control Data for Week 3 and 4
Weight of Aiptasia (g)
.1596
.1113g
Zooxanthellae Density (zoox/ g wet weight)
1.91 x 108 zoox/ g wet weight
2.34 x 108 zoox/ g wet weight
Chlorophyll Concentration (ng/106 cells)
23.3ng/106 cells
7.46ng/106 cells
Protein Concentration
(mg/g)
mg/g
mg/g

4. Bradford Assay Curve Week 3: y = 1.2611x + 0.0666 R2 = 0.9939 Week 4:

5. Aiptasia compared to Xenia
Week 3
Week 4
Zooxanthallaeper g wet weight
1.91 x 10^8
2.34 x 10^8
Chlorophyll Concentration
23.3 ng/106 cells
7.46 ng/106 cells
Protein Concentration
mg/g
mg/g
Week 1
Week 2
4.31 x 10^8
5.26 x 10^8
ng/106 cells
29.56 ng/106 cells
.095 mg/g

6. Conclusions
There is a larger zooxanthellae density in Xenia as opposed to Aiptasia.
There is a larger chlorophyll concentration in Xenia than in Aptasia.
There is a much larger protein concentration in Aptasia than in Xenia.

7. How does UV radiation affect coral bleaching over time?
Our Question
How does UV radiation affect coral bleaching over time?

8. Experimental Design Filled tank with 8L of deionized H2O
Added 8.35g of salt so the salinity would be 35 ppu
Inserted 4 Aiptasia anemones into the tank
Set the UV lamp over the tank
Temperature: 25°C

9. Our Controls The control tank will remain unexposed to UV light
The experimental tank will remain under a constant, unchanging amount of UV light of approximately 358nm
Every day we will measure our tank’s salinity and temperature in order to account for extraneous variables

10. Hypothesis
Exposure of Aiptasia to UV light over an extended period of time will cause coral bleaching.
We will test this by measuring protein concentrations as well as zoox g/wet weight and chlorophyll concentration of anemones in the control tank and the experimental tank and then compare them.

11. An additional test
We also want to test for MAA- microsporidian amino acids released by zooxanthallae when exposed to UV radiation
Method: methanol extraction**
1. Create methanol extraction fluid with 1 part 100% methanol and 1 part water
2. Add 40mL methanol extraction liquid to 10mL homogenate
3. Pour filtered extract into clean tube
4. The extract can then be used with Jellett Rapid test for ASP
We can compare the production of these amino acids to other factors such as zooxanthallae density and protein concentration to see if there is any realation.
**Source:

12. Time to Experiment!!!