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

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

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) 3.72215mg/g 1.60329mg/g

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

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 3.72215 mg/g 1.60329 mg/g Week 1 Week 2 4.31 x 10^8 5.26 x 10^8 221.55 ng/106 cells 29.56 ng/106 cells .095 mg/g

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.

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

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

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

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.

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: http://www.jellett.ca/methanol_extraction.htm

Time to Experiment!!!