1. Chemical Mechanisms of Disease Resistance in Hawai’ian Corals Deborah Gochfeld 1, Greta Aeby 2, Jesse Miller 1 1 National Institute of Undersea Science and Technology, University of Mississippi 2 Hawaii Institute of Marine Biology

2. Prevalence of Hawaiian coral diseases Variability among genera Variability among sites What accounts for these differences in susceptibility?

3. Objectives 1) Assess differences in antimicrobial activity among three dominant Hawai’ian coral genera. 2) Assess population-level variability in antimicrobial activity of common coral species from three different sites. 3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species. MontiporaPoritesPocillopora

4. General Methods 1) Collect replicate pieces of three common coral genera (Montipora, Pocillopora, Porites) from three sites on Oahu 2) Extract corals is aqueous solvents 3) Test extracts in antimicrobial assays to determine genus- and population-level variability in antimicrobial activity

5. Species of bacteria used to test coral extracts

6. Objectives 1) Assess differences in antimicrobial activity among dominant Hawai’ian coral genera. 2) Assess population-level variability in antimicrobial activity of common coral species from different sites. 3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species.

7. Overall antimicrobial activity in Hawaiian corals

8. Aurantimonas coralicida Clostridium perfringens Klebsiella pneumoniae Pseudomonas nautica Serratia marcescens Vibrio agarivorans Vibrio coralyticus Vibrio shiloi Yersinia enterocolitica 0 10 20 30 40 50 60 70 80 90 100 % active assays Montipora capitata stimulatory Porites lobata inhibitory Pocillopora meandrina Selective activity among coral genera

9. Objectives 1) Assess differences in antimicrobial activity among dominant Hawai’ian coral genera. 2) Assess population-level variability in antimicrobial activity of common coral species from different sites. 3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species.

10. Site differences in antimicrobial activity of corals

12. Montipora capitata BacteriaWaianaeMaile BeachKaneohe Bay Aurantimonas coralicida -4/+1-3 Vibrio corallyticus -5 Vibrio shiloi -1/+3+4 Serratia marcescens -5 Yersinia enterocolitica -5-3-4 Clostridium perfringens -5-3-4 Pseudomonas nautica +3+5+4 Vibrio agarivorans -5-3 Klebsiella pneumoniae -5-2-3 antimicrobial activity (%) 77.867.668.4

13. Objectives 1) Assess differences in antimicrobial activity among dominant Hawai’ian coral genera. 2) Assess population-level variability in antimicrobial activity of common coral species from different sites. 3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species. Montipora white syndrome

14. Comparison of antimicrobial activity in healthy vs. diseased colonies Healthy neighbor Higher levels of antimicrobials?Induced defenses? Control Healthy Diseased

15. Comparison of antimicrobial activity between healthy & diseased colonies

16. Comparison of antimicrobial activity between healthy & diseased tissue

17. Chemical Fingerprinting Injected extracts onto a High Performance Liquid Chromatography system Compared areas under the curve for each peak in the spectrum

18. peak 7peak 14peak 15 0 40000 80000 120000 160000 200000 240000 Area under curve * * peak 7peak 14peak 15 0 10000 20000 30000 40000 50000 60000 70000 Control Healthy Diseased 7 14 15 Differences in specific chemical components

19. Conclusions Hawaiian corals exhibit high levels of antimicrobial activity Highly selective against a diversity of known coral pathogens and potential pathogens from human sources Some stimulatory effects Variation among genera, among sites, among individual colonies Healthy colonies have higher levels of antimicrobial activity against certain bacteria Potential induction of defenses in diseased regions of affected colonies Differences in chemical profiles reflect this variation in defenses

20. Acknowledgements Collections (Hawaii Institute of Marine Biology) Sample Processing and Assays (University of Mississippi) Shellie Bailey Amy Katzenmeyer Lauren Wheeler Megan Colwin Lian Zigterman Casper Lazet Dr. Fenny Cox Dr. Frank Stanton