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G.M. Cook, J. P. Rothenberger, M. Sikaroodi, E. Peters, P.M. Gillevet, R. B. Jonas

A Comparison of Bacterial Communities Between Geographically Separated Corals Infected with White Plague Type II. G.M. Cook, J. P. Rothenberger, M. Sikaroodi, E. Peters, P.M. Gillevet, R. B. Jonas Department of Environmental Science and Policy, George Mason University Tetra Tech, Inc.

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G.M. Cook, J. P. Rothenberger, M. Sikaroodi, E. Peters, P.M. Gillevet, R. B. Jonas

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  1. A Comparison of Bacterial Communities Between Geographically Separated Corals Infected with White Plague Type II G.M. Cook, J. P. Rothenberger, M. Sikaroodi, E. Peters, P.M. Gillevet, R. B. Jonas Department of Environmental Science and Policy, George Mason University Tetra Tech, Inc.

  2. Introduction • Acquired samples of apparently healthy and diseased corals (Montastraea annularis) from the U.S. Virgin Islands and the Bahamas • Compared the microbial community composition of healthy and infected coral pairs • Microbial culturing, LH-PCR, molecular fingerprinting, and 16S rRNA sequencing

  3. M. faveolata, Andros Island, Bahamas. July 29, 2002 Photo by: Dr. Robert Jonas

  4. 13.5 cm M. faveolata, Andros Island, Bahamas. August 18, 2002 (20 days after initial observation) Photo by: Dr. Robert Jonas

  5. M. faveolata, Andros Island, Bahamas. August 1, 2003 (one year after initial observation) Photo by: Dr. Robert Jonas

  6. Hypotheses • That the causative agents are opportunistic pathogens normally present in the host or its environs rather than a novel, obligate pathogen; • That corals exhibiting WPII disease signs from different geographical regions harbor differing microbial communities in normal tissue and diseased tissue; • The WPII disease process is the result of a broad shift in the microbial community (dysbiosis).

  7. Methodology Photo by: Dr. Robert Jonas Divers sampling a colony of M. faveolata, LSI, Exumas, Bahamas

  8. Lee Stocking Island, Exumas, Bahamas ggggg X X X Photo by: Dr. Robert Jonas

  9. St. Croix, U.S. Virgin Islands X Photo by: Dr. Robert Jonas

  10. VC2D & VC2HD

  11. VC7H

  12. Experimental Design • Two cores used for histological investigations • Three cores used for microbial & molecular investigations 1.6 cm Photo By: Dr. Esther Peters Photo By: Dr. Robert Jonas

  13. OTU 1 * OTU 2 * OTU 3 * Relative Intensity Size(bp) Length Heterogeneity PCR Fingerprinting (LH-PCR) 27F 355R Peak area ~ Abundance

  14. Results

  15. Fingerprint Histogram * Active WPII

  16. Fingerprint Histogram * Inactive WPII

  17. Results/Conclusions • The Virgin Island Control/Healthy community is not nearly as diverse as the Bahamas Control/Healthy community. • There is a shift from a low complexity (H, HD) to high complexity community (D) in VI samples. • There is a shift from a high complexity (H, HD) to low complexity community (D) in the Bahamas samples.

  18. The clustering of diseased tissue (D) is different from healthy tissue (HD) and the control tissue (H). The clustering of the healthy and control tissue from the VI (●) is different from the healthy and control tissue from the Bahamas (▲). Axis 2

  19. Results/Conclusions • WPII was inactive in VI samples. • Possibleetiological agent fingerprint, Aurantimonascoralicida (313.1 bp), was found only in the diseased tissue of Bahamian samples. • Control and healthy on diseased microbial community differs with geographical separation. • We’re not finished yet!

  20. Acknowledgements Special thanks to: J. Paige Rothenberger Masoumeh Sikaroodi Roslyn Cress and the Department of ESP Perry Institute of Marine Science (PIMS) National Underwater Research Program (NURP) American Type Culture Collection (ATCC) U.S. Fish and Wildlife The people of St. Croix and the Bahamas Photo by: Dr. Robert Jonas

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