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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 E. Peters, P.M. Gillevet, R. B. Jonas Department of Environmental Science and Policy, George Mason University Tetra Tech, Inc.
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Introduction Acquired samples of apparently healthy and diseased corals (Montastraea annularis) from the U.S. Virgin Islands and the Bahamas 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 Compared the microbial community composition of healthy and infected coral pairs Microbial culturing, LH-PCR, molecular fingerprinting, and 16S rRNA sequencing Microbial culturing, LH-PCR, molecular fingerprinting, and 16S rRNA sequencing
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July 29, 2002 Photo by: Dr. Robert Jonas M. faveolata, Andros Island, Bahamas.
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Photo by: Dr. Robert Jonas August 18, 2002 (20 days after initial observation) M. faveolata, Andros Island, Bahamas. 13.5 cm
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Photo by: Dr. Robert Jonas August 1, 2003 (one year after initial observation) M. faveolata, Andros Island, Bahamas.
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Hypotheses 1. That the causative agents are opportunistic pathogens normally present in the host or its environs rather than a novel, obligate pathogen; 2. That corals exhibiting WPII disease signs from different geographical regions harbor differing microbial communities in normal tissue and diseased tissue; 3. The WPII disease process is the result of a broad shift in the microbial community (dysbiosis).
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Methodology Divers sampling a colony of M. faveolata, LSI, Exumas, Bahamas Photo by: Dr. Robert Jonas
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Lee Stocking Island, Exumas, Bahamas ggggg X X Photo by: Dr. Robert Jonas X
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St. Croix, U.S. Virgin Islands Photo by: Dr. Robert Jonas X
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VC2D & VC2HD
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VC7H
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1.6 cm Experimental Design Two cores used for histological investigations Two cores used for histological investigations Three cores used for microbial & molecular investigations Three cores used for microbial & molecular investigations Photo By: Dr. Esther Peters Photo By: Dr. Robert Jonas
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Length Heterogeneity PCR Fingerprinting(LH-PCR) OTU 1 * OTU 2 * OTU 3 *27F 355R Peak area ~ Abundance Size(bp) Relative Intensity
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Results
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Fingerprint Histogram * Active WPII
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Fingerprint Histogram * Inactive WPII
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Results/Conclusions The Virgin Island Control/Healthy community is not nearly as diverse as the Bahamas Control/Healthy community. 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 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. There is a shift from a high complexity (H, HD) to low complexity community (D) in the Bahamas samples.
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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
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WPII was inactive in VI samples. WPII was inactive in VI samples. Possible etiological agent fingerprint, Aurantimonas coralicida (313.1 bp), was found only in the diseased tissue of Bahamian samples. Possible etiological agent fingerprint, Aurantimonas coralicida (313.1 bp), was found only in the diseased tissue of Bahamian samples. Control and healthy on diseased microbial community differs with geographical separation. Control and healthy on diseased microbial community differs with geographical separation. We’re not finished yet! We’re not finished yet! Results/Conclusions
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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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