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

A Comparison of Bacterial Communities A Comparison of Bacterial Communities Between Geographically Separated Corals Between Geographically Separated Corals

Infected with White Plague Type IIInfected with White Plague Type II

G.M. Cook, J. P. Rothenberger, M. Sikaroodi,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 UniversityDepartment of Environmental Science and Policy, George Mason UniversityTetra Tech, Inc.Tetra Tech, Inc.

IntroductionIntroduction

Acquired samples of apparently healthy and Acquired samples of apparently healthy and diseased corals (diseased corals (Montastraea annularisMontastraea annularis) from ) from the U.S. Virgin Islands and the Bahamasthe U.S. Virgin Islands and the Bahamas

Compared the microbial community composition Compared the microbial community composition of healthy and infected coral pairsof healthy and infected coral pairs

Microbial culturing, LH-PCR, molecular Microbial culturing, LH-PCR, molecular fingerprinting, and 16S rRNA sequencingfingerprinting, and 16S rRNA sequencing

July 29, 2002

Photo by: Dr. Robert Jonas

M. faveolata, Andros Island, Bahamas.


August 18, 2002 (20 days after initial observation)


13.5 cm


August 1, 2003 (one year after initial observation)


HypothesesHypotheses

1.1. That the causative agents are opportunistic pathogens That the causative agents are opportunistic pathogens normally present in the host or its environs rather than normally present in the host or its environs rather than a novel, obligate pathogen;a novel, obligate pathogen;

2.2. That corals exhibiting WPII disease signs from different That corals exhibiting WPII disease signs from different geographical regions harbor differing microbial geographical regions harbor differing microbial communities in normal tissue and diseased tissue; communities in normal tissue and diseased tissue;

3.3. The WPII disease process is the result of a broad shift The WPII disease process is the result of a broad shift in the microbial community (dysbiosis).in the microbial community (dysbiosis).

MethodologyMethodology

Divers sampling a colony of M. faveolata, LSI, Exumas, Bahamas


Lee Stocking Island, Exumas, BahamasLee Stocking Island, Exumas, Bahamas

ggggg

XX


X

St. Croix, U.S. Virgin IslandsSt. Croix, U.S. Virgin Islands


X

VC2D & VC2HD

1.6 cm

Experimental DesignExperimental Design

Two cores used for Two cores used for histological histological investigationsinvestigations

Three cores used for Three cores used for microbial & molecular microbial & molecular investigationsinvestigations Photo By: Dr. Esther Peters

Photo By: Dr. Robert Jonas

Length Heterogeneity PCR Length Heterogeneity PCR FingerprintingFingerprinting

(LH-PCR)(LH-PCR)

OTU 1 *OTU 2 *OTU 3 *

27F27F

355R355R

Peak area ~ Abundance

Size(bp)

Re

lativ

e In

tens

ity

ResultsResults

Fingerprint HistogramFingerprint Histogram* Active WPII

Fingerprint HistogramFingerprint Histogram* Inactive WPII

Results/ConclusionsResults/Conclusions

The Virgin Island Control/Healthy community is not The Virgin Island Control/Healthy community is not nearly as diverse as the Bahamas Control/Healthy nearly as diverse as the Bahamas Control/Healthy community.community.

There is a shift from a low complexity (H, HD) to high There is a shift from a low complexity (H, HD) to high complexity community (D) in VI samples.complexity community (D) in VI samples.

There is a shift from a high complexity (H, HD) to low There is a shift from a high complexity (H, HD) to low complexity community (D) in the Bahamas samples.complexity community (D) in the Bahamas samples.

PCO case scores (Euclidean)

Disease BI

Healthy BI

Control BI

Disease VI

Healthy VI

Control VI

Axis 2

Axis 1

-0.1

-0.2

-0.3

-0.4

-0.5

0.1

0.2

0.3

0.4

0.5

-0.1-0.2-0.3-0.4-0.5 0.1 0.2 0.3 0.4 0.5

The clustering of diseased tissue The clustering of diseased tissue (D)(D) is different from is different from healthy tissue healthy tissue (HD)(HD) and the control tissue and the control tissue (H)(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

WPII was inactive in VI samples. WPII was inactive in VI samples.

PossiblePossible etiological agent fingerprint,etiological agent fingerprint, Aurantimonas Aurantimonas coralicidacoralicida (313.1 bp), was found only in the diseased tissue (313.1 bp), was found only in the diseased tissue of Bahamian samples.of Bahamian samples.

Control and healthy on diseased microbial community differs Control and healthy on diseased microbial community differs with geographical separation.with geographical separation.

We’re not finished yet!We’re not finished yet!

Results/ConclusionsResults/Conclusions

AcknowledgementsAcknowledgements

Special thanks toSpecial thanks to::

J. Paige RothenbergerJ. Paige Rothenberger

Masoumeh SikaroodiMasoumeh Sikaroodi

Roslyn Cress and the Department of ESPRoslyn Cress and the Department of ESP

Perry Institute of Marine Science (PIMS)Perry Institute of Marine Science (PIMS)

National Underwater Research Program (NURP)National Underwater Research Program (NURP)

American Type Culture Collection (ATCC)American Type Culture Collection (ATCC)

U.S. Fish and WildlifeU.S. Fish and Wildlife

The people of St. Croix and the Bahamas The people of St. Croix and the Bahamas


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

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