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Applied and Environmental Microbiology, September 2006, p. 5963-5973, Vol. 72, No. 9
0099-2240/06/$08.00+0     doi:10.1128/AEM.00843-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Microbial Communities in the Surface Mucopolysaccharide Layer and the Black Band Microbial Mat of Black Band-Diseased Siderastrea siderea

Raju Sekar,1* DeEtta K. Mills,1,2 Elizabeth R. Remily,1 Joshua D. Voss,1 and Laurie L. Richardson1

Department of Biological Sciences, Florida International University, Miami, Florida 33199,1 International Forensic Research Institute, Florida International University, Miami, Florida 331992

Received 10 April 2006/ Accepted 3 July 2006

Microbial community profiles and species composition associated with two black band-diseased colonies of the coral Siderastrea siderea were studied by 16S rRNA-targeted gene cloning, sequencing, and amplicon-length heterogeneity PCR (LH-PCR). Bacterial communities associated with the surface mucopolysaccharide layer (SML) of apparently healthy tissues of the infected colonies, together with samples of the black band disease (BBD) infections, were analyzed using the same techniques for comparison. Gene sequences, ranging from 424 to 1,537 bp, were retrieved from all positive clones (n = 43 to 48) in each of the four clone libraries generated and used for comparative sequence analysis. In addition to LH-PCR community profiling, all of the clone sequences were aligned with LH-PCR primer sequences, and the theoretical lengths of the amplicons were determined. Results revealed that the community profiles were significantly different between BBD and SML samples. The SML samples were dominated by {gamma}-proteobacteria (53 to 64%), followed by ß-proteobacteria (18 to 21%) and {alpha}-proteobacteria (5 to 11%). In contrast, both BBD clone libraries were dominated by {alpha}-proteobacteria (58 to 87%), followed by verrucomicrobia (2 to 10%) and 0 to 6% each of {delta}-proteobacteria, bacteroidetes, firmicutes, and cyanobacteria. Alphaproteobacterial sequence types related to the bacteria associated with toxin-producing dinoflagellates were observed in BBD clone libraries but were not found in the SML libraries. Similarly, sequences affiliated with the family Desulfobacteraceae and toxin-producing cyanobacteria, both believed to be involved in BBD pathogenesis, were found only in BBD libraries. These data provide evidence for an association of numerous toxin-producing heterotrophic microorganisms with BBD of corals.

* Corresponding author. Mailing address: Department of Biological Sciences, Florida International University, Miami, FL 33199. Phone: (305) 348-7812. Fax: (305) 348-1986. E-mail: rajus{at}fiu.edu.

Applied and Environmental Microbiology, September 2006, p. 5963-5973, Vol. 72, No. 9
0099-2240/06/$08.00+0     doi:10.1128/AEM.00843-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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