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Applied and Environmental Microbiology, July 2000, p. 3031-3036, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Penetration of the Coral-Bleaching Bacterium Vibrio shiloi into Oculina patagonica

E. Banin,1 T. Israely,1 A. Kushmaro,2 Y. Loya,2 E. Orr,3 and E. Rosenberg1,*

Department of Molecular Microbiology and Biotechnology1 and Department of Zoology,2 The George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel, and Department of Genetics, University of Leicester, Leicester LE1 7RH, United Kingdom3

Received 18 January 2000/Accepted 11 April 2000

Inoculation of the coral-bleaching bacterium Vibrio shiloi into seawater containing its host Oculina patagonica led to adhesion of the bacteria to the coral surface via a beta -D-galactose receptor, followed by penetration of the bacteria into the coral tissue. The internalized V. shiloi cells were observed inside the exodermal layer of the coral by electron microscopy and fluorescence microscopy using specific anti-V. shiloi antibodies to stain the intracellular bacteria. At 29°C, 80% of the bacteria bound to the coral within 8 h. Penetration, measured by the viable count (gentamicin invasion assay) inside the coral tissue, was 5.6, 20.9, and 21.7% of the initial inoculum at 8, 12, and 24 h, respectively. The viable count in the coral tissue decreased to 5.3% at 48 h, and none could be detected at 72 h. Determination of V. shiloi total counts (using the anti-V. shiloi antibodies) in the coral tissue showed results similar to viable counts for the first 12 h of infection. After 12 h, however, the total count more than doubled from 12 to 24 h and continued to rise, reaching a value 6 times that of the initial inoculum at 72 h. Thus, the intracellular V. shiloi organisms were transformed into a form that could multiply inside the coral tissue but did not form colonies on agar medium. Internalization of the bacteria was accompanied by the production of high concentrations of V. shiloi toxin P activity in the coral tissue. Internalization and multiplication of V. shiloi are discussed in terms of the mechanism of bacterial bleaching of corals.


* Corresponding author. Mailing address: Department of Molecular Microbiology & Biotechnology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel. Phone: 972 3 640 9838. Fax: 972 3 642 9377. E-mail: eueqene{at}ccsg.tau.ac.il.


Applied and Environmental Microbiology, July 2000, p. 3031-3036, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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