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Applied and Environmental Microbiology, July 2000, p. 3031-3036, Vol. 66, No. 7
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
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
-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.
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