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Appl Environ Microbiol, January 1998, p. 279-286, Vol. 64, No. 1
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Production of Respirable Vesicles Containing Live
Legionella pneumophila Cells by Two
Acanthamoeba spp.
Sharon G.
Berk,1,*
Rebecca S.
Ting,1
Glenn W.
Turner,2,
and
Rebecca
J.
Ashburn1
Center for the Management, Utilization and
Protection of Water Resources, Tennessee Technological University,
Cookeville, Tennessee 38505,1 and
Biology Department, Middle Tennessee State University,
Murfreesboro, Tennessee 371322
Received 23 June 1997/Accepted 31 October 1997
Two Acanthamoeba species, fed at three temperatures,
expelled vesicles containing living Legionella pneumophila
cells. Vesicles ranged from 2.1 to 6.4 µm in diameter and
theoretically could contain several hundred bacteria. Viable L. pneumophila cells were observed within vesicles which had been
exposed to two cooling tower biocides for 24 h. Clusters of
bacteria in vesicles were not dispersed by freeze-thawing and
sonication. Such vesicles may be agents for the transmission of
legionellosis associated with cooling towers, and the risk may be
underestimated by plate count methods.
*
Corresponding author. Mailing address: Center for the
Management, Utilization and Protection of Water Resources, P.O. Box 5033, Tennessee Technological University, Cookeville, TN 38505. Phone:
(931) 372-3451. Fax: (931) 372-6346. E-mail: SBERK{at}TNTECH.EDU.

Present address: Institute of Biological Chemistry, Washington,
State University, Pullman, WA 99164-6340.
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