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Applied and Environmental Microbiology, August 1998, p. 3025-3028, Vol. 64, No. 8
Department of Biology, The University of
North Carolina at Charlotte, Charlotte, North Carolina 28223
Received 4 February 1998/Accepted 8 May 1998
Vibrio vulnificus is an estuarine bacterium capable of
causing a rapidly fatal infection in humans. Because of the low
nutrient levels and temperature fluctuations found in the organism's
natural habitat, the starvation state and viable but nonculturable
(VBNC) state are of particular interest. A randomly amplified
polymorphic DNA (RAPD) PCR protocol was developed previously for the
detection of V. vulnificus strains grown in rich media and
has been applied to starved and VBNC cells of V. vulnificus
in the present study. As cells were subjected to starvation in
artificial seawater, changes in the RAPD profile were detected as early
as 15 min into the starvation period. Most noticeable was a uniform
loss of RAPD amplification products. By 4 h of starvation, the
cells were undetectable by the RAPD method. Cells that had been starved
for up to 1 year again became detectable by the RAPD method when
nutrients were added to the starvation microcosm. The same loss of
signal, but at a lower rate, was also seen as cells entered the VBNC
state. VBNC cells were resuscitated by a temperature upshift and were once again detectable by the RAPD method. The addition of
chloramphenicol prevented the RAPD signal from being lost in both the
starvation and VBNC states. This suggests that DNA binding proteins
produced during starvation and entrance into the VBNC state may be
responsible for the inability of the RAPD method to amplify V. vulnificus DNA in these states.
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Randomly Amplified Polymorphic DNA Analysis of
Starved and Viable but Nonculturable Vibrio vulnificus
Cells
*
Corresponding author. Mailing address: 9201 University-City Blvd., Biology Department, Charlotte, NC 28223. Phone: (704) 547-4049. Fax: (704) 547-3457. E-mail:
jdoliver{at}email.uncc.edu.
Applied and Environmental Microbiology, August 1998, p. 3025-3028, Vol. 64, No. 8
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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