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Differential enumeration of intact and damaged marine
planktonic
bacteria based on cell membrane integrity
Takeshi Naganuma
Faculty of Applied Biological Science, Hiroshima University,
1-4-4 Kagamiyama,
Higashi-Hiroshima 739, Japan
Received 10 September 1995; accepted
in final form 10 June 1996
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Abstract
"Viable but non-culturable" (VBNC) bacteria
are a much discussed issue in microbial ecology. Quantitative aspects
are not understood, due mostly to the lack of suitable techniques.
A widely accepted approach is dependent on the integrity of cell
membranes. Recently developed fluorescence dyes differ in permeability
with respect to the integrity of membranes: one dye permeates the
intact membranes, while another permeates those which are damaged.
Although the dyes were developed originally for determining the
viability of cultured bacteria, here they are used to enumerate
live and dead bacterial cells (designated as having intact and damaged
membranes, respectively) in natural environments. Preliminary results
from coastal waters of Seto Inland Sea, Japan, were:
1) the sums of the intact and damaged cells were very similar in
each case to the total number of acridine orange- stained cells;
and 2) about 50-60% of the total bacteriaoplankton populations are
intact with respect to membrane integrity.
Keywords: bacterioplankton, viability, eutrophication
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