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Scientific
Publications - Work Done by Microbiology Reader
Galina V. Mukamolova, Nataliya D. Yanopolskaya, Douglas B. Kell and Arseny
S. Kaprelyants, On resuscitation from the dormant state of Micrococcus
luteus, Antonie van Leeuwenhoek -
International Journal
of General and Molecular Microbiology, 73 (3): 237-243, April 1998 ABSTRACT
It has been found previously that a significant number of Micrococcus luteus
cells starved in a prolonged stationary phase (up to 2 months) and then held
on the bench at room temperature without agitation for periods of up to a
further 2–7 months can be resuscitated in
liquid media which contained (statistically) no initially-viable
(colony-forming) cells but which were fortified with sterile supernatant
from the late logarithmic phase of batch growth. Here it was found that such
resuscitation can be done only within a defined time period after taking the
first sample from such cultures, necessarily involving agitation of the
cells. The duration of this period depends on the age of the starved
culture: cells kept on the bench for 3 months possess a 2 month period of
resuscitability while cells starved for 6 months can be resuscitated only
within 10 days after the beginning of sampling. It is suggested that the
input of oxygen to the starved cultures while they are agitated may exert a
negative influence on the cells, since cultures stored in anaerobic
conditions (under nitrogen) had a more prolonged ’survival‘
time. The cells which experienced between 10 and 60 days of starvation on
the bench could be resuscitated, although the number of resuscitable cells
depended strongly on the concentration of yeast extract in the resuscitation
medium. This concentration for cells stored on the bench for more than 2
months was 0.05% while ’1-month-old‘
cells displayed a maximum resuscitability in the presence of 0.01% of yeast
extract. Application of the fluorescent probe propidium iodide revealed the
formation of cells with a damaged permeability barrier if resuscitation was
performed by using concentrations of yeast extract of 0.1% and above. Thus
the successful resuscitation of bacterial cultures under laboratory
conditions may need rather strictly defined parameters if it is to be
successfully performed for the majority of cells in a population.
Keywords
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