When stationary phase batch cultures of Campylobacter jejuni were stored in sealed flasks under static conditions, viable numbers declined from 2 x 109c.f.u. ml-1to around 103-106c.f.u. ml-1within 4-6 weeks. When the aged cultures were sparged with a microaerobic gas mixture, there was a rapid increase in viable numbers accompanied by a change from predominantly coccoid to vibrioid morphology. The most probable number (MPN) technique was used to distinguish resuscitation of injured or dormant cells from multiplication of residual viable cells. MPN estimates using fresh Brucella broth containing 0.2% mucin revealed that plate counts underestimated the true viable count by up to 23-fold. The experiments clearly demonstrated that a proportion of surviving cells in aged cultures were in an injured or latent state that prevented growth on agar plates. It is possible that the size of this fraction is greater than was demonstrated and that much higher recoveries would be obtained under other recovery conditions. Nevertheless, from presently available evidence, it must be concluded that the size of the latent fraction is quite small and that most of the increase in count that occurs on regassing a spent culture comes from multiplication of residual viable cells.
AonoR.,
YamasakiM.,
TamuraG.1978; Changes in composition of envelope proteins in adenylate cyclase- or cyclic AMP receptor protein-deficient mutants of Escherichia coli. J Bacteriol 136:812–814
BoucherS. N.,
SlaterE. R.,
ChamberlainA. H. L.,
AdamsM. R.1994; Production and viability of coccoid forms of Campylobacter jejuni. J Appl Bacteriol 77:303–307
BovillR. A.,
ShallcrossA. M.,
MackeyB. M.1994; Comparison of the fluorescent redox dye 5-cyano-2,3-ditolyltetrazolium chloride with p-iodonitrotetrazolium violet to detect metabolic activity in heat-stressed Listeria monocytogenes cells. J Appl Bacteriol 77:353–358
ColwellR. R.,
BraytonP. R.,
GrimesD. J.,
RoszakD. B.,
HuqS. A.,
PalmerL. M.1985; Viable but non-culturable Vibrio cholerae and related pathogens in the environment : implications for release of genetically engineered microorganisms. Bio/Technology 3:817–820
HazelegerW. C.,
JanseJ. D.,
KoenraadP. M. F. J.,
BeumerR. R.,
RomboutsF. M.,
AbeeT.1995; Temperature-dependent membrane fatty acid and cell physiology changes in coccoid forms of Campylobacter jejuni. Appl Environ Microbiol 61:2713–2719
HumphreyT. J.1989; An appraisal of the efficacy of preenrichment for the isolation of Campylobacter jejuni from water and food. J Appl Bacteriol 66:119–126
JacobJ.,
MartinW.,
HöllerC.1993; Characterization of viable but nonculturable stage of C. coli, characterized with respect to electron microscopic findings, whole cell protein and lipooligosaccharide (LOS) patterns. Z Mikrobiol 148:3–10
KaprelyantsA. S.,
MukamolovaG. V.,
KellD. B.1994; Estimation of dormant Micrococcus luteus cells by penicillin lysis and by resuscitation in cell-free spent culture medium at high dilution. FEMS Microbiol Lett 115:347–352
KingL. K.,
ParkerB. C.1988; A simple, rapid method for enumerating total viable and metabolically active bacteria in ground water. Appl Environ Microbiol 54:1630–1631
LangeR.,
Hengge-AronisR.1991; Growth phase regulated expression of bolA and morphology of stationary phase Escherichia coli cells are controlled by the novel sigma factor Sigma-S. J Bacteriol 173:4474–4481
MackeyB. M.,
DerrickC. M.1982; A comparison of solid and liquid media for measuring the sensitivity of heat-injured Salmonella typhimurium to selenite and tetrathionate media, and the time needed to recover resistance. J Appl Bacteriol 53:233–242
MedemaG. J.,
SchetsF. M.,
van de GiessenA. W.,
HavelaarA. H.1992; Lack of colonization of 1 day old chicks by viable, non-culturable Campylobacter jejuni. J Appl Bacteriol 72:512–516
MoranA. P.,
UptonM. E.1987a; Effect of medium supplements, illumination and superoxide dismutase on the production of coccoid forms of Campylobacter jejuni ATCC 294228. J Appl Bacteriol 62:43–51
MoranA. P.,
UptonM. E.1987b; Factors affecting production of coccoid forms by Campylobacter jejuni on solid media during incubation. J Appl Bacteriol 62:527–537
MukamolovaG. V.,
KaprelyantsA. S.,
KellD. B.1995; Secretion of an antibacterial factor during resuscitation of dormant cells in Micrococcus luteus cultures held in an extended stationary phase. Antonie Leeuwenhoek 67:289–295
OliverJ. D.,
NilssonL.,
KjellebergS.1991; Formation of nonculturable Vibrio vulnificus cells and its relationship to the starvation state. Appl Environ Microbiol 57:2640–2644
OliverJ. D.,
HiteF.,
McDougaldD.,
AndonN. L.,
SimpsonL. M.1995; Entry into, and resuscitation from, the viable but nonculturable state by Vibrio vulnificus in an estuarine environment. Appl Environ Microbiol 61:2624–2630
RasmussenL.,
ChristensenS. T.,
SchousboeP.,
WheatleyD. N.1996; Cell survival and multiplication. The overriding need for signals : from unicellular to multicellular systems. FEMS Microbiol Lett 137:123–128
RodriguezG. G.,
PhippsD.,
IshiguroK.,
RidgwayH. F.1992; Use of a fluorescent redox probe for direct visualization of actively respiring bacteria. Appl Environ Microbiol 58:1801–1808
RollinsD. M.,
ColwellR. R.1986; Viable but nonculturable stage of Campylobacter jejuni and its role in survival in the natural aquatic environment. Appl Environ Microbiol 52:531–538
VotyakovaT. V.,
KaprelyantsA. S.,
KellD. B.1994; Influence of viable cells on the resuscitation of dormant cells in Micrococcus luteus cultures held in an extended stationary phase : the population effect. Appl Environ Microbiol 60:3284–3291
WeichartD.,
KjellebergS.1996; Stress resistance and recovery potential of culturable and viable but nonculturable cells of Vibrio vulnificus. Microbiology 142:845–853