Journal of Water and Health Vol 06 No 4 pp 565–568 © IWA Publishing 2008 doi:10.2166/wh.2008.131
Resuscitation of eleven-year VBNC Citrobacter
Amel Dhiaf, Amina Bakhrouf and Karl-Paul Witzel
Cabinet Dr Allaya, Ouled Salah 5116, Ksour, Essef, Tunisia Tel.: 00216098560041 Fax: 00216073461000 E-mail: dhiafamel@yahoo.fr
Cabinet Dr Allaya, Ouled Salah 5116, Ksour, Essef, Tunisia
Department of Evolutionary Genetics, Max-Planck Institute for Evolutionary Biology, Ploen, Germany
ABSTRACT
Citrobacter freundii strain WA1 was stressed by incubation in seawater microcosms for eleven years. After two years of starvation, no culturable strain was observed. Incubation of samples in nutrient-rich broth medium not supplemented with growth factors, however, allowed resuscitation of VBNC cells so that subsequent plating yielded observable colonies for significantly extended periods of time. Recovery of VBNC Citrobacter freundii was obtained by incubation in nutrient broth even after eleven years of starvation. To see whether the samples contained the same strain of Citrobacter freundii inoculated 11 years ago. The complete 16S rRNA gene was PCR amplified and sequenced from initial, stressed and revived strains of Citrobacter freundii strain WA1.The 16S rRNA gene sequences from eleven-year stressed strains were homologous with a high degree of similarity to the GenBank reference strain and were identical to each other.
Keywords: Citrobacter freundii strain WA1; resuscitation; seawater; stress
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