Annals of Microbiology

, Volume 60, Issue 1, pp 157–160 | Cite as

Resuscitation of 20-year starved Salmonella in seawater and soil

Original Article


Salmonella enterica serovar Typhimurium was stressed by incubation in seawater and soil microcosms for 20 years. Recovery of viable but nonculturable Salmonella was obtained by incubation in nutrient broth and buffered peptone water medium for 2 months, and also in a mouse model when administered orally but not by intra-peritoneal injection. The 16S rDNA gene sequences from 20-year stressed strains were homologous, with a high degree of similarity, and all were identical to the respective initial strain.


Salmonella Seawater Stress Soil Viable but nonculturable Resuscitation 


  1. Barer MR, Gribbon LT, Harwood CR, Nwoguh CE (1993) The viable but nonculturable hypothesis and medical bacteriology. Rev Med Microbiol 4:183–191Google Scholar
  2. Colwell RR, Brayton P, Herrington B, Tall B, Huq A, Levine MM (1996) Viable but nonculturable Vibrio cholerae 01 revert to a cultivable state in the human intestine. World J Microbiol Biotechnol 12:28–31CrossRefGoogle Scholar
  3. Dhiaf A, Bakhrouf A (2004) Recovery in embryonated chicken eggs of viable but non-culturable Salmonella. J Food Agric Environ 2:104–107Google Scholar
  4. Dhiaf A, Bakhrouf A (2008) Resuscitation of eight-year VBNC Providencia stuartii. J Food Agric Environ 6:447–450Google Scholar
  5. Dhiaf A, Junier P, Bakhrouf A, Witzel K-P (2008) Resuscitation of ten-year VBNC Klebsiella oxytoca. Cah Biol Mar 49:1–6Google Scholar
  6. Jones DM, Sutcliffe EM, Curry A (1991) Recovery of viable but nonculturable Campylobacter jejuni. J Gen Microbiol 137:2477–2482PubMedGoogle Scholar
  7. Medema GJ, Schets FM, Van de Giessen AW, Havelaar A (1992) Lack of colonization of one day old chicks by viable, non-culturable Campylobacter jejuni. J Appl Bacteriol 72:512–516PubMedGoogle Scholar
  8. Muyzer G, De Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700PubMedGoogle Scholar
  9. Oliver JD, Bockian R (1995) In vivo resuscitation, and virulence towards mice, of viable but nonculturable cells of Vibrio vulnificus. Appl Environ Microbiol 61:2620–2623PubMedGoogle Scholar
  10. Oliver JD, Kjelleberg S, Kjelleberg S (1993) Formation of viable but nonculturable cells starvation in bacteria. Plenum, New York, pp 239–272Google Scholar
  11. Pommepuy M, Butin M, Derrien A, Gourmelon M, Colwell RR, Cormier M (1996) Retention of enteropathogenicity by viable but nonculturable Escherichia coli exposed to seawater and sunlight. Appl Environ Microbiol 62:4621–4626PubMedGoogle Scholar
  12. Rahman I, Shahamat M, Kirchman PA, Russek-Cohen E, Colwell RR (1994) Methionine uptake and cytopathogenicity of viable but nonculturable Shigella dysenteriae type 1. Appl Environ Microbiol 60:3573–3578PubMedGoogle Scholar
  13. Rahman I, Shahamat M, Chowdhury MAR, Colwell RR (1996) Potential virulence of viable but nonculturable Shigella dysenteriae type 1. Appl Environ Microbiol 62:115–120PubMedGoogle Scholar
  14. Saha SK, Saha S, Sanyal SC (1991) Recovery of injured Campylobacter jejuni cells after animal passage. Appl Environ Microbiol 57:3388–3389PubMedGoogle Scholar
  15. Steinert M, Emödy L, Amann R, Hacker J (1997) Resuscitation of viable but nonculturable Legionella pneumophila Philadelphia JR32 by Acanthamoeba castellanii. Appl Environ Microbiol 63:2047–2053PubMedGoogle Scholar
  16. Sylvester DM, Taylor R, LaHann TR (2001) Viable but nonculturable bacteria: a public health threat? Infect Dis Rev 3:70–82Google Scholar
  17. Van de Giessen AW, Heuvelman CJ, Abee A, Hazeleger WC (1996) Experimental studies on the infectivity of non-culturable forms of Campylobacter spp. in chicks and mice. Epidemiol Infect 117:463–470CrossRefPubMedGoogle Scholar
  18. Walsh PS, Metzger DA, Higuchi R (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10:506–513PubMedGoogle Scholar

Copyright information

© Springer-Verlag and the University of Milan 2010

Authors and Affiliations

  • Amel Dhiaf
    • 1
    • 2
  • Fethi Ben Abdallah
    • 1
  • Amina Bakhrouf
    • 1
  1. 1.Laboratoire d’Analyse, Faculté de PharmacieTraitement et Valorisation des Polluants de l’Environnement et des ProduitsMonastirTunisie
  2. 2.Cabinet Dr AllayaKsour EssefTunisia

Personalised recommendations