The term “nonculturable” was invoked by Xu et al. in 1982 (47) to describe starved but viable cells in a survival or dormant state of existence, observed to occur after an actively growing culture of Vibrio cholerae O1 or Escherichia coli was placed in a nutrient-free microcosm, e.g., a saline solution free of nutrient and incubated at low temperature. The cells were grown in a nutrient broth under optimal conditions, harvested by centrifugation, washed, and placed in sterile artificial seawater (15‰ salinity). The cells were enumerated directly, using both acridine orange and fluorescent-antibody staining, and viewed by epifluorescent microscopy. They were also plated using media optimized for their growth. It was determined by direct microscopic observation that the total number of cells did not decrease with time. However, as incubation continued over several days, it was found that colonies no longer formed when the same samples were plated on media that had been optimized for growth of the culture. To determine whether such cells were viable or dead, a microscopic technique developed by Kogure et al. (25) was employed to examine the cultures that yielded no colonies on transfer to solid media and no growth in liquid media. The Kogure et al. (25) method, a direct-viable-count procedure, when applied to these cultures revealed that nearly all of the cells enumerated by acridine orange direct counting (AODC) were metabolically active, even though they could not be recovered on any plating medium or in broth. Hence, the phrase “viable but nonculturable” (VBNC) was coined to describe these bacteria that were apparently in a dormant state.


Interstitial Cystitis Dormant State Vibrio Cholerae VBNC State VBNC Cell 
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Copyright information

© ASM Press, Washington, D.C. 2000

Authors and Affiliations

  • Rita R. Colwell
    • 1
  • D. Jay Grimes
    • 2
  1. 1.Center of Marine Biotechnology, Columbus CenterUniversity of Maryland Biotechnology InstituteBaltimoreUSA
  2. 2.Institute of Marine SciencesUniversity of Southern MississippiOcean SpringsUSA

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