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Characterization of Viable But Nonculturable Condition of Escherichia coli Induced with Copper  

Ku, Hyung-Keun (Health Metrology Center, Division of Metrology for Quality Life, Korea Research Institute of Standards and Science)
Park, Sang-Ryoul (Health Metrology Center, Division of Metrology for Quality Life, Korea Research Institute of Standards and Science)
Kim, Sook-Kyung (Health Metrology Center, Division of Metrology for Quality Life, Korea Research Institute of Standards and Science)
Publication Information
Microbiology and Biotechnology Letters / v.36, no.3, 2008 , pp. 209-214 More about this Journal
Abstract
VBNC (Viable but nonculturable) state is an adaptive response of cells in adverse environments, which lead cell not grow on routine nutrient agar. In this study, we induced VBNC in Escherichia coli using copper and verify the characterization of it. After treatment of copper, we didn't detect any cells via plate cultivation, namely, colony forming unit (CFU) was zero. However, we identified the existence of VBNC by staining live cells with Live/Dead BacLight bacterial viability kit and counting them through flow cytometry. Then we isolated genomic DNA and RNA from VBNC-induced cells and analyzed the stability of them. Degradation of RNA is more severe than that of DNA and RNA is degraded as specific fragments. In addition, we showed the morphology of VBNC cell by Bio-Transmission Electron Microscope (Bio-TEM). VBNC cell showed impaired periplasmic space and inner and outer membrane were separated and the amount of cytosol were significantly decreased.
Keywords
VBNC; Escherichia coli; copper; Bio-TEM;
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1 Grey, B. and T. R. Steck. 2001. Concentrations of copper thought to be toxic to Escherichia coli can induce the viable but nonculturable condition. Appl. Environ. Microbiol. 67: 5325-5327   DOI   ScienceOn
2 Gupte, A. R., C. L. De Rezende, and S. W. Joseph. 2003. Induction and resuscitation of viable but nonculturable Salmonella enterica serovar typhimurium DT104. Appl. Environ. Microbiol. 69: 6669-6675   DOI
3 Heim, S., M. M. Lleo, B. Bonato, C. A. Guzman, and P. Canepari. 2002. The viable but nonculturable state and starvation are different stress responses of Enterococcus faecalis, as determined by proteome analysis. J. Bacteriol. 184: 6739-6745   DOI
4 Jones, D. M., E. M. Sutcliffe, and A. Curry. 1991. Recovery of viable but non-culturable Campylobacter jejuni. J. Gen. Microbiol. 137: 2477-2482   DOI   PUBMED   ScienceOn
5 Ordax, M., E. Marco-Noales, M. M. Lopez, and E. G. Biosca. 2006. Survival strategy of Erwinia amylovora against copper: induction of the viable-but-nonculturable state. Appl. Environ. Microbiol. 72: 3482-3488   DOI   ScienceOn
6 Wai, S. N., Y. Mizunoe, A. Takade, and S. Yoshida. 2000. A comparison of solid and liquid media for resuscitation of starvation-and low-temperature-induced nonculturable cells of Aeromonas hydrophila. Arch. Microbiol. 173: 307-310   DOI
7 Xu, H. S., N. Roberts, F. L. Singleton, R. W. Attwell, D. J. Grimes, and R. R. Colwell. 1982. Survival and viability of nonculturable Escherichia coli and Vibrio cholera in the estuarine and marine environment. Microb. Ecol. 8: 313-323   DOI   ScienceOn
8 Muela, A., C. Seco, E. Camafeita, I. Arana, M. Orruno, J. A. Lopez, and I. Barcina. 2008. Changes in Escherichia coli outer membrane subproteome under environmental conditions inducing the viable but nonculturable state. FEMS Microbiol. Ecol. 64: 28-36   DOI   ScienceOn
9 Arana, I., M. Orruno, D. Perez-Pascual, C. Seco, A. Muela, and I. Barcina. 2007. Inability of Escherichia coli to resuscitate from the viable but nonculturable state. FEMS Microbiol. Ecol. 62: 1-11   DOI   ScienceOn
10 Vora, G. J., C. E. Meador, M. M. Bird, C. A. Bopp, J. D. Andreadis, and D. A. Stenger. 2005. Microarray-based detection of genetic heterogeneity, antimicrobial resistance, and the viable but nonculturable state in human pathogenic Vibrio spp. Proc. Natl. Acad. Sci. USA. 102: 19109-19114
11 Oliver, J. D., L. Nilsson, and S. Kjelleberg. 1991. Formation of nonculturable Vibrio vulnificus cells and its relationship to the starvation state. Appl. Environ. Microbiol. 57: 2640-2644   PUBMED
12 Vattakaven, T., P. Bond, G. Bradley, and C. B. Munn. 2006. Differential effects of temperature and starvation on induction of the viable-but-nonculturable state in the coral pathogens Vibrio shiloi and Vibrio tasmaniensis. Appl. Environ. Microbiol. 72: 6508-6513   DOI   ScienceOn
13 Lleo, M. M., S. Pierobon, M. C. Tafi, C. Signoretto, and P. Canepari. 2000. mRNA detection by reverse transcription-PCR for monitoring viability over time in an Enterococcus faecalis viable but nonculturable population maintained in a laboratory microcosm. Appl. Environ. Microbiol. 66: 4564-4567   DOI
14 Whitesides, M. D. and J. D. Oliver. 1997. Resuscitation of Vibrio vulnificus from the Viable but Nonculturable State. Appl. Environ. Microbiol. 63: 1002-1005   PUBMED
15 Day, A. P. and J. D. Oliver. 2004. Changes in membrane fatty acid composition during entry of Vibrio vulnificus into the viable but nonculturable state. J. Microbiol. 42: 69-73   PUBMED
16 Smith B. and J. D. Oliver. 2006. In situ and in vitro gene expression by Vibrio vulnificus during entry into, persistence within, and resuscitation from the viable but nonculturable state. Appl. Environ. Microbiol. 72: 1445-1451   DOI   ScienceOn
17 Keep, N. H., J. M. Ward, G. Robertson, M. Cohen-Gonsaud, and B. Henderson. 2006. Bacterial resuscitation factors: revival of viable but non-culturable bacteria. Cell. Mol. Life Sci, 63: 2555-2559   DOI
18 Beumer, R. R., J. de Vries, and F. M. Rombouts, 1992. Campylobacter jejuni non-culturable coccoid cells. Int. J. Food Microbiol. 15: 153-163   DOI   ScienceOn
19 Signoretto, C., M. M. Lleo, and P. Canepari. 2002. Modification of the peptidoglycan of Escherichia coli in the viable but nonculturable state. Curr. Microbiol. 44: 125-131   DOI
20 Alexander, E., D. Pham, and T. R. Steck. 1999. The viable-but-nonculturable condition is induced by copper in Agrobacterium tumefaciens and Rhizobium leguminosarum. Appl. Environ. Microbiol. 65: 3754-3756   PUBMED
21 Pujol, M., E. Badosa, and E. Montesinos. 2007. Epiphytic fitness of a biological control agent of fire blight in apple and pear orchards under Mediterranean weather conditions. FEMS Microbiol. Ecol. 59: 186-193   DOI   ScienceOn
22 Porter, J., C. Edwards, and R. W. Pickup. 1995. Rapid assessment of physiological status in Escherichia coli using fluorescent probes. J. Appl. Bacteriol. 79: 399-408   DOI