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Appl Environ Microbiol. 1996 January; 62(1): 115–120.
PMCID: PMC167780

Potential virulence of viable but nonculturable Shigella dysenteriae type 1.


We examined a virulent strain of Shigella dysenteriae type 1 after induction into the viable but nonculturable (VBNC) state for its ability to (i) maintain the Shiga toxin (stx) gene; (ii) maintain biologically active Shiga toxin (ShT); and (iii) adhere to intestinal epithelial cells (Henle 407 cell line). PCR was used to amplify the stx gene from VBNC cells of S. dysenteriae type 1, thereby establishing its presence even when cells are in the VBNC state. VBNC S. dysenteriae type 1 ShT was monitored by the enzyme-linked immunosorbent assay with mouse monoclonal antibodies against the B subunit of ShT and affinity-purified rabbit polyclonal antibodies against ShT. We used the Henle 407 cell line to study the adhesive property of VBNC S. dysenteriae type 1 cells in a series of tissue culture experiments. Results showed that VBNC S. dysenteriae type 1 not only maintained the stx gene and biologically active ShT but also remained capable of adhering to Henle 407 cells. However, S. dysenteriae type 1 cells lost the ability to invade Henle 407 cells after entering the VBNC state. From results of the study, we conclude that VBNC cells of S. dysenteriae type 1 retain several virulence factors and remain potentially virulent, posing a public health problem.

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Selected References

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  • Brauns LA, Hudson MC, Oliver JD. Use of the polymerase chain reaction in detection of culturable and nonculturable Vibrio vulnificus cells. Appl Environ Microbiol. 1991 Sep;57(9):2651–2655. [PMC free article] [PubMed]
  • Gentry MK, Dalrymple JM. Quantitative microtiter cytotoxicity assay for Shigella toxin. J Clin Microbiol. 1980 Sep;12(3):361–366. [PMC free article] [PubMed]
  • Hale TL. Genetic basis of virulence in Shigella species. Microbiol Rev. 1991 Jun;55(2):206–224. [PMC free article] [PubMed]
  • Hobbie JE, Daley RJ, Jasper S. Use of nuclepore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol. 1977 May;33(5):1225–1228. [PMC free article] [PubMed]
  • Islam MS, Hasan MK, Miah MA, Sur GC, Felsenstein A, Venkatesan M, Sack RB, Albert MJ. Use of the polymerase chain reaction and fluorescent-antibody methods for detecting viable but nonculturable Shigella dysenteriae type 1 in laboratory microcosms. Appl Environ Microbiol. 1993 Feb;59(2):536–540. [PMC free article] [PubMed]
  • Jackson MP. Detection of Shiga toxin-producing Shigella dysenteriae type 1 and Escherichia coli by using polymerase chain reaction with incorporation of digoxigenin-11-dUTP. J Clin Microbiol. 1991 Sep;29(9):1910–1914. [PMC free article] [PubMed]
  • Kogure K, Simidu U, Taga N. A tentative direct microscopic method for counting living marine bacteria. Can J Microbiol. 1979 Mar;25(3):415–420. [PubMed]
  • Lee K, Ruby EG. Symbiotic Role of the Viable but Nonculturable State of Vibrio fischeri in Hawaiian Coastal Seawater. Appl Environ Microbiol. 1995 Jan;61(1):278–283. [PMC free article] [PubMed]
  • Linder K, Oliver JD. Membrane fatty acid and virulence changes in the viable but nonculturable state of Vibrio vulnificus. Appl Environ Microbiol. 1989 Nov;55(11):2837–2842. [PMC free article] [PubMed]
  • Medema GJ, Schets FM, van de Giessen AW, Havelaar AH. Lack of colonization of 1 day old chicks by viable, non-culturable Campylobacter jejuni. J Appl Bacteriol. 1992 Jun;72(6):512–516. [PubMed]
  • Morgan JA, Rhodes G, Pickup RW. Survival of nonculturable Aeromonas salmonicida in lake water. Appl Environ Microbiol. 1993 Mar;59(3):874–880. [PMC free article] [PubMed]
  • Nilsson L, Oliver JD, Kjelleberg S. Resuscitation of Vibrio vulnificus from the viable but nonculturable state. J Bacteriol. 1991 Aug;173(16):5054–5059. [PMC free article] [PubMed]
  • O'Neill KR, Jones SH, Grimes DJ. Seasonal incidence of Vibrio vulnificus in the Great Bay estuary of New Hampshire and Maine. Appl Environ Microbiol. 1992 Oct;58(10):3257–3262. [PMC free article] [PubMed]
  • Qadri F, Haq S, Hossain SA, Ciznar I, Tzipori S. The association of haemagglutination and adhesion with lipopolysaccharide of Shigella dysenteriae serotype 1. J Med Microbiol. 1991 May;34(5):259–264. [PubMed]
  • Rahman I, Shahamat M, Kirchman PA, Russek-Cohen E, Colwell RR. Methionine uptake and cytopathogenicity of viable but nonculturable Shigella dysenteriae type 1. Appl Environ Microbiol. 1994 Oct;60(10):3573–3578. [PMC free article] [PubMed]
  • Rollins DM, Colwell RR. Viable but nonculturable stage of Campylobacter jejuni and its role in survival in the natural aquatic environment. Appl Environ Microbiol. 1986 Sep;52(3):531–538. [PMC free article] [PubMed]
  • Roszak DB, Colwell RR. Survival strategies of bacteria in the natural environment. Microbiol Rev. 1987 Sep;51(3):365–379. [PMC free article] [PubMed]
  • Shahamat M, Mai U, Paszko-Kolva C, Kessel M, Colwell RR. Use of autoradiography to assess viability of Helicobacter pylori in water. Appl Environ Microbiol. 1993 Apr;59(4):1231–1235. [PMC free article] [PubMed]
  • Siegele DA, Kolter R. Life after log. J Bacteriol. 1992 Jan;174(2):345–348. [PMC free article] [PubMed]
  • Turpin PE, Maycroft KA, Rowlands CL, Wellington EM. Viable but non-culturable salmonellas in soil. J Appl Bacteriol. 1993 Apr;74(4):421–427. [PubMed]
  • Weichart D, Oliver JD, Kjelleberg S. Low temperature induced non-culturability and killing of Vibrio vulnificus. FEMS Microbiol Lett. 1992 Dec 15;100(1-3):205–210. [PubMed]
  • Zimmermann R, Iturriaga R, Becker-Birck J. Simultaneous determination of the total number of aquatic bacteria and the number thereof involved in respiration. Appl Environ Microbiol. 1978 Dec;36(6):926–935. [PMC free article] [PubMed]

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