Abstract
A well-studied, long-term survival mechanism employed by Gram-positive bacteria is formation of endospores. For Gram-negative bacteria, the assumption has been that a survival state does not exist. However, a dormancy state has been described for Gram-negative bacteria and designated as the viable but nonculturable (VBNC) strategy of nonspore-forming cells. A variety of environmental factors are involved in induction of the viable but nonculturable state and Vibrio cholerae provides a useful paradigm for the VBNC phenomenon. It is now accepted that plate counts cannot be relied upon to enumerate or detect VBNC cells. Therefore, direct methods employing fluorescent staining, molecular genetic probes, and other molecular methods have proven both useful and reliable in detecting and enumerating both culturable and nonculturable cells. A predictive model for cholera has been developed, based on ground truth data gathered using these molecular methods and combining them with data obtained by remote sensing, employing satellites. It is clear that microbiology in the twenty-first century has been enhanced by these new tools and paradigms.
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References
Alam M, Sultana M, Nair GB, Siddique AK, Hasan NA, Sack RB, Sack DA, Ahmed KU, Sadique A, Watanabe H, Grim CJ, Huq A, Colwell RR (2007) Viable but nonculturable Vibrio cholerae 01 in biofilms in the aquatic environment and their role in cholera transmission. Proc Natl Acad Sci U S A 104(45):17801–17806
Arana I, Ortuno M, Seco C, Muela A, Barcina I (2008) Is Urografin density gradient centrifugation suitable to separate nonculturable cells from Escherichia coli populations? Antonie Van Leeuwenhoek 93(3):249–257
Asakura H, Ishiwa A, Arakawa E, Makino S, Okada Y, Yamamoto S, Igimi S (2006) Gene expression profile of Vibrio cholerae in the cold stress-induced viable but non-culturable state. Environ Microbiol 9(4):869–879
Azevedo NF, Almeida C, Cerqueira L, Dias S, Keevil CW, Vieira MJ (2007) Coccoid form of Helicobacter pylori as a morphological manifestation of cell adaptation to the environment. Appl Environ Microbiol 73(10):3423–3427
Blake P (1994) Endemic cholera in Australia and the United States. In: Wachsmuth I, Blake P, Olsvik O (eds) Vibrio cholerae and cholera: molecular to global perspectives. American Society for Microbiology, Washington, DC, pp 309–320
Boyle M, Sichel C, Fernández-Ibáñez P, Arias-Quiroz GB, Iriarte-Puña M, Mercado A, Ubomba-Jaswa E, McGuigan KG (2008) Bactericidal effect of solar water disinfection under real sunlight conditions. Appl Environ Microbiol 74(10):2997–3001
Brayton PR, Colwell RR (1987) Fluorescent antibody staining method for the enumeration of viable but nonculturable V. cholerae. J Microbiol Methods 6:309–314
Byrd JJ, Colwell RR (1990) Maintenance of plasmids pBR322 and pUC8 in nonculturable Escherichia coli in the marine environment. Appl Environ Microbiol 56(7):2104–2107
Cappelier JM, Besnard V, Roche SM, Velge P, Federighi M (2007) Avirulent viable but non culturable cells of Listeria monocytogenes need the presence of an embryo to be recovered in egg yolk and regain virulence after recovery. Vet Res 38:573–583
Chaiyanan S, Chaiyanan S, Grim C, Maugel T, Huq A, Colwell R (2007) Ultrastructure of coccoid viable but non-culturable Vibrio cholerae. Environ Microbiol 9(2):393–402
Chowdhury M, Xu B, Montilla R, Hasan J, Huq A, Colwell R (1995) A simplified immunoflourescence technique for detection of viable cells of V. cholerae O1 and O139. J Microbiol Methods 24:165–170
Chowdhury MAR, Huq A, Xu B, Colwell RR (1997) Effect of Alum on free living and copepod associated V. cholerae O1 and O139. Appl Environ Microbiol 63:3323–3326
Colwell RR (1996) Global climate and infectious disease: the cholera paradigm. Science 274:2025–2031
Colwell RR, Grimes DJ (2000) Nonculturable microorganisms in the environment. American Society for Microbiology, Washington, DC, p 354
Colwell R, Huq A (1994) Environmental reservoir of Vibrio cholerae. The causative agent of cholera. In: Wilson M, Levins R, Spielman A (eds) Disease in evolution: global changes and emergence of infectious diseases. Ann N Y Acad Sci 740:44–54
Colwell RR, Huq A (2001) Marine Ecosystem and Cholera. In The Ecology and Etiology of Newly Emerging Marine Diseases. Porter JW (ed). Kluner Academic Publisher, Dordrecht, Netherland, pp. 141–145
Colwell R, Seidler R, Kaper J, Joseph S, Garges S, Lockman H, Maneval D, Bradford H, Roberts N, Remmers R, Huq I, Huq A (1981) Occurrence of V. cholerae serotype O1 in Maryland and Louisiana estuaries. Appl Environ Microbiol 41:555–558
Colwell R, Brayton P, Grimes D, Roszak D, Huq S, Palmer L (1985) Viable but non-culturable Vibrio cholerae and related environmental pathogens in the environment: implications for release of genetically engineered microorganisms. Bio-Technology 3:817–820
Colwell R, Brayton P, Huq A, Tall B, Harrington P, Levine M (1996) Viable but non-culturable Vibrio cholerae O1 revert to a culturable state in the human intestine. World J Microbiol Biotechnol 12:28–31
Constanin de Magny, Guillaume, Raghu Murtugudde, Mathew RP Sapiano, Azhar Nizam, Christopher W Brown, Antonio J Busalacchi, Mohammad Yunus, Balakrish Nair G, Ana I Gil, Claudio F Lanata, John Calkins, Byomkesh Manna, Krishnan Rajendran, Mihir Kumar Bhattacharya, Anwar Huq, Bradley Sack R, Colwell RR (2008) Environmental signatures associated with cholera epidemics. Proc Nat’l Acad Sci 105(46):17676–17681
Du M, Chen J, Zhang X, Li A, Li Y, Wang Y (2007) Retention of virulence in a viable but nonculturable Edwardsiella tarda isolate. Appl Environ Microbiol 73(4):1349–1354
Fera MT, Maugeri TL, Gugliandolo C, La Camera E, Lentini V, Favaloro A, Bonanno D, Carbone M (2008) Induction and resuscitation of viable nonculturable Arcobacter butzleri cells. Appl Environ Microbiol 74(10):3266–3268
Folk R (1996) In defense of nanobacteria. Science 274:1288
Hobbie J, Daley R, Jasper J (1977) Use of nucleopore filters for counting bacteria by fluorescent microscopy. Appl Environ Microbiol 33:1225–1228
Huq A, Colwell R, Rahman R, Ali A, Chowdhury M, Parveen S, Sack D, Russek-Cohen E (1990) Detection of Vibrio cholerae O1 in the aquatic environment by fluorescent monoclonal antibody and culture method. Appl Environ Microbiol 56:2370–2373
Huq A, Rivera I, Colwell R (2000) Epidemiological significance of viable but nonculturable microorganisms. In: Colwell R, Grimes D (eds) Nonculturable microorganisms in the environment. ASM, Washington, DC, pp 301–323
Huq A, Sack R, Nizam A, Longini I, Nair G, Ali A, Morris G, Huda M, Siddique A, Yunus M, Albert J, Sack D, Colwell R (2005) Critical factors influencing the occurrence of Vibrio cholerae in the environment of Bangladesh. Appl Environ Microbiol 71:4645–4654
Knight I (2000) Molecular genetic methods for detection and identification of viable but nonculturable microorganisms. In: Colwell R, Grimes D (eds) Nonculturable microorganisms in the environment. ASM, Washington, DC, pp 77–85
Koch R (1884) An address on cholera and its bacillus. Br Med J 2:403–407, 453–459
Kogure K, Simidu U, Taga N (1979) A tentative direct microscopic method for counting living marine bacteria. Can J Microbiol 25:415–420
Kurath G, Morita Y (1983) Starvation survival and physiological studies of a marine Pseudomonas spp. Appl Environ Microbiol 45:1206–1211
Lee K, Ruby E (1995) Symbiotic role of the viable but nonculturable state of Vibrio fischeri in Hawaiian coastal seawater. Appl Environ Micbrobiol 61:278–283
Lewis K (2007) Persister cells, dormancy and infectious disease. Nature 5(1):48–56
Liu Y, Gilchrist A, Zhang J, Li X-F (2008) Detection of viable but nonculturable Escherichia coli O157:H7 bacteria in drinking water and river water. Appl Environ Microbiol 74(5):1502–1507
Lobitz B, Beck L, Huq A, Wood B, Fuchs G, Faruque A, Colwell R (2000) Climate and infectious disease: use of remote sensing for detection of Vibrio cholerae by indirect measurement. Proc Natl Acad Sci 97:1438–1443
McFeters G, LeChevallier M (2000) Chemical disinfection and injury of bacteria in water. In: Colwell R, Grimes D (eds) Nonculturable microorganisms in the environment. ASM, Washington, DC, pp 255–275
Novitsky J, Morita R (1976) Morphological characterization of small cells resulting from nutrient starvation of a psychrophilic marine vibrio. Appl Environ Microbiol 32:617–622
Nystul TG, Goldmark JP, Padilla PA, Roth MB (2003) Suspended animation in C. elegans requires the spindle checkpoint. Science 302:1038–1041
Pommepuy M, Utin MB, 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(12):4621–4626
Postgate J (1967) Viability, measurements, and survival of microbes under minimum stress. Adv Microb Physiol 1:1–24
Postgate J (1969) Viable counts and viability. In: Norris J, Ribbons D (eds) Methods in microbiology. Academic, London, pp 611–628
Postgate JR, Hunter JR (1962) The survival of starved bacteria. J Gen Microbiol 29:233–263
Roszak D, Grimes D, Colwell R (1984) Viable but non-recoverable stage of Salmonella enteritidis in aquatic systems. Can J Microbiol 30:334–338
Sack RB, Siddique AK, Longini I, Nizam A, Yunus M, Islam S, Morris JG, Ali A, Huq A, Nair GB, Qadri F, Faruque SM, Sack DA, Colwell RR (2003) A 4-year study of the epidemiology of Vibrio cholerae in four rural areas of Bangladesh. J Infect Dis 187:96–101
Sochard MR, Wilson DF, Austin B, Colwell RR (1979) Bacteria associated with the surface and gut of marine copepods. Appl Environ Microbiol 37:750–759
Valentine RC, Bradfield JRG (1954) The urea method for bacterial viability counts with electron microscope and its relation to other viability counting methods. J Gen Microbiol 11:349–357
Vital M, Fuchslin HP, Hammes F, Egli T (2007) Growth of Vibrio cholerae O1 Ogawa Eltor in freshwater. Microbiology 153(7):1993–2001
Xu H-S, Roberts N, Singleton FL, Attwell RW, Grimes DJ, Colwell RR (1982) Survival and viability of nonculturable Escherichia coli and Vibrio cholerae in the estuarine and marine environment. Microb Ecol 8:313–323
Xu HS, Roberts N, Singleton FL, Attwell RW, Grimes DJ, Colwell RR (1984) An indirect fluorescent antibody staining procedure for detection of Vibrio cholerae serovar O1 cells in aquatic environmental samples. J Microb Methods 2:221–231
Acknowledgments
This work was supported by National Institutes of Health Grant No. R01A139129-01 and National Oceanic and Atmospheric Administration Grant No. S0660009 of the Oceans and Human Health Initiative (OHHI).
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Colwell, R.R. (2009). Viable but Not Cultivable Bacteria. In: Epstein, S. (eds) Uncultivated Microorganisms. Microbiology Monographs, vol 10. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-85465-4_1
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