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Volume 152, Issue 6

Flow-cytometric study of vital cellular functions in during solar disinfection (SODIS) Free

Abstract

The effectiveness of solar disinfection (SODIS), a low-cost household water treatment method for developing countries, was investigated with flow cytometry and viability stains for the enteric bacterium . A better understanding of the process of injury or death of during SODIS could be gained by investigating six different cellular functions, namely: efflux pump activity (Syto 9 plus ethidium bromide), membrane potential [bis-(1,3-dibutylbarbituric acid)trimethine oxonol; DiBAC(3)], membrane integrity (LIVE/DEAD BacLight), glucose uptake activity (2-[-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy--glucose; 2-NBDG), total ATP concentration (BacTiter-Glo) and culturability (pour-plate method). These variables were measured in K-12 MG1655 cells that were exposed to either sunlight or artificial UVA light. The inactivation pattern of cellular functions was very similar for both light sources. A UVA light dose (fluence) of <500 kJ m was enough to lower the proton motive force, such that efflux pump activity and ATP synthesis decreased significantly. The loss of membrane potential, glucose uptake activity and culturability of >80 % of the cells was observed at a fluence of ∼1500 kJ m, and the cytoplasmic membrane of bacterial cells became permeable at a fluence of >2500 kJ m. Culturable counts of stressed bacteria after anaerobic incubation on sodium pyruvate-supplemented tryptic soy agar closely correlated with the loss of membrane potential. The results strongly suggest that cells exposed to >1500 kJ m solar UVA (corresponding to 530 W m global sunlight intensity for 6 h) were no longer able to repair the damage and recover. Our study confirms the lethal effect of SODIS with cultivation-independent methods and gives a detailed picture of the ‘agony’ of when it is stressed with sunlight.

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Keyword(s): 2-NBDG, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose , DiBAC4(3), bis-(1,3-dibutylbarbituric acid)trimethine oxonol , EB, ethidium bromide , PEP, phosphoenolpyruvate , PEP-PTS, phosphoenolpyruvate-phosphotransferase system , PI, propidium iodide and SODIS, solar disinfection
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2006-06-01
2024-04-19
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References

  1. Acra A, Raffoul Z, Karahagopian Y. 1984 Solar Disinfection of Drinking Water and Oral Rehydration Solutions New York: United Nations Children's Fund;
     [Google Scholar]
  2. Aldsworth T. G, Sharman R. L, Dodd C. E. 1999; Bacterial suicide through stress. Cell Mol Life Sci 56:378–383 [CrossRef]
     [Google Scholar]
  3. Arami S, Hada M, Tada M. 1997a; Reduction of ATPase activity accompanied by photodecomposition of ergosterol by near-UV irradiation in plasma membranes prepared from Saccharomyces cerevisiae . Microbiology 143:2465–2471 [CrossRef]
     [Google Scholar]
  4. Arami S, Hada M, Tada M. 1997b; Near-UV-induced absorbance change and photochemical decomposition of ergosterol in the plasma membrane of the yeast Saccharomyces cerevisiae . Microbiology 143:1665–1671 [CrossRef]
     [Google Scholar]
  5. Barer M. R, Harwood C. R. 1999; Bacterial viablility and culturability. Adv Microb Physiol 41:93–137
     [Google Scholar]
  6. Berney M, Weilenmann H, Ihssen J, Bassin C, Egli T. 2006; Specific growth rate determines the sensitivity of E. coli to thermal, UVA and solar disinfection. Appl Environ Microbiol 72:2586–2593 [CrossRef]
     [Google Scholar]
  7. Bromberg R, George S. M, Peck W. 1998; Oxygen sensitivity of heated cells of Escherichia coli O157: H7. J Appl Microbiol 85:231–237 [CrossRef]
     [Google Scholar]
  8. Colwell R. R. 2000; Viable but nonculturable bacteria: a survival strategy. J Infect Chemother 6:121–125 [CrossRef]
     [Google Scholar]
  9. Czechowicz S. M, Santos O, Zottola E. A. 1996; Recovery of thermally-stressed Escherichia coli O157: H7 by media supplemented with pyruvate. Int J Food Microbiol 33:275–284 [CrossRef]
     [Google Scholar]
  10. Dodd C. E. R, Sharman R. L, Bloomfield S. F, Booth I. R, Stewart G. S. A. B. 1997; Inimical processes: bacterial self-destruction and sub-lethal injury. Trends Food Sci Tech 8:238–241 [CrossRef]
     [Google Scholar]
  11. Downes A. 1886; On the action of sunlight on micro-organisms, &c., with a demonstration of the influence of diffused light. Proc R Soc 40:14–22 [CrossRef]
     [Google Scholar]
  12. Fewtrell L, Kaufmann R. B, Kay D, Enanoria W, Haller L, Colford J. M. Jr 2005; Water, sanitation, and hygiene interventions to reduce diarrhoea in less developed countries: a systematic review and meta-analysis. Lancet Infect Dis 5:42–52 [CrossRef]
     [Google Scholar]
  13. George S. M, Richardson L. C, Pol I. E, Peck M. W. 1998; Effect of oxygen concentration and redox potential on recovery of sublethally heat-damaged cells of Escherichia coli O157: H7, Salmonella enteritidis and Listeria monocytogenes . J Appl Microbiol 84:903–909 [CrossRef]
     [Google Scholar]
  14. Haugland R. P. 2002 Handbook of Fluorescent Probes and Research Products, 9th edn.. Edited by Gregory J. Eugene, OR: Molecular Probes;
     [Google Scholar]
  15. Hewitt C. J, Nebe-Von-Caron G. 2004; The application of multi-parameter flow cytometry to monitor individual microbial cell physiological state. Adv Biochem Eng Biotechnol 89:197–223
     [Google Scholar]
  16. Hewitt C. J, Nebe-Von Caron G, Nienow A. W, McFarlane C. M. 1999; Use of multi-staining flow cytometry to characterise the physiological state of Escherichia coli W3110 in high cell density fed-batch cultures. Biotechnol Bioeng 63:705–711 [CrossRef]
     [Google Scholar]
  17. Hinrichsen D, Bryant Robey M. A, Upadhyay U. D. 1997 Solutions for a Water-Short World. Population Reports Baltimore: Johns Hopkins School of Public Health; http://www.infoforhealth.org/pr/m14edsum.shtml
     [Google Scholar]
  18. Hobbins M. 2004 Home-based water purification through sunlight: from promotion to health effectiveness. Doctoral thesis Swiss Tropical Institute, University of Basel;
     [Google Scholar]
  19. Jagger J. 1981; Near-UV radiation effects on microorganisms. Photochem Photobiol 34:761–768 [CrossRef]
     [Google Scholar]
  20. Joyce T. M, McGuigan K. G, Elmore-Meegan M, Conroy R. M. 1996; Inactivation of fecal bacteria in drinking water by solar heating. Appl Environ Microbiol 62:399–402
     [Google Scholar]
  21. Kaprelyants A. S, Kell D. B. 1993; Dormancy in stationary-phase cultures of Micrococcus luteus : flow cytometric analysis of starvation and resuscitation. Appl Environ Microbiol 59:3187–3196
     [Google Scholar]
  22. Khaengraeng R, Reed R. H. 2005; Oxygen and photoinactivation of Escherichia coli in UVA and sunlight. J Appl Microbiol 99:39–50 [CrossRef]
     [Google Scholar]
  23. Kobayashi H, Miyamoto T, Hashimoto Y, Kiriki M, Motomatsu A, Honjoh K, Iio M. 2005; Identification of factors involved in recovery of heat-injured Salmonella enteritidis . J Food Prot 68:932–941
     [Google Scholar]
  24. Lakchaura B. D, Fossum T, Jagger J. 1976; Inactivation of adenosine 5′-triphosphate synthesis and reduced-form nicotinamide adenine dinucleotide dehydrogenase activity in Escherichia coli by near-ultraviolet and violet radiations. J Bacteriol 125:111–118
     [Google Scholar]
  25. Lonnen J, Kilvington S, Kehoe S. C, Al-Touati F, McGuigan K. G. 2005; Solar and photocatalytic disinfection of protozoan, fungal and bacterial microbes in drinking water. Water Res 39:877–883 [CrossRef]
     [Google Scholar]
  26. Martin-Dominguez A, Alarcon-Herrera M. T, Martin-Dominguez I. R, Gonzalez-Herrera A. 2005; Efficiency in the disinfection of water for human consumption in rural communities using solar radiation. Solar Energy 78:31–40 [CrossRef]
     [Google Scholar]
  27. Midgley M. 1987; An efflux system for cationic dyes and related compounds in Escherichia coli . Microbiol Sci 4:125–127
     [Google Scholar]
  28. Miller J. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  29. Mukamolova G. V, Yanopolskaya N. D, Kell D. B, Kaprelyants A. S. 1998; On resuscitation from the dormant state of Micrococcus luteus . Antonie Van Leeuwenhoek 73:237–243 [CrossRef]
     [Google Scholar]
  30. Natarajan A, Srienc F. 1999; Dynamics of glucose uptake by single Escherichia coli cells. Metab Eng 1:320–333 [CrossRef]
     [Google Scholar]
  31. Natarajan A, Srienc F. 2000; Glucose uptake rates of single E. coli cells grown in glucose-limited chemostat cultures. J Microbiol Methods 42:87–96 [CrossRef]
     [Google Scholar]
  32. Nebe-von-Caron G, Stephens P. J, Hewitt C. J, Powell J. R, Badley R. A. 2000; Analysis of bacterial function by multi-colour fluorescence flow cytometry and single cell sorting. J Microbiol Methods 42:97–114 [CrossRef]
     [Google Scholar]
  33. Nystrom T. 2001; Not quite dead enough: on bacterial life, culturability, senescence, and death. Arch Microbiol 176:159–164 [CrossRef]
     [Google Scholar]
  34. Porter J, Deere D, Hardman M, Edwards C, Pickup R. 1997; Go with the flow cytometry in environmental microbiology. FEMS Microbiol Ecol 24:93–101 [CrossRef]
     [Google Scholar]
  35. Postgate J. R. 1967; Viability measurements and the survival of microbes under minimum stress. Adv Microb Physiol 1:2–23
     [Google Scholar]
  36. Postgate J. R. 1989; A microbial way of death. New Scientist 122:43–47
     [Google Scholar]
  37. Postgate J. R, Hunter J. R. 1964; Accelerated death of Aerobacter aerogenes starved in the presence of growth-limiting substrates. J Gen Microbiol 34:459–473 [CrossRef]
     [Google Scholar]
  38. Reed R. H. 1997; Solar inactivation of faecal bacteria in water: the critical role of oxygen. Lett Appl Microbiol 24:276–280 [CrossRef]
     [Google Scholar]
  39. Smith R. J, Kehoe S. C, McGuigan K. G, Barer M. R. 2000; Effects of simulated solar disinfection of water on infectivity of Salmonella typhimurium . Lett Appl Microbiol 31:284–288 [CrossRef]
     [Google Scholar]
  40. Wegelin M, Canonica S, Mechsner K, Fleischmann T, Pesaro F, Metzler A. 1994; Solar water disinfection: scope of the process and anlysis of radiation experiments. J Water SRT-Aqua 43:154–169
     [Google Scholar]
  41. White D. 1995 The Physiology and Biochemistry of Procaryotes New York: Oxford University Press;
     [Google Scholar]
  42. World Health Organization 1996 Guidelines for Drinking-Water Quality , vol. 2, Health Criteria and Other Supporting Information, 2nd edn.. Geneva: World Health Organization;
     [Google Scholar]
  43. World Health Organization/United World Nations Children's Fund 2005 Water for Life: Making it Happen Geneva: World Health Organization;
     [Google Scholar]
  44. Yoshioka K, Saito M, Oh K. B, Nemoto Y, Matsuoka H, Natsume M, Abe H. 1996; Intracellular fate of 2-NBDG, a fluorescent probe for glucose uptake activity, in Escherichia coli cells. Biosci Biotechnol Biochem 60:1899–1901 [CrossRef]
     [Google Scholar]
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Flow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS)
Microbiology 152, 1719 (2006); https://doi.org/10.1099/mic.0.28617-0
/content/journal/micro/10.1099/mic.0.28617-0
/content/journal/micro/10.1099/mic.0.28617-0
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