Archives of Microbiology

, Volume 195, Issue 1, pp 63–74 | Cite as

Wavelength dependence of biological damage induced by UV radiation on bacteria

  • Ana L. Santos
  • Vanessa Oliveira
  • Inês Baptista
  • Isabel Henriques
  • Newton C. M. Gomes
  • Adelaide Almeida
  • António Correia
  • Ângela Cunha
Original Paper


The biological effects of UV radiation of different wavelengths (UVA, UVB and UVC) were assessed in nine bacterial isolates displaying different UV sensitivities. Biological effects (survival and activity) and molecular markers of oxidative stress [DNA strand breakage (DSB), generation of reactive oxygen species (ROS), oxidative damage to proteins and lipids, and the activity of antioxidant enzymes catalase and superoxide dismutase] were quantified and statistically analyzed in order to identify the major determinants of cell inactivation under the different spectral regions. Survival and activity followed a clear wavelength dependence, being highest under UVA and lowest under UVC. The generation of ROS, as well as protein and lipid oxidation, followed the same pattern. DNA damage (DSB) showed the inverse trend. Multiple stepwise regression analysis revealed that survival under UVA, UVB and UVC wavelengths was best explained by DSB, oxidative damage to lipids, and intracellular ROS levels, respectively.


UV radiation Bacteria Inactivation Oxidative stress 



The authors would like to thank the anonymous reviewers and editors who provided helpful criticism and suggestions which greatly contributed to improve the original manuscript. Acknowledgments are due to Francisco Coelho and Abel Ferreira for assistance in UV intensity measurements and to Prof. Rosário Correia (Physics Department, Universiy of Aveiro) for reviewing the manuscript. Financial support for this work was provided by CESAM (Centre for Environmental and Marine Studies, University of Aveiro) and the Portuguese Foundation for Science and Technology (FCT) in the form of a PhD grant to A. L. Santos (SFRH/BD/40160/2007) and a post-Doctoral grant to I. Henriques (SFRH/BPD/63487/2009).


  1. Abboudi M, Surget SM, Rontani JF, Sempéré R, Joux F (2008) Physiological alteration of the marine bacterium Vibrio angustum S14 exposed to simulated sunlight during growth. Curr Microbiol 57:412–417PubMedCrossRefGoogle Scholar
  2. Alonso-Sáez L, Gasol JM, Lefort T, Hofer J, Sommaruga R (2006) Effect of natural sunlight on bacterial activity and differential sensitivity of natural bacterioplankton groups in Northwestern Mediterranean coastal waters. Appl Environ Microbiol 72:5806–5813PubMedCrossRefGoogle Scholar
  3. Anderl JN, Zahller J, Roe F, Stewart PS (2003) Role of nutrient limitation and stationary-phase existence in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother 47:1251–1256PubMedCrossRefGoogle Scholar
  4. Arrieta JM, Weinbauer MG, Herndl GJ (2000) Interspecific variability in sensitivity to UV radiation and subsequent recovery in selected isolates of marine bacteria. Appl Environ Microbiol 66:1468–1473PubMedCrossRefGoogle Scholar
  5. Bauermeister A, Bentchikou E, Moeller R, Rettberg P (2009) Roles of PprA, IrrE, and RecA in the resistance of Deinococcus radiodurans to germicidal and environmentally relevant UV radiation. Arch Microbiol 191:913–918PubMedCrossRefGoogle Scholar
  6. Beers RFJ, Sizer IW (1952) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–140PubMedGoogle Scholar
  7. Berney M, Weilenmann H-U, Egli T (2006a) Gene expression of Escherichia coli in continuous culture during adaptation to artificial sunlight. Environ Microbiol 8:1635–1647PubMedCrossRefGoogle Scholar
  8. Berney M, Weilenmann HU, Ihssen J, Bassin C, Egli T (2006b) Specific growth rate determines the sensitivity of Escherichia coli to thermal, UVA, and solar disinfection. Appl Environ Microbiol 72:2586–2593PubMedCrossRefGoogle Scholar
  9. Berney M, Weilenmann HU, Simonetti A, Egli T (2006c) Efficacy of solar disinfection of Escherichia coli, Shigella flexneri, Salmonella typhimurium and Vibrio cholerae. J Appl Microbiol 101:828–836PubMedCrossRefGoogle Scholar
  10. Bosshard F, Bucheli M, Meur Y, Egli T (2010a) The respiratory chain is the cell’s Achilles’ heel during UVA inactivation in Escherichia coli. Microbiology 156:2006–2015PubMedCrossRefGoogle Scholar
  11. Bosshard F, Riedel K, Schneider T, Geiser C, Bucheli M, Egli T (2010b) Protein oxidation and aggregation in UVA-irradiated Escherichia coli cells as signs of accelerated cellular senescence. Environ Microbiol 12:2931–2945PubMedCrossRefGoogle Scholar
  12. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  13. Cadet J, Courdavault S, Ravanat JL, Douki T (2005) UVB and UVA radiation-mediated damage to isolated and cellular DNA. Pure Appl Chem 77:947–961CrossRefGoogle Scholar
  14. Chamberlain J, Moss SH (1987) Lipid peroxidation and other membrane damage produced in Escherichia coli K1060 by near-UV radiation and deuterium oxide. Photochem Photobiol 45:625–630PubMedCrossRefGoogle Scholar
  15. Chun H, Kim J, Chung K, Won M, Song KB (2009) Inactivation kinetics of Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Campylobacter jejuni in ready-to-eat sliced ham using UV-C irradiation. Meat Sci 83:599–603PubMedCrossRefGoogle Scholar
  16. Coohill TP, Sagripanti J-L (2008) Overview of the inactivation by 254 nm ultraviolet radiation of bacteria with particular relevance to biodefense. Photochem Photobiol 84:1084–1090PubMedGoogle Scholar
  17. Coohill TP, Sagripanti J-L (2009) Bacterial inactivation by solar ultraviolet radiation compared with sensitivity to 254 nm radiation. Photochem Photobiol 85:1043–1052PubMedCrossRefGoogle Scholar
  18. De La Vega UP, Rettberg P, Douki T, Cadet J, Horneck G (2005) Sensitivity to polychromatic UV-radiation of strains of Deinococcus radiodurans differing in their DNA repair capacity. Int J Radiat Biol 81:601–611CrossRefGoogle Scholar
  19. Di Capua C, Bortolotti A, Farías ME, Cortez N (2011) UV-resistant Acinetobacter sp. isolates from Andean wetlands display high catalase activity. FEMS Microbiol Lett 317:181–189PubMedCrossRefGoogle Scholar
  20. Dillon JG, Tatsumi CM, Tandingan PG, Castenholz RW (2002) Effect of environmental factors on the synthesis of scytonemin, a UV-screening pigment, in a cyanobacterium (Chroococcidiopsis sp.). Arch Microbiol 177:322–331PubMedCrossRefGoogle Scholar
  21. Dodson ML, Michaels ML, Lloyd RS (1994) Unified catalytic mechanism for DNA glycosylases. J Biol Chem 269:32709–32712PubMedGoogle Scholar
  22. Eisenstark A (1998) Bacterial gene products in response to near-ultraviolet radiation. Mutat Res Fund Mol M 422:85–95CrossRefGoogle Scholar
  23. Fernández Zenoff V, Siñeriz F, Farías ME (2006) Diverse responses to UV-B radiation and repair mechanisms of bacteria isolated from high-altitude aquatic environments. Appl Environ Microbiol 72:7857–7863PubMedCrossRefGoogle Scholar
  24. Friedberg EC, Walker GC, Siede W (1995) DNA repair and mutagenesis. American Society of Microbiology Press, Washington, DCGoogle Scholar
  25. Garcia-Pichel F (1994) A model for internal self-shading in planktonic organisms and its implications for the usefulness of ultraviolet sunscreens. Limnol Oceanogr 39:1704–1717CrossRefGoogle Scholar
  26. Girotti AW (1998) Lipid hydroperoxide generation, turnover, and effector action in biological systems. J Lipid Res 39:1529–1542PubMedGoogle Scholar
  27. Gomes AA et al (2005) Reactive oxygen species mediate lethality induced by far-UV in Escherichia coli cells. Redox Rep 10:91–95PubMedCrossRefGoogle Scholar
  28. He YY, Häder DP (2002) UV-B-induced formation of reactive oxygen species and oxidative damage of the cyanobacterium Anabaena sp.: protective effects of ascorbic acid and N-acetyl-l-cysteine. J Photochem Photobiol, B 66:115–124CrossRefGoogle Scholar
  29. Hernandez EA, Ferreyra GA, Mac Cormack WP (2006) Response of two Antarctic marine bacteria to different natural UV radiation doses and wavelengths. Antarct Sci 18:205–212CrossRefGoogle Scholar
  30. Hobbie JE, Daley RJ, Jasper S (1977) Use of nuclepore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol 33:1225–1228PubMedGoogle Scholar
  31. Hörtnagl P, Pérez MT, Sommaruga R (2011) Contrasting effects of ultraviolet radiation on the growth efficiency of freshwater bacteria. Aquat Ecol 45:125–136PubMedCrossRefGoogle Scholar
  32. Ikehata H et al (2008) UVA1 genotoxicity is mediated not by oxidative damage but by cyclobutane pyrimidine dimers in normal mouse skin. J Invest Dermatol 128:2289–2296PubMedCrossRefGoogle Scholar
  33. Imlay JA (2006) Iron-sulphur clusters and the problem with oxygen. Mol Microbiol 59:1073–1082PubMedCrossRefGoogle Scholar
  34. Jacobs JL, Sundin GW (2001) Effect of solar UV-B radiation on a phyllosphere bacterial community. Appl Environ Microbiol 67:5488–5496PubMedCrossRefGoogle Scholar
  35. Jagger J (1985) Solar UV actions on living cells. Praeger Publishing, New YorkGoogle Scholar
  36. Joux F, Jeffrey WH, Lebaron P, Mitchell DL (1999) Marine bacterial isolates display diverse responses to UV-B radiation. Appl Environ Microbiol 65:3820–3827PubMedGoogle Scholar
  37. King B, Kesavan J, Sagripanti J-L (2011) Germicidal UV sensitivity of bacteria in aerosols and on contaminated surfaces. Aerosol Sci Tech 45:645–653CrossRefGoogle Scholar
  38. Krisko A, Radman M (2010) Protein damage and death by radiation in Escherichia coli and Deinococcus radiodurans. Proc Natl Acad Sci USA 107:14373–14377PubMedCrossRefGoogle Scholar
  39. Matallana-Surget S, Meador JA, Joux F, Douki T (2008) Effect of the GC content of DNA on the distribution of UVB-induced bipyrimidine photoproducts. Photoch Photobio Sci 7:794–801CrossRefGoogle Scholar
  40. Matallana-Surget S, Joux F, Raftery MJ, Cavicchioli R (2009a) The response of the marine bacterium Sphingopyxis alaskensis to solar radiation assessed by quantitative proteomics. Environ Microbiol 11:2660–2675PubMedCrossRefGoogle Scholar
  41. Matallana-Surget SM, Douki T, Cavicchioli R, Joux F (2009b) Remarkable resistance to UVB of the marine bacterium Photobacterium angustum explained by an unexpected role of photolyase. Photoch Photobio Sci 8:1313–1320CrossRefGoogle Scholar
  42. Matallana-Surget S, Joux F, Wattiez R, Lebaron P (2012) Proteome analysis of the UVB-resistant marine bacterium Photobacterium angustum S14. PLoS ONE 7:e42299PubMedCrossRefGoogle Scholar
  43. McCord JM, Fridovich I (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244:6049–6055PubMedGoogle Scholar
  44. Misiaszek R, Crean C, Joffe A, Geacintov NE, Shafirovich V (2004) Oxidative DNA damage associated with combination of guanine and superoxide radicals and repair mechanisms via radical trapping. J Biol Chem 279:32106–32115PubMedCrossRefGoogle Scholar
  45. Mitchell DL, Karentz D (1993) The induction and repair of DNA photodamage in the environment. In: Young AR, Bjorn LO, Moan J, Nultsch W (eds) Environmental UV photobiology. Plenum Press, New York, pp 345–377Google Scholar
  46. Moan J, Peak MJ (1989) Effects of UV radiation on cells. J Photochem Photobiol, B 4:21–34CrossRefGoogle Scholar
  47. Moeller R et al (2010) Genomic bipyrimidine nucleotide frequency and microbial reactions to germicidal UV radiation. Arch Microbiol 192:521–529PubMedCrossRefGoogle Scholar
  48. Murphy TM, Huerta AJ (1990) Hydrogen peroxide formation in cultured rose cells in response to UV-C radiation. Physiol Plantarum 78:247–253CrossRefGoogle Scholar
  49. Ordoñez O, Flores M, Dib J, Paz A, Farías M (2009) Extremophile culture collection from Andean Lakes: extreme pristine environments that host a wide diversity of microorganisms with tolerance to UV radiation. Microb Ecol 58:461–473PubMedCrossRefGoogle Scholar
  50. Pattison DI, Davies MJ (2006) Actions of ultraviolet light on cellular structures. EXS 96:131-157Google Scholar
  51. Pérez JM et al. (2007) Bacterial toxicity of potassium tellurite: Unveiling an ancient enigma. PLoS ONE 2:e211Google Scholar
  52. Pfeifer GP (1997) Formation and processing of UV photoproducts: effects of DNA sequence and chromatin environment. Photochem Photobiol 65:270–283PubMedCrossRefGoogle Scholar
  53. Pizarro RA, Orce LV (1988) Membrane damage and recovery associated with growth delay induced by near-UV radiation in Escherichia coli K-12. Photochem Photobiol 47:391–397PubMedCrossRefGoogle Scholar
  54. Qiu X, Sundin GW, Chai B, Tiedje JM (2004) Survival of Shewanella oneidensis MR-1 after UV radiation exposure. Appl Environ Microbiol 70:6435–6443PubMedCrossRefGoogle Scholar
  55. Qiu X, Sundin GW, Wu L, Zhou J, Tiedje JM (2005) Comparative analysis of differentially expressed genes in Shewanella oneidensis MR-1 following exposure to UVC, UVB, and UVA radiation. J Bacteriol 187:3556–3564PubMedCrossRefGoogle Scholar
  56. Rünger TM, Farahvash B, Hatvani Z, Rees A (2012) Comparison of DNA damage responses following equimutagenic doses of UVA and UVB: a less effective cell cycle arrest with UVA may render UVA-induced pyrimidine dimers more mutagenic than UVB-induced ones. Photoch Photobio Sci 11:207–215CrossRefGoogle Scholar
  57. Santos AL et al (2011) Diversity in UV sensitivity and recovery potential among bacterioneuston and bacterioplankton isolates. Lett Appl Microbiol 52:360–366PubMedCrossRefGoogle Scholar
  58. Santos AL et al (2012a) The UV responses of bacterioneuston and bacterioplankton isolates depend on the physiological condition and involve a metabolic shift. FEMS Microbiol Ecol 80:646–658PubMedCrossRefGoogle Scholar
  59. Santos AL et al (2012b) Effects of UV-B radiation on the structural and physiological diversity of bacterioneuston and bacterioplankton. Appl Environ Microbiol 78:2066–2069PubMedCrossRefGoogle Scholar
  60. Schenk M, Raffellini S, Guerrero S, Blanco GA, Alzamora SM (2011) Inactivation of Escherichia coli, Listeria innocua and Saccharomyces cerevisiae by UV-C light: study of cell injury by flow cytometry. LWT Food Sci Technol 44:191–198CrossRefGoogle Scholar
  61. Semchyshyn H, Bagnyukova T, Storey K, Lushchak V (2005) Hydrogen peroxide increases the activities of soxRS regulon enzymes and the levels of oxidized proteins and lipids in Escherichia coli. Cell Biol Int 29:898–902PubMedCrossRefGoogle Scholar
  62. Shick JM, Dunlap WC (2002) Mycosporine-like amino acids and related gadusols: biosynthesis, accumulation, and UV-protective functions in aquatic organisms. Annu Rev Physiol 64:223–262PubMedCrossRefGoogle Scholar
  63. Smith DC, Azam F (1992) A simple, economical method for measuring bacterial protein synthesis rates in seawater using tritiated-leucine. Mar Microb Food Webs 6:107–114Google Scholar
  64. Sundin GW, Jacobs JL (1999) Ultraviolet radiation (UVR) sensitivity analysis and UVR survival strategies of a bacterial community from the phyllosphere of field-grown peanut (Arachis hypogeae L.). Microb Ecol 38:27–38PubMedCrossRefGoogle Scholar
  65. Ubomba-Jaswa E, Navntoft C, Polo-López MI, Fernandez-Ibáñez P, McGuigan KG (2009) Solar disinfection of drinking water (SODIS): an investigation of the effect of UV-A dose on inactivation efficiency. Photoch Photobio Sci 8:587–595CrossRefGoogle Scholar
  66. Visser PM, Poos JJ, Scheper BB, Boelen P, Van Duyl FC (2002) Diurnal variations in depth profiles of UV-induced DNA damage and inhibition of bacterioplankton production in tropical coastal waters. Mar Ecol Prog Ser 228:25–33CrossRefGoogle Scholar
  67. Warnecke F, Sommaruga R, Sekar R, Hofer JS, Pernthaler J (2005) Abundances, identity, and growth state of Actinobacteria in mountain lakes of different UV transparency. Appl Environ Microbiol 71:5551–5559PubMedCrossRefGoogle Scholar
  68. Zeeshan M, Prasad SM (2009) Differential response of growth, photosynthesis, antioxidant enzymes and lipid peroxidation to UV-B radiation in three cyanobacteria. S Afr J Bot 75:466–474CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Ana L. Santos
    • 1
  • Vanessa Oliveira
    • 1
  • Inês Baptista
    • 1
  • Isabel Henriques
    • 1
  • Newton C. M. Gomes
    • 1
  • Adelaide Almeida
    • 1
  • António Correia
    • 1
  • Ângela Cunha
    • 1
  1. 1.Department of Biology & CESAMUniversity of AveiroAveiroPortugal

Personalised recommendations