Abstract
Highly resistant endospores may cause severe problems in medicine as well as in the food and packaging industries. We found that bacterial endospores can be inactivated quickly with reactive oxygen species (ROS) that were generated by a new generation of flavin photosensitizers. Flavins like the natural compound vitamin B2 are already known to produce ROS but they show a poor antimicrobial photodynamic killing efficacy due to the lack of positive charges. Therefore we synthesized new flavin photosensitizers that have one (FLASH-01a) or eight (FLASH-07a) positive charges and can hence attach to the negatively charged surface of endospores. In this study we used standardized Bacillus atrophaeus endospores (ATCC 9372) as a biological surrogate model for a proof-of-concept study of photodynamic inactivation experiments using FLASH-01a and FLASH-07a. After incubation of spores with different flavin concentrations, the flavin derivatives were excited with blue light at a light dose of 70 J cm−2. The inactivation of spores was investigated either in suspension or after attachment to polyethylene terephthalate (PET) surfaces. Incubation of spores suspended in Millipore water with 4 mM FLASH-01a for 10 seconds and irradiation with blue light for 10 seconds caused a biologically relevant decrease of spore survival of 3.5 log10 orders. Using FLASH-07a under the same conditions we achieved a decrease of 4.4 log10 orders. Immobilized spores on PET surfaces were efficiently killed with 7.0 log10 orders using 8 mM FLASH-07a. The total treatment time (incubation + irradiation) was as short as 20 seconds. The results of this study show evidence that endospores can be fastly and effectively inactivated with new generations of flavin photosensitizers that may be useful for industrial or medical applications in the future.
Article PDF
Similar content being viewed by others
References
I. Baldea, A. G. Filip, J. Physiol. Pharmacol. 2012 63 109–118.
T. G. St Denis, T. Dai, L. Izikson, C. Astrakas, R. R. Anderson, M. R. Hamblin, G. P. Tegos, Virulence 2011 2 509–520.
K. L. Brown, Br. Med. Bull. 2000 56 158–171.
P. Setlow, J. Appl. Microbiol. 2006 101 514–525.
W. L. Nicholson, P. Fajardo-Cavazos, R. Rebeil, T. A. Slieman, P. J. Riesenman, J. F. Law, Y. Xue, Antonie Van Leeuwenhoek 2002 81 27–32.
Y. J. Li, N. Zhu, H. Q. Jia, J. H. Wu, Y. Yi, J. C. Qi, J. Zhejiang Univ., Sci. B 2012 13 254–260.
J. V. Rogers, Y. W. Choi, W. R. Richter, D. C. Rudnicki, D. W. Joseph, C. L. Sabourin, M. L. Taylor, J. C. Chang, J. Appl. Microbiol. 2007 103 1104–1112.
N. Sudhaus, M. C. Pina-Perez, A. Martinez, G. Klein, Foodborne Pathog. Dis. 2012 9 442–452.
F. Barbut, D. Menuet, M. Verachten, E. Girou, Infect. Control Hosp. Epidemiol. 2009 30 507–514.
G. Jori, C. Fabris, M. Soncin, S. Ferro, O. Coppellotti, D. Dei, L. Fantetti, G. Chiti, G. Roncucci, Lasers Surg. Med. 2006 38 468–481.
M. Wainwright, J. Antimicrob. Chemother. 1998 42 13–28.
T. Maisch, Lasers Med. Sci. 2007 22 83–91.
T. Maisch, F. Spannberger, J. Regensburger, A. Felgenträger, W. Bäumler, J. Ind. Microbiol. Biotechnol. 2012 39 1013–1021.
T. N. Demidova, M. R. Hamblin, Appl. Environ. Microbiol. 2005 71 6918–6925.
J. S. Dickson, M. Koohmaraie, Appl. Environ. Microbiol. 1989 55 832–836.
L. M. He, B. M. Tebo, Appl. Environ. Microbiol. 1998 64 1123–1129.
F. Cieplik, A. Späth, J. Regensburger, A. Gollmer, L. Tabenski, K. A. Hiller, W. Bäumler, T. Maisch, G. Schmalz, Free Radical Biol. Med. 2013 65C, 477–487.
A. Felgenträger, T. Maisch, D. Dobler, A. Späth, BioMed Res. Int. 2013 2013 482167.
A. Eichner, F. P. Gonzales, A. Felgenträger, J. Regensburger, T. Holzmann, W. Schneider-Brachert, W. Bäumler, T. Maisch, Photochem. Photobiol. Sci. 2013 12 135–147.
A. Späth, C. Leibl, F. Cieplik, K. Lehner, J. Regensburger, K. A. Hiller, W. Bäumler, G. Schmalz, T. Maisch, J. Med. Chem. 2014 57 5157–5168.
I. Banerjee, K. K. Mehta, J. S. Dordick, R. S. Kane, J. Appl. Microbiol. 2012 113 1461–1467.
R. N. da Silva, A. C. Tome, J. P. Tome, M. G. Neves, M. A. Faustino, J. A. Cavaleiro, A. Oliveira, A. Almeida, A. Cunha, Microbiol. Immunol. 2012 56 692–699.
A. Knak, J. Regensburger, T. Maisch, W. Bäumler, Photochem. Photobiol. Sci. 2014 13, 5, 820–829.
W. Bäumler, J. Regensburger, A. Knak, A. Felgenträger, T. Maisch, Photochem. Photobiol. Sci. 2012 11 107–117.
J. Regensburger, A. Knak, T. Maisch, M. Landthaler, W. Bäumler, Exp. Dermatol. 2012 21 135–139.
J. Baier, T. Maisch, M. Maier, E. Engel, M. Landthaler, W. Bäumler, Biophys. J. 2006 91 1452–1459.
J. L. Sagripanti, M. Carrera, J. Insalaco, M. Ziemski, J. Rogers, R. Zandomeni, J. Appl. Microbiol. 2007 102 11–21.
J. Regensburger, T. Maisch, A. Felgenträger, F. Santarelli, W. Bäumler, J. Biophotonics 2010 3 319–327.
T. Maisch, A. Eichner, A. Späth, A. Gollmer, J. Regensburger, B. König, W. Bäumler, PLoS One 2014 10.1371/journal.pone.0111792.
E. Alves, L. Costa, C. M. B. Carvalho, J. P. C. Tome, M. A. Faustino, M. G. P. M. S. Neves, A. C. Tome, J. A. S. Cavaleiro, A. Cunha, A. Almeida, BMC Microbiol. 2009 9 70.
R. Kuhn, F. Weygand, Ber. Dtsch. Chem. Ges. A/B 1935 68 1282–1288.
B. Neises, W. Steglich, Angew. Chem., Int. ed. Engl. 1978 17 522–524.
A. A. Miles, S. S. Misra, J. O. Irwin, J. Hygiene 1938 38 732–749.
T. Mosmann, J. Immunol. Methods 1983 65 55–63.
J. M. Boyce, D. Pittet, Am. J. Infect. Control 2002 30 S1–46.
VDMA, Hygienische Abfüllmaschinen der Klasse IV nach VDMA für flüssige und pastöse Nahrungsmittel.
FDA, Class II Special Controls Guidance Document: Medical Washers and Medical Washer-Disinfectors. Guidance for the Medical Device Industry and FDA Review Staff.
P. W. Morrison, C. J. Connon, V. V. Khutoryanskiy, Mol. Pharm. 2013 10 756–762.
J. Schoenen, J. Jacquy, M. Lenaerts, Neurology 1998 50 466–470.
W. D. Lienhart, V. Gudipati, P. Macheroux, Arch. Biochem. Biophys. 2013 535 150–162.
T. Maisch, J. Baier, B. Franz, M. Maier, M. Landthaler, R. M. Szeimies, W. Bäumler, Proc. Natl. Acad. Sci. U. S. A. 2007 104 7223–7228.
T. Maisch, J. Wagner, V. Papastamou, H. J. Nerl, K. A. Hiller, R. M. Szeimies, G. Schmalz, J. Appl. Microbiol. 2009 107 1569–1578.
R. T. Kashiwabuchi, Y. Khan, F. R. Carvalho, F. Hirai, M. S. Campos, P. J. McDonnell, Arq. Bras. Oftalmol. 2012 75 423–426.
P. S. Thakuri, R. Joshi, S. Basnet, S. Pandey, S. D. Taujale, N. Mishra, Nepal Med. Coll. J. 2011 13 281–284.
I. Ahmad, and F. Vaid, in Comprehensive Series in Photochemistry and Photobiology, ed. D. P. Häder and G. Jori, RSC Publishing, http://pubs.rsc.org/en/content/chapter/bk9780854043316-00013/978-0-85404-331-6#!divabstract, 2006.
E. Tagliaferri, R. Sieber, U. Bütikofer, P. Eberhard, J. O. Bosset, Mitteilungen aus dem Gebiet der Lebensmittel-Untersuchung und -Hygiene 1992 83 467–491.
F. Vatansever, C. Ferraresi, M. V. de Sousa, R. Yin, A. Rineh, S. K. Sharma, M. R. Hamblin, Virulence 2013 4 796–825.
G. Pesce, G. Rusciano, A. Sasso, R. Isticato, T. Sirec, E. Ricca, Colloids Surf., B 2014 116C, 568–575.
W. H. Coleman, D. Chen, Y. Q. Li, A. E. Cowan, P. Setlow, J. Bacteriol. 2007 189 8458–8466.
S. E. Fiester, S. L. Helfinstine, J. C. Redfearn, R. M. Uribe, C. J. Woolverton, Int. J. Microbiol. 2012 2012 579593.
J. D. Hemmer, M. J. Drews, M. LaBerge, M. A. Matthews, J. Biomed. Mater. Res., Part B 2007 80 511–518.
A. Mohan, J. Dunn, M. C. Hunt, C. E. Sizer, J. Food Sci. 2009 74 M411–M417.
L. A. Dauphin, B. R. Newton, M. V. Rasmussen, R. F. Meyer, M. D. Bowen, Appl. Environ. Microbiol. 2008 74 4427–4433.
A. Oliveira, A. Almeida, C. M. Carvalho, J. P. Tome, M. A. Faustino, M. G. Neves, A. C. Tome, J. A. Cavaleiro, A. Cunha, J. Appl. Microbiol. 2009 106 1986–1995.
A. Harimawan, S. Zhong, C. T. Lim, Y. P. Ting, J. Colloid Interface Sci. 2013 405 233–241.
A. Andersson, U. Ronner, P. E. Granum, Int. J. Food Microbiol. 1995 28 145–155.
G. Lopez-Carballo, P. Hernandez-Munoz, R. Gavara, M. J. Ocio, Int. J. Food Microbiol. 2008 126 65–70.
Z. Luksiene, I. Buchovec, E. Paskeviciute, J. Appl. Microbiol. 2010 109 1540–1548.
Z. Luksiene, E. Paskeviciute, J. Photochem. Photobiol., B 2011 105 69–74.
N. Tortik, A. Spaeth, K. Plaetzer, Photochem. Photobiol. Sci. 2014 13 1402–1409.
P. Udompijitkul, M. Alnoman, D. Paredes-Sabjaa, M. R. Sarker, Food Microbiol. 2013 34 328–336.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Eichner, A., Gollmer, A., Späth, A. et al. Fast and effective inactivation of Bacillus atrophaeus endospores using light-activated derivatives of vitamin B2. Photochem Photobiol Sci 14, 387–396 (2015). https://doi.org/10.1039/c4pp00285g
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1039/c4pp00285g