Czech J. Food Sci., 2016, 34(3):204-210 | DOI: 10.17221/528/2015-CJFS

Biofilm formation by Pseudomonas aeruginosa and disinfectant susceptibility of planktonic and biofilm cellsFood Microbiology and Safety

Magdalena A. Olszewska, Aleksandra M. Kocot, Aleksandra Stanowicka, Łucja Łaniewska-Trokenheim
Chair of Industrial and Food Microbiology, Faculty of Food Science, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland

Epifluorescence microscopy (EFM) was used to study the biofilm formation of Pseudomonas aeruginosa after 6, 24, 30, 48, 54, 72, 78, and 96 h growth in a chamber slide system. For this purpose, the biofilm was stained with the Live/Dead BacLight, wherein live and dead cells were visualised based on the cell membrane integrity. With the use of EFM we described 8- of 9-stage biofilm characteristics after 78 h of growth, since the majority of microscopic fields were fully covered with attached cells. However, the 96-h growth resulted in the cell detachment and revealed 30% of dead cells of all those cells that remained on the surface. The susceptibility testing of planktonic and biofilm cells to two disinfectants, chlorine-based and quaternary ammonium compound-based, revealed that biofilm cells were more tolerant to a chlorine-based sanitiser than planktonic counterparts. P. aeruginosa was inhibited by lower concentrations of the quaternary ammonium compound-based sanitiser than the chlorine-based sanitiser, which on the other hand was more effective in cell inactivation, as both the MIC/MBC (inhibitory/bactericidal) measurement and the CFDA/PI (carboxyfluorescein diacetate/propidium iodide) staining indicated.

Keywords: mode of existence; disinfection; fluorescence microscopy

Published: June 30, 2016  Show citation

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Olszewska MA, Kocot AM, Stanowicka A, Łaniewska-Trokenheim Ł. Biofilm formation by Pseudomonas aeruginosa and disinfectant susceptibility of planktonic and biofilm cells. Czech J. Food Sci. 2016;34(3):204-210. doi: 10.17221/528/2015-CJFS.
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    References

    1. Arnold J.W., Bailey G.W. (2000): Surface finishes on stainless steel reduce bacterial attachment and early biofilm formation: scanning electron and atomic force microscopy study. Poultry Science, 79: 1839-1845. Go to original source... Go to PubMed...
    2. Bagge D., Hjelm M., Johansen C., Huber I., Gram L. (2001): Shewanella putrefaciens adhesion and biofilm formation on food processing surfaces. Applied and Environmental Microbiology, 67: 2319-2325. Go to original source... Go to PubMed...
    3. Bjarnsholt T., Kirketerp-Moller K., Kristiansen S., Phipps R., Nielsen A.K., Jensen P.Ø., Høiby N., Givskov M. (2007): Silver against Pseudomonas aeruginosa biofilms. Acta Pathologica, Microbiologica et Immunologica Scandinavica, 115: 921-928. Go to original source... Go to PubMed...
    4. Cochran W.L., McFeters G.A., Stewart P.S. (2000): Reduced susceptibility of thin Pseudomonas aeruginosa biofilms to hydrogen peroxide and monochloramine. Journal of Applied Microbiology, 88: 22-30. Go to original source... Go to PubMed...
    5. Eginton P.J., Holah J., Allison D.G., Handley P.S., Gilbert P. (1998): Changes in the strength of attachment of micro-organisms to surfaces following treatment with disinfectants and cleaning agents. Letters in Applied Microbiology, 27: 101-105. Go to original source... Go to PubMed...
    6. Fuster-Valls N., Hernández-Herrero M., Marín-de-Mateo M., Rodríguez-Jerez J.J. (2008): Effect of different environmental conditions on the bacteria survival on stainless steel surfaces. Food Control, 19: 308-314. Go to original source...
    7. Greene A.K., Few B.K., Serafini J.C. (1993): A comparison of ozonation and chlorination for the disinfection of stainless steel surfaces. Journal of Dairy Science, 76: 3617-3620. Go to original source... Go to PubMed...
    8. Hendry E.R., Worthington T., Conway B.R., Lambert P.A. (2009): Antimicrobial efficacy of eucalyptus oil and 1,8-cineole alone and in combination with chlorhexidine digluconate against microorganisms in planktonic and biofilm cultures. Journal of Antimicrobial Chemotherapy, 64: 1219-1225. Go to original source... Go to PubMed...
    9. Jurcisek J.A., Dickson A.C., Bruggeman M.E., Bakaletz L.O. (2011): In vitro biofilm formation in an 8-well chamber slide. Journal of Visualized Experiments, 47: e2481. doi: 10.3791/2481 Go to original source... Go to PubMed...
    10. Kim J., Hahn J.S., Franklin M.J., Stewart P.S., Yoon J. (2009): Tolerance of dormant and active cells in Pseudomonas aeruginosa PA01 biofilm to antimicrobial agents. Journal of Antimicrobial Chemotherapy, 63: 126-135. Go to original source... Go to PubMed...
    11. Kim J., Pitts B., Stewart P.S., Camper A., Yoon J. (2008): Comparison of the antimicrobial effect of chlorine, silver ion, and tobramycin on biofilm. Antimicrobial Agents and Chemotherapy, 52: 1446-1456. Go to original source... Go to PubMed...
    12. Kubota H., Senda S., Nomura N., Tokuda H., Uchiyama H. (2008): Biofilm formation by lactic acid bacteria and resistance to environmental stress. Journal of Bioscience and Bioengineering, 106: 381-386. Go to original source... Go to PubMed...
    13. Le Thi T.T., Prigent-Combaret C., Dorel C., Lejeune P. (2001): First stages of biofilm formation: characterization and quantification of bacterial functions involved in colonization process. Methods in Enzymology, 336: 152-159. Go to original source... Go to PubMed...
    14. Myszka K., Białas W., Czaczyk K. (2005): The kinetics of bacterial biofilm formation on some technical materials depending on the availability of nutrients. Zywnosc Nauka Technolologia Jakosc, 3: 127-137.
    15. Nosyk O., Haseborg E., Metzger U., Frimmel F.H. (2008): A standardized pre-treatment method of biofilm flocs for fluorescence microscopic characterization. Journal of Microbiological Methods, 75: 449-456. Go to original source... Go to PubMed...
    16. Rault A., Beal C., Ghorbal S., Ogier S., Ogier J.C., Bouix M. (2007): Multiparametric flow cytometry allows rapid assessment and comparison of lactic acid bacteria viability after freezing and during frozen storage. Cryobiology, 55: 35-43. Go to original source... Go to PubMed...
    17. Simões M., Carvalho H., Pereira M.O., Vieira M.J. (2003): Studies on the behaviour of Pseudomonas fluorescens biofilms after ortho-phthalaldehyde treatment. Biofouling, 19: 151-157. Go to original source...
    18. Simões M., Pereira M.O., Vieira M.J. (2005): Effect of mechanical stress on biofilms challenged by different chemicals. Water Research, 39: 5142-5152. Go to original source... Go to PubMed...
    19. Simões M., Simões L.C., Vieira M.J. (2010): A review of current and emergent biofilm control strategies. LWT-Food Science and Technology, 43: 573-583. Go to original source...
    20. Simões M., Simões L.C., Vieira M.J. (2009): Species association increases biofilm resistance to chemical and mechanical treatments. Water Research, 43: 229-237. Go to original source... Go to PubMed...
    21. Smith K., Hunter I.S. (2008): Efficacy of common hospital biocides with biofilms of multi-drug resistant clinical isolates. Journal of Medical Microbiology, 57: 966-973. Go to original source... Go to PubMed...
    22. Stewert P.S., Costerton J.W. (2001): Antibiotic resistance of bacteria in biofilms. The Lancet, 358: 135-138. Go to original source... Go to PubMed...
    23. Surdeau N., Laurent-Maquin D., Bouthers S., Gelle M.P. (2006): Sensitivity of bacterial biofilms and planktonic cells to a new antimicrobial agent. Journal of Hospital Infection, 62: 487-493. Go to original source... Go to PubMed...
    24. Takenaka S., Iwaku M., Hoshino E. (2001): Artificial Pseudomonas aeruginosa biofilms and confocal laser scanning microscopic analysis. Journal of Infection and Chemotherapy, 7: 87-93. Go to original source... Go to PubMed...
    25. Takeuchi K., Frank J.F. (2000): Penetration of Escherichia coli O157:H7 into lettuce tissues as affected by inoculum size and temperature and the effect of chlorine treatment on cell viability. Journal of Food Protection, 63: 434-440. Go to original source... Go to PubMed...
    26. Takeuchi K., Frank J.F. (2001): Expression of red-shifted green fluorescent protein by Escherichia coli O157:H7 as a marker for the detection of cells on fresh produce. Journal of Food Protection, 64: 298-304. Go to original source... Go to PubMed...
    27. Takeuchi K., Matute C.M., Hassan A.N., Frank J.F. (2000): Comparison of the attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas fluorescens to lettuce leaves. Journal of Food Protection, 63: 1433-1437. Go to original source... Go to PubMed...
    28. Toté K., Horemans T., Berghe D.V., Maes L., Cos P. (2010): Inhibitory effect of biocides on the viable masses and matrices of Staphylococcus aureus and Pseudomonas aeruginosa biofilms. Applied and Environmental Microbiology, 76: 3135-3142. Go to original source... Go to PubMed...
    29. Tsuneda S., Aikawa H., Hayashi H., Yuasa A., Hirata A. (2003): Extracellular polymeric substances responsible for bacterial adhesion onto solid surface. FEMS Microbiology Letters, 223: 287-292. Go to original source... Go to PubMed...
    30. Whitchurch C.B., Tolker-Nielsen T., Ragas P.C., Mattick J.S. (2002): Extracellular DNA required for bacterial biofilm formation. Science, 295: 1487-1487. Go to original source... Go to PubMed...
    31. Wirtanen G., Salo S., Helander I.M., Mattila-Sandholm T. (2001): Microbiological methods for testing disinfectant efficiency on Pseudomonas biofilm. Colloids and Surfaces B: Biointerfaces, 20: 37-50. Go to original source... Go to PubMed...

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