Comparison of Gamma Irradiation and Sodium Hypochlorite Treatments to Inactivate Staphylococcus aureus and Pseudomonas aeruginosa Biofilms on Stainless Steel Surfaces

  • Kim, Jang-Ho (Bio Technology Regional Innovation Center, Youngdong University) ;
  • Jo, Cheo-Run (Department of Animal Science and Technology Chungnam National University) ;
  • Rho, Yong-Taek (Bio Technology Regional Innovation Center, Youngdong University) ;
  • Lee, Chun-Bok (Department of Biology, Kyungsung University) ;
  • Byun, Myung-Woo (Radiation Food Science and Biotechnology Team, Advanced Radiation Technology Institute)
  • Published : 2007.04.30


Biofilm formation on various surfaces is a well-known phenomenon and it has caused pollution problems, health and safety hazards, and substantial economic loss in many areas including the food industry. In the present study, Gamma irradiation at a dose of 2.0 kGy reduced the bacterial counts of Staphylococcus aureus and Pseudomonas aeruginosa suspensions by 6.7 and >6.5 log CFU/mL, respectively, and 30 ppm of sodium hypochlorite effectively reduced the counts of both bacterial suspensions to below the limit of detection ($<2\;log\;CFU/cm^2$). However, in bacterial biofilms attached to stainless steel, gamma irradiation at a dose of 10.0 kGy reduced the counts of S. aureus attached fur 1 hr and overnight by ${\geq}5.1\;and\;5.0\;log\;CFU/cm^2$, respectively. Gamma irradiation at a dose of 1.0 kGy reduced the counts of P. aeruginosa counts to below the limit of detection ($<2\;log\;CFU/cm^2$). On the contrary, S. aureus and P. aeruginosa cells attached to stainless steel chips were difficult to eliminate using sodium hypochlorite. Four hundred ppm of sodium hypochlorite reduced the counts of S. aureus and P. aeruginosa attached for 1 hr by 2.5 and $3.3\;log\;CFU/cm^2$, respectively.


  1. Costerton JW, Geesey GG, Cheng KJ. How bacteria stick. Sci. Am. 238: 86-95 (1978)
  2. Brackett RE. Shelf stability and safety of fresh produce as influenced by sanitation and disinfection. J. Food Protect. 55: 808-814 (1992)
  3. Lindsay D, Geomaras I, von Holly A. Biofilms associated with poultry processing equipment. Microbios 86: 105-116 (1996)
  4. Gu JD, Roman M, Esselman T, Mitchell R. The role of microbial biofilms in deterioration of space station candidate materials. Int. Biodeter. Biodegr. 41: 25-33 (1998)
  5. Wong H-C, Chung Y-C, Yu J-A. Attachment and inactivation of Vibrio parahaemolyticus on stainless steel and glass surface. Food Microbiol. 19: 341-350 (2002)
  6. Jay JM. Mordem Food Microbiology. 6th ed. Aspen Publishers, Gaithersburg, MD, USA. pp. 322-326 (2000)
  7. Lomander A, Schreuders P, Russek-Cohen E, Ali L. Evaluation of chlorines, impact on biofilms on scratched stainless steel surfaces. Bioresource Technol. 94: 275-283 (2004)
  8. Dychdala GR. Chlorine and chlorine compounds. pp. 157-172. In: Disinfection, Sterilization, and Preservation. Block SS (ed). 3rd ed. Lea & Febiger, Philadelphia, PA, USA (1983)
  9. Kwon JH. Effects of gamma irradiation and methyl bromide fumigation on the qualities of fresh chestnuts during storage. Food Sci. Biotechnol. 14: 181-184 (2005)
  10. Sommer P, Martin-Rouas C, Mettler E. Influence of the adherent population level on biofilm population, structure and resistance to chlorination. Food Microbiol. 16: 503-515 (1999)
  11. William I, Venables WA, Lloyd D, Paul F, Critchley I. The effects of adherence to silicone surfaces on antibiotic susceptibility in Staphylococcus aureus. Microbiology 143: 2407-2413 (1997)
  12. Somers EB, Schoeni JL, Wong ACL. Effect of trisodium phosphate on biofilm and planktonic cells of Campylobacter jejuni, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium. Int. J. Food Microbiol. 22: 269-276 (1994)
  13. Joseph B, Otta SK, Karunasagar I, Karunasagar I. Biofilm formation by Salmonella spp. on food contact surfaces and their sensitivity to sanitizers. Int. J. Food Microbiol. 64: 367-372 (2001)
  14. James GA, Beaudette L, Costerton JW. Interspecies bacterial interactions in biofilms. J. Ind. Microbiol. Biot. 15: 257-262 (1995)
  15. APHA. Standard methods for the examination of water and wastewater. 18th ed. Method 4500-CIB. American Public Health Association, Washington, DC, USA (1992)
  16. SAS Institute, Inc. SAS User's Guide. Statistical Analysis System Institute, Cary, NC, USA (1990)
  17. Sheehan E, McKenna J, Mulhall KJ, Marks P, McCormack D. Adhesion of Staphylococcus to orthepaedic metals, an in vivo study. J. Orthop. Res. 22: 39-43 (2004)
  18. Sinde E, Caballo J. Attachment of Salmonella spp. and Listeria monocytogenes to stainless steel, rubber, and polytetrafluorethylene:the influence of free energy and the effect of commercial sanitizers. Food Microbiol. 17: 439-447 (2000)
  19. Luppens SBI, Reji MW, van der Heijden RWL, Rombouts FM, Abee T. Development of a standard test to assess the resistance of Staphylococcus aureus biofilm cells to disinfectants. Appl. Environ. Microb. 68: 4194-4200 (2002)
  20. Hamilton WA. Sulphate reducing bacteria and anaerobic corrosion. Annu. Rev. Microbiol. 39: 195-217 (1985)
  21. Flemming HC. Biofouling and microbiologically influenced corrosion (MIC)-an economical and technical overview. pp. 5-14. In: Microbial Deterioration of Materials. Heitz E, Sand W, Flemming HC (eds). Springer, Heidelberg, Germany (1996)
  22. Kausar T, Kwon JH, Kim HK. Comparative effect of gamma irradiation and fumigation on total phenol content and biological activities of different teas (Camellia sinessis). Food Sci. Biotechnol. 13: 672-675 (2004)
  23. Suarez B, Ferreiros CM, Criado MT. Adherence of psychrotropic bacteria to dairy equipment surfaces. J. Dairy Res. 59: 381-388 (1992)
  24. Demirer S, Gecim IE, Aydinuraz K, Ataoglu H, Yerdel MA, Kuterdem E. Affinity of Staphylococcus epidermis to various prosthetic graft materials. J. Surg. Res. 99: 70-74 (2001)
  25. Wirtanen G, Salo S, Helander IM, Mattila-Sandhohn T. Microbiological methods for testing disinfectant efficacy on Pseudomonas biofilm. Colloid. Surface B 20: 37-50 (2001)
  26. Ingraham A, Fleischer TM. Disinfectants in laboratory animal science: what are they and who says they work? Lab Animal 32: 36-40 (2003)
  27. Gu JD, Lu C, Thorp K, Crasto A, Mitchell R. Fibre-reinforced polymeric composite materials are susceptible to microbial degradation. J. Ind. Microbiol. Biot. 18: 364-369 (1997)
  28. Bal'a MFA, Jamilah lD, Marshall DL. Attachment of Aeromonas hydrophilia to stainless steel surface. Dairy Food Environ. Sanit. 18: 642-649 (1998)
  29. Hecker M, Engehnann S, Cordwell JS. Proteomics of Staphylococcus aureus-current state and future challenges. J. Chromatogr. B 787: 179-195 (2003)
  30. Niemira BA. Irradiation of fresh and minimally processed fruits, vegetables, and juices. pp. 279-300. In: The Microbial Safety of Minimally Processed Foods. Novak JS, Sapers GM, Juneja VK (eds). CRC Press, Boca Raton, FL, USA (2003)
  31. Davis D. Understanding biofilm resistance to antibacterial agents. Nat. Rev. Microbiol. 2: 114-122 (2003)
  32. Niemira BA, Solomon EB. Sensitivity of planktonic and biofilm associated Salmonella spp. to ionizing radiation. Appl. Environ. Microb. 71: 2732-2736 (2005)
  33. Beech lB. Corrosion of technical materials in the presence of biofilms-current understanding and state-of-the art methods of study. Int. Biodeter. Biodegr. 53: 177-183 (2004)
  34. Gross RA, Gu JD, Eberiel DT, Nelson M, McCarthy SP. Cellulose acetate biodegradability in simulated aerobic composting and anaerobic bioreactor environments as well as by a bacteria isolate derived from compost. pp. 257-279. In: Biodegradable Polymers and Packaging. Ching C, Kaplan DL, Thomas EL (eds). Technomic, Lancaster, PA, USA (1993)
  35. Sabin C, Mitchell EP, Pokoma M, Gautier C, Utille JP, Wimmerova M, Imberty A. Binding of different monosaccharides by lectin PAIlL from Pseudomonas aeruginosa: Thermodynamics data correlated with X-ray structures. FEBS Lett. 580: 982-987 (2006)
  36. Gibson H, Taylor JH, Hall KE, Holah JT. Effectiveness of cleaning techniques used in the food industry in terms of the removal of bacterial biofilms. J. Appl. Microbiol. 87: 41-48 (1999)
  37. Austin JW, Bergeron G. Development of bacterial biofilms in dairy processing lines. J. Dairy Res. 62: 509-519 (1995)
  38. Le Magrex-Debar E, Lemoine J, Gelle MP, Jacquelin LF, Choisy C. Evaluation of biohazards in dehydrated biofilms on foodstuff packaging. Int. J. Food Microbiol. 55: 239-243 (2000)
  39. Buret A, Ward KH, Olson ME, Costerton JW. An in vivo model to study the pathobiology of infectious biofilms on biomaterial surfaces. J. Biomed. Mater. Res. 25: 865-874 (2004)
  40. Simmons A. Sterilization of Medical Devices (Business Briefing: Medical Device Manufacturing & Technology 2004). Touch Briefings, London, UK. pp. 45-46 (2004)
  41. de Beer D, Srinivasan R, Stewart PS. Direct measurement of chlorine penetration into biofilms during disinfection. Appl. Environ. Microb. 60: 4339-4344 (1994)
  42. Characklis WG, Marshall KC. Biofilms. Wiley, New York, NY, USA. pp. 195-231 (1990)
  43. Gu JD, Ford T, Mitchell R. Microbial deterioration of fiber reinforced composite polymeric materials. pp. 16-17. In: Corrosion/ 95 Research in Progress Symposium. March 27, Orlando, FL, USA. National Association of Corrosion Engineering, Houston, TX, USA (1995)
  44. Buys EM, Nortje GL, Jooste PJ, Von Holy A. Bacterial populations associated with bulk packaged beef supplemented with dietary vitamin E. Int. J. Food Microbiol. 56: 239-244 (2000)
  45. Tsuji K. Low-dose cobalt 60 irradiation for reduction of microbial contamination in raw materials for animal health products. Food Technol.-Chicago 37: 48-54 (1983)