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Applied and Environmental Microbiology, September 2002, p. 4194-4200, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4194-4200.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Development of a Standard Test To Assess the Resistance of Staphylococcus aureus Biofilm Cells to Disinfectants

Suzanne B. I. Luppens,1,2 Martine W. Reij,1 Rob W. L. van der Heijden,1 Frank M. Rombouts,1 and Tjakko Abee1*

Food Hygiene and Microbiology Group,1 Consumer Technology and Product Use Group, Department of Agrotechnology and Food Sciences, Wageningen University and Research Center, 6700 EV Wageningen, The Netherlands2

Received 10 December 2001/ Accepted 30 May 2002

A standardized disinfectant test for Staphylococcus aureus cells in biofilms was developed. Two disinfectants, the membrane-active compound benzalkonium chloride (BAC) and the oxidizing agent sodium hypochlorite, were used to evaluate the biofilm test. S. aureus formed biofilms on glass, stainless steel, and polystyrene in a simple system with constant nutrient flow that mimicked as closely as possible the conditions used in the current standard European disinfectant test (EN 1040). The biofilm that was formed on glass contained cell clumps and extracellular polysaccharides. The average surface coverage was 60%, and most (92%) of the biofilm cells were viable. Biofilm formation and biofilm disinfection in different experiments were reproducible. For biofilms exposed to BAC and hypochlorite the concentrations needed to achieve 4-log killing were 50 and 600 times higher, respectively, than the concentrations needed to achieve this level of killing with the European phase 1 suspension test cells. Our results show that a standardized disinfectant test for biofilm cells is a useful addition to the current standard tests.

* Corresponding author. Mailing address: Food Hygiene and Microbiology Group, Department of Agrotechnology and Food Sciences, Wageningen University and Research Center, P.O. Box 8129, 6700 EV Wageningen, The Netherlands. Phone: 31 317 484981. Fax: 31 317 484893. E-mail: Tjakko.Abee{at}micro.fdsci.wau.nl.

Applied and Environmental Microbiology, September 2002, p. 4194-4200, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4194-4200.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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