Quantification of the ease of removal of bacteria from surfaces

Abstract

This paper describes a technique which reproducibly quantifies the ease of removal of microorganisms from surfaces. Tiles (22 mm×22 mm) of various materials were colonised withStaphylococcus epidermidis NCTC 11047,Escherichia coli K12 HB101 orPseudomonas aeruginosa PaWH, by submersion, for various times (2 min–48 h), in inoculated Tryptone Soya broth (37°C). Colonised tiles were blotted onto a Tryptone Soya agar plate for 1 min and the process was repeated through a succession of agar plates. The final plate contained tetrazolium salts (0.05% w/v) and was incubatedin situ with the tile. Tetrazolium plates indicated that very few organisms remained on the tiles after 15 successive blots. In all instances, the number of recovered colonies per plate decreased exponentially with plate succession number, according to the relationship, CFU-A.10^−kN, where CFU is the number of colonies transferred,k is the removal exponent, A is the intercept and N is the plate succession number. Removal exponents differed significantly between organisms (P>0.95), depended on the nature of the test surface, and decreased as the inital attachment and colonisation time was increased from 2 min–48 h. Intercept values (A) but not the gradients were dependent upon the initial numbers of bacteria in suspension. These data indicate that the gradients derived from counting recoverable viable cells from successive blots of test tiles onto agar is a measure of the strength of attachment of the organisms to the surface.