Rapid monitoring method to assess efficacy of sanitizers against Pseudomonas putida biofilms

Abstract

Biofilms of luminescent Pseudomonas putida were developed on rubber surfaces by incubation in brain heart infusion (BHI) broth. Scanning electron microscopy (SEM) and epifluorescence microscopy (EFM) were used to examine biofilm formation. To test the efficacy of two sanitizers commonly employed in dairy plants for CIP (cleaning in place) procedures, a novel bioluminescence method and aerobic plating were used to enumerate cells. Immediately after the sanitizer treatments an apparent 5-log reduction of biofilm-associated cells was determined. However, when the samples were resuscitated for 18 h in BHI broth, high numbers of cells were detected which reached levels close to those of nontreated controls. The results demonstrated that neither sanitizer could completely eliminate biofilm-associated P. putida. The microbial bioluminescence method proved to be the best way for assessing effectiveness of sanitizers against microbial biofilms.