Biofilm-Forming Ability of Pathogenic Bacteria Isolated from Retail Food in Poland

Abstract

Biofilms have a significant impact on food safety in the food industry. Many foodborne outbreaks have been associated with pathogenic bacterial strains that can form a biofilm. The present study was conducted under the Official Control and Monitoring Program in Poland to examine the ability of pathogenic bacteria collected from retail food samples to form biofilms. Biofilm formation was assessed by qualitative detection of extracellular polymeric substances on Congo red agar, by adherence to glass with the tube method, by the crystal violet biofilm (CV) assay, and by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. A total of 40 isolates from food samples (10 strains each of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, and Bacillus cereus) were examined. The strains were classified as adherent, slightly adherent, or nonadherent; biofilm production was classified as weak (WBP), moderate (MBP), or strong (SBP); and metabolic activity was classified as weak (WMA), moderate (MMA), or high (HMA). The incubation conditions and time influenced the amount of biofilm formed as well as did the growth medium. In the test tubes with Luria-Bertani broth (LBB), 22.5% of the strains were adherent and 77.5% were slightly adherent. Stronger adhesion was obtained in brain heart infusion (BHI) with 2% sucrose; 60% of the isolates were classified as adherent. With the CV assay with LBB, SBP was noted for 7.5% of the strains after 24 h of incubation and for 37.5% of the strains after 48 h. In BHI plus 2% sucrose, SBP was noted for 42.5 and 37.6% of the strains after 24 and 48 h, respectively. With the MTT assay with LBB, HMA was found for 15% of the strains after 24 h of incubation and for 25% of the strains after 48 h. In BHI plus 2% sucrose, 70 and 85% of the incubated strains were classified as HMA after 24 and 48 h, respectively.